Early detection of neurological conditions is critical for timely diagnosis and treatment. Identifying cellular-level changes is essential for implementing therapeutic interventions prior to symptomatic disease onset. However, monitoring brain tissue directly through biopsies is invasive and high-risk. Bodily fluids such as blood or cerebrospinal fluid contain information in many forms, including proteins and nucleic acids. Particularly, cell-free DNA (cfDNA) has potential as a versatile neurological biomarker. Yet, our knowledge of cfDNA released by brain tissue and how cfDNA changes in response to deleterious events within the brain is incomplete. Mapping changes in cfDNA to specific cellular events is difficult in vivo, where many tissues contribute to circulating cfDNA. Organoids are tractable systems for examining specific changes consistently in a human background. However, few studies have investigated cfDNA released from organoids. Here, we examined cfDNA isolated from cerebral organoids. We found that cerebral organoids release quantities of cfDNA sufficient for downstream analysis with droplet-digital PCR and whole-genome sequencing. Further, gene ontology analysis of genes aligning with sequenced cfDNA fragments revealed association with terms related to neurodevelopment and autism spectrum disorder. We conclude that cerebral organoids hold promise as tools for the discovery of cfDNA biomarkers related to neurodevelopmental and neurological disorders.

We investigated the development of a simple and sensitive assay for the identification of minute amounts of animal and human biological material. The suggested system uses isothermal amplification of DNA in a rapid assay format, which returns the results in as little as 40 minutes from sampling. The assay used for the proof of concept employs the novel handheld SaLux19 device (Max-Planck Innovation prototype, Germany). This well-established infield detection system could also be utilized for forensic scenarios. The results presented here demonstrated that the assay is sufficiently specific and sensitive and can detect the presence of Sus scrofa nucleic acids.

Peanut (Arachis hypogaea) contains allergenic proteins, which make it harmful to the sensitive population. The presence of peanut in foods must be indicated on label, to prevent accidental consumption by allergenic population. The development of suitable analytical methodologies to detect this allergen in processed foods is advisable. Real Time PCR allowed a specific and accurate amplification of allergen sequences. The optimal genome targets for specific and sensitive detection are those that show interspecific variation and high copy number. Some food processing methods could induce structural and/or conformational changes in proteins and produce the fragmentation and/or degradation of genomic DNA. In this work, we use chloroplast markers for specifically detection of peanut by Real Time PCR, in order to increase the sensitivity. Three different protocols of DNA isolation were evaluated, for total and organelle-DNA extraction. Binary mixtures of raw and processed peanut flour in wheat were performed at concentrations ranging from 100000 to 0.1 ppm. DNA isolation from peanut, mixtures and other legumes was carried out following three protocols for obtaining genomic and chloroplast-enrich DNA. Quantity and quality of DNA was evaluated, obtaining better results for protocol 2. Specificity and sensitivity of the method has been assayed with specific primers for three chloroplast markers (mat k, rpl16 and trnH-psbA) and Ara h 6 peanut allergen-coding region was selected as nuclear low-copy target and TaqMan probes. Efficiency and linear correlation of calibration curves were within the adequate ranges. Moreover, the influence of pressure and thermal processing on the peanut detectability was analyzed.

The spread of the infection caused by the new coronavirus SARS-CoV-2 (COVID-19) became pandemic on March 11, 2020. From the time of the first cases (in November 2019, Wuhan, China), to date, a large number of COVID-19 observations have been accumulated in different age groups of patients both in China and abroad. Published scientific data allows us to conclude that children suffer from COVID-19 much less often than adults and tolerate the disease in a milder form, often appear to be asymptomatic. There is currently no final answer why children are less susceptible to this virus; however, scientists are increasingly inclined to consider a complex effect of the immune response and components of the renin-angiotensin system (RAS), which according to recent studies affects not only the cardiovascular system, but is also responsible for the activation of inflammatory reactions. A hypothesis of genetic predisposition to the development of severe forms of COVID-19 has recently been made. We conducted a search for publications in the databases and showed current scientific ideas about COVID-19 pathogenesis and factors influencing the disease development in childhood. Childhood immunity may have several protective features against SARS-CoV-2: immaturity of particular elements of the innate immune response, constitutional lymphocytosis with a shift towards anti-inflammatory Th2-response, as well as "trained" immunity. The influence of renin-angiotensin system reactions in this review is shown from two perspectives: expression of ACE2 receptors and polymorphisms of certain genes of this system. It was established that ACE2 transmembrane protein is not only the entry point for the virus but also plays a regulatory role, turning the pro-inflammatory vasoconstrictor angiotensin II into anti-inflammatory angiotensin (1-7), which has vasodilating properties. Higher ACE2 content in children compared with adults helps maintain balance in the renin-angiotensin system and prevents the development of complications. It was also shown that the presence of certain genetic polymorphisms (AGTR1, AGTR2, ACE2, ACE) could determine the imbalance inside the RAS, leading to more pronounced reactions of alveolocytes, vascular endothelium and smooth muscle fibers in response to SARS-CoV-2 infection due to a shift towards vasoconstrictor, proliferative and profibrotic mechanisms.

Sunflower is one of the oil crops and its planting areas was listed behind Soybean, peanut and rapeseeds in China. It will encounter different kind of diseases during planting, thus limited the seeds yield. Fusarium species are the major threat to different crops nowadays, it is very important to identify the species caused diseases on different hosts, so that the potential control measures may be explored. In Inner mongolia, Sunflower Disk Rot (SDR) was found in different sunflower planting field, the average infection ratio around 8.50%, typically in some verieties, its infection ratio reach to 11.67%. The diseased receptacle were collected from different locations and Koch’s postulate was conducted. The results showed that Fusarium verticillioides was the main causal agent caused SDR. Fungicide toxicity tests shown that Fludioxonil and Flutolanil were screened out from six fungicides for controlling F. verticillioides, with EC50 values of 0.05 µg/mL (R = 0.9825) and 0.96 µg/mL (R = 0.9964) , respectively. This is the first report of SDR caused by F. verticillioides, and it will alert sunflower reseachers to include SDR in the disease list, so as to control sunflower diseases with integrated management strategy successfully.

This work discusses seismic isolation technologies and analyses the concept of geotechnical seismic isolation (GSI) system, which is characterized by new principles of operation, using geomaterial with modified properties to reduce inertial seismic loads on buildings and structures. The aim of the study is to develop a geotechnical seismic isolation system in the form of vertical barriers, with different geometrical characteristics, using rubber-soil mixture (RSM). The novelty of the work consists in determination of effective constructive and technical solutions of vertical seismic barriers from RSM, which are characterized by reliability in providing seismic isolation of construction objects. Soil and superstructure interactions are modeled in Plaxis software, using the Northridge earthquake accelerogram. The results confirm the positive impact of using RSM as an effective material for GSI vertical barriers. The present numerical and experimental results show that the GSI system using RSM can significantly reduce the seismic hazard. The results of computational-experimental modeling will become a research, analytical and design basis for the development of new ways to ensure earthquake resistance of construction objects.

Loneliness is becoming prevalent across Japan in spite of its a dense population and communal culture. Surveys show 40% of adults report isolation across demographics. The contradiction emerges due to deteriorating social structure with modernization, economic parity, urbanization, and fading conventional setup. The continually social withdrawals, called “hikikomori." Relief seekers have embraced AI based emotive robots, digital avatars, and immersive VR as low-stakes social surrogates despite limited capacity to cultivate human intimacy. To stabilise government attempts awareness through reforms, scheme and support services. But a concrete transformation can be brough in with work-life balance, gender equality and rewire the metrics for self-esteem. Even a perfectly personalized AI cannot replace the human affection lacking in the society. Lasting solutions demand technological aids be balanced by confronting and reinventing community purpose and bring in structural change. Overall Japan’s endemic isolation reveals the societal fraying when modernization privileges independence over interdependence. The path forward rests on restoring cultural foundations to reverse loneliness and social isolation.

Several bio-actors are involved in the occurrence of leishmaniasis, which is made too complex for prevention and control options. Currently, all forms of leishmaniasis are being treated with chemical drugs, which have limitations and adverse effects on patients. Searching the bioactive molecules from plant sources allows the development of a novel drug. The essential oil and solvent extract of the roots of E. kebericho showed antileishmanial activity. But looking for biological activities of the isolated active ingredient from tubers of E. kebericho was scant. Thus, the isolation of the leishmanicidal compound from the roots of E. kebericho through a bioassay-guided technique was an aim of this work. Dehydrocostus lactone showed doubled potent than the standard drug miltefosine on leishmanicidal activities against L. major and L. tropica. Besides, the broad-spectrum biological activities of dehydrocostus lactone could be a potential drug candidate for co-infected patients caused by an association of leishmania and another pathogen.

Quite a number of cottonseed varieties are existed. The seeds of each variety produce a different amount of gossypol as a natural toxic compound. The increased interest in this natural product and its toxic effects were enhanced the demand for establishing simple silica-gel-based methods for its isolation, purification, and characterization. Historically, silica gel plays a significant role in isolation and purification of many different natural products. Silica gel is variants and can intro-duce a lab, simple tool for tracking a compound in certain plants during its purification process. In this study, gossypol was isolated and purified in the form of yellow crystals as gossypol acetic ac-id. Several methods and tools were used for its characterization including TLC, preparative TLC, silica gel column, UV/IR spectrophotometer and HPLC (Robust spherical silica gel). A compara-tive study between its amounts in both Egyptian and Chinese varieties were performed. Under the experimental condition, the Egyptian cottonseeds contain 8.705 gm/kg, while the Chinese contains 5.395 gm/kg. Thin Layer Chromatography (TLC) had been flourished the science of the phytochemistry and still popular till nowadays and is recommended for fast, accurate, and inex-pensive evaluation and quantification of gossypol. It is recommended to isolate and purify gossy-pol as gossypol acetic acid using optimized combined methods used in this study. Using Silica gel different protocols will assist and facilitate the parallel comparison and save the compact column from any unexpected contaminants during the purification steps. During each step the silica gel itself could be simply removed by paper filtration.

Sea cucumber peptides are small molecular peptides with biological function obtained from fresh sea cucumber as feedstock, which were hydrolyzed by protease, separated ,and purified. As the main product of sea cucumber refinement, it more accurately expresses the nutritional effect of sea cucumber and has a wide range of potential applications. It has been demonstrated that sea cucumber has a certain value-added effect on bone mineral density regulation, but the differences in bone mineral density regulation of extracts from different parts of sea cucumber and the techniques contributing to the efficient isolation of bone sea cucumber peptides have not been reported. Firstly, the aim of this study was to reveal the differences in osteogenic activities of enzymatic peptides from three major parts of sea cucumber: the body wall, the egg protein, and the intestinal protein. Then, the bone-enhancing sea cucumber peptides were isolated based on the characteristics of the dif-ferences in molecular weight distribution, hydrophilicity, and ionisation degree of sea cucumber, and to explore the possible mechanisms of their action. The findings indicated that sea cucumber peptides extracted from three distinct sites exhibited a dose-dependent enhancement of osteoblast proliferation and differentiation. Upon evaluating the three methods of sea cucumber peptide separation based on molecular weight distribution, hydrophilicity, and ionization, it was observed that the fraction isolated by hydrophobicity and

The increase in the frequency of droughts in Africa has increased the interest in crops with drought escape mechanisms like grain Amaranth (Amaranthus hypochondriacus). However, the white-seeded grain Amaranth is easily cross-pollinated by its black seeded wild relatives such as smooth pigweed (Amaranthus hybridus). A study to investigate a minimum isolation distance enough to prevent seed contamination study was carried out at three sites namely Gweru and Bindura (2020/2021 rainy season) and Shamva (2021/2022 rainy season). Smooth pigweed was planted at distances of 3 to 150 m from pure grain Amaranth. Close monitoring of phenology and measurement of meteorological conditions such as temperature, wind speed and direction were done during the anthesis period. Cross-pollination in grain Amaranth was influenced by distance from the pollen source. A maximum of 12.9 % contamination rate was measured within the 10 m radius from the pollen donor plot and a minimum outcrossing rate of 0.45 % was recorded at the widest distance of 150 m. It can be concluded that complete removal of wild relatives of grain Amaranth is important within the field at as far as 150 m from the crop.

(1) Background: In most cases, passive isolation control methods are commonly used for the seismic isolation design of bridge engineering. However, for passive seismic isolation devices, due to their non-adjustable performance parameters, it is challenging to achieve a good seismic isolation effect for structures under random and variable seismic loads in the wide frequency range of 0 Hz to 20 Hz; (2) Methods: the sensitivity of the design parameters of the seismic isolation bearing was analyzed using the optimization center gradient method, and an improved genetic algorithm was employed to quickly optimize and obtain the optimal design parameters; (3) Results: the effectiveness of the three-dimensional seismic isolation bearing was validated through experiments. (4) Conclusions: The multi-factor sensitivity analysis approach used in this study for designing novel isolation bearings is applicable not only to seismic design in bridges but also serves as a reference for parameter design in isolation bearings requiring medium to high precision in seismic performance.

With the fast growth of smartphones and digital cameras, massive images are generated every day in the world. They are easily transmitted on the insecure channel-Internet. It has been a hot spot for protecting sensitive images during communication. A new cryptosystem is proposed using a six dimensional (6D) hyperchaotic system and DNA computing. The hash value is obtained with the function of SHA-512. It keeps the encrypted result closely connected with the original image. The initial values of the cryptosystem are produced with the hash value and the secret key. The pixel is divided into four parts, and a large matrix is formed. Scrambling is performed on the new image. DNA coding, modern DNA complementary rules, DNA computing, and DNA decoding are performed on the new image. Diffusion is also executed, and ciphered image is achieved finally. The experimental result reveals the outcome of the proposed algorithm. Security analysis shows that the designed algorithm has a huge secret key space, low correlation, and high sensitivity. It also signifies that the designed algorithm could protect against common attacks and is more secure than some existing methods.

Many types of banana (Musa spp.)—one of the most economically important horticultural crops—exist, and their ploidy (diploid, triploid, and tetraploid, usually) and genome types (containing A or/and B genome in most cases) differ. At present, observation and genome type detection are commonly used to identify banana germplasm resources. However, the former is tedious, while the latter cannot distinguish categories below genome types. It is therefore urgent that a simple and effective method for identifying banana germplasm resources is established. We sequenced and analyzed the ribosomal DNA internal transcribed spacer (ITS) sequences of 62 banana germplasm and found that the sequencing peaks, especially the 20 bp region near the 420 bp position (referred to as the “420 bp region”), exhibited relatively recognizable and repeatable polymorphism characteristics. Using the “420 bp region” as a marker, we were able to quickly distinguish bananas belonging to different genome type groups (comprising the AA, AB, AAA, ABB, AAB, ABBB, AAAB, and AAAA groups) or different subgroups in the same genome type group (for example, the Cavendish, Gros Michel, Red, Lakatan, and Ibota Bota subgroups in the AAA group; and the Pisang Raja, Plantain, Pome and Silk subgroups in the AAB group). Moreover, it appeared that Sanger sequencing of ITS could be used for identifying banana hybrid offspring. These results demonstrated that the banana ITS region contains rich genetic information that is useful for genotyping. In general, ITS sequencing simplifies the classification of banana germplasm resources and has potential application in several areas of Musa improvement.

