The aim of this study was to examine pathologic complete response (pCR) and overall survival (OS) of patients diagnosed with non-metastatic inflammatory breast cancer (IBC). A total of N=8,550 cases undergoing surgery were identified between 2004-2013, using the National Cancer Database (NCDB). Patients were grouped into 4 biologic subtypes (HR+/HER2-, HR+/HER2+, HR-/HER2+, HR-/HER2-). The median age at diagnosis was 56 years. On average, women were followed for 3.7 years [interquartile range=3.0]. The majority were white (80%), had private health insurance (50%), and presented with poorly differentiated tumors (57%). Approximately 46% of the cancers were >5cm. Most patients underwent mastectomy (94%) and received radiotherapy (71%). Differences by biologic subtypes were observed for grade, lymph node invasion, race, and tumor size (p<.0001). Compared with non-pCR (54%), patients experiencing pCR had superior 5-year survival (77%) (p<.0001). Survival was poor for triple-negative (TN) tumors (37%) vs. other biologic subtypes (60%) (p<.0001). On multivariable analysis, TN-IBC, positive margins, and not receiving either chemotherapy, hormonal therapy or radiotherapy were independently associated with poor 5-year survival (p<.0001). In this large multicentric analysis of IBC, categorized by biologic subtypes, we observed significant differential tumor, patient and treatment characteristics, and OS.

Background: Pathologic complete response (pCR) of breast cancer after neoadjuvant chemotherapy (NAC) is highly related to molecular subtypes. Patients who achieved tumor pCR after NAC have a better prognosis. However, despite of better prognosis, pCR patients have a potential for recurrence. There is little evidence of risk factors of recurrence in patients with pCR. We aim to analyze factors associated with tumor recurrence in patients who achieved pCR. Methods: This study retrospectively reviewed the data of patients diagnosed with breast cancer who achieved pCR after receiving NAC between January 2009 and December 2018 in Samsung Medical Center. pCR was defined as no residual invasive cancer in the breast and axillary nodes even if there is residual ductal carcinoma in situ (ypT0 or ypTis with ypN0). Breast cancers are classified into 4 subtypes based on hormone receptors (HR) and human epithelial growth factor receptor 2 (HER2) status. Patients who had bilateral breast cancer, inflammatory breast cancer, distant metastasis, unknown subtype, and histologically unique case were excluded from the study. Results: In total 483 patients were included in this study except for patients who corresponded to the exclusion criteria. The median follow-up duration was 59.0 months (range, 0.5-153.3 months). Breast cancer recurred in 4.1% of patients (20 of 483). There was a significant difference in clinical T (p = .004) and clinical N (p = .034) stage in the Kaplan-Meier curve for disease-free survival. Molecular subtypes (p = .573), Ki67 (p = 1.000), and breast surgery type (p = .574) were not associated with tumor recurrence in patients who achieved pCR after NAC. In the clinical T stage and clinical N stage, there was a significant difference between recurrence and no-recurrence groups (clinical T stage; p = .045, clinical N stage; p = .002). Univariable Cox regression revealed statistical significance in the clinical T stage (p = .049) and clinical N stage (p = .010), while multivariable Cox regression demonstrated non-significance in the clinical T stage (p = .320) and clinical N stage (p = .073). Conclusion: Results in this study showed that clinical T, clinical N stage, and molecular subtypes were not statistically significant predictors of recurrence in patients who achieved pCR after NAC. It is hypothesized that a multicenter-based study would lead to the identification of factors as predictors of recurrence after achieving pCR.

Our study aimed to identify predictors for the effectiveness of tumor regression in lung cancer patients undergoing neoadjuvant treatment and cancer resections. Patients admitted between 2016 and 2022 were included in the study. Based on the histology of the tumor, patients were categorized into lung adenocarcinoma group (LUAD), and squamous cell carcinoma group (SQCA). Ninety-five patients with non-small cell lung cancer were included in the study. Fifty-eight (61.1%) and 37 (38.9%) patients were included in the LUAD and SQCA groups, respectively. Nine (9.5%), 56 (58.9%), and 30 (31.6%) patients were categorized with a tumor regression score of I, II, and III respectively. In multivariable analyses, histology of the primary tumor (SQCA), lymph node size in the preoperative CT scan (>1.7cm) and absolute tumor size reduction after neoadjuvant treatment (>2.6 cm) independently predict effectiveness of tumor regression (OR, [95% Confidence interval, p-value] of 6.88 [2.40-19.77, p70 years, extended resection >one lobe, and tumor recurrence or metastasis were identified as significant independent predictors of reduced overall survival. Assessment of tumor size before and after neoadjuvant treatment might help to identify high-risk patients with decreased survival and to improve patient management and care.

The Chikungunya virus (CHIKV) poses a significant global public health concern, especially in Africa. Since its first isolation in Tanzania in 1953, CHIKV has caused recurrent outbreaks, challenging healthcare systems in low-resource settings. Recent outbreaks in Africa highlight the dynamic nature of CHIKV transmission and the challenges of underreporting and underdiagnosis. Here, we review literature and analyze publicly available cases, outbreaks, and genomic data providing insights into the epidemiology, genetic diversity, and transmission dynamics of CHIKV in Africa. Our analyses reveal the circulation of geographically distinct CHIKV genotypes, with certain regions experiencing a disproportionate burden of disease. Phylogenetic analysis of sporadic outbreaks in West Africa suggests repeated emergence of the virus through enzootic spillover, which is markedly different from inferred transmission dynamics in East Africa, where the virus is often introduced from Asian outbreaks, including the recent reintroduction of the Indian Ocean lineage from the Indian subcontinent to east Africa. Furthermore, there is limited evidence of viral movement between these two regions. Understanding the history and transmission dynamics of outbreaks is crucial for public health planning. Despite advances in surveillance and research, diagnostic and surveillance challenges persist. This review and secondary analysis highlight the importance of ongoing surveillance, research, and collaboration to mitigate the burden of CHIKV in Africa and improve public health outcomes.

Tumour microenvironment, composed of pro- and anti-tumour immune cells, affects cancer cells behaviour. We aimed to evaluate if tumour-infiltrating lymphocyte (TIL) density and TILs subtypes in core biopsies at diagnosis of breast cancer patients could predict pathologic complete response (pCR; ypT0/is ypN0) from neoadjuvant systemic therapy (NST). The TIL subtypes were determined based on the proportions of presumably anti-tumour (CD8+, CXCL13+) and pro-tumour (PD-1+, FOXP3+) immune cells. A prospective, non-interventional study including 171 participants undergoing NST was performed. The median TIL density for the entire cohort was 10% (IQR: 3.5-23.8), and 59 (35%) patients achieved pCR. TIL density was positively associated with pCR (univariately and multivariably). In the multivariable logistic regression model, TIL density was an independent predictor of pCR (p=0.012, OR 1.27; 95% CI 1.05-1.54) when controlled for age (p=0.232), Ki-67 (p=0.001), node-negative status (p=0.024), and HER2+/triple negative vs luminal B-like subtype (p&lt;0.001). In our sample, higher proportions of PD-1+ TILs and FOXP3+ TILs were associated with a higher probability of pCR but the association was not statistically significant. In the exploratory multivariable analysis, we showed that only higher CD8+ TILs were associated with pCR. In conclusion, TIL density and its subtypes are associated with pCR.

There is currently no effective treatment strategy for the recurrent/metastatic adenoid cystic carcinoma (R/M ACC). Furthermore, recent single-agent and combination immunotherapy trials have failed in unselected ACC cohorts unlike non-ACC salivary gland cancers. Genomic profiling revealed no actionable targets but NOTCH1 and KDM6A frameshift and CTCF splice site mutations (no MYB/L fusion) with a low TMB, MSS and negative PD-L1. We recommended anti-PD-1 plus anti-CTLA-4 combination based on TMB 2-fold greater than median TMB in ACC, tumor harboring multiple immunogenic frameshift or splice site mutations, and PD-L1 negativity. Accordingly, we achieved a complete response in a radiotherapy and chemotherapy-refractory patient with locally recurrent lacrimal gland ACC and lung metastasis following personalized immunotherapy in combination with integrative therapeutics. Therefore, it is crucial to assess not only conventional immune biomarkers but also patient-specific parameters.

The novel COVID-19 pandemic prompted an unprecedented Institutional reaction to aggregate existing capacity from silos of research laboratories to establish a multidisciplinary research laboratory for COVID19 testing. In less than two weeks, resources were mobilized from the community to strengthen public health response and epidemic control. Such strengthening of institutional research capacity to support public health response contributes to a natural knowledge transfer, facilitates collaboration, and generates new frontiers for knowledge production that should ultimately lead to professional development and retention of skilled human resources. This report describes the pre-established mechanisms and involvement of the authors that made it possible to set-up a multidisciplinary laboratory in a remarkably short period of time. We also discuss the opportunities and sustainability of multidisciplinary laboratory research post-COVID19. Existing institutional capacity can be repurposed to establish multidisciplinary research laboratories to support the strengthening of basic and clinical translational research capacity in resource limited settings and impact on public health and scientific knowledge for socioeconomic development.

We investigated humoral immune responses in 222 unvaccinated Japanese people after recovery from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in 2021. Anti-spike protein IgG antibody levels and neutralizing antibody titers were measured in serum samples obtained within 20–180 days after diagnosis. The geometric mean of antibody titers was 1555 ELU/mL (95% confidence interval [CI] = 1257-1923), and the neutralizing activities (50% inhibitory dilution) was 253 (95% CI = 204-313). The antibody titer and neutralizing activity both increased with increasing disease severity, and both values were approximately 4-fold higher for hospitalized patients than for non-hospitalized patients. However, these differences were smaller in older patients. The humoral immune response, which increased with increasing disease severity, gradually decreased over time after SARS-CoV-2 infection. Most patients with mild or moderate symptoms sustained neutralizing activity for up to 180 days after the infection, the decay of the neutralizing activity in the patients with asymptomatic was rather faster than other groups. 11.7% (26/222) of patients had very low neutralizing activity, and half of these were in the age of 20s. Our study results show the importance of measuring the neutralizing activity to confirm the immune status and also estimate the timing of vaccine.

Depression is the most common chronic mental illness and is characterized by low mood, insomnia, and affective disorders. However, the pathological mechanisms of depression remain unclear. Numerous studies have suggested that the ghrelin/GHSR system may be involved in the pathophysiologic process of depression. Ghrelin plays a dual role in the experimental animals, increasing depressed behavior and decreasing anxiety. By combining several neuropeptides and traditional neurotransmitter systems to construct neural networks, it modifies signals connected to depression. The present review emphasizes the role of ghrelin in neuritogenesis, astrocyte protection, inflammatory factor production, and endocrine disruption in depression. Furthermore, ghrelin/GHSR can activate multiple signaling pathways to produce antidepressant effects, including cAMP/CREB/BDNF, PI3K/Akt, Jak2/STAT3, and p38-MAPK. Given the antidepressant effects of ghrelin/GHSR1a system, it is expected to become a potential therapeutic target for the treatment of depression.

Surface water plays a crucial role in climate change, human production, and life, making accurate monitoring and observation of surface water particularly important. However, due to the significant diversity and complexity of water distribution in surface space, accurate mapping of surface water faces considerable challenges. When extracting water bodies from medium-resolution satellite remote sensing images, CNN methods may suffer from limitations in receptive fields and insufficient context modeling capabilities, leading to the loss of water body boundary details and poor fusion of multiscale features. Currently, there is relatively little research on this issue; therefore, it is necessary to explore new combinations of deep learning networks to address these challenges. The purpose of this study is to address the above issues. We propose a new combination of deep learning networks that fully utilize multiscale information to enhance water features. Specifically, we first combine deformable convolutions with the Swin Transformer to increase effective receptive fields while better integrating global semantic information. This combination can capture features of water bodies at different scales, improve the accuracy and integrity of water extraction, and provide reliable technical support for detailed water body extraction. We tested the newly constructed model using Sentinel-2 satellite images. Our model achieved results of over 90%, with an average accuracy of 97.89%, average precision of 94.98%, average recall of 90.05%, and an average F1 score of 92.33%. In addition, our model achieved an accuracy of 98.03% in mountainous areas. Our experiments and results validate the potential of combining the Swin Transformer and deformable convolutions in detailed water body extraction.