Roof and roofings built to protect the housing and homeowner from undesirable rainwater, have been seen an important structural element throughout history in the entire of housing. In this research, structural and spatial relationships between building envelope with isolation from water in traditional housing architecture are analyzed. Roof, eaves and rain gutters which are isolation from water items were investigated according to shapes, slope direction, materials, carrier systems and covering materials. The qualification of the roofing, eave and rain gutter elements which are equipment elements related to the comfort of the building envelope and the extent of changes in these elements were examined at the traditional housing level. Exemplary, the traditional housing in the city of Trabzon in Turkey were selected. During the completion process of the working, detailed aims related to structure-health reinforcements were specified with the relevant literature review. The chart form has been developed for compilation of information about the general characteristics of the housing in the research area and the building-health reinforcements within the aim of the research. Exemplary houses were selected by external observation technique and the data obtained as a result of observations, interview and determinations related to general characteristics and building-health reinforcements of these housing were handled into a chart form. By evaluating the data obtained; harmony and contrasts in system level practices, the dimensional properties of the elements at the spatial level, the greatness and accuracy levels preferred in various fittings, the harmony and contrasts in the materials, dimensions and practices of the elements at the system and spatial level, harmony and contrasts of applications of new opportunities at the level of building-health reinforcements in traditional housing, conclusions were drawn to identify special and general situations at the level of building-health reinforcements and to obtain information about the comfort requirements of traditional housing. Nowadays, where the health of the building, which is mostly ignored, is gaining importance day by day, concrete and applicable solution data are presented and interpreted for new designs to be made in this direction.

The uterine leiomyoma is the most common benign tumor in women of childbearing age. It may lead to problem of conception or complications during gestational period. The methods of treatment can be surgical (myomectomy and hysterectomy, embolization of arteries) and therapeutic treatment (Ulipristal acetate, Leuprolide acetate, Cetrorelix, Goserelin, Mifeprestone). Both approaches are efficient, but are incompatible with pregnancy planning. Therefore, there is a call for medical practice in developing therapeutical means of preventing leiomyoma onset in patients planning on pregnant. Based on the analysis of GWAS data on the search for mononucleotide polymorphisms associated with the risk of leiomyoma, meta-transcriptomic and meta-methylomic studies, target proteins have been proposed. Prospective therapeuticals of leiomyoma may be based on chemical compounds, humanized recombinant antibodies, vaccines based on markers of the uterine leiomyoma cells that are absent in the adult organism, DNA and RNA preparations. Three different nosological forms of the disease associated with driver mutations in the MED12, HMGA2 and FH genes should be considered when developing or prescribing drugs. E.g. synthetic inhibitors and vaccines based on matrix metalloproteinases MMP11 and MMP16 are expected to be effective only for the prevention of the occurrence of MED12-dependent nodules.

Dried blood spot (DBS) testing on confetti is one of the most common methods used to collect and extract DNA from field samples for molecular epidemiology of malaria studies. Here, we investigated whether ethanol or other solutions used to clean scissors are a source of sample contamination. DBS were prepared from 3 drops of blood from two Plasmodium falciparum cultures spotted on confetti, and 3 solutions (water, ethanol and DNase) were used to clean scissors between spots during DNA extraction. For each cleaning solution, two blank confetti were used as negative controls. Samples were analyzed by PCR-based genotyping of merozoite surface proteins 1 & 2 (msp-1 and msp-2), and P. falciparum chloroquine resistance transporter (Pfcrt). A total of 15 samples were analyzed. Based on msp-1 and msp-2 amplification, P. falciparum was detected on blank confetti when scissors were washed with ethanol. Based on Pfcrt amplification, DNA of P. falciparum was detected on blank confetti when scissors were cleaned with ethanol or water. No P. falciparum genes were detected on blank confetti when scissors were cleaned with DNase. Scissors cleaned with DNase prevented cross-contamination between samples during processing of dried blood spots, whereas ethanol (which is commonly used) fails to avert cross-contamination.

Background: Breast cancer and other neoplasms, such as acute lymphocytic leukaemia, are a severe hazard to public health globally. L-glutamine-degrading enzymes are required for the treatment of malignancies such as acute lymphocytic leukemia and breast cancer. L-glutaminase, an amidohydrolase, has been shown to be an effective chemotherapeutic tool for treating a variety of malignancies. The aim of the study: Molecular characterization is used in the study and production of bacterial L- glutaminase as an anticancer agent from a variety of Egyptian soil locales. The type of the study: Screening experimental study. Methodology: In the current study, many bacterial isolates were tested on mineral L-glutamine agar selective medium (MGA) to evaluate if they generated L-glutaminase. The most effective positive bacterial isolates that produce L-glutaminase, however, were discovered by morphological and biochemical analysis. The primary positive isolate producing L-glutaminase was also identified utilizing molecular detection by 16S rRNA sequencing homology technique. The properties of bacteria-produced L- glutaminase were investigated. The anticancer activity was assessed with an MTT assay. Results: Sole bacterial isolates that only needed L-glutamine for metabolic nitrogen grew well on MGA. The optimal conditions for producing positive bacterial isolates were pH 7.4 and 37 ̊C. The morphological and biochemical investigations of soil samples from diverse soil conditions in Egypt revealed that Bacillus subtilis subsp. niger (ATCC 9372) was the primary positive bacterial isolate producing L-glutaminase. At a pH of 7.4 and a temperature of 37 °C, the activators KCL, ZnSO4, FeSO4, K2HPO4, and MgSO4 offered ideal conditions for defining L-glutaminase production. L-glutaminase from Bacillus subtilis was detected to be a highly bioavailable and efficient anticancer treatment. The original yield [productivity] from MGA was 6.3 U/ml, but invitro coupled transcription and translation production raised it to 89± 1 U/ml. L-glutaminase had a molecular mass of approximately 36 kDa. The purification of L-glutaminase resulted total activity of 21,883 ± 15.67 (U), specific activity of 379.14 ± 7.35 (U/mg of protein), and purification fold of 1.82 ± 3.06. The enzymatic activators Ni2+ and Mg 2+ increased L-glutaminase activity by 20%, 27%, respectively. The compound showed significant anticancer activity (IC50 = 38.72, 8.6, 6.41, 19.02, 49.83, and 9.45 μg/ml) against cancer cell lines from the liver (HepG-2), colon (HCT- 116), breast (MCF-7), lung (A-549), lymphocytic (CCL-120), and cervical (Hela). The kinetic parameters Km and Vmax were 15.2^ 10 -3 M and 119.86 μmol/ml/min, indicating a higher affinity for the substrate. The enzyme demonstrated high efficacy at pH 6.5 and temperature 35. The enzyme was thermostable at 58 ̊C for 1 hour. Conclusion: Acute lymphocytic leukemia and breast cancer are two instances of auxotrophic malignancies for which L-glutamine is the sole metabolic source, and Bacillus subtilis- derived L-glutaminase was a suitable enzyme in their treatment.

As the fast growth of information technology (IT), the safety of image transmission and the storing of images are increasingly concerned. Traditional image encryption algorithms have certain limitations in transmission and security, so there is an urgent need for a secure and reliable image encryption algorithm. A universal compression sensing (CS) image encryption (IE) algorithm based on DNA-triploid mutation (DTM) is presented in this paper. Firstly, by using CS algorithm an image is compressed while obtaining a range of chaotic sequences by iteration of chaotic map. Then DNA sequences are generated by encoding the image, and based on the DTM, new mutant DNA sequences are generated according to specific rules. Next, the chaotic sequences are operated at the DNA level to do confusion and diffusion operations on the image to ensure the security of the data. At last, DNA decoding is carried out to obtain the compressed-encrypted image. Both simulation experiments and performance tests fully show that a high level of security and reliability of the proposed algorithm in protecting image privacy is achieved.

Abstract: Congenital cytomegalovirus (cCMV) infection poses significant risks to fetal development, particularly affecting the nervous system. This study reports a fetal autopsy case, examining cCMV infection and focusing on CMV DNA measurements in various fetal organs before formalin fixation, a novel approach for comprehensive CMV DNA evaluations in fetal organs affected by cCMV. A 20-week-old male fetus was diagnosed with cCMV following the detection of CMV DNA in ascites obtained via abdominocentesis in utero. After termination of pregnancy, multiple organs of the fetus, including the cerebrum, thyroid gland, heart, lungs, liver, spleen, kidneys, and adrenal glands, were extracted and examined for CMV DNA loads using real-time polymerase chain reaction. Histopathological examination involved hematoxylin-eosin and CMV-specific immunostaining. A correlation was found between CMV DNA loads and pathology, with higher CMV-infected cell numbers observed in organs positively identified with both staining methods, exhibiting CMV DNA levels of ≥1.0 × 104 copies/mL, compared to those detected solely by CMV-specific immunostaining, where CMV DNA levels ranged from 1.0 x 103 to 1.0 x 104 copies/mL. These results highlight a quantifiable relationship between organ infection extent and CMV DNA concentration, providing insights into cCMV pathogenesis and potentially informing future diagnostic and therapeutic strategies for cCMV infection.

The usage of Quantum Similarity through the equation Z = {∀θ ∈ Z → ∃s ∈ S ∧ ∃t ∈ T : θ = (s, t)}, represents a way to analyze the way communication works in our DNA. Being able to create the object set reference for z being (s, t) in our DNA strands, we are able to set logical tags and representations of our DNA in a completely computational form. This will allow us to have a better understanding of the sequences that happen in our DNA. With this approach, we can also utilize mathematical formulas such as the Euler–Mascheroni constant, regression analysis, and computational proofs to answer important questions on Quantum biology, Quantum similarity, and Theoretical Physics.

Consumption of refined foods and beverages high in sugar make the teeth susceptible to the formation of biofilm, and lead to dental caries and diseases of the oral cavity such as periodontitis and periodontal disease. The aim of the present study was to determine the ability of selected probiotics to inhibit growth and biofilm formation by the cariogenic bacterium Streptococcus mutans in vitro. We screened strains of latic acid bacteria (LAB) (n=120) from the Bioresources Collection and Research Center (BCRC), saliva of healthy adults and infant stool. The antimicrobial activity of LAB in vitro was evaluated by agar spot culture and co-culture of the S. mutans strains. We determined the effect of heating and dilution factors (2- and 4-fold) on the antagonistic activity of LAB. Antagonistic substances in the spent culture suspensions (SCS) of LAB were precipitated by extraction with ammonium sulphate and chloroform to characterise the protein and lipophilic fractions. Results of co-culturing show that the SCS of the three LAB strains (Lactobacillus pentosus 13-1, 13-4 and Lactobacillus crispatus BCRC 14618) subjected to heat treatment showed significantly high antimicrobial activity. We found that substances produced by L. pentosus 13-4 which have the potential to exhibit antimicrobial properties might be lipophilic proteins. Additionally, we infer that the mechanism of reducing biofilm formation by Lactobacillus strains is associated with sucrose-dependent cell–cell adhesion and the gtfC level of glucosyltransferases (Gtfs) in the biofilm. Native LAB strains screened in our study may be used in chewing gums and other processed foods for preventing tooth decay.

Hepatocellular carcinoma (HCC) development is associated with altered modifications in DNA methylation, changing transcriptional regulation. Emerging evidence indicates that DNA methyltransferase 1 (DNMT1) plays a key role in carcinogenesis process. This study aimed to investigate how pirfenidone (PFD) modifies this pathway and the effect generated by the association between c-Myc expression and DNMT1 activation. Rats F344 were used for HCC development using 50 mg/kg of diethylnitrosamine (DEN) and 25 mg/kg of 2-aminofluorene (2-AAF). HCC/PFD group received simultaneous doses of 300 mg/kg of PFD. All treatments lasted 12 weeks. On the other hand, HepG2 cells were used to evaluate the effects of PFD in restoring DNA methylation in the presence of the inhibitor 5-Aza. Histopathological, biochemical, immunohistochemical, and western blot analysis were carried out and our findings showed that PFD treatment reduced the amount and size of tumors along with decreased Glipican-3, β-catenin, and c-Myc expression in nuclear fractions. Also, this treatment improved lipid metabolism by modulating PPARγ and SREBP1 signaling. Interestingly, PFD augmented DNMT1 and DNMT3a protein expression, which restores global methylation, both in our in vivo and in vitro models. In conclusion, our results suggest that PFD could slow down HCC development by controlling DNA methylation.

Context: The global escalation of conflict, violence, and human rights violations sets a pressing backdrop for examining the resilience of forcibly displaced youth (FDY) in Canada. This study aims to unpack the multifaceted challenges and resilience mechanisms of FDY, focusing on their health, well-being, and integration into host communities. It seeks to identify current models of resilience, understand the factors within each model, and highlight gaps and limitations. Methodology: Using a university librarian-supported structured search strategy, this exploratory rapid review searched literature from Ovid Medline and Open Source databases, published in English between January 2019 and January 2024, that fit specific inclusion criteria. The eligible articles (N=12 out of 4,096) were charted and analyzed by two student researchers with the PI. Charted data were analyzed thematically. Results: The selected studies captured diverse geographical perspectives, resilience models (such as Ungar's ecological perspective and Masten's resilience developmental models), as well as protective and promotive frameworks. Key findings indicate the complexity of resilience influenced by individual, familial, societal, and cultural factors. Each model offers insights into the dynamic interplay of these influences on FDY's resilience. However, these models often fall short of addressing the nuances of cultural specificity, the impact of trauma, and the intersectionality of FDY's identities. Conclusion: Recognizing the diverse and evolving nature of FDY's coping mechanisms, this study advocates for a culturally appropriate approach to resilience that integrates an intersectionality framework of individual attributes and culturally sensitive models.

Zucchini (Cucurbita pepo L.) is an important crash crop for producing seeds in Inner Mongolia, China. In August 2021, the harvest season of seed-producing zucchini, fruit rot symptom (cv. Jindi No.1) was found in the Wuyuan region. The incidence of infected fruits of zucchini ranged from 10% to 30% and symptom consisted in 4 to 5 cm diameter lesions where the fruit was in contact with the soil. Ten isolations from six symptomatic fruits were preliminary classified as Fusarium species based on its morphological characters in potato dextrose agar. Phylogenetic tree was constructed by concatenating TEF-1α and RPB2 gene and exhibited the isolates clustered together with the Fusarium solani f. sp. cucurbitae sequences. Pathogenicity tests revealed that two isolates caused over 3 cm diameter lesions on the fruits while negative-controls remained asymptomatic. The colonies reisolated from the leasions were morphologically identical to the original isolates, thus fulfilling Koch’s postulates. Results demonstrated that F. solani f. sp. cucurbitae was the causal agent of fruit rot observed in the field. To our knowledge, this is the first report of fruit rot of seed-producing zucchini caused by F. solani f. sp. cucurbitae in Inner Mongolia, China.

The environment significantly impacts the life of bees and their feeding. This study aimed to investigate bee foraging using melissopalynological analysis and DNA metabarcoding in ​​intensive farming, reserved, and urbanized areas. The highest alpha diversity was observed in the reserved and in the intensive farming areas. The urbanized area has less diversity. In the intensive farming area predominated: Sinapis, Helianthus, and Fagopyrum; in the reserved area: Melilotus, Helianthus, and Brassica. In the urbanized area garden plants: radish (Raphanus sativus) and cucumber (Cucumis hystrix) and agricultural plants: soybean (Glycine max) and melon (Cucumis melo) were often found. The most significant agreement was between the rbcL and the melissopalynological analysis. The ITS2 revealed equal matches with both rbcL and melissopalynology but this marker missed or underestimated some genera. Trifolium pretense and Brassica nigra, were identified simultaneously by three analyses. Species Convolvulus arvensis, Melilotus officinalis, Echium vulgare, Brassica rapa, Helianthus divaricatus, and Onobrychis viciifolia were found in all ecological areas. Imperfect databases put some limits for identification of some taxa using metabarcoding. Further research and expansion of plant databases is needed. Studying the food preferences of bees in different environmental conditions and landscapes is necessary to develop measures to preserve their population.

This study analyzes the modeling and dynamics of a novel passive in Multi-Degree-of-Freedom (MDOF) vibration isolation platform which can achieve significant isolation effect. Symmetrical Scissor-Like structures (SLSs) are utilized in the proposed MDOF isolation platform as the supporting and isolation elastic components. Based on the mathematical modeling and theoretical analysis of the MDOF vibration isolation system with SLSs, the effect of structural parameter and joint friction on stiffness and damping properties is investigated. It is shown that due to geometric relations within the SLSs, the natural frequencies can be reduced via adjusting structural parameters of the SLS for different direction vibration isolation. Theoretical and experimental results show that the SLS isolation platform can achieve much better loading capacity and vibration isolation performance simultaneously by only using linear passive components because of the MDOF adjustable stiffness property. Therefore, with low costing and energy consumption, the proposed novel isolation platform can provide the improvement of vibration suppression in various engineering practices.

Environmental DNA (eDNA) sampling has emerged as a powerful approach for monitoring biodiversity in the context of diverse terrestrial and aquatic ecosystems, and has broad potential for both environmental surveys and species management. eDNA studies focusing on plant and animal species have capitalized on analytical methodologies developed originally for metagenomic studies of microorganisms, and on technological improvements in sample collection. However, while eDNA studies have proliferated rapidly, efforts to standardize sampling and analytical methods are still nascent. Critically, fundamental data concerning the distribution and persistence of eDNA in different ecological contexts are lacking, complicating both confidence in species detection and comparability between studies. We propose that key deficits could be addressed by using well-designed synthetic DNA standards or tracers, which are widely used in fields ranging from medical diagnostics to hydrology. In-field application of a standardized mixture of barcoded synthetic DNA fragments of varying lengths and concentrations that are recovered concommitantly with eDNA sampling should provide critical yet missing data on environmental DNA distribution and turnover, as well as an internal molecular standard to enhance both species detection and comparability between eDNA studies.

Adenoviruses (Ads) have been extensively manipulated for the development of cancer selective replication, leading to cancer cell death or oncolysis. Clinical studies using E1-modified oncolytic Ads have shown that this therapeutic platform was safe, but with limited efficacy, indicating the necessity of targeting other viral genes for manipulation. To improve the therapeutic efficacy of oncolytic Ads, we treated the entire Ad genome repeatedly with UV-light and have isolated AdUV which efficiently lyses cancer cells as reported previously [1]. In this report, we show that no mutations were observed in the early genes (E1 or E4) of AdUV while several mutations were observed within the Ad late genes which have structural or viral DNA packaging functions. This study also reported the increased release of AdUV from cancer cells. In this study, we found that AdUV inhibits tumor growth following intratumoral injection. These results indicate the potentially significant role of the viral late genes, in particular the DNA packaging genes, to enhance Ad oncolysis.

Mango (Mangifera indica L.) is an allotetraploid (2n = 4X= 40) drupe fruit and has high nutritional value belongs to genus Mangifera and family Anacardiaceae. Mango cultivars are used with worldwide acceptance to pharmacological, ethnomedical, and phytochemical industries. Assessment of the genetic distinctiveness of a cultivar through morphological descriptors is an important tool for both the registration and the protection. New mango genotypes have been improved using valuable diverse germplasm resources to ensure food security. DNA fingerprinting based simple sequence repeats (SSR)-markers have been the most broadly used, effective and accurate in evaluation of genetic characterization of a cultivar. Molecular breeding is an effective source of genetic gain after improvement of fruit trees using marker assisted genomic selection. Total genomic DNA (gDNA) was generated using CTAB method from each cultivar. The most effective 50 hyper-variable SSR markers were selected. Highly specific DNA fingerprints were identified in the candidate line ‘Azeem Chaunsa’ compared with three standard cultivars using SSR-PCR. An agglomerative hierarchical clustering method was used to construct dendrogram based on the UPGMA clustering method. Cultivar identification diagram (CID) was constructed to evaluate association among standard cultivars and Azeem Chaunsa. Our results showed that SSR markers could efficiently assess genetic diversity in mango. The genetic similarity coefficients were recorded between the cultivars of mango ranged from 0.49 to 0.67. CID results concluded that cultivar ‘Azeem Chaunsa’ varied significantly from the check cultivar, Sindhri (46.2%), S.B Chaunsa (45%) and Sufaid Chaunsa (46.7%). The results obtained in this study will orient cultivar identification strategies for a successful future.

Model-based fault diagnosis serves as a powerful technique for addressing fault detection and isolation issues in control systems. However, diagnosing faults in closed-loop control systems is more challenging due to their inherent robustness. This paper aims to detect and isolate actuator and sensor faults in the cascade electro-hydraulic control system of a turbofan engine. Based on the fault characteristics, we design a robust unknown perturbation decoupling residual generator and an optimal fault observer specifically for the inner and outer control loops to detect potential faults. To locate the faults, we analyze the steady-state propagation laws of actuator and sensor faults within the loops using the final value theorem. Based on this, we establish the minimal-dimensional fault influence distribution matrix specific to the cascade turbofan engine control system. Subsequently, we construct the normalized residual vectors and monitor its vector angles against each row of the fault influence distribution matrix to isolate faults. Experiments conducted on an electro-hydraulic test bench demonstrate that our proposed method can accurately locate four typical faults of actuators and sensors within the cascade electro-hydraulic control system. This study enriches the existing fault isolation methods for complex dynamic systems, and lays the foundation for guiding component repair and maintenance.

DNA origami has emerged in recent years as a powerful technique for designing and building 2D and 3D nanostructures. While the breadth of structures that have been produced is impressive, one of the remaining challenges, especially for DNA origami structures intended to carry out useful biomedical tasks in vivo, is to endow them with the ability to detect and respond to molecules of interest. Target molecules may be disease indicators or cell surface receptors, and the responses may include conformational changes leading to release of therapeutically relevant cargo. Nucleic acid aptamers are ideally suited to this task and are beginning to be used in DNA origami designs. In this review we consider examples of uses of DNA aptamers in DNA origami structures and summarise what is currently understood regarding aptamer-origami integration. We review three major roles for aptamers in such applications: protein immobilisation, triggering of structural transformation, and cell targeting. Finally, we consider future perspectives for DNA aptamer integration with DNA origami.

Early weaning management followed by energy supplementation can lead to metabolic alterations in the calf that exert long-term effects on the animal’s health and performance. It is believed that the main molecular basis underlying these metabolic adaptations are epigenetic mechanisms that regulate, activate, or silence genes at different stages of development and/or in response to different environmental stimuli. However, little is known about postnatal metabolic programming in Bos indicus. Therefore, this study aimed to compare the DNA methylation profile of Nellore animals submitted to conventional and early weaning and to correlate the findings with genes differentially expressed in Longissimus thoracis skeletal muscle of Bos indicus cattle. A total of 481 differentially methylated regions were identified, with 52% (250) being hypermethylated and 48% (231) hypomethylated. Functional enrichment analysis of 53 differentially methylated and differentially expressed genes was performed. The main enriched terms and pathways were associated with 3’-5’-cyclic adenosine monophosphate (cAMP) signaling which present the upregulated adenylate cyclase 3 (ADCY3) gene and significatively hypomethylated in promoter region. Alterations in cAMP signaling are involved in numerous processes, many of them related to lipid metabolism. The relative differential expression of key genes of this pathway demonstrates the relationship between cAMP signaling and de novo lipogenesis. These findings suggest an important role of postnatal metabolic programming through DNA methylation mechanisms in determining fat deposition in beef.

A hybrid material, consisting of commercially available nitrocellulose (NC) membrane non-covalently modified with amino-polyethylene glycol functionalized reduced graphene oxide (NH2-PEG-rGO) nanoparticles, was successfully synthesized for oligonucleotide extraction. Fourier Transform Infrared Spectroscopy (FTIR) confirmed the modification of the NC membrane, revealing characteristic peaks of both compounds NC and NH2-PEG-rGO. Scanning Electron Microscopy (SEM) exhibited morphological changes in the NC/NH2-PEG-rGO hybrid membrane, marked by the introduction of NH2-PEG-rGO particles, resulting in a distinctly smothered surface compared to the porous surface of the NC control membrane. Wettability assays revealed hydrophobic behavior for the NC/NH2-PEG-rGO hybrid membrane, with a water contact angle exceeding 90°, contrasting with the hydrophilic behavior characterized by a 16.7° contact angle in the NC membrane. The performance of the NC/NH2-PEG-rGO hybrid membrane was evaluated for the extraction of ssDNA with fewer than 50 nucleotides from solutions containing various ionic species (MnCl2, MgCl2, and MnCl2/MgCl2). The NC/NH2-PEG-rGO hybrid membrane exhibited optimal performance when incubated in MgCl2, presenting the highest fluorescence emission at 525 relative fluorescence units (r.f.u.). This corresponds to the extraction of approximately 610 pg (≈13%) of the total oligo-DNA, underscoring the efficacy of the pristine material, which extracts 286 pg (≈6%) of oligo-DNA in complex solutions.

Optical tweezers offer a non-contact method for selecting single cells and translocating them from one microenvironment to another. We have characterized the optical tweezing of yeast S. cerevisiae and can manipulate single cells at velocities up to 0.77 mm/s using laser powers of 40 mW from a 785 nm diode laser. We have fabricated and tested three cell isolation devices; a micropipette, a PDMS chip and laser machined fused silica chip and we have isolated single bacteria, yeast and cyanobacteria cells. The most effective isolation was achieved in PDMS chip, where single yeast cells were grown and observed for 18 hours without contamination. The duration of budding in S. cerevisiae was not affected by the laser parameters used, but the time from tweezing until the first budding event began increased with increase laser energy (laser power x time). Cells tweezed using 25 mW for 1 minute were viable after isolation. We have constructed a micro-consortium of yeast cells, and a co-culture of yeast and bacteria, using optical tweezers in combination with the PDMS network of channels and isolation chambers, which may impact on both industrial biotechnology and understanding pathogen dynamics.

A simulation is performed to analyze the forces that arise in a conducting DNA-like double helix in the field of a microwave electromagnetic wave under half-wave resonance conditions. The helix is comprised of twenty-and-a-half turns and has geometric parameters proportional to the size of an actual DNA molecule. The forces acting on the strands of a double helix, both in the central region and at the edges of the helix, are investigated. It has been demonstrated that the afore-mentioned forces induce a change in the shape of the helix, specifically mutual repulsion of the strands, as well as their stretching and twisting in the field of electromagnetic waves. As opposed to a pair of parallel straight conductors, there is a radial repulsion of strands in all regions of a double DNA-like helix, including the center and the edges. This unique interaction of currents in a double DNA-like helix can be attributed to its pitch angle, a characteristic that has been estab-lished through empirical evidence. Consequently, exposure to an electromagnetic wave under half-wave resonance can damage the double helix. Conversely, the impact of electromagnetic waves has the potential to introduce novel avenues for controlling the shape of the double helix. These nonlinear effects must be considered for both actual DNA molecules and double DNA-like helices that serve as components of metamaterials and metasurfaces.

Wheat is threatened by global warming. This study aimed to characterize based on DNA profile and identify heat-tolerant wheat genotypes. Fifteen wheat genotypes were sown in Nov 21 and Dec 21 and evaluated within 13 SSR markers. A dendrogram classified the genotypes into two main groups. The highest genetic distances (0.93) were drawn between BAW 1290 and BARI Gom 28, and the lowest one between Nadi 2 and BAW 1147. Heat susceptibility index (HSI) values for 1000-grain weight (TGW) and grain yield indicated moderate heat tolerance in Cluster I genotypes (HSI 0.50-1.00), closely matching Group A's Cluster II members. In Group B, Cluster III genotypes had higher HSI (1.04-1.23) and significant reductions in TGW and grain yield under heat stress, indicating heat susceptibility. The varieties BARI Gom 25, 26, 27, 28, 29, and 30 were identified as heat-stress tolerant. This research provides insights for future heat stress-molecular breeding programs.

Chromothripsis is a mutational mechanism leading to complex and relatively clustered chromosomal rearrangements resulting in diverse phenotypic outcomes depending on the involved genomic landscapes. It may occur both in the germ and the somatic cells resulting in congenital and developmental disorders and cancer, respectively. Asymptomatic individuals may be carriers of chromotriptic rearrangements and experience recurrent reproductive failures when two or more chromosomes are involved. Several mechanisms are postulated to underly chromothripsis. The most attractive hypothesis involves chromosome pulverization in micronuclei followed by incorrect reassembly of fragments through DNA repair to explain the clustered nature of the observed complex rearrangements. Moreover, exogenous or endogenous DNA damage induction and dicentric bridge formation may be involved. Chromosome instability is commonly observed in the cells of patients with DNA-repair disorders, such as ataxia telangiectasia, Nijmegen breakage syndrome and Bloom syndrome. In addition, germline variations of TP53 have been associated with chromothripsis in Sonic-Hedgehog medulloblastoma and acute myeloid leukemia. In the present review, we focus on the underlying mechanisms of chromothripsis and the involvement of defective DNA-repair genes resulting in chromosome instability and chromothripsis-like rearrangements.