Locally advanced rectal cancer requires multimodal treatment. Radiotherapy is being explored for intensification to improve rates of pathological complete responses (ypCR-rates) that correlates with better outcomes. This study reports a comparison between standard versus escalated dose in preoperative scenario. YpCR-rates, toxicity, postoperative complications, disease-free and overall survival at 5 years have been described. From 2012 to 2019, 99 patients were analyzed retrospectively: standard arm (mean 47.5Gy) vs. dose-escalated arm (mean 54.3Gy). All patients were treated with 3DRT in 25 fractions, concomitantly capecitabine and surgery performed according to total mesorectal excision principles in both arms. YpCR was reported by “College of American Pathologist grades”; gastrointestinal (GI) and genitourinary (GU) toxicity using the “Common Terminology Criteria for Adverse Events” (CTCAE 4.0). YpCR-rates were higher in the dose-escalated group (25% vs. 10.64%; p = 0.07) with a lower rate of non-treatment response (61.36% vs. 38.64%; p = 0.11). No statistical differences between arms were found in terms of oncological outcomes, all postoperative complications (p = 0.15), second surgeries (p = 0.62), or deaths (p = 0.62). CTCAE acute-GI and GU toxicity were grade I or II in both arms. Our study shows that the dose-escalated scheme is safer and obtains higher ypCR-rates.

G-protein-coupled receptors (GPCR) belong to a large family of molecules eliciting different responses to a variety of signaling molecules. These receptors participate in various physiologic pathways such as metabolism, growth, immune responses, inflammation, vision, taste, olfaction, neurotransmission and even and pathologic responses including chronic inflammatory and vascular diseases. Receptors contributing to the biological responses of renin-angiotensin system (RAS) are members of GPCR family. COVID-19-induced inflammatory cascade has been attributed to acute ACE2 downregulation and imbalance of proinflammatory ACE/AngII/AT1R and anti-inflammatory ACE2/angiotensin (1-7)/Mas axes in favor of the former. Some of the receptors contributing to activities of proteins in RAS including AT1R, AT2R and Mas receptors are members of GPCR family. It is notable that these receptors induce their effects both through G protein and β-arrestin pathway; the former exerts temporary and the latter more sustained effects. In addition to the imbalance of GPCR responses contributing to RAS activities, it has been suggested that SARS-CoV2 pathogenesis might be attributed to the activation of GPCRs or modulating G-proteins involved in adenosine-CFTR regulation system and epithelial Na channel function.This article includes a minireview about the physiological functions of GPCRs and their contribution to COVID-19.

The article presents the case of a 27-year-old female patient with idiopathic congenital complete heart block who was referred by her primary care physician for cardiological evaluation. The conduction disturbance was recognized at the age of 6 years and was asymptomatic. The disqualification from pacemaker implantation included detailed history of a patient’s symptoms, echocardiographic assessment of the heart, exercise testing and ECG Holter monitoring. The aid of salbutamol administered orally was also useful.

Salmonella is a foodborne pathogen associated with localized outbreaks that potentially pose a huge risk to the public health in some countries such as Africa and India, especially in the regions lacking health care measures. The focus of the present study is the molecular detection method of salmonella. The study was conducted to identify Salmonella strains by PCR targeting flagellin specific gene from suspected patient’s clinical blood samples. Serologically, Widal test results showed that all serum samples except one patient (ID 7) were positive for O antigen. The serum of three patients (ID 4, 5 and 10) were found to be positive for H and O antigens while one patient (ID 9) showed agglutination for antigen H. The PCR approach used in this study was successful for fast and precise detection of clinically related Salmonella Typhi. Seven samples out of 10 were positive for Salmonella Typhi as fragments of 458-497bp were observed on the gel-agarose 0.8% corresponds to flagellin gene of S. Typhi. Relatively, smaller amplicons of 366-343bp were observed utilizing nested PCR as well, which seems more sensitive than the conventional PCR. Therefore, we recommend the PCR approach performed in this study to be used as a fast, cost-effective, and time-consuming tool for diagnostic purposes of Salmonella.

Metastatic pancreatic ductal adenocarcinoma pancreatic (PDAC) is characterized by poor prognosis and short survival. Today, the use of new polytherapeutic regimens increases clinical outcome of these patients opening new clinical scenario. A crucial issue related to the actual improvement achieved with these new regimens is represented by the occasional possibility to observe a radiological complete response of metastatic lesions in patients with synchronous primary tumor. What could be the best therapeutic management of these patients? Could surgery represent an indication? Herein we reported a case of a patient with a PDAC of the head with multiple liver metastasis, who underwent first line chemotherapy with mFOLFIRINOX. After 10 cycles, he achieved a complete radiological response of liver metastases and a partial response of pancreatic lesion. A, duodenocephalopancreasectomy was performed. Due to liver a lung metastases after 8 months from surgery, a second line therapy was started with a disease free survival and overall survival of 8 months and 45 months, respectively. Improvement in the molecular characterization of PDAC could help in the selection of patients suitable for multimodal treatments.

Currently, there are few effective methods to monitor patients with gynecologic malignancies, including ovarian and endometrial cancers. The ‘tumor fraction’ of circulating tumor DNA may offer a minimally-invasive method to evaluate tumor aggressiveness via simple blood draw. To date, there is limited data on utility of ctDNA TFx interrogation in gynecologic malignancies, although there is evidence that TFx may be low or undetectable in malignancies primarily confined to the peritoneal cavity. The objective was a descriptive assessment of ctDNA TFx in a cohort of patients with ovarian and endometrial cancer. The association of TFx to continuous and categorical baseline clinicopathologic factors and progression-free survival was assessed. In a large cohort comprised of 210 plasma samples from 78 patients with biopsy-proven ovarian and endometrial cancer, plasma samples collected between 4/2018 and 4/2020 were subjected to shallow whole genome sequencing with determination of TFx. Overall, we demonstrate that TFx was low for most samples with mean TFx for ovarian cancer 5.5% and endometrial cancer 2.4%, yet can be quantified above lower limit of detection in many patients particularly when evaluating serial samples. There was no significant association between TFx level and clinical (primary vs recurrent, best response) or pathologic (e.g. histology, grade) features. Our data demonstrates low TFx levels in ovarian and endometrial cancers likely reflect that tumor types are ‘low ctDNA shed’, suggesting that higher sensitivity assessments may be required for clinical utility.

There is no routine approach to identify patients with gynecologic malignancies at high risk for chemotherapy-resistance. Circulating tumor DNA (ctDNA) tumor fraction (TFx) represents a minimally-invasive approach to tumor profiling, with as yet limited data on utility in gynecologic malignancies. The objective was to investigate the use of ctDNA TFx in a cohort of patients with ovarian and endometrial cancer. Plasma samples from patients with biopsy-proven ovarian or endometrial cancer collected between 4/2018 and 4/2020 were subjected to shallow whole genome sequencing with determination of TFx via ichorCNA package. The association of TFx to continuous and categorical baseline clinicopathologic factors and progression-free survival was assessed. 210 plasma samples from 78 patients with gynecologic cancers were analyzed. Mean TFx for ovarian cancer was 5.5% and endometrial cancer 2.4% and there was no significant difference in TFx among histology either for endometrial or ovarian cancers. Grade was associated with significant difference in ‘sentinel’ TFx among ovarian cancers but not endometrial cancers. ctDNA TFx dynamics over time demonstrated rapid clearance of ctDNA in most patients with ovarian cancer, while endometrial cancer had consistently low TFx. ctDNA TFx is feasible in ovarian and endometrial cancers and may be a valuable important tool for prognostication.

A mathematical procedure enabling the transformation of an arbitrary tessellation of a surface into a bi-colored complete graph is introduced. Polygons constituting the tessellation are represented by vertices of the graphs. Vertices of the graphs are connected by two kinds of links/edges, namely, by a green link, when polygons have the same number of sides, and by a red link, when the polygons have a different number of sides. This procedure gives rise to a semi-transitive, complete, bi-colored Ramsey graph. The Ramsey number was established as R_trans (3,3)=5. Shannon entropies of the tessellation and graphs are introduced. Ramsey graphs emerging from random Voronoi and Poisson Line tessellations were investigated. The limits ζ=lim┬(N→∞)⁡〖N_g/N_r 〗, where N is the total number of green and red seeds, N_g and N_r, were found ζ=0.272±0.001 (Voronoi) and ζ= 0.47±0.02 (Poisson Line). The Shannon Entropy for the random Voronoi tessellation was calculated as S= 1.690±0.001 and for the Poisson line tessellation as S =1.265±0.015.

Using a scaling symmetry, it is shown how to compute polynomial conservation laws, generalized symmetries, recursion operators, Lax pairs, and bilinear forms of polynomial nonlinear partial differential equations thereby establishing their complete integrability. The Gardner equation is chosen as the key example for it comprises both the Korteweg-de Vries and modified Korteweg de Vries (mKdV) equations. The Gardner and Miura transformations which connect these equations are also computed using the concept of scaling homogeneity. Exact solitary wave solutions and solitons of the Gardner equation are derived using Hirota’s method and other direct methods. The nature of these solutions depends on the sign of the cubic term in the Gardner equation and the underlying mKdV equation. It is shown that flat (table-top) waves of large amplitude only occur when the sign of the cubic nonlinearity is negative (defocusing case) whereas the focusing Gardner equation has the standard elastically colliding solitons. The paper’s aim is to provide a review of integrability properties and solutions of the Gardner equation and illustrate the applicability of the scaling symmetry approach. The methods and algorithms used in this paper have been implemented in Mathematica but can be adapted for major computer algebra systems.

Synchronous electrical cardioversion is a relatively common procedure in the emergency department (ED), often performed for unstable supraventricular tachycardia (SVT) or unstable ventricular tachycardia (VT). However, it is also used for stable cases resistant to drug therapy, which carry a risk of deterioration. In addition to the inherent risks linked with procedural sedation, there is a possibility of malignant arrhythmias or bradycardia, which could potentially result in cardiac arrest following this procedure. Here, we present a case of complete heart block unresponsive to transcutaneous pacing and positive inotropic and chronotropic drugs for 90 minutes, resulting in multiple cardiac arrests. The repositioning of the transcutaneous cardio-stimulation electrodes, one of them placed in the left latero-sternal position and the other at the level of the apex, led to immediate stabilization of the patient. Extubation of the patient was performed the following day, while full recovery and discharge within 7 days, after insertion of a permanent pacemaker.

The Svenonius theorem establishes the correspondence between definability of relations in a countable structure and automorphism groups of these relations in the extension of the structure. This correspondence may help in finding the description of the definability lattice constituted by all definability spaces (reducts) of the original structure. However, the major difficulty here is the necessity to consider the extensions which generally are obscure and hardly amenable to classification. Because of that results on definability lattices were obtained only for $\omega$-categorical structures (i.\,e. those in which all elementary extensions are isomorphic to the structure itself). All known definability lattices for such structures proved to be finite. In this work we introduce the concept of upwards complete structure as such in which all its extensions are isomorphic. Further we define upwards completions structures. For such structures Galois correspondence between definability lattice and the lattice of closed supergroups of automorphism group of the structure is an anti-automorphism. These lattices could be infinite in general. We describe the natural class of structures which have upwards completion, that is discretely homogeneous graphs, present the explicit constrcution of their completion and automorphism groups of completions. We establish the general \emph{localness} property of discretely homogeneous graphs and present the examples of completable structures and their completions.

Achieving effective control over microbial contamination necessitates the precise and concurrent identification of numerous pathogens. In this research, we have devised a remarkably sensitive duplex droplet digital PCR (dddPCR) reaction system to simultaneously detect Pseudomonas aeruginosa (P. aeruginosa) and Pseudomonas fragi (P. fragi). Employing comparative genomics, we identified four genes of P. fragi. By specific analysis, RS22680 gene was selected as the detection target of P. fragi and LasR gene was chosed as P. aeruginosa, which were applied to construct a dddPCR reaction. In terms of specificity, sensitivity and anti-interference ability, the constructed dddPCR detection system was verified and analyzed. The assay showed excellent sensitivity and applicability, as evidenced by a limit of detection of 100 CFU/mL. When the concentration of natural background bacteria in milk or fresh meat was 100 times that of the target detection bacteria, the method was still capable of completing the absolute quantification. In the simulation of actual sample contamination, P. aeruginosa could be detected after 3 h of enrichment culture, and P. fragi could be detected after 6 h. The established ddPCR detection system exhibits exceptional performance, serving as a foundation for the simultaneous detection of various pathogenic bacteria in food products.

The P versus NP problem is a fundamental question in computer science. It asks whether problems whose solutions can be quickly verified can also be quickly solved. Here, "quickly" refers to computational time that grows proportionally to the size of the input (polynomial time). While the problem's roots trace back to a 1955 letter from John Nash, its formalization is attributed to Stephen Cook and Leonid Levin. Despite extensive research, a definitive answer remains elusive. Closely tied to this is the concept of NP-completeness. If a single NP-complete problem could be solved efficiently, it would imply that all problems in NP can be solved efficiently, proving that P equals NP. This work posits that MONOTONE ONE-IN-THREE 3SAT, a notoriously difficult NP-complete problem, can be solved efficiently, thereby establishing the equivalence of P and NP.