Lumpy skin disease (LSD), caused by a virus within the Poxviridae family and Capripoxvirus genus, induces nodular skin lesions in cattle, spreading through direct contact and insect vectors, significantly affecting global cattle farming. Despite available vaccines, their effectiveness is limited by poor bioavailability and adverse effects. Our study aimed to identify potential inhibitors targeting LSD-associated DNA polymerase protein, selecting LSDV039 for further investigation through comprehensive analysis and computational methods. Virtual screening revealed rhein and taxifolin as potent binders among 380 phytocompounds, with respective affinities of -8.965 and -7.195 kcal/mol. Canagliflozin and tepotinib exhibited strong affinities (-9.858 and -8.856 kcal/mol) among 718 FDA-approved antiviral drugs. Molecular dynamics simulations of canagliflozin, tepotinib, rhein, and taxifolin highlighted taxifolin's superior stability and binding energy. Rhein displayed compactness in RMSD and RMSF but fluctuations in Rg and SASA, while canagliflozin demonstrated stability compared to tepotinib. The study highlights the promising potential of repurposed drugs and phytocompounds as LSD therapeutics. However, extensive validation through in-vitro, in-vivo, and clinical trials is crucial for their practical application.

Extracellular vesicles (EVs) carry diverse cargo of DNA, RNA, proteins, and lipids. They are crucial mediators for regulating various functions through interactions with the host. Recent investigations suggest that EVs or their constituent proteins derived from probiotics could improve the effectiveness of probiotic strains in disease management compared to probiotic strains themselves. This enhanced effec-tiveness arises from their innate antimicrobial properties, which can specifically target pathogenic mol-ecules or cells, in addition to their significant ability to regulate immune responses. Significantly, previous studies have highlighted the inhibitory potential of Lactobacillus crispatus PMC201 against Mycobacterium tuberculosis. In this study, we focused on optimizing isolation conditions for studying extracellular vesicles and proteins from anti-TB potent strains. We provide analysis results based on food-grade media (FGM) optimization and isolation methods. Nanoparticle tracking analysis (NTA) was used to determine se-cretion patterns. Proteomic analysis was performed by liquid chromatography and tandem mass spec-trometry (LC-MS/MS). A total of 64 proteins were identified in the L. crispatus PMC201 strain, particularly enriched in s-layer and splX. Questions remain about their relevance to anti-TB. This anti-tuber strain has potential as a therapeutic agent, given its significant role in intercellular killing. Further studies are needed to investigate its mechanisms of action.

Non-viral mediated plasmid DNA transfection by electroporation (EP) is an established method for gene transfection. In this study, the usefulness of direct EP at an intradermal (i.d.) site (D_EP) with implanted electrodes to achieve a high protein expression level was investigated. In addition, D_EP application with various intervals with a low application voltage was also evaluated to confirm its effect on protein expression. A higher protein expression level was observed after green fluorescent protein (GFP)- and luciferase-encoding DNA were delivered by i.d. injection followed by D_EP application. When luciferase expression was observed with an in vivo imaging system, continuous expression was confirmed over 21 days after i.d. injection followed by D_EP at 100 V. This approach provided increased gene expression levels compared with conventional EP methods via the stratum corneum layer. In addition, the effect of application voltage on luciferase expression was investigated; two-time applications (repeated D_EP) at 20 V with 5 min intervals showed similar luciferase expression level to single D_EP application with 100 V. Histological observations showed the skin became thicker after a single D_EP at 100 V, whereas no apparent thickness changes was confirmed after repeated D_EP at 20 V with 5 min intervals. This study revealed that direct i.d. voltage application achieved high protein expression levels even at low voltages. Skin is a promising administration site for DNA vaccines, so this approach may be effective in DNA vaccine delivery into skin tissue.

Bats are of significant interest as reservoirs for various zoonotic viruses with high diversity. During the past two decades, many herpesviruses have been identified in various bats worldwide by genetic approaches, whereas there have been few reports on the isolation of infectious herpesviruses. Herein, we report the prevalence of herpesvirus infection of bats captured in Zambia and genetic characterization of novel gammaherpesviruses isolated from Macronycteris vittatus bats. By our PCR screening, herpesvirus genomes were detected in 29.2% (7/24) of Rousettus aegyptiacus, 78.1% (82/105) of Macronycteris vittatus, and 100% (1/1) of Hipposideros caffer bats in Zambia. Phylogenetic analyses of the detected herpesvirus genomes revealed that the Zambian bat herpesviruses were divided into seven betaherpesvirus groups and five gammaherpesvirus groups. Two infectious strains of a novel gammaherpesvirus, tentatively named Macronycteris gammaherpesvirus 1 (MaGHV1), were successfully isolated from Macronycteris vittatus bats, and their complete genomes were sequenced. The genome of MaGHV1 encoded 79 open reading frames, and phylogenic analyses of the DNA polymerase and glycoprotein B demonstrated that MaGHV1 formed an independent lineage sharing a common origin with other bat-derived gammaherpesviruses. Our findings provide new information regarding the genetic diversity of herpesviruses maintained in African bats.

The following study reports on the first thraustochytrid isolates identified from Iceland. They were collected from three different locations off the northern coast of the country (Location A, Skagaströnd; Location B, Hveravík; and Location C, Eyjafjörður). Using 18S rDNA sequence analysis, isolates from Locations A and B were identified within the Thraustochytrium kinnei species while other isolates within the Sicyoidochytrium minutum species when compared to other known strains. Cells isolated from Locations A (2.10 ± 0.70 g/L) and B (1.54 ± 0.17 g/L) produced more biomass than the ones isolated from Location C (0.43 ± 0.02 g/L). This study offers the first-time examination of the utility of byproducts from fisheries as a nitrogen source in media formulation for thraustochytrids. Experiments showed that isolates produced more biomass (per unit of substrate) when cultured on nitrogen of marine (2.55 ± 0.74 g/L) as compared to of commercial origin (1.06 ± 0.57 g/L). Glycerol (2.43 ± 0.56 g/L) was a better carbon source than glucose (1.84 ± 0.57 g/L) in growth studies. Fatty acid (FA) profiles showed that the isolates from Location C (S. minutum) had low ratios of monounsaturated (4.21 ± 2.96%) and omega-6 (0.68 ± 0.59%) FAs. However, the isolates also had high ratios of docosahexaenoic acid (DHA; 35.65 ± 1.73%) and total omega-3 FAs (40.39 ± 2.39%), indicating that they could serve as a source of marine oils for human consumption and in aquaculture feeds. The T. kinnei isolates from Location A could be used in biodiesel production due to their high ratios of monounsaturated (18.38 ± 6.27%) long chain (57.43 ± 8.27%) FAs.

The efficient preparation of single-stranded DNA (ssDNA) rings, as a macromolecular construction approach with topological features, has aroused much interest due to their numerous applications in biotechnology and DNA nanotechnology. However, an extra splint is essential for enzymatic circularization, and by-products of multimers are usually present at high concentrations. Here, we proposed a simple and robust strategy of using permuted precursor (linear ssDNA) for circularization by forming an intramolecular dynamic nick using a part of the linear ssDNA substrate itself as the template. After the simulation of the secondary structure for desired circular ssDNA, the linear ssDNA substrate is designed to have its ends on the duplex part (≥5 bp). By using this permuted substrate with 5'-phosphate, the splint-free circularization is simply carried out by T4 DNA ligase. Very interestingly, formation of only several base pairs (2–4) flanking the nick is enough for ligation, although they form only instantaneously under ligation conditions. More significantly, the 5 bp intramolecular duplex part commonly exists in genomes or functional DNA, demonstrating the high generality of our approach. Our findings are helpful for understanding the mechanism of enzymatic DNA ligation from the viewpoint of substrate binding.

Natural phytochemicals in foods, including anthocyanins, can play an important role in human health. Anthocyanins have been reported to cause many various useful effects, such as reducing cancer cell proliferation, regulating blood pressure, preventing tumour formation, improving eyesight, and preventing diabetes. In this study, we aimed to reveal the qualitative anthocyanin content, antiproliferative and antimicrobial effects of different extracts derived from Vaccinium myrtillus, V. corymbosum, Sambucus nigra and Aronia melanocarpa. The anthocyanin content of the plants mentioned in the study was characterized after the freeze-drying process. MTT assay was used to determine the antiproliferative effect of extracts on cancer cells. Antimicrobial effects of extracts were studied on typical and clinical strains of 5 different bacteria. As a result, the anthocyanin content in the extracts obtained was determined to be quite good with the freeze-drying method, and it was also determined that the extracts had various levels of antiproliferative and antibacterial effects.

(1) Background: Group A rotaviruses (RVAs) are the primary cause of severe intestinal diseases in piglets. Porcine rotaviruses (PoRVs) are widely prevalent in Chinese farms, resulting in significant economic losses to the livestock industry. However, isolation of PoRVs is challenging, and their pathogenicity in piglets is not well understood. (2) Methods: We conduct clinical testing on a farm in Jiangsu Province, China, and isolate strains of porcine rotavirus by continuously passaging on MA104 cells. Subsequently, the pathogenicity of the isolated strain in piglets was investigated.The piglets of the PoRV-infection group were orally inoculated with 1 mL of 1.0 × 10⁶ TCID50 PoRV, whereas those of the mock-infection group were fed with an equivalent amount of DMEM. (3) Results: Fecal samples were collected from 28 piglets, all of which tested positive for RVAs. A G5P[23] genotype porcine rotavirus strain was successfully isolated from one of the positive samples and named RVA/Pig/China/JS/2023/G5P[23](JS). The genomic constellation of this strain was G5-P[23]-I5-R1-C1-M1-A8-N1-T1-E1-H1. Sequence analysis revealed that the genes VP3, VP7, NSP2, and NSP4 of the JS strain were closely related to human RVAs, whereas the remaining gene segments were closely related to porcine RVAs, indicating a reassortment between porcine and human strains. Furthermore, infection of 15-day-old piglets with the JS strain resulted in a diarrheal rate of 100% (8 of 8) and mortality rate of 37.5% (3 of 8).(4) Conclusion: The isolated G5P [23] genotype rotavirus strain, which exhibited strong pathogenicity in piglets, may have resulted from recombination between porcine and human strains. It may serve as a potential candidate strain for developing vaccines, and its immunogenicity can be tested in future studies.

Objectives: One of the home-made herbal remedy being traditionally used against gout is evaluated to identify any new and characteristic marker compound which may be used as characteristic analytical marker to standardize this polyherbal remedy. Materials and method: A distinct and remarkable peak in the HPLC chromatogram of remedy was identified which was further isolated by fraction collector. This fraction was then evaporated and characterized for structure elucidation using set of various techniques such as RP-HPLC,UV, FTIR, TLC, EIMS and NMR. Results: The compound characterized was non polar compound, ester of betulin and palmitoleic acid. Although the remedy has been standardized using set of four markers as discussed in literature but this newly developed marker compound can be used to further standardize this remedy and other herbal preparations comprising of this marker compound.Conclusion: Hence the isolated compound can be used as novel analytical marker in the standardization of this herbal remedy and the other herbal preparations containing such marker compound

Terminalia arjuna possesses significant cardio protective, antidiabetic and antioxidant properties as these properties are described in Ayurveda. In the present study, three flavonoids were isolated through the separation and chromatographic purification of the whole plant material of T. arjuna. Spectroscopic characterization identified one of them as a new flavonoid “Terminalone A (1)” and two known flavonoids i.e. 6-hydroxy-2-(4-hydroxyphenyl)-7-methoxy-4H-chromen-4-one (2) and 2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-4H-chromen-4-one (3). The bioactivity studies showed considerable antibacterial and antioxidant (DPPH was used as a scavenger) potential for all the three compounds 1-3 where the compound 1 showed strong antibacterial and antioxidant activity.

Ochna kibbiensis (Family: Ochnaceae) has been employed in ethnomedicine for the treatment of malaria, and inflammation among others. The aim of this study was to isolate and characterize prophylactic antimalarial agents from the leaves of O. kibbiensis against Plasmodium berghei, in vivo and in silico. The median lethal dose (LD50) of the methanol extract and its fractions (n-hexane, dichloromethane, ethylacetate and n-butanol) was determined according to Lorke’s method while the antimalarial effect of the extract and its fractions was investigated according to the method described by Peters prophylactic test using Chloroquine-sensitive Plasmodium berghei (NK65). All the extract/fractions exhibited LD50 values ≥ 5000 mg/kg with the exception of the n-butanol fraction (1702.94 mg/kg) which indicate that the plant is non-toxic. Dichloromethane fraction exhibited significant (p<0.05) and dose-dependent prophylactic effect with 47.62, 85.12 and 100.0 % prophylaxis (at 500, 250 & 125 mg/kg) while the least effect was observed by the n-butanol fraction with percentage prophylaxis of 64.29 and 76.19, respectively; the standard drug, pyrimethamine had 95.24 % prophylaxis. Based on the result obtained, dichloromethane fraction of O. kibbiensis was subjected to chromatographic purification which led to the isolation of a mixture of two compounds identified as stigmasterol and β-sitosterol by analysis of the NMR spectral data and comparison with existing literature; the compounds exhibited good binding affinities (- 5.129 and - 4.889 kcal/mol) against pfLDH and favorable ADMET profile. In conclusion, the leaf of O. kibbiensis have demonstrated a significant prophylactic antimalarial activity and the two known steroids (stigmasterol and β-sitosterol) isolated from the dichloromethane fraction for the first time.

The order Mucorales, the largest in number of species within the Mucoromycotina, comprises typically fast-growing saprotrophic fungi. During a study of the fungal diversity of undiscovered taxa in Korea, two novel mucoralean strains, CNUFC-GWD3-9 and CNUFC-EGF1-4, were isolated from specific habitats including freshwater and fecal samples, respectively, in Korea. On the basis of their morphological characteristics and sequence analyses of internal transcribed spacer (ITS) and large subunit (LSU) of 28S ribosomal DNA regions, the CNUFC-GWD3-9 and CNUFC-EGF1-4 isolates were confirmed to be Gilbertella persicaria and Pilobolus crystallinus, respectively. It is ecologically, pathologically and mycologically significant to find such rare zygomycetous fungi in such specific habitats.

This paper describes a novel thruster fault-tolerant control system (FTC) for open-frame remotely operated vehicles (ROVs). The proposed FTC consists of two subsystems: a model-free thruster fault detection and isolation subsystem (FDI) and a fault accommodation subsystem (FA). The FDI subsystem employs fault detection units (FDUs), associated with each thruster, to monitor their state. The robust, reliable and adaptive FDUs use a model-free pattern recognition neural network (PRNN) to detect internal and external faulty states of the thrusters in real time. The FA subsystem combines information provided by the FDI subsystem with predefined, user-configurable actions to accommodate partial and total faults and to perform an appropriate control reallocation. Software-level actions include penalisation of faulty thrusters in solution of control allocation problem and reallocation of control energy among the operable thrusters. Hardware-level actions include power isolation of faulty thrusters (total faults only) such that the entire ROV power system is not compromised. The proposed FTC system is implemented as a LabVIEW virtual instrument (VI) and evaluated in virtual (simulated) and real-world environments. The proposed FTC module can be used for open frame ROVs with up to 12 thrusters: eight horizontal thrusters configured in two horizontal layers of four thrusters each, and four vertical thrusters configured in one vertical layer. Results from both environments show that the ROV control system, enhanced with the FDI and FA subsystems, is capable of maintaining full 6 DOF control of ROV in the presence of up to 6 simultaneous total faults in the thrusters. With the FDI and FA subsystems in place the control energy distribution of the healthy thrusters is optimised so that the ROV can still operate in difficult conditions under fault scenarios.

Rice is currently the most important food crop in the world and we are only just beginning to study the bacterial associated microbiome. It is of importance to perform screenings of the core rice microbiota and also to develop new plant-microbe models and simplified communities for increasing our understanding about the formation and function of its microbiome. In order to begin to address this aspect, we have performed the isolation of hundreds bacterial isolates obtained from endorhizosphere of two rice cultivars from Venezuela. The validation of plant-growth promoting bacterial activities in vitro has led us to select and characterize 15 isolates for in planta studies such as germination test, endophytism ability and plant growth promotion. Consequently, a set of 10 isolates was selected for the set-up of an endophytic consortium as a simplified model of the natural rice bacterial endomicrobiota. Upon inoculation, the colonization and abundance of each strain within the rice roots was tracked by a culture-independent technique in gnotobiotic conditions in a 30 days period. Four strains belonging to Pseudomonas, Agrobacterium and Delftia genera have shown a promising capacity for colonizing and coexistence in root tissues. On the other hand, a bacterial community taxonomic profiling of the rhizosphere and the endorhizosphere of both cultivars were obtained and are discussed. This study is part of a growing body of research on core crops microbiome and simplified microbiomes, which strengthens the formation process of the endophytic community leading to a better understanding of the rice microbiome.

At the end of mitosis the Mitotic Exit Network (MEN) pathway triggers complex tasks which mainly include the spindle disassembly and the nuclear envelopes assembly. In the course of telophase, which often lasts less than an hour and corresponds to only about 2% of the entire cell cycle’s duration, spatial and temporal cues are integrated to ensure that cytokinesis occurs after the genome has partitioned between mother and daughter cells. From the end of anaphase through telophase, sequential components of a Ras-like GTPase signaling pathway are controlled by a set of different spatial and temporal signals. Successful propagation of these signals through multi-step transduction requires a remarkable sequential coordination. By considering that cells lacking proper MEN function fail to exit from mitosis, I argue that in a hybrid genome impaired coordination between two diverged MENs is prone to result in critical mitotic defects, from late anaphase through telophase. The so-called HyMEN model of hybrid incompatibility depicted here can be regarded as an extension of the Bateson-Dobzhansky-Muller model of speciation, centered on the MEN.

Much research has been conducted to determine how hair regeneration is regulated, as this could provide therapeutic, cosmetic, and even psychological interventions for hair loss. The current study focused on the hair growth effect and effective utilization of fatty oil obtained from Bryde's whales through high-throughput DNA microarray approach in conjunction with immunohistochemical observations. The research also examined the mechanisms and factors involved in hair growth. In an experiment using female C57BL/6J mice, the vehicle control group (VC: propylene glycol: ethanol: water), the positive control group (MXD: 3% minoxidil), and the experimental group (WO: 20% Whale Oil) were topically applied to the back of the mouse. The results showed that 3% MXD and 20% WO were more effective than VC in promoting hair growth, especially 20% WO. Furthermore, in hematoxylin and eosin-stained skin tissue, an increase in the number of hair follicles and subcutaneous tissue thickness was observed with 20% WO. Whole-genome transcriptomic analysis also showed that 20% WO may have lower stress- and inflammation-related responses than 3% MXD. Therefore, whale oil can be expected to be used as a safe hair growth agent.

The study titled "Molecular Choreography of DNA: Insights and Innovations" explores the intricate dance of DNA, shedding light on its structure, function, and regulation. This comprehensive review delves into the historical significance of DNA research, from Watson and Crick's double helix discovery to recent innovations in genomics and molecular biology. The review highlights DNA's roles in replication, repair, gene regulation, and chromatin dynamics, emphasizing the precision and complexity of its molecular choreography. Innovations like CRISPR-Cas9 gene editing and advanced sequencing technologies have revolutionized our understanding of DNA, opening new avenues for scientific exploration. This study aims to provide readers with a deeper appreciation of DNA's central role in life's choreographed processes.

Abstract: There are a large number of cross-linked chemical bonds such as tight disulfide bonds, hydrogen bonds,and salt bonds in keratin, which makes it difficult to break the structural stability of the protein using conventional methods. The degradation of feather waste using keratinases produced by microorganisms has the advantage of being efficient and environmentally friendly. In this study, 23 strains capable of producing transparent circles were obtained via preliminary screening with skim milk as the sole carbon and nitrogen source, and 9 strains capable of degrading feathers were obtained via rescreening with feathers as the sole carbon and nitrogen source. Finally, through the determination of enzyme activity, the strain CY-A with a high ability to degrade feathers was obtained. The strain was identified as Bacillus tequilensis via morphological observation, physiological and biochemical identification, and 16 S rRNA molecular biology. The keratinase gene (bta) of Bacillus tequila was cloned, and its full-length gene sequence was 1110 bp, containing an open reading frame of 1089, encoding 369 amino acids (GeneBank accession number OR733336). The physicochemical properties, secondary structure, and tertiary structure of the recombinant protein were analyzed using bioinformatics tools such as ProtParm, SOPMA, MEGA, and SWISS-MODELS. It was found that the relative molecular mass was 37.953 kD and that the isoelectric point was 8.48. The protein was a hydrophilic protein. The keratinase bta gene was ligated to the expression vector pET28a(+) using T4 ligase, and the recombination expression plasmid pET28-bta was constructed and transformed into E. coli BL21(DE3) to obtain recombinant engineered bacteria. The expressed keratinase was purified using a Ni2+ affinity chromatography column to obtain recombinant bta keratinase bta with a molecular weight of 37.953 kD. The identification and characterization of this keratinase gene provides some theoretical basis for the further enhancement of keratinase activity by means of genetic engineering.

The extraction of cell cultures from fresh tissue is a process that requires significant computa-tional resources and is associated with substantial overheads. The objective of our research is to de-vise a rapid, straightforward, and efficient technique for the cultivation of neuronal cells derived from frozen biological material. We utilized freshly isolated hippocampus and cerebral cortex tissue from rats, in addition to whole brain tissue. This tissue underwent digestion and processing to yield a cellular suspension. This suspension was subsequently cryopreserved using a variety of cryopro-tectants: CTS™ Synth-a-Freeze™, SpermFreeze®, and a medium supplemented with 10% DMSO. CTS™ Synth-a-Freeze™ is a commercially available cryoprotectant typically employed in the preservation of cellular cultures, while the addition of 10% DMSO to the medium is a conventional technique for the cryopreservation of cells to mitigate damage. The innovative aspect of our meth-odology is the application of SpermFreeze®, a product used for the cryopreservation of sperm in in vitro clinics. We postulated that SpermFreeze® might enhance the viability and quality of the cryo-preserved neuronal cells, given the delicate and energetically demanding nature of both sperm and neuronal cells. The ultimate objective was to ascertain whether a cryoprotectant formulated for sperm could augment the post-thawing efficacy of our nervous system cells. This investigation intro-duces an innovative methodology for the optimization of neuronal cell isolation and cryopreserva-tion from animal-derived tissues.

MicroRNAs (miRNAs) within the exosomes are short non-coding RNAs that are associated with many diseases including cancers and are crucial players in regulating gene expression and regula-tion. Owing to their potential role as emerging biomarkers, efficient isolation, and quantification methods of miRNA from the exosomes in the cell’s supernatant are complicated and challenging. The quantitative PCR method is the gold standard for miRNA identification and estimation. Nucleic acid-based assays are expensive, labour-intensive, and require expertise. In this work, we demon-strated an aptamer-based magnetic separation of the exosomes, and quantification of the miRNA using a fluorescence switching assay. The fluorescence assay relies on the fluorescence of the fluorophore labelled cDNA quenched by carbon nanomaterials coated on magnetic beads (OFF state). When the target miRNA210 is introduced to the system, the cDNA detaches from the quenching surface. As a result, the fluorescence intensity increases (ON state) with increasing miRNA concentration within the dynamic range of 0-100 nM. This method can detect miR210 as low as 5 pM. This method does not exhibit any cross-reactivity with other closely related miRNAs. The method was validated by spiking miRNA with the standard RT-PCR method using SYBR green as a fluorescent probe. This method can be used for the minimally invasive detection of cancer biomarkers in breast cancer patients.

Faba bean (Vicia faba L.) is a partially outcrossing species requiring an isolation distance to maintain genetic purity when more than one variety is grown in field conditions. This information is crucial for seed growers and faba bean breeders. A study was conducted at the University of Sydney’s Plant Breeding Institute, Narrabri over two years to examine the extent of natural outcrossing using a creamy white flower characteristic as a morphological marker, which is controlled by a single recessive gene. The white flowered genotype (IX225c) was grown in paired rows of 150 m length in four directions from a central 480 m2 plot of the normal flowered genotype PBA Warda. A beehive was placed in the central plot at flowering time and natural pollination was allowed. At maturity, seed samples were taken from the white flowered genotype at designated intervals along each axis and 100 seeds from each sample were grown in the glasshouse/birdcage to the 4-5 leaf stage and the proportion of plants displaying stipule spot pigmentation (normal flower colour and spotted stipule are linked) was used to determine the percentage of outcrossing. Maximum outcrossing of 2.28% occurred where both genotypes were grown side by side (0 m) and the degree of outcrossing decreased as the distance along each axis from the central plot increased. At 6 m distance the outcrossing was less than 1%, however on occasion it increased to 1% beyond a distance of 100 m, indicating the volatile and unpredictable nature of bee flights. Distance had the major effect on outcrossing, but direction and its interaction had no effect. The results suggest that to limit outcrossing to below 0.5%, a distance of more than 150 m between plots of different faba beans cultivars would be required. It also indicated that Australian faba bean genotypes are mostly self-fertile and a relatively narrow isolation distance will ensure self-fertilization in seed production and breeding programs.

Proso millet starch (PMS) as an unconventional and underutilized cereal starch is becoming increasingly popular worldwide due to its health-promoting properties. This review summarizes research progress in the isolation, characterization, modification, and applications of PMS. PMS can be isolated from proso millet grains by acidic, alkaline, or enzymatic extraction. PMS exhibits typical A-type polymorphic diffraction patterns and shows polygonal and spherical granular structures with a granule size of 0.3-17 µm. PMS is modified by chemical, physical, and biological methods. The native and modified PMS are analyzed for swelling power, solubility, pasting properties, thermal properties, retrogradation, freeze-thaw stability, and in vitro digestibility. The improved physicochemical, structural, and functional properties and digestibility of modified PMS are discussed in terms of their suitability for specific applications. The potential applications of native and modified PMS in food and non-food products are presented. Future prospects for research and commercial use of PMS in the food industry are also highlighted.

Structural DNA nanotechnology provides an excellent foundation for diverse nanoscale shapes that can be used in various bioapplications and materials research. From all existing DNA assembly techniques, DNA origami has proven to be the most robust one for creating custom nanoshapes. Since its invention in 2006, building from the bottom up using DNA has drastically advanced, and therefore, more and more complex DNA-based systems have become accessible. So far, vast majority of the demonstrated DNA origami frameworks are static by nature, but interestingly, there also exist dynamic DNA origami devices that are increasingly coming into view. In this review, we discuss DNA origami nanostructures that perform controlled translational or rotational movement triggered by predefined DNA strands, various molecular interactions and/or other external stimuli such as light, pH, temperature and electromagnetic fields. The rapid evolution of such dynamic DNA origami tools will undoubtedly have a significant impact on molecular scale precision measurements, targeted drug delivery and diagnostics, but they can also play a role in development of optical/plasmonic sensors, nanophotonic devices and nanorobotics for numerous different tasks.

The initiation reactions of DNA synthesis are central processes during human chromosomal DNA replication. They are separated into two main processes: the initiation events at replication origins, the start of the leading strand synthesis for each replicon, and the numerous initiation events taking place during lagging strand DNA synthesis. In addition, a third mechanism is the re-initiation of DNA synthesis after replication fork stalling, which takes place when DNA lesions hinder the progression of DNA synthesis. The initiation of leading strand synthesis at replication origin is regulated at multiple levels, from the origin recognition to the assembly and activation of replicative helicase, the Cdc45-MCM2-7-GINS (CMG) complex. In addition, the multiple interactions of the CMG complex with the eukaryotic replicative DNA polymerases such as DNA polymerase α-primase (Pol α), DNA polymerase δ and ε at replication forks play pivotal roles in the mechanism of initiation reactions including the initiation of signalling unperturbed and stalled replication forks, ‘replication stress’ events, via ATR (ATM-Rad 3-related protein kinase). All these processes are essential for the accurate transfer of the cells’ genetic information to their daughters. Failures and dysfunctions in these processes gives rise to genome instability followed by genetic diseases including cancer. Thus, in their review ‘Hallmarks of Cancer: New Dimensions‘ Hanahan and Weinberg (2022) called genome instability a fundamental function in the development process of cancer cells. In recent years, the understanding of the initiation processes and mechanisms of human DNA replication has made substantial progress at all levels which will be discussed in the review.