A sparse language is a formal language such that the number of strings of length $n$ is bounded by a polynomial function of $n$. We create a class with the opposite definition, that is a class of languages that are dense instead of sparse. We define a dense language on $m$ as a formal language (a set of binary strings) where there exists a positive integer $n_{0}$ such that the counting of the number of strings of length $n \geq n_{0}$ in the language is greater than or equal to $2^{n - m}$ where $m$ is a real number and $0 < m \leq 1$. We call the complexity class of all dense languages on $m$ as $DENSE(m)$. We prove that there exists an $\textit{NP--complete}$ problem that belongs to $DENSE(m)$ for every possible value of $0 < m \leq 1$.

Garcinia indica (Thouars) Choisy (commonly known as Kokum) is an important medicinal plant in Ayurvedic medicine and belonging to the Clusiaceae family. Kokum is a highly traded medicinal plant species of India, dried rind of fruits and butter from seeds are traded in the markets. This study aimed to assemble the complete chloroplast (cp) genome of G. indica and compare it with the previously published cp genome within the Garcinia taxa to identify the potential species-specific barcode marker for Kokum authentication. The assembled cp genome, with a size of 1,56,891 bp, exhibits a typical quadripartite structure. The large single-copy (LSC) region spans 85,580 bp, and the small single-copy (SSC) region spans 17,181 bp, together comprising 64.5% of the genome. The pair of inverted repeats (IRA and IRB) are each 27,065 bp, covering the remaining 35.5% of the genome. A total of 126 unique genes were identified, including 86 protein-coding genes, 32 tRNA genes, and 8 rRNA genes. Phylogenetic analysis using complete cp genomes from 12 species in the Clusiaceae family indicated that 10 Garcinia species form a sister clade. Notably, the ycf1 gene in the LSC region was more divergent within the Garcinia species. This study presents the first report on the chloroplast genome of G. indica. This chloroplast genome resource provides a basis for identifying new DNA barcode marker as well as species-specific marker for herbal drug authentication of Kokum, and species delineation within the Garcinia genus.

A popular account of the mixing patterns for the three generations of quarks and leptons is through the characters $\kappa$ of a finite group $G$. Here we introduce a $d$-dimensional Hilbert space with $d=cc(G)$, the number of conjugacy classes of $G$. Groups under consideration should follow two rules, (a) the character table contains both two- and three-dimensional representations with at least one of them faithful and (b) there are minimal informationally complete measurements under the action of a $d$-dimensional Pauli group over the characters of these representations. Groups with small $d$ that satisfy these rules coincide in a large part with viable ones derived so far for reproducing simultaneously the CKM (quark) and PNMS (lepton) mixing matrices. Groups leading to physical $CP$ violation are singled out.

Butirosins are naturally occurring aminoglycoside (AG) antibiotics featuring a 4,5-disubstituted 2-deoxystreptamine (2-DOS) with a (2S)-4-amino-2-hydroxybutyrate (AHBA) side chain. This side chain has been shown to confer resistance against AG modifying enzymes, leading to ongoing studies on the butirosin biosynthetic pathway and the corresponding enzymes. Butirosin is produced by Niallia (formerly Bacillus) circulans and Bacillus vitellinus, with most research focused on the former strain. To date, no whole genome analysis has been performed on B. vitellinus. In this study, we sequenced the complete genome of B. vitellinus NBRC 13296 and performed a comparative analysis of different butirosin biosyntheric gene clusters (BGCs), including those from N. circulans. The complete genome of B. vitellinus NBRC 13296 comprises a 6,331,192 base circlular chromosome with a GC content of 52.68%. The annotation revealed the presence of 5,605 CDSs, 70 tRNA genes, 30 rRNA genes, and 3 ncRNA genes in NBRC 13296. The highest dDDH and ANI values between NBRC 13296 and the most closely related type strain, Paenibacillus chitinolyticus KCCM 41400, were 97.8% and 98.66%, respectively. Based on these genome-based comparative analyses, we propose reclassifying B. vitellinus NBRC 13296 as P. chitinolyticus. Genome mining revealed 18 gene clusters encoding the biosynthesis of diverse secondary metabolites in the genome of B. vitellinus NBRC 13296, indicating the enormous biosynthetic potential of this strain. The predicted structural diversity of secondary metabolites includes aminoglycoside, PKS, NRPS, PKS-NRPS hybrids, metallophores, phosphonates, terpenes, β-lactones, and RiPP peptides. We then comparatively characterized butorosin BGCs previously studied in several N. circulans strains. Additionally, comparative genome analysis revealed complete butirosin BGCs identified from P. chitinolyticus KCCM 41400, P. chitinolyticus NRRL B-23119, P. chitinolyticus NRRL B-23120, P. chitinolyticus B-14908, P. chitinolyticus YSY-3.1, P. chitinolyticus JMW06, Paenibacillus sp. GbtcB18, Paenibacillus sp. HGH0039, Paenibacillus sp. MZ04-78.2. Finally, we identified the core region consisting of BtrS, BtrN, BtrM, BtrL, BtrA, BtrB, BtrC, BtrD, BtrD, BtrE, BtrF, BtrG, BtrH, BtrI, BtrI, BtrJ, BtrK, BtrO, BtrP, and BtrV, followed by an upstream region organizing BtrQ, BtrW, BtrX, BtrY, and BtrZ in the same transcriptional direction and sequential genetic arrangement, and a downstream region organizing various proteins based on BtrT, BtrR2, BtrU, and BtrR1. Our study provided insights into the reclassification of B. vitellinus NBRC 13296 to P. chitinolyticus and suggested the need for continued studies on butirosin biosynthesis from an enzymatic perspective.

Background and Objectives: Renal cell carcinomas and upper tract urothelial carcinomas are types of malignancies that originate in the kidneys. Each of these examples shows an increasing trend in the frequency and the mortality rate. This study aims to comprehensively define carcinomas by analysing clinical, paraclinical, and histological aspects to predict aggressiveness and mortality. Materials and Methods: We conducted a retrospective investigation on a group of patients suspected with kidney cancers. Results: We identified 188 cases. We observed a higher mortality rate and older age in individuals with urothelial carcinomas. Anemia, acute kidney injury, hematuria and perineural invasion were the main risk factors that predicted their mortality. Tumour size in renal cell carcinomas correlates with the presence of necrosis and sarcomatoid areas. Factors that indicate a higher rate of death are older age, exceeding of the renal capsule, the lesion that includes the entire kidney, lymphovascular invasion, acute kidney injury, and anemia. Conclusions: Even if they originate at the renal level, and the clinical-paraclinical picture is similar, the histopathological characteristics make the difference. In addition, to these are added the previously mentioned common parameters that can represent important prognostic factors. In conclusion, the characteristics commonly identified in one type of cancer may act as risk factors for the other tumor. The detected data include threshold values and risk factors, making a significant contribution to the existing literature.

Droplet digital polymerase chain reaction (ddPCR) is a method used to detect and quantify nu-cleic acids even when present in exceptionally low numbers. While it has proven to be valuable for clinical studies, it has failed to be widely adopted for environmental and applied studies. Due to the complexity of the chemical and biological composition of environmental samples, protocols tailored to clinical studies are not appropriate, and results are difficult to interpret. We used en-vironmental DNA samples originating from field studies to determine a protocol for environ-mental samples. Samples included field soils which had been inoculated with the soil fungus Rhizophagus irregularis (environmental positive control), field soils that had not been inoculat-ed and the targeted fungus was not naturally present (environmental negative control), and root samples from both field categories. To control for the effect of soil inhibitors, we also in-cluded DNA samples of an organismal control extracted from pure fungal spores (organismal positive control). Finally, we included a no-template control consisting only of the PCR reaction reagents and nuclease free water instead of template DNA. Using original data, we examined which factors contribute to poor resolution in root and soil samples and propose best practises to ensure accuracy and repeatability. Furthermore, we evaluated manual and automatic threshold determination methods and we propose a novel protocol based on multiple controls that is more appropriate for environmental samples.

This study examined the efficiency of a combined diagnostic approach in which bioamplification of Arizona 861 S1 citron was integrated with RT‒PCR to detect Citrus tristeza virus (CTV) and Hop Stunt Viroid (HSVd) in Citrus plants. The capacity of the combined method to diagnose mixed infections of CTV and HSVd, along with early single infections of both pathogens, was assessed by evaluating the incubation periods and differential detection capabilities between the combined and direct methods. The bioamplification method exhibited high sensitivity (100%) and specificity (100%) for CTV, enabling early detection even in patients with low-titer infections compared to direct RT‒PCR. However, for HSVd, the combined method achieved 50% sensitivity in early infections, highlighting the need for additional replicates to enhance diagnostic reliability. The relative concentrations of pathogens in mixed and simple infections were evaluated; the analysis revealed no significant differences, suggesting minimal interference between CTV and HSVd. This approach allows early detection of CTV but requires further refinement for HSVd diagnosis. These findings underscore the complementary roles of the cost-effectiveness of direct RT‒PCR and the sensitivity of the combined method in implementing effective disease control strategies in citrus crops.

Sleep is still considered a mystery, despite intense scientific investigation. Here we present the first complete biological theory of sleep. The role of sleep is to restore the optimal homeostatic state, which is essential for tissue performance and health. Non-rapid eye movement sleep (NREMS) restores cortical and most other brain neurons, via relaxed global activity managed by thalamocortical circuits. The role of REM sleep is to restore acetylcholine (ACh) neurons, which support focused responses and hence cannot participate in global oscillations. Sleep enhances learning and memory via state restoration and ACh-affected paths. NREMS induces a lack of consciousness because global synchronous activity prevents focused responses, which are essential for consciousness. Dreams result from focused neural firing during sharp-wave ripples and REMS, and have a sense of reality because they involve the same neurons representing focused perceptual responses during wake. Anesthetics utilize a variety of mechanisms that prevent focused responses.

Tungiasis is a neglected tropical disease caused by skin-penetrating female Tunga penetrans fleas. Although tungiasis causes severe health problems, its ecology is poorly understood and morphological descriptions of larvae are unavailable. To identify T. penetrans immature stages and sites where they develop, diagnostic PCRs are required. However, flea larvae feed on soil organic matter rich in PCR inhibitors. Here, three DNA preparation methods, a soil DNA kit removing inhibitors, a simple ammonium acetate precipitation approach (AmAcet) and a crude lysate of larvae (CL), were combined with amplification by the highly processive FIREPol® Taq or the inhibitor-resistant Phusion® polymerase. Independent of the polymerase used, frequency of successful amplification, Cq values and PCR efficacies for the low-cost CL and AmAcet methods were superior to the commercial kit for amplification of a 278 bp partial internal transcribed spacer-2 (ITS-2) and a 730 bp pan-Siphonaptera cytochrome oxidase I PCR. For the CL method combined with Phusion® polymerase, costs were approximately 20-fold lower than for methods based on the soil DNA kit, which is a considerable advantage in resource-poor settings. The ITS-2 PCR did not amplify Ctenocephalides felis genomic or Tunga trimammilata ITS-2 plasmid DNA allowing it to be used to specifically identify T. penetrans.

Emergency Medical Response Systems (EMRS) play a vital role in delivering medical aid during disasters. In Serbia, these systems encounter several obstacles due to economic limitations, scarce resources, and ongoing healthcare reforms, all of which affect their performance. This study focuses on examining the risk factors and effectiveness of Serbia's EMRS, identifying systemic issues, and proposing enhancements to improve emergency preparedness for disasters. A survey was conducted across healthcare institutions in Serbia to gather comprehensive insights. The survey explored various aspects, including organizational structures, resource distribution, communication systems, and the speed of emergency responses. Statistical methods such as Pearson's correlation, multivariate regression analysis and chi-square tests were utilized to interpret data collected from 172 healthcare institutions. The multivariate regression analysis of EMS organization identified that factors such as the structuring of working hours and shift patterns are significant predictors, contributing to small but meaningful variations in various EMS outcomes. In contrast, other factors, such as financial resources and the availability of ambulance vehicles, showed more targeted significance depending on the specific EMS aspect being analyzed. Institutions with written mass casualty plans and procedures demonstrate superior effectiveness in EMS activities, excelling in resource allocation, operational continuity, and emergency preparedness, particularly through enhanced coordination and specialized systems for managing large-scale emergencies. The findings highlighted notable disparities in how EMRS are organized, with only 46% of institutions having specific emergency departments. Resource limitations were evident, as many facilities operated with minimal staff and outdated equipment. The communication systems revealed weaknesses, with just 54% having established protocols for handling emergency calls. Response times frequently surpassed international standards, which negatively affected patient outcomes. Additionally, the study found training deficiencies, with 68% of participants expressing a need for further education. Serbia’s EMRS are significantly impacted by systemic issues such as insufficient funding, disjointed service delivery, and inadequate training. Overcoming these challenges necessitates a comprehensive approach that includes boosting financial support, implementing standardized protocols, and fostering enhanced collaboration across different sectors.