Cell-free DNA (cfDNA) analysis is a pivotal tool in non-invasive diagnostics, including cancer monitoring and prenatal testing. However, the preanalytical phase, particularly the choice of anticoagulant, significantly impacts cfDNA integrity and yield. This study aims to compare cfDNA yield, stability, and DNase activity across plasma-citrate, plasma-heparin, plasma-EDTA, and serum, to identify the most suitable specimen for cfDNA analysis among the tested anticoagulants. Blood samples from 15 healthy volunteers were collected in four types of tubes (citrate, heparin, EDTA, and serum). cfDNA was extracted and quantified using qPCR, and endogenous DNase activity was assessed through hydrolysis probe assays. Samples were incubated at 37°C for 24 hours to evaluate cfDNA degradation rates. Statistical analyses were performed using repeated measures ANOVA and Sidak's multiple comparison tests. Baseline cfDNA levels were highest in serum, followed by heparin-plasma, while EDTA and citrate-plasma exhibited the lowest levels. DNase activity was fully inhibited in EDTA-plasma, partially inhibited in citrate-plasma, and active in both heparin-plasma and serum. cfDNA degradation was most pronounced in heparin-plasma (85.3%) and serum (55.6%), compared to minimal degradation in EDTA (8%) and citrate-plasma (13.3%). Among the tested anticoagulants, EDTA-plasma proved to be the most suitable specimen for cfDNA analysis due to its low DNase activity and minimal gDNA contamination. Citrate-plasma, with intermediate DNase inhibition, could be an alternative if the citrate concentration were increased. Heparin-plasma and serum showed significant cfDNA degradation, rendering them unsuitable for cfDNA analysis. These findings highlight the importance of anticoagulant selection in preserving cfDNA integrity for diagnostic applications.

Tdp1 and Tdp2 are DNA repair enzymes that repair DNA damages caused by various agents, including anticancer drugs. Thus, these enzymes resist anticancer therapy and could be the reasons of resistance to such widely used drugs as topotecan and etoposide. In the present work, we found compounds capable of inhibiting both enzymes among derivatives of (-)-usnic acid. Both (+)- and (-)-enantiomers of compounds act equally effectively against Tdp1, and only (-)-enantiomers inhibited Tdp2. Surprisingly, the compounds protect HEK293FT wild type cells from the cytotoxic effect of etoposide but potentiate it against Tdp2 knockout cells. We assume that the sensitizing effect of the compounds in the absence of Tdp2 is associated with effective inhibition of Tdp1, which could take over the functions of Tdp2.

In E. coli DNA replication forks stall on average once per cell cycle. When this occurs, replisome components disengage from the DNA, exposing an intact, or nearly intact fork. Consequently, the fork structure must be regressed away from the initial impediment so repair can occur. Regression is catalyzed by the powerful, monomeric DNA helicase, RecG. During this reaction, the enzyme couples unwinding of fork arms to rewinding of duplex DNA resulting in the formation of a Holliday junction. RecG works against large opposing forces enabling it to clear the fork of bound proteins. Following subsequent processing of the extruded junction, the PriA helicase mediates reloading of the replicative helicase DnaB leading to the resumption of DNA replication. The single-strand binding protein (SSB) plays a key role in mediating PriA and RecG functions at forks. It binds to each enzyme via linker/OB-fold interactions and controls fork loading sites in a substrate-dependent manner that involves helicase remodeling. Finally, it is displaced by RecG during fork regression. The intimate and dynamic SSB-helicase interactions play key roles in ensuring fork regression and DNA replication restart.

Water samples for bacterial microbiome studies undergo a biomass concentration, DNA extraction and taxonomic identification steps. Through benchmarking, we studied the applicability of Skimmed milk flocculation (SMF) for bacterial enrichment, an adapted in-house DNA extraction protocol, and six 16S rRNA databases (16S-DBs). Surface water samples from two rivers were treated with SMF and Vacuum filtration (VF) and subjected to amplicon or shotgun metagenomics. A mock community standard underwent five DNA extraction protocols, taxonomical identification with six different 16S-DBs and evaluated by the Measurement Integrity Quotient (MIQ) score. In SMF samples, the skimmed milk was metabolized by members of lactic acid bacteria or genera such as Polaromonas, Macrococcus, and Agitococcus, resulting in increased relative abundance (p < 0.5) up to 5.0 log fold change compared to VF, rendering SMF inapplicable for bacterial microbiome studies. The best performing DNA extraction protocols were FastSpin Soil, the in-house method and EurX. All 16S-DBs yielded comparable MIQ scores within each DNA extraction kit, ranging from 61-66 (ZymoBIOMICs) up to 80-82 (FastSpin). DNA extraction kits exert more bias to the composition than the 16S-DBs. This benchmarking study presented valuable insights to inform future water metagenomic study designs.

Malassezia species are fastidious and slow-growing yeasts whose isolation from polymicrobial samples is hampered by fast-growing microorganisms. Malassezia selective culture media are needed because Malassezia are resistant to cycloheximide, but some fungi, including the chief human commensal Candida albicans resist to this compound. This study aimed to test whether the macrolide rapamycin could be used in combination with cycloheximide to develop a Malassezia-selective culture medium. Rapamycin susceptibility testing was performed via microdilution assays in modified Dixon against M. furfur and five Candida spp. The MIC was the lowest concentration producing 90% growth inhibition. Rapamycin medium ± cycloheximide 500 mg/L was also added to FastFung solid and yeast suspensions were inoculated and incubated for 72h. Rapamycin MICs against Candida spp. ranged from 0.5 to 2 mg/L, except for C. krusei whose MIC was >32 mg/L. M. furfur stains were rapamycin resistant. Rapamycin and cycloheximide supplementation of the FastFung medium effectively inhibited the growth of non-Malassezia yeast, including the cycloheximide-resistant C. albicans and C. tropicalis. Based on our findings, we recommend using this “MalaSelect” medium for Malassezia isolation and culture from polymicrobial samples.

The genus Naegleria, of free-living amoeba (FLA) group, has been investigated mainly due to its human health impact resulting in deadly infections and their worldwide distribution on freshwater systems. Naegleria fowleri, colloquially known as the “brain-eating amoeba”, is the most studied Naegleria species because it causes Primary Amoebic Meningoencephalitis (PAM) of high lethality. The assessment of FLA biodiversity is fundamental to evaluate the presence of pathogenic species and the possibility of human contamination. However, the knowledge of FLA distribution in Brazil is unknown, and to rectify this situation we present a research on identifying Naegleria spp. in the Monjolinho River, as a model study. The river is a public Brazilian freshwater source that crosses the city of São Carlos. Five distinct sampling sites were examined through limnological features, trophozoites culturing and PCR against internal transcribed spacers (ITS) regions and 5.8S rRNA sequence. The results identified N. philippinensis, N. canariensisi, N. australiensis, N.gruberi, N. dobsoni sequences, as well as a Vahlkampfia sequence. The methodology delineated here represents the first Brazilian Naegleria spp. study on a freshwater system. Our result stresses the urgency of a large scale evaluation of the presence of free-living amoebas in Brazil.

Autism spectrum disorder (ASD) is a complex neurodevelopmental disorder, with a highly variable expression of phenotypes (restricted interest or activity and repetitive behavior in communication and social interactions), genes (mutation), markers (alteration of transcription) and pathways. Loss of function of the CC2D1A gene appears to primarily affect the brain, leading to a range of behavioral problems in humans. In our study published in 2020; we found that the expressions of miR-19a-3p, miR-361-5p, miR-150-5p, miR-3613-3p, miR-126-3p and miR-499a-5p were downregulated in the serum samples of autistic patients and their families. Here, acquired non-Mendelian hereditary character in a genetically defined mouse model of autism (Cc2d1a +/-) correlates with the transcriptional alteration of five miRNAs. We seek to test the hypothesis that miRNA levels are varying by changes in RNA/DNA structure during development, thereby creating transcription alteration. Behavioral tests were conducted on the offspring of Cc2d1a +/- mutant and control mice, such as novel object, social interaction, marble burying and tail suspension behavior. Two RNA fractions were isolated from mouse hippocampal tissues and sperm cells by standard TRIzol extraction: total RNA and fraction of RNA bound to DNA in the form of a DNA/RNA hybrid (R-loop). The expression levels of miR-19a-3p, miR-361-5p, miR-150-5p, miR-126-3p and miR-499a-5p were investigated by quantitative real-time RT-PCR. We report differences in the distribution of five miRNAs in the hippocampus between male and female mice in RNA fractions, particularly in colonies of Cc2d1a +/- mice. Furthermore, the number of miRNAs engaged in the DNA/RNA hybrid fraction is generally higher in the mutant pedigree than in the control group. On the other hand, in sperm both fractions are at lower levels than in controls. R-loops contribute to the physiology and pathology of organisms including human disease. Here we report a variation in five miRNA levels between gender and tissue. Our results suggest that the transcription levels of these five miRNAs are directly regulated by their RNA.

The population of Bifidobacterium spp in faecal samples from breast-fed and formula-fed infants were investigated and developed antibiogram. The common bacterial growth media such as Man Rogos Sharpe (MRS), Brain Heart Infusion (BHI), Luria Bertani (LB) broth and Bifidobacteria agar were evaluated by the enumeration of Bifidobacterium spp. in faecal material. The results demonstrated that the population of Bifidobacterium spp. in the faecal samples of breast-fed infants is numerically high and, low in formula-fed infants. Phylogenetic analysis of the 16S rRNA, (Xylose/Fructose-6-phosphate phosphoketolase) xfp, and RFLP map of Bifidobacteria isolates suggested a presence of possible new and different Bifidobacterium spp. The intensity of the reddish brown colour formed during the f6ppk (Fructose-6-phosphate phosphoketolase) assay confirms the presence of variable Bifidobacteria. The highest recovery and diversity of Bifidobacteria were obtained from Bifidobacteria agar media. Growth in various media resulted in the formation of small (SCV) and big colony variants (BCV). We understand that the various antibiotic minimum inhibitory concentration (MIC) values obtained by applying different media, growth conditions, after bile salt treatment, and at low pH can vary. This study concludes that the variation of microbiological conditions differentiating resistant and susceptible strains is influenced by the test conditions as well.

The soil and groundwater of Bhagobangola I block of Murshidabad district, West Bengal, India is severely arsenic contaminated. A bacterium was isolated from the garden soil of Mahishasthali village, which could tolerate 36.49 mM arsenic (III), 280.44 mM arsenic (V) and 63 mM chromium (III), which makes it arsenic (III & V) and chromium (III) hyper-tolerant bacterium. The growth pattern of this bacterium does not show much alteration in presence of 10 mM arsenic (III) and chromium (III), emphasizing its resistance to these heavy metals. Scanning electron microscopy depicted the size of this bacterium to be ~1.45 µm. 16S rDNA sequencing followed by subsequent phylogenetic analysis established the identity of this bacterium as Microbacterium paraoxydans. The bacterium is capable of bioremediation of arsenic and showed 30.8% & 22.6%, and 35.2% and 30.5% of bioremediation over a period of 24 and 48 hours in 1 mM and 4 mM arsenite, respectively. Microbacterium paraoxydans also exhibit plant growth promoting properties like nitrogen fixation, phosphate solubilization, indole-3-acetic acid production and production of siderophores. Therefore, the heavy metal resistance, bioremediation potential and plant growth promoting potential of the bacterium could be utilized not only for reduction of arsenic toxicity in soil and groundwater but also for plant growth promotion by using it as a biofertilizer.

Theoretical and experimental approaches have been applied to study the polymer physics under-lying compaction of DNA in the bacterial nucleoid. Knowledge of the compaction mechanism is necessary to obtain a mechanistic understanding of the process of segregation of replicating chromosome arms (replichores) during the cell cycle. In the first part of this review light microscope observations are discussed that have demonstrated that the nucleoid has a lower refractive index and thus a lower density than the cytoplasm. A polymer-physical explanation for this phenomenon was given by the theory of Theo Odijk pro-posed in 1998. By assuming a phase separation between nucleoid and cytoplasm and by imposing equality of osmotic pressure and chemical potential between the two phases, a minimal energy situation is obtained in which soluble proteins are depleted from the nucleoid, thus explaining its lower density. The theory is compared with recent views for DNA compaction that are based on mere exclusion of polyribosomes from the nucleoid or on the transcriptional activity of the cell. These new views raise the question of whether they can still explain the lower refractive index or density of the nucleoid. In the second part of this review the question is discussed how DNA segregation occurs in Esche-richia coli, in the absence of the so-called active ParABS system, present in the majority of bacteria. How is entanglement prevented of nascent chromosome arms, generated at the origin in the pa-rental network of the E. coli nucleoid? The observations of the groups of Sherratt and Hansen in 2006, that the four nascent chromosome arms synthesized in the initial replication bubble, segre-gate to opposite halves of the sister nucleoids implies that extensive intermingling of daughter strands does not occur. Based on the hypothesis that leading and lagging replichores synthesized in the replication bubble, fold into micro-domains that do not intermingle, a passive "four-excluding-arms model" for segregation is proposed. This model implies that the key for seg-regation already lies in the structure of the replication bubble at the very start of DNA replication; it explains the different patterns of chromosome arms as well as the segregation distances between replicated loci as experimentally observed.

Transcription factors (TFs) and microRNAs (miRNAs) are co-actors in genome-scale decoding and regulatory networks, often targeting common genes. In this paper, we describe the algebraic geometry of both TFs and miRNAs thanks to group theory. In TFs, the generator of the group is a DNA-binding domain while, in miRNAs, the generator is the seed of the sequence. For such a generated (infinite) group $\pi$, we compute the $SL(2,\mathbb{C})$ character variety, where $SL(2,\mathbb{C})$ is simultaneously a \lq space-time' (a Lorentz group) and a \lq quantum' (a spin) group. A noteworthy result of our approach is to recognize that optimal regulation occurs when $\pi$ looks like a free group $F_r$ ($r =1$ to $3$) in the cardinality sequence of its subgroups, a result obtained in our previous papers. A non free group structure features a potential disease. A second noteworthy result is about the structure of the Groebner basis $\mathcal{G}$ of the variety. A surface with simple singularities (like the well known Cayley cubic) within $\mathcal{G}$ is a signature of a potential disease even when $G$ looks like a free group $F_r$ in its structure of subgroups. Our methods apply to groups with a generating sequence made of two to four distinct DNA/RNA bases in $\{A,T/U,G,C\}$. Several human TFs and miRNAs are investigated in detail thanks to our approach.

Drug abuse is a major issue worldwide. Methylphenidate (MPH) and alprazolam (ALZ) are commonly prescribed drugs for the treatment of ADHD and anxiety disorders, respectively. The limited studies suggest that abusers primarily use benzodiazepines to counteract adverse effects associated with methylphenidate usage. The main aim of this study was to investigate the interaction of drugs with DNA using spectroscopic methods. Female Wistar rats were administered with MPH (10, 20, 40 mg/kg) and ALZ (5, 10, 20 mg/kg) alone and in combination for a period of 28 days. The FT-IR and UV results reveal some spectral changes in a dose-dependent manner, which indicates interactions of drugs with DNA. Thus, the changes in spectral peaks provide some insight into the mechanism of the interaction of drugs with DNA.

Toxoplasma gondii is a widespread protozoan parasite that infects most warm-blooded animals, and felids can serve as intermediate and definitive hosts. Pathological diagnosis and serological and etiological investigations were conducted on a captive caracal (Caracal caracal) carcass collected from China in 2022. Pathological diagnosis revealed that cardiac insufficiency, pulmonary edema, hepatic failure, and renal insufficiency were the causes of the caracal’s death. A modified agglutination test (cut-off: 1:25) revealed that IgG antibodies against T. gondii were present in the myocardium juice, hydroperitoneum, and hydropericardium (≥1:800). A viable T. gondii (TgCaracalCHn2) strain was isolated from the tissue samples (heart, brain, spleen, and skeletal muscle) of this caracal using a mouse bioassay. The genotype of TgCaracalCHn2 was ToxoDB#5 (Type II variant), as determined by RFLP-PCR. The strain was avirulent in Swiss mice and matched the prediction of ROP18 and ROP5 gene alleles of TgCaracalCHn2 (2/2). Many tissue cysts (203 ± 265) were observed in mice brains after inoculation with TgCaracalCHn2 tachyzoites. ToxoDB#5 is the dominant genotype in North American wildlife, and this is the first documented isolation of T. gondii ToxoDB#5 from China. This indicates that caracals can accelerate the transmission of T. gondii genotypes.

Extracellular vesicles (EVs) are enclosed by a lipid-bilayer membrane and secreted by all types of the cells under various physiological conditions. EVs play important roles in intercellular communication and crosstalk between tissues in the body. They are classified into three groups, such as apoptotic bodies, microvesicles, and exosomes. Exosomes were isolated from biofluids including blood, urine, milk, and cell culture media. Exosomes have significant potential for drug delivery and diagnosis. However, the method of isolation affects the physical and biological properties of exosomes. Several methods based on different principles have been developed for exosome isolation. These include ultrafiltration, precipitation, ultracentrifugation, size-exclusion chromatography, and microfluidics. In this study, we applied three common methods, such as ultrafiltration, precipitation, and ultracentrifugation, to isolate exosomes from the cell culture medium and investigated the effects of these different isolation methods on the size distribution and quality of the isolated exosomes. Field emission scanning electron microscopy (FESEM) images, size distribution, total protein content, and the effect of exosomes on the viability of hypoxic cells were analyzed in this study. The analysis revealed that compared to other methods, the ultracentrifugation method can isolate exosomes with smaller diameter (ranging from 20 to 80 nm), lower total protein content (50 µg/ml), and causing the increased viability of the hypoxic cells. The precipitation method does not require special equipment and is inexpensive, if the quality and purity of this method are solved, and it can be used as the best method for exosome isolation. This study can serve as a guide for choosing the best exosome isolation method for applications in medicine according to the needs, time, cost, and equipment.

Fermented milk product "Laban" in Libya is one of the most a traditional fermented milk product consumed a refreshing drink, particularly in the warm season The average values of the physicochemical including titratable acidity, pH, total solids, and fat were 0.73%, 4.16, 8.12%, and 1.54% respectively. Coliform, yeast and mold counts were 21×10⁴, 39×10⁴, and 41 ×10³ cfu/ ml., respectively. Most strains of coliform bacteria were Serratia odorifera, Escherichia coli 1, E. coli 2. and Klebsiella oxytoca. The average Lactococcus, Streptococcus, Mesophilic Lactobacillus / Leuconostoc and Thermophilic Lactobacillus counts were 99 ×10⁷, 96 ×10⁷, 93 ×10⁷ and 15 ×10⁷ cfu / ml. respectively. A total of 142 lactic acid bacteria (LAB) isolates were identified to the genus level as Lactobacillus (48.59%), Lactococcus (43.66%), Streptococcus (4.93%) and Leuconostoc (2.82%). Sugar fermentation tests revealed the most frequent Lactobacillus species found to be Lactobacillus delbrueckii ssp. lactis (62.32%), followed by Lactobacillus plantarum (31.88%). Furthermore, other selected LAB isolates were identified by API 50 CH test as Lactococcus lactis ssp. lactics, Lactobacillus pentosus, Lactobacillus brevis, and Leuconostoc mesenteroides ssp. cremoris. Thus, our research documented the lactic acid bacteria strains and will provides fundamental basic and useful information for further studies of strain selection starter culture, with regard to the industrial production of fermented dairy milk products.

Bacterial Calcium-carbonate Precipitation (BCP) has been studied for multiple applications such as remediation, consolidation and cementation. Isolation and screening of strong calcifying bacteria is the main task of BCP-technique. In this paper we studied CaCO₃ precipitation by different bacteria isolated from a rhizospheric soil in both solid and liquid media. It has been found, through culture-depending studies, that bacteria belonging to Actinobacteria, Gammaproteobacteria and Alphaproteobacteria are the dominant bacteria involved in CaCO₃ precipitation in this environment. Pure and mixed cultures of selected strains were applied for sand biocementation experiments. Scanning Electron Microscopy (SEM) and Energy Dispersive X-ray Spectroscopy (EDS) analyses of the biotreated samples revealed the biological nature of the cementation and the effectiveness of the biodeposition treatment by mixed cultures. X-ray diffraction (XRD) analysis confirmed that all the calcifying strains selected for sand biocementation precipitated CaCO₃, mostly in the form of calcite. In this study Biolog® Eco-plate is evaluated as a useful method for a more targeted choice of the sampling site with the purpose of obtaining interesting candidates for BCP applications. Furthermore, ImageJ software was investigated, for the first time to our knowledge, as a potential method to screen high CaCO₃ producer strains.

The medical care and welfare of domestic dogs are now respected worldwide at the same level as that of humans. We previously established a test method to determine whole mitochondrial DNA (mtDNA) using oral mucosal DNA that may be useful for the medical care and welfare of dogs. However, the sample types tested in dogs are not limited to those obtained from the oral mucosa. Therefore, in the present study, we attempted to establish a test method to determine whole mtDNA sequences using feces, which represents the least invasive specimen. Two Japanese domestic dogs were used in the present study. DNA was extracted from approximately 100 mg of fresh feces from each dog, and PCRs were performed using four primer-pairs that can amplify whole mtDNA. Following PCR, amplicons were pooled to create a DNA library using an experimental robot with an original program. Data were then acquired via NGS and data analysis was performed. The results showed that the whole mtDNA sequence of the two dogs was determined with high accuracy. Our results suggest that feces can be adapted for mitochondrial disease and individual identification testing and could serve as a useful testing method for the future medical care and welfare of domestic dogs.

Cholangiocarcinoma is an epithelial malignancy arising in the region between the intrahepatic bile ducts and the ampulla of Vater at the distal end of the common bile duct. The effect of current chemotherapy regimens against cholangiocarcinoma is limited, and the prognosis of patients with cholangiocarcinoma is poor. Aberrant DNA methylation and histone modification induce silencing of tumor suppressor genes and chromosomal instability during carcinogenesis. Studies have shown that the tumor suppressor genes and microRNAs (miRNAs) including MLH1, p14, p16, DAPK, miR-370 and miR-376c are frequently methylated in cholangiocarcinoma. Silencing of these tumor suppressor genes and miRNAs plays critical roles in the initiation and progression of cholangiocarcinoma. In addition, recent studies have demonstrated that DNA methylation inhibitors induce expression of endogenous retroviruses and exert the anti-tumor effect of via an anti-viral immune response. Aberrant DNA methylation of tumor suppressor genes and miRNAs could be a powerful biomarker for diagnosis and treatment of cholangiocarcinoma. Epigenetic therapy with DNA methylation inhibitors hold considerable promise for the treatment of cholangiocarcinoma through re-activation of tumor suppressor genes and miRNAs as well as induction of an anti-viral immune response.

The present study investigates the role of a simple inerter in supplemental devices for possible implementation in the matured seismic base isolation technique. Firstly, the response of the base-isolated structure with an optimally tuned mass damper-inerter (TMDI) is investigated to see the tuning effects. The time required to tune the TMDI was found to be significantly longer than the duration of a strong-motion earthquake. There was still a reduction in the response of the isolated structure, which is primarily due to the added damping and stiffness (ADAS) of TMDI and not because of the tuning effects. Hence, it is proposed that the corresponding ADAS of the TMDI be directly added to the isolation device. Secondly, the response of the base-isolated structures with the fluid inerter damper (FID) is studied. It was observed that the inerter of the FID does not influence the displacement variance of an isolated structure under broadband earthquake excitation. It implies that the response of the isolated structure with FID is primarily controlled by its counterpart fluid damper (FD). The performance of optimal TMDI, ADAS, FID, and FD to mitigate the seismic response of the flexible multi-story base-isolated structure under real earthquake excitations is also investigated. In terms of suppressing the displacement and acceleration responses of the isolated structure, it has been found that TMDI and ADAS perform similarly. Comparing the response of the isolated structure with FID and FD demonstrated that the inerter in the FID has detrimental effects on the isolated structures, in which the top floor's acceleration and base shear are substantially increased.

Reproductive isolating barriers are a crucial element in the speciation process. There are various types of barriers, and within those that act early in the speciation process, the behavioral one can play an important role in isolating populations or closely related species. It has been shown that the high diversity of some taxa correlates with the variation of sexual signals, which facilitate isolation. Here it is explored whether chemical signals may contribute to the high diversity of Liolaemus lizards, which has more than 290 species. Population chemical discrimination was investigated in two co-distributed species, L. tenuis and L. lemniscatus. The studied populations were closely located and likely belonged to the same genetic lineages. Lizards of both species discriminated between scents of their own and different populations, and in most cases, scents from their population were preferred. Results suggest a fast evolution of scents, and their recognition, and there for the ethological barriers associated with the chemical modality may evolve fast. However, because the studied species differed in the strength of the exhibited population chemical discrimination, ethological barriers evolve at different rates across species. It can be concluded that ethological barriers involving the chemical modality may disrupt species' cohesion, contributing to Liolaemus diversification, in synergy or not with geographical barriers.

Microalgae are known as a rich source of bioactive compounds which exhibit different biological activities. Increased demand for sustainable biomass for production of important bioactive components with various potential especially therapeutic applications has resulted in noticeable interest in algae. Utilization of microalgae in multiple scopes has been growing in various industries ranging from harnessing renewable energy to exploitation of high-value products. The focuses of this review are on the isolation of bioactive compounds from microalgae regarding different metabolites which are currently used and new possible applications of the compounds in industries and future prospects. Moreover, this work discusses the advantage, potential new beneficial strains, applications, limitations, research gaps and future prospect of microalgae in industry.

The study aimed to isolate and characterize bacteriocin-producing Enterococcus species from Ethiopian traditional fermented foods and beverages. The bacteriocin bioassay was tested against food-borne pathogens at various temperatures, pH, solvents, additives, and enzymes. A solvent extraction method was established to concentrate bacteriocins from Enterococcus species. The study found that crude extracts with proteinase K, pepsin, and trypsin deactivated antimicrobial activity, confirming their proteinaceous nature. Organic solvents were more effective at extracting bacteriocins from CFS, with chloroform alone and in combination providing rapid and efficient recovery. The optimal bacteriocin production and strong inhibition activity were achieved at 37°C with a pH range of 5.5 to 6.5 incubated for 16-18 hours. The six Enterococcus faecium species were the best bacteriocin-producing isolates. Optimizing bacteriocin production and changing environmental growing circumstances can enhance the application of bacteriocins as food preservation agents. Traditional fermented products are potential sources of bacteriocin-producing LAB.

Investigation of the methanol extract of the poroid fungus Fuscoporia torulosa resulted in the isolation of a novel triterpene, fuscoporic acid (1) together with inoscavin A and its previously undescribed Z isomer (2 and 3), 3,4-dihydroxy-benzaldehide (4), osmundacetone (5), senexdiolic acid (6), natalic acid (7), and ergosta-7,22-diene-3-one (8). The structures of fungal compounds were determined on the basis of NMR and MS spectroscopic analysis, as well as molecular modelling studies. Compounds 1, 6-8 were examined for their antibacterial properties on resistant clinical isolates, and cytotoxic activity on human colon adenocarcinoma cell lines. Compound 8 was effective against Colo 205 (IC50 11.65±1.67 µM), Colo 320 (IC50 8.43±1.1 µM) and MRC-5 (IC50 7.92±1.42 µM) cell lines. Potentially synergistic relationship was investigated between 8 and doxorubicin, which revealed a synergism between the examined compounds with a combination index (CI) at the 50% growth inhibition dose (ED50) of 0.521±0.15. Several compounds (1, and 6-8) were tested for P‐glycoprotein modulatory effect in Colo 320 resistant cancer cells, but none of the compounds proved to be effective in this assay. Fungal metabolites 2-5 were evaluated for their antioxidant activity using the oxygen radical absorbance capacity (ORAC) and DPPH assays. Compounds 4 and 5 proved to possess considerable antioxidant effect with EC50 0.25±0.01 (DPPH) and 12.20±0.92 mmol TE/g (ORAC). The current article provides valuable information on both chemical and pharmacological profiles of Fuscoporia torulosa, paving the way for future studies with this species.

Major depressive disorder (MDD) is a prevalent mental health condition with substantial social and economic consequences. This study focuses on the interplay between genetic and environmen-tal factors in MDD, with particular emphasis on epigenetic mechanisms such as stochastic epige-netic mutations (SEMs), epigenetic age acceleration, and epigenetic drift. We conducted a meta-analysis of DNA methylation patterns across six datasets (n=1125 MDD cases, 398 controls in whole blood; n=95 MDD cases, 96 controls in brain tissues) from the Gene Expression Omnibus. The analysis did not reveal significant global methylation differences between MDD cases and controls. However, a brain-specific probe (cg25801113) near the SHF gene consistently exhibited hypomethylation, suggesting its potential relevance to MDD. SEMs were assessed, revealing a dis-tinct burden at the gene level in MDD cases, though no significant findings emerged for epigenetic age acceleration, indicating it may not play a central role in MDD. The study also identified rare epivariations in specific genes unique to MDD cases. In blood tissue, 51 genes were associated with these unique epivariations, while 1 gene was highlighted in the brain tissue. These results un-derscore the importance of exploring rare epivariations in understanding the molecular mecha-nisms underlying MDD and offer potential targets for further investigation.

In a software-defined radar (SDR) system, most of the signal processing usually implemented in hardware is implemented by software, thus allowing for higher flexibility and modularity compared to conventional radar systems. However, the majority of SDR demonstrators and proofs-of-concept reported in the open literature so far have been based on simple antenna systems. As a result, the full potentialities of an SDR approach have not been completely exploited yet. In this work, we propose a flexilbe antenna module to be integrated into an active electronically scanning array (AESA) with controlled sidelobe level over a wide angular range, exhibiting polarization reconfigurability with low cross-polarization level and high isolation. For this purpose, analytical and numerically efficient techniques for the synthesis of the aperture distribution and the correct evaluation of the radiating features (e.g., beamwidth, pointing angle, sidelobe levels, etc.) are presented in order to grant a real-time control of the digital beamforming network. A sub-array module demonstrator is fabricated and measured to corroborate the concept.