Hox gene clusters are crucial in Embryogenesis. It was observed that some Hox genes are located in order along the telomeric to centromeric direction of the DNA sequence: Hox1, Hox2, Hox3…. These genes are expressed in the same order in the ontogenetic units of the Drosophila embryo along the Anterior-Posterior axis. The two entities (genome and embryo) differ significantly in linear size and in-between distance. This strange phenomenon was named Spatial Collinearity (SP). Later, it was observed that, particularly in the Vertebrates, a Temporal Collinearity (TC) coexists: first is Hox1 expressed, later Hox2 and even later Hox3,…,. According to a Biophysical Model (BM), pulling forces act at the anterior end of the cluster while a cluster fastening applies at the posterior end. Hox clusters are irreversibly elongated along the force direction. During Evolution, the elongated Hox clusters are broken at variable lengths thus split clusters may be created. An Empirical Rule was formulated distinguishing development due to a complete Hox cluster from development due to split Hox clusters. BM can ‘explain’ this Empirical Rule. In a spontaneous mutation where the cluster fastening is dismantled, a weak pulling force automatically shifts the cluster inside the Hox activation domain. This cluster translocation can probably explain the absence of Temporal Collinearity in Drosophila.

Background: Gender differences characterize the prevalence and attitudes toward weight management. Despite limited evidence suggesting greater weight loss in women with anti-obesity pharmacotherapy, gender-specific analysis remains underexplored. This retrospective study aimed to evaluate the gender-specific response to liraglutide 3.0 mg treatment in people with obesity without type 2 diabetes (T2D). Methods: Data were collected from 47 patients (31 women, 16 men), with age > 18 years, BMI ≥ 30 kg/m^2, absence of T2D, and with exclusion of prior anti-obesity treatment, comorbidities, or bariatric surgery. Only patients who maintained the liraglutide 3.0 mg dose for at least 6 months were included. Results: Both sexes showed significant reductions in weight and BMI at 3 and 6 months. Men achieved greater weight loss (WL), BMI reduction, % WL, WL >5% and >10% than women, and they also showed more significant improvements in metabolic parameters (total and LDL cholesterol, Fibrosis-4 Index FIB-4). No significant gender differences were observed in glucose metabolism or renal function. Conclusion: This study showed a greater therapeutic effect of liraglutide 3.0 mg in men. Given men's higher risk of cardiovascular disease (CVD), and underrepresentation in clinical weight loss programs, these findings may increase male engagement and improve their CVD risk.

A Infectious diseases account for about 3 million deaths per year. The advent of molecular techniques has led to an enormous improvement in their diagnosis, both in terms of sensitivity and specificity and in terms of the speed with which a clinically useful result can be obtained. Digital PCR, or 3rd generation PCR, is based on a series of technical modifications that result in more sensitive techniques, more resistant to the action of inhibitors and capable of direct quantification, without the need for standard curves.This review presents the main applications that have been developed for the diagnosis of viral, bacterial and parasitic infections, and the potential prospects for the clinical use of this technology.

We present a first formal analysis of specific and complete local integration. Complete local integration was previously proposed as a criterion for detecting entities or wholes in distributed dynamical systems. Such entities in turn were conceived to form the basis of a theory of emergence of agents within dynamical systems. Here, we give a more thorough account of the underlying formal measures. The main contribution is the disintegration theorem which reveals a special role of completely locally integrated patterns (what we call ι-entities) within the trajectories they occur in. Apart from proving this theorem we introduce the disintegration hierarchy and its refinement-free version as a way to structure the patterns in a trajectory. Furthermore we construct the least upper bound and provide a candidate for the greatest lower bound of specific local integration. Finally, we calculate the i-entities in small example systems as a first sanity check and find that ι-entities largely fulfil simple expectations.

In this paper we study the properties of a complete quadrangle in the Euclidean plane. Similarly, as it was done in our previous work the proofs are based on using rectangular coordinates symmetrically on four vertices and four parameters a,b,c,d. Here, we gather many properties of the complete quadrangle known from earlier but proved by the same method together with some new results given.

Robin criterion states that the Riemann Hypothesis is true if and only if the inequality $\sigma(n) < e^{\gamma } \times n \times \log \log n$ holds for all $n > 5040$, where $\sigma(n)$ is the sum-of-divisors function and $\gamma \approx 0.57721$ is the Euler-Mascheroni constant. We prove that the Robin inequality is true for all $n > 5040$ which are not divisible by any prime number between $2$ and $953$. Using this result, we show there is a contradiction just assuming the possible smallest counterexample $n > 5040$ of the Robin inequality. In this way, we prove that the Robin inequality is true for all $n > 5040$ and thus, the Riemann Hypothesis is true.

The arsRBC operon encodes a 3-protein arsenic resistance system. ArsR regulates the transcription of the operon, while ArsB and ArsC are involved in exporting trivalent arsenic and reducing pentavalent arsenic respectively. Previous research in Agrobacterium tumefaciens 5A demonstrated ArsR has regulatory control over a wide range of metal related proteins and metabolic pathways. We hypothesized that ArsR has broad regulatory control in other Gram-negative bacteria and set out to test this. Here, we use differential proteomics to investigate changes caused by the presence of the arsR gene in human microbiome-relevant Escherichia coli during arsenite (AsIII) exposure. We show that ArsR has broad-ranging impacts such as the expression of TCA cycle enzymes during AsIII stress. Additionally, we found that the Isc [Fe-S] cluster and molybdenum cofactor assembly proteins are upregulated regardless of the presence of ArsR under these same conditions. An important finding from this differential proteomics analysis was the identification of response mechanisms that were strain, ArsR, and arsenic specific, providing new clarity to this complex regulon. Given the widespread occurrence of the arsRBC operon, these findings should have broad applicability across microbial genera, including sensitive environments such as the human gastrointestinal tract.

With the rapid development of wind power generations, the inertial response of wind turbines (WTs) are widely concerned recently, which is important for grid frequency dynamic and stability. This paper recognizes and understands the inertial response of type-3 and type-4 WTs from the view of equivalent internal voltage, in analogy with typical synchronous generators (SGs). Due to the dynamic of the equivalent inertial voltage different from SGs, the electromechanical inertia of WTs is completely hidden. The rapid power control loop and synchronization control loop is the main reasons that the WT's inertial response is disenabled. On the basis of the equivalent internal voltage's dynamic, the existing inertia control method for WTs are reviewed and summarized as three approaches from the view of WT's control, i.e. optimizing the power control or synchronization control or both. At last, the main challenges and issues of these inertia controls are attempted to explain and address.

Background: The aim of this study was to investigate a series of complete blood cell count – based biomarkers of systemic inflammation as predictors of clinical outcomes in patients who underwent first-line chemoimmunotherapy for advanced NSCLC. Methods: Consecutive patients with pathologically diagnosed stage III/IV NSCLC who underwent first-line chemoimmunotherapy were retrospectively enrolled. The clinical outcomes used for biomarker evaluation were Objective Response Rate (ORR) and Overall Survival (OS). Results: Non-responders had significantly higher values of neutrophil to lymphocyte ratio (NLR, median: 5.36; IQR: 2.78–10.82 vs. 3.31; IQR: 2.15–4.12, p = 0.019), neutrophil to monocyte ratio (NMR, median: 14.00; IQR: 8.82–21.20 vs. 9.20; IQR: 7.45–11.20, p = 0.013), and systemic inflammation index (SII, median: 1,395; IQR: 929–3,334 vs. 945; IQR: 552–1,373, p = 0.025), but only NLR and NMR remained independently related with clinical response in multivariate logistic regression. In univariate analysis white blood cells (OR:1.2202; 95% CI: 1.0339 - 1.4400, p=0.019), neutrophils (OR:1.2916; 95% CI: 1.0692 - 1.5604, p=0.008), NLR (OR:1.3601: 95% CI: 1.0949 - 1.6896, p=0.005) and NMR (OR:1.2159; 95% CI: 1.00396 - 1.4221, p=0.015) were significantly associated with survival; Cox regression models confirmed that neutrophils, NLR and MLR were independently associated with survival; NLR at a cut-off value of 4.0 showed the better AUC (0.749) in predicting survival. Conclusions: Baseline complete blood cell count biomarkers, especially the NLR, can predict clinical outcomes in patients with advanced NSCLC treated with first-line chemoimmunotherapy.

The Cyber Security Incident Response Team executes its critical duties in the Centralized Security Operation Centers. Its primary target is to protect organizational resources from all types of Advanced Persistent Threats (APT) and attacks while making the correct judgments at the right time. Despite the great efforts and the efficient tools, the incident response mechanism faces two significant challenges. The generation of large amounts of alerts with huge rates of false positives per unit of time and the time-consuming manual expert engagement in the post-alert decision phase. The main aim of this research study is to investigate the potential of Machine Learning techniques in solving the above challenges. This study proposes six event detection modules to identify 14 different types of malicious behavior using Splunk. The available APT dataset is extended by adding more alert types. Then, the machine learning techniques i.e., Random Forest, and XGBoost, are applied to improve the decision process and reduce the false positive alerts of the Security Information and Event Management system. The proposed model outperforms the state-of-the-art techniques by predicting an accuracy of 99.6%.

Canine distemper virus (CDV) poses a substantial threat to diverse carnivores, leading to sys-temic and often fatal diseases. Accurate and prompt diagnosis is paramount for effective man-agement and curbing further transmission. This study evaluates the diagnostic efficacy of drop-let digital PCR (ddPCR) in comparison to conventional reverse transcription (RT-PCR) and quantitative reverse transcription real-time PCR (RT-qPCR). Seventy-six clinical samples were collected from dogs with CDV symptoms diagnosed by specialized veterinarians, and sixteen samples from asymptomatic individuals. Conventional PCR, quantitative real-time PCR, and ddPCR were deployed, and their diagnostic capabilities were meticulously assessed. DdPCR ex-hibited heightened analytical sensitivity, reaching a detection limit of 3 copies/μL, whereas RT-qPCR had a detection limit of 86 copies/μL. The comparative analysis between clinical diagnosis and molecular techniques, including RT-PCR and RT-qPCR, demonstrated low concordance, with Kappa coefficients of 0.268 and 0.324, respectively. In contrast, ddPCR, showed a moderate concordance, with a Kappa coefficient of 0.477. Sensitivity was 42.39% for RT-PCR, 57.89% for RT-qPCR and 72.37% for ddPCR, with 100% of specificity for all methods. This study under-scores ddPCR's superior sensitivity and concordance with clinical CDV diagnosis, even at low viral concentrations, suggesting it as a promising alternative for CDV diagnosis.

Aspergillosis is a fungal disease caused by the inhalation of fungal spores of the genus Aspergillus spp. This fungus mainly affects the lungs but can spread and infect the maxillofacial region through the bloodstream. The disease has nonspecific clinical manifestations, which hinders early diagnosis. Even though the Polymerase Chain Reaction (PCR) technique holds promising potential to aid in diagnosing aspergillosis, anatomopathological analysis services have not yet routinely adopted the method. Therefore, the present study aims to evaluate the applicability and standardize the techniques of preparation of biological samples and PCR for the detection of the species Aspergillus niger, Aspergillus fumigatus and Aspergillus flavus. Six samples of formalin-fixed, paraffin-embedded tissue (FFPE), with histopathological diagnosis suggestive of aspergillosis, were investigated by PCR. Five of the six samples evaluated were positive for the fungus Aspergillus spp, 2/5 of which were co-infected by A. fumigatus and A. flavus species, and the others were positive for A. flavus species. The findings of this study suggest that PCR may be an auxiliary method in diagnosing aspergillosis. However, the expansion of the sample size and the evaluation of PCR in comparison with other diagnostic tests for aspergillosis is essential to determine the accuracy of the method.

A total of five protoclones were successfully cultured on PDA medium out of regenerated twenty two colonies of Termitomyces protoplast and further studied. Liquid MYG grown mycelial tissue is used for protoplast isolation by enzymatic digestionin a mixture containing Lysing enzyme 2% and Cellulase R10 2% in 0.6 M mannitol. The incubation conditions like temperature, shaking and time were standardized at 24ºC, 60 rpm and 10 hours, respectively for healthy protoplasts liberation. The purified protoplasts showed an average yield of 1.2 × 10⁷ cells/gm tissue with 31.60 ± 9.31% regeneration efficiency on specific medium and 77.12 ± 2.72% viability by FDA test. Four ISSR primers were used in this study resulting a total of 27 reproducible bands with mean value of 6.75. They showed similar banding pattern in all the lines with zero percent polymorphism ranged from 280 bp–2700 bp. The amplified rRNA-ITS gene showed ~600 bp size in gel and found a single restriction site for enzyme HaeIII in all the protoclones and parent with similar fragment size in all.

(1) Background: A growing number of communities are re-discovering the value of their streets as important public spaces for many aspects of daily life, thus creating the need for a transformation in the quality of streets. An emerging concept is to accommodate all users of the transportation system, a concept that has been labelled ‘complete streets’. (2) Methods: In this paper, we present sustainable complete streets design criteria that integrate complete streets by adding in socio-environmental design criteria related to aesthetics, environment, liveability, and safety. (3) Results: Proposed design criteria provide a street network which provides improvements in aesthetics, to recover historical urban character and realize historical area planning goals; environment, to increase permeable surfaces, reduce the heat island, and absorb traffic-related air pollution; liveability, to create a public space destination in the urban landscape; and safety, to improve the safety of all road users. and (4) Conclusions: The case study of the urban rehabilitation of the “Mostra d’Oltremare” area and of its cultural and architectural assets in Naples, Italy, highlights the practical application of the proposed criteria and the possibility of using these criteria in other urban contexts.

As a member of smart materials, electrorheological elastomers (EREs) formed by pre-treating active electrorheological particles are attracting more and more attention. In this work, four Mg-doped strontium titanate particles with spherical, dendritic, flake-like and pinecone-like morphorlogies were obtained by hydrothermal and low-temperature co-precipitation. XRD, SEM, Raman and FT-IR were used to characterize the obtained products. The results showed that the sizes of Mg-doped strontium titanates are about 1.5‒2.0 μm, and the phase structure are dominated by cubic crystal. These Mg-doped strontium titanates were dispersed in a hydrogel composite elastic medium. Then, Mg-doped strontium titanate/glycerol/gelatin electrorheological composite hydrophilic elastomers were obtained with or without electric field. The electric field response properties of Mg-doped strontium titanate composite elastomers were investigated. It concluded that dendritic Mg-doped strontium titanate composite elastomers is high-performance EREs. And the maximum value of energy storage modulus of Mg-doped strontium titanate composite elastomers has achieved 8.70 MPa. This high electrorheological performance of the products is helpful for their applications in vibration control, force transducers, smart structures, dampers, and other fields.

The paper is dedicated to the numerical and experimental study of nonlinear oscillations exhibited by the Vilnius chaotic generator. The need for practical applicability of the mentioned generator in chaotic communication systems defined the necessity to study the operation of the circuit in a wide parameter range. The bifurcation maps and corresponding one-parameter diagrams, obtained and analysed utilising the Method of Complete Bifurcation Groups, reveal the complex nonlinear dynamics and regions of robust chaotic oscillations. Laboratory experiments allow the verification of the numerical results and identification of the most practically relevant phenomena.

We find that the degeneracies and many peculiarities of the DNA genetic code may be described thanks to two closely related (fivefold symmetric) finite groups. The first group has signature $G=\mathbb{Z}_5 \rtimes H$ where $H=\mathbb{Z}_2 . S_4\cong 2O$ is isomorphic to the binary octahedral group $2O$ and $S_4$ is the symmetric group on four letters/bases. The second group has signature $G=\mathbb{Z}_5 \rtimes GL(2,3)$ and points out a threefold symmetry of base pairings. For those groups, the representations for the $22$ conjugacy classes of $G$ are in one-to-one correspondence with the multiplets encoding the proteinogenic amino acids. Additionally, most of the $22$ characters of $G$ attached to those representations are informationally complete. The biological meaning of these coincidences is discussed.

Students studying dentistry and nursing experience anxiety daily, which affects both their general well-being and academic achievement. Anxiety is exacerbated by several circumstances, such as stressors in one's personal life, clinical environment exposure, and academic expectations. To effectively regulate anxiety and support students' achievement, treatments that take these elements into account must be designed. In this study, 271 students from the National and Kapodistrian University of Athens, Greece's departments of dentistry and nursing were polled. Being the following resources, participants filled out an electronic questionnaire that was uploaded to Google Forms and assessed their levels of stress, anxiety, depression, resilience, hope, and spiritual well-being: a) the Depression, Anxiety, Stress Scale (DASS-21), b) the Functional Assessment of Chronic Illness Therapy-Spiritual Well-Being Scale (FACIT-12), c) the Adult Hope Scale (AHS), d) the Resilience Assessment Questionnaire (RAQ8), and e) the Brief Resilience Scale (BRS). Additionally, demographic data was gathered, such as gender, educational attainment, family income, and country of origin. Statistical analyses were performed to investigate the relationships between the variables, including hierarchical multiple linear regression and t-tests. The sample consisted of 271 students from the Department of Nursing (n=126, 46.5%) and Dentistry (n=145, 53.5%), with 68.60% of them being female, 80.10% being undergraduate students, and 48.30% being from Athens, the country's capital. Comparatively speaking, 74.50 percent of nursing students were female, while 62.80% of dental students were. Compared to dental students (2.10%), nurses (40.50%) participated in postgraduate studies at a higher rate. Compared to dental students (44.80%), nursing students reported a household income of just 13.90%, which is much less than €35,000. Anxiety was reported by 51.30% of participants overall, with dental students reporting higher anxiety than nursing students. Higher family wealth was associated with reduced stress levels, while female undergraduate students reported higher levels of anxiety than their male counterparts. Hope was a strong predictor of resilience, but stress and worry had a negative correlation. Promoting students' well-being and academic success requires effective stress-reduction and resilience-building techniques aimed to not only improve student performance but also support future healthcare professionals' personal sustainability and holistic growth.

Megaselia scalaris and Senotainia tricuspis are two parasitoid flies of the honeybee Apis mellifera and are known to infest apiaries of different European and Mediterranean countries. However, their prevalence is still little known and no systematic control of their presence and countermeasures are currently adopted. To achieve a better knowledge about their distribution in apiaries, quantitative PCR (qPCR) based methods for their rapid detection directly in hive matrices were developed in this study. These were applied to samples of adult honeybees and hive debris collected in the summer season in Central Italy and Kosovo. In total 16 samples per type per region were analyzed. Both flies were detected in the two geographical contexts but more frequently in Central Italy. The positive apiaries in Central Italy were re-sampled one year later by capturing viable honeybees and isolating emerging flies after differentiation into adults. These were genotyped by sequencing regions of the mitochondrial cytochrome oxidase subunit I gene COI for M. scalaris and cytochrome B gene cytB for S. tricuspis amplified with specific primers designed in this study. Different genotypes of M. scalaris were detected and these were related to biotypes from distant nations. Some fly isolates were shown to harbor DNA or RNA of honeybee microbial pathogens. Results suggested to implement efficient control against these parasites in apiaries to limit their detrimental effects. The qPCR tests that have been developed can facilitate larger screening studies and early detection.

For the in-house validation of a droplet digital PCR method, a factorial experimental design was implemented. This design serves different purposes. On the one hand, it is an efficient design in relation to the workload involved in achieving a desirable level of reliability of variance estimates. On the other hand, it allows a partitioning of total variance into different components, thus providing information regarding the dominant sources of random variation. The statistical modelling reflects the actual measurement mechanism, establishing relationships between nominal target DNA copies per well, the range of variation of copy numbers per droplet, probability of detection values, and estimated numbers of copies.

Bicarbonate and CO2 are essential substrates for carboxylation reactions in bacterial central metabolism. In Staphylococcus aureus, the bicarbonate transporter, MpsABC (membrane potential-generating system) is the only carbon concentrating system. A mpsABC deletion mutant can hardly grow in ambient air. In this study, we investigated the changes that occur in S. aureus when it suffers from CO2/bicarbonate deficiency. Electron microscopy revealed that mpsABC has a twofold thicker cell wall thickness compared to the parent strain. The mutant was also substantially inert to cell lysis induced by lysostaphin and the non-ionic surfactant Triton X-100. Mass spectrometry analysis of muropeptides revealed the incorporation of alanine into the pentaglycine interpeptide bridge, which explains the mutant’s lysostaphin resistance. Flow cytometry analysis of wall teichoic acid (WTA) glycosylation patterns revealed a significantly lower -glycosylated and higher ß-glycosylated WTA, explaining the mutant’s increased resistance towards Triton X-100. Comparative transcriptome analysis showed altered gene expression profiles. Autolysin-encoding genes such as sceD, a lytic transglycosylase encoding gene, were upregulated, while genes related to cell wall-anchored proteins, secreted proteins, transporters, and toxins were downregulated. Overall, we demonstrate that bicarbonate deficiency is a stress response that causes changes in cell wall composition and global gene expression resulting in increased resilience to cell wall lytic enzymes and detergents.

Sepsis is a life-threatening syndrome triggered by infection and accompanied by high mortality, with antimicrobial resistances (AMRs) further escalating clinical challenges. Rapid and reliable detection of causative pathogens and AMRs are key factors for fast and appropriate treatment, in order to improve outcome in septic patients. However, current sepsis diagnostics based on blood culture is limited by low sensitivity and specificity while current molecular approaches fail to enter clinical routine. Therefore, we developed a Suppression PCR-based selective enrichment sequencing approach (SUPSETS), providing a molecular method combining multiplex suppression PCR with Nanopore sequencing to identify most common sepsis-causative pathogens and AMRs using plasma cell-free DNA. Applying only 1 mL of plasma, we target eight pathogens across three kingdoms and ten AMRs in a proof-of-concept study. SUPSETS was successfully tested on first ten clinical samples and revealed comparable results to clinical metagenomics while clearly outperforming blood culture. Several clinically relevant AMRs could be detected additionally. Furthermore, SUPSETS provided first pathogen and AMR-specific sequencing reads within minutes of starting sequencing, thereby potentially decreasing time to results to 11 - 13 hours and suggesting diagnostic potential in sepsis.

Viral contamination in the aquaculture industry is a significant challenge that requires further research. Emerging evidence suggests that long noncoding RNAs (lncRNAs) play a role in regulating various biological processes, including immunity. However, most studies have focused on model organisms, and little is known about non-coding RNAs in the immune response to viral hemorrhagic septicemia (VHS) in rainbow trout. In this study, we investigated the listing of non-coding RNAs (ncRNAs) in rainbow trout challenged with VHS virus. We explored their potential role in defense-like responses based on transcriptomic analysis of the virus-treated and control groups. As a result of this research, 4207 lncRNAs were identified, of which 182 lncRNAs were differentially expressed between treated and untreated samples. Among 7753 lncRNA transcripts, 1920 were novel intergenic (class code “u”), and 3748 transcripts were potentially novel isoform of lncRNAs. Gene ontology analysis revealed biological pathways associated with the stress response and immune signaling. Pairs of genes such as TP63-MSTRG.62, IL10-MSTRG.9783, IL10-MSTRG.9784, TK1-MSTRG.22826, and MSTRG.17881-GLI1 showed cis interactions, while DOCK10-MSTRG.30015, MSTRG.36357-LOC101268921, and MSTRG.34190-ENPP2 showed trans interactions. In this study, the role of lncRNAs in regulating the expression of the target genes was clearly identified. This finding is consistent with the regulatory processes of immune system responses mediated by cytokines, as well as the regulation of oxygen levels in response to viral infection. The information presented here highlights the various regulatory mechanisms of lncRNAs in the expression of immune system genes in fish treated with VHS virus.

This paper presents a compact wideband circularly polarized cross dipole antenna with low-pass filter response. It consists of two pairs of folded crossed dipole arms printed separately on both sides of the top substrate, and the two dipole arms on the same surface are connected by an annular phase-shifting delay line to generate circular polarization. A bent metal square ring and four small metal square rings around the cross dipoles are employed to introduce new resonant frequencies, effectively extending the impedance and axial ratio bandwidth. Four square patches printed on the middle substrate are connected to the ground plane by the vertical metal plates, in order to reduce the antenna height. Thus, a compact wideband circularly polarized antenna is realized. In addition, a transmission zero can be introduced at the upper frequency stopband by the bent metal square rings, without using extra filter circuits. For verification, the proposed model is implemented and tested. The overall size of the model is 90mm×90mm×33mm(0.37λ0×0.37λ0×0.14λ0, λ0 denotes the center operating frequency). The measured impedance bandwidth and 3 dB axial ratio bandwidth are 53.3% and 41%, respectively. More than 15 dB upper-band radiation suppression level is realized, indicating a good low-pass filter response.