DNA barcoding has emerged as a powerful tool in the field of taxonomy, transforming the way we identify and classify species. This review article aims to provide an up-to-date overview of the advancements in DNA barcoding techniques, their applications, and their impact on taxonomy and biodiversity studies. We will explore the evolution of DNA barcoding, the development of standardized barcode regions, and the challenges and limitations associated with this methodology. Additionally, we will delve into the diverse range of taxonomic groups for which DNA barcoding has been applied, highlighting key case studies and their contributions to our understanding of biodiversity. Furthermore, we will discuss the integration of DNA barcoding with other molecular and bioinformatic approaches, such as metabarcoding and machine learning, to enhance species identification and biodiversity assessment. This review aims to provide a comprehensive perspective on how DNA barcoding is reshaping the field of taxonomy and its implications for conservation, ecology, and evolutionary biology.

Systematics plays the most crucial role in biodiversity conservation which is at stake due to anthropogenic activities and environmental degradations. The ever-increasing decline of classical taxonomic expertise drives the need to develop molecular marker-based tools for quick, efficient, and reliable detection of organisms, to assess their ecological impacts for deepening our understanding of systematic and evolutionary relationships between organisms which is central to biology. The pace of alpha taxonomy has quickened by its integration with an increasingly fashionable and novel concept called DNA barcoding which utilizes a short genetic marker or barcode to categorize species for enhanced biodiversity assessment. As a supplementary but not complete alternative of systematics research, DNA barcoding, however, not error-free, brings precision in identification by solving existing problems of classical taxonomy and phylogenetics, irrespective of the growth stage of organisms, particularly for known taxa rather than unknown ones. Mitochondrial gene Cytochrome C Oxidase 1 (COI) serves as a universal animal barcode but there is no such universal barcode for plants and developing a suitable one is more challenging. With the recent advancement of Next Generation Sequencing (NGS), DNA metabarcoding technology is advancing rapidly. Still, ambiguity and error prevail with the correct identification of species due to some problems. After extensive analysis of the existing DNA barcoding papers, this review discusses commonly used DNA barcodes in plants and animals, their roles, advantages, limitations to solve existing problems of conservation biology and add the author’s views and recommendations.

DNA methylation, one of the major epigenetic mechanisms, plays critical roles in regulating gene expression, genomic stability and cell lineage commitment. Establishment and maintenance of DNA methylation in mammals is achieved by two groups of DNA methyltransferases: DNMT3A and DNMT3B, which are responsible for installing DNA methylation patterns during gametogenesis and early embryogenesis, and DNMT1, which is essential for propagating DNA methylation patterns during replication. Both groups of DNMTs are multi-modular proteins, containing a large N-terminal regulatory region in addition to the C-terminal methyltransferase domain. Recent structure-function investigations of the individual domains or large fragments of DNMT1 and DNMT3A have revealed the molecular basis for their substrate recognition and specificity, intramolecular domain-domain interactions, as well as their crosstalk with other epigenetic mechanisms. These studies highlight a multifaceted regulation for both DNMT1 and DNMT3A/3B, which is essential for the precise establishment and maintenance of lineage-specific DNA methylation patterns in cells. This review summarizes current understanding of the structure and mechanism of DNMT1- and DNMT3A-mediated DNA methylation, with emphasis on the functional cooperation between the methyltransferase and regulatory domains.

Agrobacterium tumefaciens (A. tumefaciens) is a plant pathogenic bacterium that causes crown gall disease, leading to significant economic losses in various agriculturally important crops, including apple, pear, peach, and almond. The difficulty of controlling A. tumefaciens with conventional pesticides underscores the need for alternative antibacterial agents such as bacteriophages. In this study, a novel lytic bacteriophage, named Agrobacterium phage PAT1 (PAT1), with high lysis potential against A. tumefaciens, was isolated from wastewater. The interaction between PAT1 and A. tumefaciens cells, investigated using Transmission electron microscopy, revealed that PAT1 efficiently adsorb, infect, and replicate on A. tumefaciens in a very short time period (i.e., ≤30 min for a complete infectious cycle). Furthermore, the turbidity assay showed that PAT1 (MOI = 1) exerted a significant inhibitory effect on A. tumefaciens for 48 hours, resulting in an 82% reduction in bacterial growth. Additionally, stability tests showed that PAT1 was highly resistant to a broad range of pH (4-10) and temperatures (4-60°C). Bioinformatics analyses of the genomic sequence suggested that PAT1 is a new phage closely related to Agrobacterium phages of the genus Atuphduovirus within the family Autographiviridae. The genome size was 45,040 base pairs with a G+C content of 54.5%, consisting of 54 coding sequences (CDS), of which the functions of 22 CDS were predicted, including two endolysins genes which could be used as antimicrobials against A. tumefaciens. Additionally, no lysogenic mediated genes or genes encoding virulence factors, antibiotic resistance, or toxins were detected in PAT1 genome. Overall, PAT1 demonstrated suitability as a potential biocontrol agent for combating A. tumefaciens infections, expanding the limited catalog of such lytic A. tumefaciens phages. However, its efficacy in controlling crown gall disease in planta remains to be evaluated.

Pulmonary fibrosis is a chronic, progressive and fatal disease of the interstitial lung， there still lack of efficient therapy to reverse the prognosis of patients currently. In this study, a fucoidan from Costaria costata was isolated and its anti-idiopathic fibrosis activity was investigated both in vitro and in vivo. The chemical composition analysis results show that Costaria costata polysaccharide (CCP) is consisting of galactose and fucose as the main monosaccharides with a sulfate group content of 18.54%. The results found that CCP could resist TGF-β1-induced epithelial-mesenchymal transition (EMT) in A549 cells by inhibiting the TGF-β/Smad and PI3K/AKT/mTOR signaling pathways. In vivo data found that CCP treatment alleviated bleomycin (BLM)-stimulated fibrosis and inflammation in mice lung tissue. This study suggests that CCP could protect the lung from fibrosis by relieving the EMT process and inflammation in lung cells.

A large body of evidence indicates that environmental agents can induce alterations in DNA methylation (DNAm) profiles. Radiofrequency electromagnetic fields (RF-EMFs) are radiations emitted by everyday devices which have been classified as “possibly carcinogenic”, however their biological effects are unclear. As aberrant DNAm of genomic repetitive elements (RE) may promote genomic instability, here we sought to determine whether exposure to RF-EMFs could affect DNAm of different classes of RE, such as long interspersed nuclear element1s (LINE-1), Alu short interspersed nuclear element and ribosomal repeats. To this purpose, we analysed DNAm profiles of cervical cancer and neuroblastoma cell lines (HeLa, BE(2)C and SH-SY5Y) exposed to 900 MHz GSM-modulated RF-EMF through an Illumina-based targeted deep bisulfite sequencing approach. Our findings showed that radiofrequency exposure did not affect DNAm of Alu elements in any of the cell lines analysed. Conversely, it influenced DNAm of LINE1 and ribosomal repeats in terms of both average profiles and organization of methylated and un-methylated CpG sites, in different ways in each of the three cell lines studied.

The ribonuclease H (RNase H) active site of HIV-1 reverse transcriptase (RT) is the only viral enzyme not targeted by approved antiretroviral drugs. Using a fluorescence-based in vitro assay, we screened 65,239 compounds at a final concentration of 10 µM to identify inhibitors of RT RNase H activity. We identified 41 compounds that exhibited 50% inhibitory concentration (i.e., IC50) values < 1.0 µM. Two of these compounds, 2-(4-methyl-3-(piperidin-1-ylsulfonyl)phenyl)benzo[d]isothiazol-3(2H)-one (1) and ethyl 2-(2-(3-oxobenzo[d]isothiazol-2(3H)-yl)thiazol-4-yl)acetate (2), which both share the same benzisothiazolone pharmacophore, demonstrate robust antiviral activity (50% effective concentrations of 1.68 ± 0.94 µM and 2.68 ± 0.54, respectively) in the absence of cellular toxicity. A limited structure-activity relationship analysis identified two additional benzisothiazolone analogs, 2-methylbenzo[d]isothiazol-3(2H)-one (3) and N,N-diethyl-3-(3-oxobenzo[d]isothiazol-2(3H)-yl)benzenesulfonamide (4), which also resulted in inhibition of RT RNase H activity and virus replication. Compounds 1, 2 and 4, but not 3, inhibited the DNA polymerase activity of RT (IC50 values ~ 1 to 6 µM). In conclusion, benzisothiazolone derivatives represent a new class of multifunctional RT inhibitors that warrants further assessment for the treatment of HIV-1 infection.

The soybean yield per unit area and soybean acreage are increased to boost overall soybean production as two effective solutions. There is a large area of saline soils in northeast China, where if soybeans could be grown with the help of isolated saline-tolerant rhizobia, the cultivation area of soybeans in China can be effectively expanded. Several rhizobia with different salt-, acid- and alkali-resistant characteristics were obtained in this research by isolating, identifying, and analyzing the nitrogen-fixing activities of indigenous rhizobia from different soybean planting soils, and further analyzed the host compliance between rhizobia and different soybean germplasm concerning 30 representative germplasm resources in the northeast region. Collectively, the results of the resistance study of HLNEAU2 isolated from the Shaanxi region are great references for guiding the application of rhizobia on soybeans.

Various factors, such as uncertainty of component parameters and uncertainty of sensor meas-urement values, contribute to the difficulty of fault detection in the landing gear retrac-tion/extension hydraulic system. In this paper, we introduce linear fractional transformation technology and uncertainty analysis theory for the construction of the diagnostic bond graph of the landing gear retraction/extension hydraulic system. In this way, interval analytical redundancy relations and fault signature matrix can be derived. Using the fault signature matrix, existing faults of the system can be preliminarily detected and isolated. Additionally, interval analytical re-dundancy relations can be used to detect system faults in detail, and cases analysis can be carried out to determine if the actuator is externally or internally leaky, and if the landing gear selector valve is reversing stuck. Compared to the traditional analytical redundancy relations, this method takes into account the negative factors of uncertainty; and compared to the traditional absolute diagnostic threshold, the interval diagnostic threshold is more accurate and sensitive.

The DNA Damage Response (DDR) network and the mitogen-activated protein kinase (MAPK) signaling pathway are crucial mechanisms for the survival of all living beings. An accumulating body of evidence suggests that there is a crosstalk between these two systems, thus favoring the appropriate functioning of multi-cellular organisms. On the other hand, aberrations within these mechanisms are thought to play a vital role in the onset and progression of several diseases, including cancer, as well as in the emergence of drug resistance. Here, we provide an overview of the current knowledge regarding alterations in the DDR machinery and the MAPK signaling pathway, as well as abnormalities in the DDR-MAPK functional crosstalk in multiple myeloma, the second most common hematologic malignancy. We also present the latest advances in the development of anti-myeloma drugs targeting crucial DDR- and MAPK-associated molecular components. These data could potentially be exploited to discover new therapeutic targets and effective biomarkers, as well as for the design of novel clinical trials. Interestingly, they might provide a new approach to increase the efficacy of anti-myeloma therapy by combining drugs targeting the DDR network and the MAPK signaling pathway.

The paper highlights the performance of the seismic isolation devices, installed on retrofitted buildings in reducing the seismic response when subjected to earthquakes. Two buildings from the beginning of the XX-th century in Bucharest are chosen from many more, monitored over the city area. We discuss the response of these base-seismic isolated structures, relying on good quality data acquired from the recent strong earthquake (03.11.2022, MW = 5.0). Elastic response spectra computed from recordings at two levels of each structure are used, placed under and right above the isolating layer. At one building the existence of previous recordings and the particularity of the sensors allow a comparison with other two relatively recent medium intensity earthquakes. The assessment is carried out in terms of maximum acceleration amax, measured at certain levels in each structure, spectral acceleration amplitude SAmax and spectral-peak corresponding period. We find that the base-isolation methodology is effective in reducing the response of the building right above the isolating layer, an observation valid for both structures, all components of the recordings, and spectral acceleration values. Moreover, the outcomes from the modal evaluation performed prior to rehabilitation and seismic isolation process are presented, by pointing out a higher newly acquired fundamental period of the isolated structures.

The DNA Damage Response (DDR) is a complex signaling network that comes into play when cells experience genotoxic stress. Upon DNA damage, cellular signaling pathways are rewired to slow down cell cycle progression and allow recovery. However, when the damage is beyond repair, cells activate complex and still not fully understood mechanisms, leading to a complete proliferative arrest or cell death. A number of conventional and novel anti-neoplastic treatments rely on causing DNA damage or on the inhibition of the DDR in cancer cells. However, the identification of molecular determinants directing cancer cells toward recovery or death upon DNA damage is still far from complete, and it is object of intense investigation. SPRY-COntaining RAN binding Proteins (Scorpins) RANBP9 and RANBP10 are evolutionarily conserved and ubiquitously expressed proteins whose biological functions are still debated. RANBP9 has been previously implicated in cell proliferation, survival, apoptosis and migration. Recent studies also showed that RANBP9 is involved in the Ataxia Telangiectasia Mutated (ATM) signaling upon DNA damage. Accordingly, cells lacking RANBP9 show increased sensitivity to genotoxic treatment. Although there is no published evidence, extensive protein similarities suggest that RANBP10 might have partially overlapping functions with RANBP9. Like RANBP9, RANBP10 bears sites putative target of PIK-kinases and high throughput studies found RANBP10 to be phosphorylated following genotoxic stress. Therefore, this second Scorpin might be another overlooked player of the DDR alone or in combination with RANBP9. This review focuses on the relatively unknown role played by RANBP9 and RANBP10 in responding to genotoxic stress.

This review focuses on the regulation and modulation of human DNA polymerase δ (Pol δ). The emphasis is on mechanisms that regulate the activity and properties of Pol δ in DNA repair and replication. The areas covered are the degradation of the p12 subunit of Pol δ, which converts it from a heterotetramer (Pol δ4) to a heterotrimer (Pol δ3), in response to DNA damage and also during the cell cycle. The biochemical mechanisms that lead to degradation of p12 are reviewed, as well as the properties of Pol δ4 and Pol δ3 that provide insights into their functions in DNA replication and repair. The second focus of the review involves the functions of two Pol δ binding proteins, PDIP46 and PDIP38, both of which are multi-functional proteins. PDIP46 is a novel activator of Pol δ4, and the impact of this function is discussed in relation to its potential roles in DNA replication. Several new models for the roles of Pol δ3 and Pol δ4 in leading and lagging strand DNA synthesis that integrate a role for PDIP46 are presented. PDIP38 has multiple cellular localizations including the mitochondria, the splicesosomes and the nucleus. It has been implicated in a number of cellular functions, including the regulation of specialized DNA polymerases, mitosis, the DNA damage response, Mdm2 alternative splicing and the regulation of the Nox4 NADPH oxidase.