Groundwater pollution in intensive horticultural areas is becoming an increasingly important problem. Therefore, this study evaluates nitrogen (N) fertilization in open field processing pepper crop in Extremadura, Spain. Previous research on the effect of N on sweet peppers grown in greenhouses is abundant, but data on outdoor cultivation, especially considering variety and site influences, are lacking. Field trials were conducted in 2020, 2021 and 2022 to determine the opti-mum N fertilizer rate for processing peppers, with the aim of reducing environmental impacts such as nitrate leaching while maintaining crop yields. Different N rates were applied in the trials, ranging from no N fertilizer to 300 kg N/ha. Meas-urements included crop yield, biomass, intercepted photosynthetically active radiation (PAR), and canopy cover. The study also developed a critical nitrogen curve (CNC) to determine the minimum N concentration required for optimal growth. The results showed that applying more than 200 kg N/ha did not significantly increase yield. Nitrogen-free treatments resulted in earlier fruit maturity, concentrating the harvest and reducing waste. In addition, excessive N application led to envi-ronmental problems such as groundwater contamination due to nitrate leaching. The study concludes that outdoor pepper crops in this region can achieve optimal yields with lower N rates (around 120 kg N/ha) compared to current practices, thereby reducing environmental risks and fertilizer costs. It also established relationships between biomass, canopy cover, and N uptake to improve fertilization strategies. These data support future crop modelling and sustainable fertili-zation practices.

LCK, a member of the Scr kinase family, is a non-receptor tyrosine kinase involved in immune cell activation, antigen recognition, tumor growth, and cytotoxic response. The enzyme has usually been linked to T lymphocyte activation upon antigen recognition. Lck activation is central to CD4, CD8, and NK activation. However, recently, it has become clearer that activating the enzyme in CD8 cells can be independent of antigen presentation and enhance cytotoxic response. The role of Lck in NK cytotoxic function has been controversial in a similar fashion as the role of the enzyme in CAR T cells. Inhibition of Lck has been mainly to treat hematologic malignancies with a high rate of success. They may be useful in treating other tumor types, and they may be useful to prevent cell exhaustion. New, more selective inhibitors have been documented, and they have shown inter-esting activities not only in tumor growth but in the treatment of autoimmune diseases, asthma, and graft vs host disease. Drug repurposing and bioinformatics can aid in solving several unsolved is-sues on the role of Lck in cancer. In summary, the role of Lck in immune response and tumor growth is not a simple event and requires more research.

Early onset neonatal sepsis is a significant contributor to neonatal morbidity and mortality. Although blood cultures remain the diagnostic gold standard, they detect pathogens in only a minority of suspected cases. This study compared the accuracy of blood cultures with a rapid multiplex PCR test. Newborns at risk for neonatal sepsis were prospectively screened as recommended per national guidelines. Evaluations included laboratory parameters (CrP, IL6, differential blood count), blood culture, and a molecular multiplex PCR test (ROCHE LightCycler SeptiFast) identifying 20 common microbial agents. Blood samples were taken simultaneously from umbilical cord or venous sources on the first day of life. Of 229 infants included, 69% were born preterm. Blood culture and multiplex PCR sensitivity were 7.4% and 11.1% respectively. Specificity, negative and positive predictive values between methods showed no significant variance, though multiplex PCR had more false positives due to contamination. The limited sensitivity of blood cultures for early onset neonatal sepsis is concerning. Despite quicker results, multiplex PCR does not enhance diagnostic accuracy or antibiotic therapy guidance, thus it can not be recommended for this indication.

Elevated carbon dioxide (eCO₂) stimulates wheat grain yield, but simultaneously reduces protein (N) concentration. Also other essential nutrients are subject to change. This study is a comprehensive synthesis of wheat experiments with eCO₂, estimating effects on N, minerals (B, Ca, Cd, Fe, K, Mg, Mn, Na, P, S, Zn), and starch. Analysis was made by i) deriving response functions for the relative effect on element concentration in relation to CO₂ concentration, ii) meta-analysis to test the magnitude and significance of observed effects, and iii) relating CO₂ effects on minerals to effects on N and grain yield. Responses range from zero to strong negative effects of eCO₂ on mineral concentration, with largest reductions for the nutritionally important elements N, Fe, S, Zn and Mg. Together with the positive but small and non-significant effect on starch concentration, the large variation in effects suggests that CO₂-induced responses cannot be explained by a simple dilution model. To explain the observed pattern, uptake and transport mechanisms may have to be considered, along with the link of different elements to N uptake. Our study shows that eCO₂ has a significant effect on wheat grain stoichiometry, with implications for human nutrition in a world of rising CO₂.

Abiotic stress, especially drought stress poses a significant threat to the terrestrial plant growth, development, and productivity. Although mulberry have great genetic diversity and extensive stress tolerant traits in agroforestry systems, only a few reports offer preliminary insight into biochemical responses of mulberry leaves under drought conditions. In this study, we performed a comparative metabolomic and transcriptomic analysis on the “drooping mulberry” (Morus alba var. Pendula Dippel) under PEG-6000-simulated drought stress. Our research revealed that drought stress significantly enhanced flavonoid accumulation and up-regulated the expression of phenylpropanoid biosynthetic genes. Furthermore, the activities of superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA) content were elevated. In vitro enzyme assays and fermentation tests indicated the involvement of flavonol synthase/flavanone 3-hydroxylase (XM_010098126.2) and anthocyanidin 3-O-glucosyltransferase 5 (XM_010101521.2) in the biosynthesis of flavonol aglycones and glycosides, respectively. The recombinant MaF3GT5 protein was found to recognize kaempferol, quercetin, and UDP-glucose as substrates, but not 3-/7-O-glucosylated flavonols and UDP-rhamnose. MaF3GT5 is capable of forming 3-O- and 7-O-monoglucoside, but not di-O-glucosides, form kaempferol. This implies its role as a flavonol 3, 7-O-glucosyltransferase. The findings from this study provided insights into the biosynthesis of flavonoids and could have substantial implications for the future diversified utilization of mulberry.

Boeing and Airbus developed a special testing procedure to investigate the compressive response of laminates that have been impacted (following standards ASTM D 7137 and DIN 65561). This study focuses on both experimental and numerical analysis of Kevlar plates subjected to compression after impact. To ensure high quality and appropriate mechanical properties, the composite plates were manufactured using autoclaving. The DIN 65561 protocol was followed for all three test systems. Initially, ultrasonic C-scanning was performed on all plates before testing to confirm they were free of any significant defects arising from the manufacturing process. Subsequently, low-energy impact testing was conducted at levels ranging from 0 to 8 Joules. Three specimens were tested at each energy level. After the impact, all specimens underwent ultrasonic C-scanning again to assess the internal delamination damage caused by the impactor. Finally, both pristine and impacted specimens were subjected to compressive testing using the special jig specified in DIN 65561. The compressive impact strength results obtained from these tests were plotted against the delamination area measured by C-scanning. This data was then compared to results obtained from specimens with artificial damage. Semi-empirical equations were used to fit both sets of curves. The same procedure (impact testing, C-scanning, and data analysis) was repeated to investigate the relationship between impact energy and total delamination area. Lastly, finite element modeling was employed to predict the buckling stresses that develop under compression in the impacted systems studied. These modelling approaches have demonstrated good accuracy in reproducing experimental results for CAI tests.

Cartilage regeneration requires a balance of anabolic and catabolic processes. This study examined the susceptibility of fibromodulin (FMOD) and lumican (LUM) to degradation by MMP-13, ADAMTS-4 and ADAMTS-5, the three major degradative proteinases in articular cartilage in osteoarthritis (OA). Immunolocalisation of FMOD and LUM in foot sections of developmental cartilages demonstrated prominent localisations in metatarsal and phalangeal foetal rudiment cartilages and growth plate. An MMP-13 neoepitope antibody (TsYG11) demonstrated localisation of MMP-13 cleaved FMOD in the hypertrophic chondrocytes of the metatarsal growth plate. FMOD was more prominently localised in the superficial cartilage of normal and fibrillated zones in OA cartilage, TsYG11 positive FMOD was located deeper in the cartilage samples. Ab TsYG11 also identified FMOD fragmentation in Western blots of extracts of normal and fibrillated cartilage and total knee replacement OA cartilage. The C-terminal anti-FMOD used in this study (PR-184) failed to identify FMOD fragmentation due to C terminal processing, an equivalent Ab to the C-terminus of LUM (pAb PR-353) identified 3 prominent LUM fragments in OA human knee cartilages. In-vitro digestion of human knee cartilage with MMP-13, ADAMTS-4 and ADAMTS-5 generated equivalently sized FMOD fragments of 54, 45 and 32kDa to those in blots of OA cartilage, LUM was not less susceptible to fragmention in in-vitro digestions however Ab PR-353 detected N-terminally processed LUM fragments of 39, 38 and 22 kDa in 65-80 year old OA knee cartilage. FMOD and LUM were differentially processed during in-vitro digestions with MMP-13, ADAMTS-4 and ADAMTS-5 with FMOD susceptible to degradation by MMP-13, ADAMTS-4 and to a lesser extent ADAMTS-5 however LUM was less susceptible to fragmentation. FMOD was processed by MMP-13 in metatarsal and phalangeal foetal rudiment developmental cartilages and growth plate indicating a role in skeletogenesis.

In this paper, we demonstrate the Collatz conjecture using the mathematical complete induction method. We show that this conjecture is satisfied for the first values of natural numbers, and in analyzing the sequence generated by odd numbers, we can deduce a formula for the general term of the Collatz sequence for any odd natural number n after several iterations. This formula is used in one case that we analyze using the mathematical complete induction method in the process of demonstrating the conjecture.

Escherichia coli phage Eco_BIFF was isolated from several laboratory stocks of E. coli K-12 MG1655 derivatives. The source of the contamination is unknown. Eco_BIFF is a lytic phage that shows effective growth inhibition of E. coli K-12. Here, we announce the complete genome sequence of Eco_BIFF, and major findings from its genome annotation.

In nature and engineering applications, flexible fiber beds covering biological surfaces can play a role in reducing resistance. These fibers deform under the action of fluids, and this deformation affects the fluid flow state, forming a complex fluid-solid interaction phenomenon. To quantitatively analyze such issues, the physical model is simplified to the deformation problem of an soft hair bed caused by Stokes flow, and the deformation problem of a single fiber caused by Stokes flow is further studied. The deformation problem of an elastic single fiber in a channel caused by Stokes flow can be described by a nonlinear integral equation. We have obtained a new series solution, which has been compared with the previous perturbation method to verify the accuracy and effectiveness of the series solution. Meanwhile, we have further provided an extended form of flexible fiber deformation through experimental fitting. This fluid-solid interaction problem involves multiple fields and is very important in many natural and engineering systems. The research in this paper can not only help us better understand complex phenomena in nature but also delve into the interaction mechanism between fluids and solids, providing a theoretical basis for future scientific research and engineering applications.

Europium is one of the most reactive lanthanides and humans use it in many different applica-tions, but we still know little about its potential toxicity and cellular response to its exposure. Two strains of the eukaryotic microorganism model Tetrahymena thermophila adapted to high concentrations of two Eu(III) compounds (EuCl3 or Eu2O3) were obtained and compared to a control strain and cultures treated with both compounds. In this ciliate, EuCl3 is more toxic than Eu2O3. LC50 values show that this microorganism is more resistant to these Eu(III) compounds than other microorganisms. Oxidative stress originated mainly by Eu2O3 is minimized by overexpression of genes encoding important antioxidant enzymes. The overexpression of metallothionein genes under treatment with Eu(III) compounds supports the possibility that this lanthanide may interact with the -SH groups of the cysteine residues from metallothi-oneins and/or displace essential cations of these proteins during their homeostasis function. Both lipid metabolism (lipid droplets that fuse with europium-containing vacuoles) and au-tophagy are involved in the cellular response to europium stress. Bioaccumulation, together with a possible biomineralization to europium phosphate, seems to be the main mechanism of Eu(III) detoxification in these cells.

Rift Valley fever (RVF) is a mosquito-borne zoonotic viral disease that infects both human and animals. This is a descriptive study summarizes investigation and response to RVF outbreak, using surveillance data. A RVF outbreak occurred in Berber locality in northern Sudan. We identified 246 cases of RVF including seven deaths among humans, 119 deaths and 82 abortions among Livestock. Our entomological surveillance confirmed the presence of Anopheles arabiensis, An. stephensi, Aedes aegypti, Ae. vexans, Culex pipiens, Cx. Quinquefasciatus, and Cx. theileri in the area during the outbreak. In a comparison with another outbreak of Rift Valley fever in nearby area while ago, a multisectoral One Health response strategy has resulted in strengthening the health system through re-innovating health facilities and centres and training about 200 healthcare and emergency responders. Additionally, it reduced cases and fatalities among humans by 78% and 63%, respectively, and loss in livestock due to deaths and/or abortions by 82%. The added-value of implementing One Health strategy was saving more lives and resources, while strengthening the health system. We recommend the systematic institutionalization of One Health strategy in the country as a core part of the national prevention and control strategy for zoonotic diseases epidemics and pandemics.

No system nor standardized analytical procedures at commercial laboratories exist to facilitate and accurately measure potential viable virus contamination in feed ingredients and complete feeds globally. As a result, there is high uncertainty of the extent of swine virus contamination in global feed supply chains. Many knowledge gaps need to be addressed to improve our ability to prevent virus contamination and transmission in swine feed. This review summarizes the current state of knowledge involving: 1) the need for biosecurity protocols to identify production, processing, storage, and transportation conditions that may cause virus contamination of feed ingredients and complete feed, 2) challenges of measuring virus inactivation, 3) virus survival in feed ingredients during transportation and storage, 4) minimum infectious doses, 5) differences between using a Food Safety Objective versus a Performance Objective as potential approaches for risk assessment in swine feed, 6) swine virus inactivation from thermal and irradiation processes, and chemical mitigants in feed ingredients and complete feed, 7) efficacy of virus decontamination strategies in feed mills, 9) benefits of functional ingredients, nutrients, and commercial feed additives in pig diets during a viral health challenge, and 10) considerations for improved risk assessment models of virus contamination in feed supply chains.

Verification of inclusivity of molecular detection methods gives indications on the reliability of viral infections diagnosis because of the tendency of viral pathogens to undergo sequence variation. This study was aimed to select inclusive probe based reverse transcription quantitative PCR (RT-qPCR) assays for the diagnosis of the most widespread and detrimental viruses infecting honeybees, namely the acute bee paralysis virus (ABPV), the black queen cell virus (BQCV), the chronic paralysis bee virus (CBPV), the deformed wing virus variants A (DWVA) and B (DWVB), and the sacbrood virus (SBV). Therefore, detection methods previously described were re-evaluated in silico for specificity and inclusivity. Based on this evaluation, selected methods were modified or new ones were designed and tested in duplex RT-qPCR reactions. The limits of detection (LODs), effect of multiplexing on sensitivity and viral RNA quantification potential in bees and hive debris were assessed. This study made available diagnostic assays able to detect an increased number of virus variants compared to previously described tests and two viral pathogens in a single PCR reaction.

It is important to be able to detect and differentiate between distinct porcine enteric coronaviruses that can cause similar diseases. However, the existence of naturally occurring recombinant coronaviruses such as swine enteric coronavirus (SeCoV) can give misleading results with currently used diagnostic methods. Therefore, we have developed and validated three duplex real-time quantitative RT-PCR assays for the simultaneous detection of, and differentiation between, porcine epidemic diarrhea virus (PEDV) and SeCoV. Transmissible gastroenteritis virus (TGEV) is also detected by two out of these three assays. In addition, a novel triplex assay was set up that was able to detect and differentiate between these alphacoronaviruses and the porcine deltacoronavirus (PDCoV). The validated assays have low limits of detection, close to 100% efficiency and were able to correctly identify the presence of PEDV and SeCoV in 55 field samples, while 20 samples of other pathogens did not give a positive result. Implementing one or more of these multiplex assays into the routine diagnostic surveillance for PEDV will ensure that the presence of SeCoV, TGEV and PDCoV will not go unnoticed.

A brown seaweed consumed worldwide, Fucus vesiculosus, has been used to prevent atherosclerosis and hypercholesterolemia, among other uses, however, the mechanisms of action that lead to these effects are not yet fully understood. An F. vesiculosus aqueous extract, prepared as a soup was characterized as rich in sev-eral bioactive compounds, mainly phlorotannin and peptides. This work aims to study the in vitro effect of this extract on the expression of different proteins involved in the synthesis and transport of cholesterol in HepG2 cells, though a proteomic analysis, western blot and qRT-PCR. The results demonstrated that, in liver cells, the extract decreases the expression of 4 proteins involved in cholesterol biosynthesis process and also decrease the expression of two important transporters proteins of cholesterol, NPC1L1 and ABCG5, as well as decrease the NPC1L1 mRNA levels. Our study demonstrates some mechanisms of action of bioactive com-pounds from F. vesiculosus that may explain its previously reported hypocholesterolemic effect.

Background and Objectives: Most individuals infected by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are asymptomatic or have mild symptoms of COVID-19, which usually resolve after few days. Regardless of symptoms, infected people can transmit the virus to others especially on the first days of infection. Quantitative reverse transcription-polymerase chain reaction (RT-qPCR) is used to confirm SARS-CoV-2 infection; some individuals show persistent PCR-positivity after recovering from COVID-19. In this study, 12 individuals who showed persistence of COVID-19 symptoms and of SARS-CoV-2 PCR-positivity were followed-up. Methods: nasopharyngeal samples were collected for SARS-CoV-2 detection by RT-qPCR; clinical and epidemiological data were analyzed. Results: that persistence of SARS-CoV-2 PCR positivity was associated with duration of symptoms (r_s 0.81338), which varied between one and 49 days, with 75% of the individuals reporting symptoms for more than two weeks; 83.33% of cases remained positive after two weeks of onset of symptoms, despite decreases in viral load. Conclusion: neither RT-qPCR test nor a symptom-based approach alone are sufficient to evaluate discontinuation of patient isolation; other factors such as viral loads and symptom severity should also be considered. Additional studies are needed to understand how RT-PCR-positivity is related to symptoms and the risk of viral transmission, and to better support isolation guidelines.

Herein, we reported a case of complete excision of the jejunum for the treatment of esophageal cancer. A 62-year-old male was admitted with complaints of “difficulty in eating for one month and chest pain for 10 days". For complete gastrectomy and colonic polyposis, we chose the jejunum. We completely excised the jejunum and its vessels, anastomosed to the esophageal tumor resection, with no significant complications. This case showed that complete excision of the jejunum with bowel and vessels is an alternative surgical method for the treatment of esophageal cancer with free bowel shortage and no torsion of blood vessels.

Rheumatoid arthritis (RA) is a chronic inflammatory disease. Despite new methods of diagnostics and treatment as well as extensive biological and immunosuppressive treatment, the etiology of RA is not fully understood. Moreover, the problem of diagnosis and treatment of RA patients is still current and affects a large group of patients. It is suggested that these ER-related features may impair adaptation to chronic stress, inferring the risk of rheumatoid arthritis. The main goal in this study was evaluation of changes in mRNA translation to determine chronic ER stress conditions in rheumatoid arthritis patients. The study group consist of 86 individuals including total 56 rheumatoid arthritis patients and 30 healthy controls. The expression level of mRNA form blood samples of RA patients as well as controls of the PERK-UPR associated genes: p-eIF2, BCL-2, PERK, ATF4, BAX were investigated by Real-Time qPCR. GAPDH expression was used as a standard control. Considering the median, the expression of PERK, BCL-2, p-eIF2, ATF4, BAX was found to be significantly increased in the blood of RA patients compared with the control group. The p-value for the PERK gene was 0.0000000036, the p-value for the BCL-2 gene was 0.000000014, the p-value for the p-eIF2 gene was 0.006948, the p-value for the ATF4 gene was 0.0000056 and the p-value for the BAX gene was 0.00019, respectively. Thus, it can be concluded that targeting of the components of the PERK-dependent UPR signaling pathway via small-molecule PERK inhibitors, may contribute to the development of novel, innovative treatment strategies against rheumatoid arthritis.

Technological advancements in the Internet of Things (IoT), unmanned aerial vehicles (UAVs), and novel technological tools have transformed disaster management practices from localized approaches to a global framework. These innovations necessitate a unified understanding and application of diverse methodologies at an international level, ensuring they are accessible and operational across different sectors. This integration is crucial not only for technical personnel but also for medical teams, who play a pivotal role in disaster response and management. By fostering a comprehensive grasp of these methods, we can enhance coordination, efficiency, and effectiveness in addressing disasters on a global scale. The authors aimed to provide a comprehensive overview of emerging techniques that are driving shifts in strategic approaches and treatment technologies. Recognized as a domain of continuous research and development, this overview, by necessity, remains an incomplete snapshot of an evolving field. It highlights the dynamic nature of technological and strategic advancements, underscoring the importance of ongoing investigation and innovation to stay abreast of current trends and future directions. The article also highlights the importance of international collaboration beyond societal and national boundaries A single paragraph of about 200 words maximum. For research articles, abstracts should give a pertinent overview of the work. We strongly encourage authors to use the following style of structured abstracts, but without headings: (1) Background: Place the question addressed in a broad context and highlight the purpose of the study; (2) Methods: briefly describe the main methods or treatments applied; (3) Results: summarize the article’s main findings; (4) Conclusions: indicate the main conclusions or interpretations. The abstract should be an objective representation of the article and it must not contain results that are not presented and substantiated in the main text and should not exaggerate the main conclusions.

The quantification of human DNA extracts from forensic samples plays a key role in the forensic genetics process. Quantification guarantees maximum efficiency and avoids repeated analyses, over amplified samples, or unnecessary examinations. Here, we present our experience using an extremely sensitive real-time-based method with the Quantifiler® Trio DNA quantification kit, which we have explored in the ISO/IEC17025 accredited test validation process conducted in our laboratory. In this study, our focus was on establishing a minimum threshold beyond which DNA typing is unnecessary, by defining the correlation of negative quantification results with the absence of genetic profiles resulting from DNA typing. We investigated the results in the range in which the technique is used (50-0.005 ng/μl). In the validation process of the internally accredited method, we used data from the commercial quantification system, based on the average values of the standard curve using a wide range of samples, including artificially degraded samples. This method was extensively used continuously for nine years to quantify the DNA extract coming from a wide range of samples from criminal cases. The validation allowed us to verify the limits of the technique, especially in the use of the minimum quantities necessary to obtain genetic profiles acceptable for sending to the Italian DNA database. This optimization of the protocol for analysing forensic samples allows us to optimise the protocol for analysing forensic samples. However, the technique is used in our laboratory with a completely manual preparative phase, and we have decided to investigate the contribution of human error more accurately. Thus, in our experience, we have found that it is preferable to use quantification ranges rather than exact limits before deciding how to analyse the extracts via PCR. In addition, here we present some preliminary data related to the analysis of certain samples that initially appeared negative during quantification, applying a concentration method for maximum yield from forensic samples of judicial interest. Our aim is to ensure that no source of evidence is lost in the reconstruction of a criminal event.

Klebsiella pneumonia (K. pneumoniae) is a Gram-negative bacterium that causes nosocomial infections in the lung, bloodstream, and urinary tract. Therefore, detecting K. pneumoniae in early time is important in preventing severe infections. However, clinical detection of K. pneumoniae requires a long time of agar plate culture. Nucleic acid detection like qPCR is precise but requires expensive equipment. Recent research reveals that collateral cleavage activity of CRISPR-LbCas12a has been applied in nucleic acid detection. In this study, PCR combined with CRISPR-LbCas12a targeting the K. pneumoniae system was established. This system showed excellent detection specificity and sensitivity in both bench work and clinical samples. Due to its advantages, its application can meet different detection requirements in health centers where qPCR is not accessible.

Numerous protocols have been published for extracting DNA from phlebotomines. Nevertheless, their small size is generally an issue in terms of yield, efficiency, and purity, for large-scale individual sand fly DNA extractions when using traditional methods. Even though this can be circumvented with commercial kits, these are generally cost-prohibitive for developing countries. We encountered these limitations when analysing parasite infection in Lutzomyia spp. by PCR [1] and, for this reason, we evaluated various modifications on a previously published protocol ([2] and Acardi personal communication). The most significant variation was the use of a different lysis buffer [3] to which added Ca2+ (buffer TESCa), because this ion protects proteinase K against autolysis, increases its thermal stability, and could have a regulatory function for its substrate-binding site [4]. Individual sand fly DNA extraction success was confirmed by amplification reactions using internal control primers that amplify a fragment of the cacophony gene [5,6]. To the best of our knowledge, this is the first time a lysis buffer containing Ca2+ has been reported for the extraction of DNA from sand flies.

A 68-year-old man with malignant metastatic melanoma in complete remission after Pembrolizumab treatment, developed 10 months after Pembrolizumab cessation due a grade 3 diarrhea, confusion, altered mental status, progressive memory loss, gait disturbance. He was admitted to the neurologic department and comprehensive neurological workup, brain magnetic resonance imaging, cerebral fluid analysis, EEG, blood test allowed the diagnosis of autoimmune encephalitis. The patient was treated with plasmapheresis, high dose of intravenous steroids and intravenous immunoglobulins. The patient improved and he is now well with a performance status 1. This case is interesting since the AE developed approximately 10 months after the cessation of immunotherapy, the underlying cancer was in complete remission and the AE showed a good response to the treatment done.

In this work, two plasmonic structures with two different periodic nanostructures (nanodisk and nanostrip arrays) separated from the gold layer by an ultrathin dielectric Al2O3 layer are proposed, and their optical properties are simulated as 3-dimensional models using COMSOL Multiphysics software tool. The optical responses, such as the amplification of the electric field and the scattering, absorption and extinction cross sections of the two structures, are investigated and compared. The results show that both structures show strong field enhancement in their gap layer, and the structure with the periodic array of metal nanodisks shows stronger field enhancement and larger absorption, extinction, and scattering cross sections in its dipole resonance wavelength, which is defined as a bright plasmonic mode. In addition, some weaker resonant modes are observed for both structures and the reason for their generation is dis-cussed. The analysis of different possible modes in such structures is crucial for their application in various fields.

Studying models of healthy aging and exceptional longevity is crucial to understand a possible longevity signature as most of them show resistance to age-related diseases. In particular, semi- and supercentenarians are a highly selected group, having survived significant adversities, including the Spanish Flu and COVID-19 pandemics, indicating distinctive immune system characteristics. This paper analyse the inflammatory scores (INFLA-score, Systemic Inflammation Response Index (SIRI)) and Aging-Related Immune Phenotype (ARIP) indicators calculated from the dataset of the DESIGN project, including 249 participants aged 19-111 years, trying to understand the immune-inflammatory (IMFLAM) role to achieve longevity. Statistical analyses, including linear regression and one-way ANOVA, were performed to explore the correlations between these parameters and age. Both INFLA-score and SIRI showed a significant increase with age. However, no statistical difference is observed when analysing the values for semi- and supercentenarians with those for other age groups, whose values are similar to that of adults and less than younger centenariansRegarding ARIP values, it is noteworthy that when comparing the CD8 Naive/Effector scores between groups, no significant differences were observed between the semi- and supercentenarian group and the other groups. These results support the idea that the control of IMFLAM response can promote extreme longevity.

The aim of the present trial was to evaluate the growth performance of buffalo calves fed diets characterized by different forage/concentrate ratio with or without the Saccharomyces Cerevisiae supplementation (CBS 493.94, Yea-Sacc®). Twenty-four male buffalo calves (mean age 145.1 ± 16.1 days; mean weight 108.0 ± 18.7) were assigned randomly to 4 groups, homogeneous for age, fed four different diets: diet 1 F:C ratio 50:50; diet 2 F:C ratio 30:70; diet 3 F:C ratio 50:50 + Yea-Sacc® and diet 4 F:C ratio 30:70 + Yea-Sacc®. Buffalo calves were individually weighted before the start of the experiment and the data were used as a covariate, monthly until the end of the trial. Dry matter intake (DMI), daily weight gain (DWG) and feed conversion ratio (FCR) were calculated. The differences in diets’ composition significantly (p

Human influenza A viruses (IAVs) cause global pandemics and epidemics, which remain serious threats to public health because of the shortage of effective means of control. To combat the surge of viral outbreaks, new treatments are urgently needed. Developing new virus control modalities requires better understanding of virus-host interactions. Here we describe how IAV infection triggers cellular apoptosis, and how this process can be exploited towards development of new therapeutics, which might be more effective than the currently available anti-influenza drugs.

Infant electrocardiograms (ECG) and heart rates (HR) are very useful biosignals for psychological research and clinical work, but can be hard to analyze properly, particularly long form (≥5 minutes) recordings taken in naturalistic environments. Infant HRs are typically much faster than adult HRs, and so some of the underlying frequency assumptions made about adult ECGs may not hold for infants. However, the bulk of publicly available ECG approaches focus on adult data. Here, existing open-source ECG approaches are tested on infant datasets. The best performing open-source method is then modified to maximize its performance on infant data (e.g., including a 15Hz high pass filter, adding local peak correction). The HR signal is then subsequently analyzed, developing an approach for cleaning data with separate sets of parameters for the analysis of cleaner and noisier HR. A Signal Quality Index (SQI) for HR is also developed, providing insight into where a signal is recoverable and where it is not, allowing for more confidence in analysis performed on naturalistic recordings. The tools developed and reported in this paper provide a base for future analysis of infant ECG and related biophysical characteristics. Of particular importance, the proposed solutions outlined here can be efficiently applied to real-world large datasets.

Tug-of-war (TOW) is an internationally played activity including professional and amateur athletes and defined as early (4000 years ago as a rope less version) in the artwork on Egyptian tomb engravings and is played as per the rules laid out by TWIF, which has 73 member countries and administrative headquarters in the USA. Typically, two teams of “pullers” participate and apply enormous contra directional forces on the pulling rope. Originally, two types of competition are used: knockout and points. This narrative review describes the scientific state of the art about of TOW. For the best of the author’s knowledge no previous information has been published. Anthropometric parameters are near 83.6, lean body mass 69.4, and body fat 16. The VO2MAX is 55.8 ml/kg/min. Relative strength, the dynamic leg power was 4659.8 N. Endurance TOW elicits minimal muscle damage. The injured strains and sprains comprised over half of all injuries: back (42%), shoulder–upper limb (23%) and knee (17%). Pulling movement in TOW contests can be divided into three phases: namely "Drop", "Hold" and "Drive" phase. The maximal pulling forces was 1041.6 ± 123.9 N. The percentage of dynamic pulling force in static maximal pulling force was 75.5 ± 14.4% and the dynamic ranged from 106.4 to 182.5%. There are two gripping styles, indoor and outdoor. The friction characteristics between surface and shoe in TOW is important to determine a suitable shoe for indoor TOW. Waist Belt might be a useful piece of equipment for TOW sport. The EMG technique in Tow described a high activity of dorsal muscle during the pulling. The factor of force vanishing was the coordination among athletes. The force vanishing percentage goes from 8.82±5.59 for 2 contenders to 19.74±2.22 for 8 athletes, 6.4 % in the sum of 2 pullers. However, in the drop phase, for female elite TOW team, only the 0.5 % of them pulling force was wasted. Future studies are need in order to understand better this historical sport activity.

Electrospun ultrathin fibers based on binary compositions of polylactide (PLA) and poly(ε-caprolactone) (PCL) with different ratios of polymer components from 0/100 to 100/0 wt. % were studied. Using a combination of thermal (DSC) and relaxation (EPR) measurements, the effect of biopolymers content on the characteristics of the crystal structure of PLA and PCL and the diffusive rotational mobility of the stable TEMPO radical in the amorphous regions of PLA/PCL compositions was shown. It was revealed that after PLA and PCL blending, significant changes in the degree of crystallinity of PLA, PCL segmental mobility, sorption capacity of the radical as well as activation energy of rotational diffusion in the amorphous regions of PLA/PCL composition are observed. The characteristic region of biopolymers’ composition from 50/50 to 30/70 % PLA/PCL blends ratio was found, where the inversion transition of PLA from dispersive medium to dispersive phase where an inversion transition is assumed when the continuous medium of the PLA transforms into a discrete phase. The performed studies made it possible for the first time to carry out a comprehensive analysis of the effect of the system component ratio on the structural and dynamic characteristics of the PLA/PCL film material at the molecular level.

Inflammation is implicated in the etiology of obesity-related diseases. Thromboxane-prostanoid receptor (TPR) is known to play a role in mediating an inflammatory response in a variety of cells. Gut-derived lipopolysaccharide (LPS), a TLR4 agonist, is elevated in obesity. Moreover, free fatty acids (FFAs) are important mediators of obesity-related inflammation. However, the role and mechanisms by which TPR regulates the inflammatory response in human immune cells remain unclear. We sought to determine the link between TPR and obesity and the role/mechanisms by which TPR alters LPS or stearic acid (SA)-induced inflammatory response in PBMCs. Cells were pre-treated with agents blocking TPR signaling, followed by treatment with LPS or stearic acid (SA). Our findings showed that TPR mRNA levels are higher in PBMCs from individuals with obesity. Blockade of TPR as well as ROCK which acts downstream of TPR, attenuated LPS and/or SA-induced pro-inflammatory response. On the other hand, TPR activation using its agonist enhanced the pro-inflammatory effects of LPS and/or SA. Of note, TPR agonist by itself elicits an inflammatory response which was attenuated by blocking TPR or ROCK. Our data suggest that TPR plays a key role in promoting an inflammatory response in human PBMCs and this effect is mediated via TLR4 and/or ROCK signaling.

Wetland ecosystems in the Qinghai-Tibet Plateau are pivotal for global ecology and regional sustainability, contributing significantly to terrestrial ecosystems by regulating runoff, mitigating floods, and enhancing water quality. This study investigates the dynamic changes in wetland ecosystems within the Chaidamu Basin and their response to drought, aiming to foster sustainable wetland utilization in the Qinghai-Tibet Plateau. Using Landsat TM/ETM/OLI data on the Google Earth Engine platform, we employed a random forest method for annual long-term land cover classification. Meteorological drought conditions were assessed using SPEI3, SPEI6, SPEI9, and SPEI12, derived from monthly precipitation and evapotranspiration data. Pearson correlation analysis examined the relationship between wetland changes and various SPEI scales. The BFASAT method evaluated the impact of SPEI12 trends on wetlands, while cross-wavelet analysis explored teleconnections between SPEI12 and atmospheric circulation factors. Our findings revealed that the land cover dataset of the Chaidamu Basin (1990-2020) exhibited diverse categories with high classification accuracy (OA: 90.27%, Kappa: 88.34%). Wetlands, including lake, glacier, and marsh types, exhibited a noticeable increasing trend. Wetland expansion occurred during specific periods (1990-1997, 1998-2007, 2008-2020), featuring extensive conversions between wetland and other types, notably from other types to wetlands. Spatially, lake and marsh wetlands predominated in the low-latitude basin, while glacier wetlands were situated at higher altitudes. The study identified significant negative correlations between SPEI at various scales and total wetland area and types, with SPEI12 exhibiting the most substantial effect between September and December (r <-0.75) on wetlands. SPEI12 displayed a decreasing trend with non-stationarity and distinct breakpoints in 1996, 2002, and 2011, indicating heightened drought severity. Atmospheric circulation indices (ENSO, NAO, PDO, AO, WP) exhibited varying resonance with SPEI12, with NAO, PDO, AO, and WP demonstrating longer resonance times and pronounced responses.The continuous growth of wetlands amidst increasing aridification emphasizes the need for thoughtful wetland development to establish a sustainable "forest-lake-grass-field-river" ecological community. These findings underscore the significance of comprehending wetland changes and drought dynamics for effective ecological management in the Chaidamu Basin of the Qinghai-Tibet Plateau.

The Herpotrichiellaceae family is an important group of dematiaceous filamentous fungi, associated with a variety of pathogenic fungal species causing Chromoblastomycosis (CBM) and Phaeohyphomycosis (PHM), both with polymorphic clinical manifestations and worldwide incidence. Currently, the identification of this family and determination of the causative agent is challenging due to the subjectivity of morphological identification methods, necessitating the use of molecular techniques to complement diagnosis. In this context, genetic sequencing of the Internal Transcribed Spacer (ITS) has become the norm due to a lack of alternative molecular tools for identifying these agents. Therefore, this study aimed to develop PCR-Multiplex methodologies to address this gap. Sequences from the ITS and Large subunit (LSU) of ribosomal DNA were used, and after manual curation and in vitro analyses, primers were synthesized for the identification of the aforementioned targets. The primers were optimized and validated in vitro, resulting in two PCR-Multiplex methodologies: one for identifying the Herpotrichiellaceae family and the bantiana clade, and another for determining the species Fonsecaea pedrosoi and Fonsecaea monophora. Ultimately, the assays developed in this study aim to complement other identification approaches for these agents, reducing the need for sequencing, improving the management of these infections, and enhancing the accuracy of epidemiological information.

Respiratory tract infections (RTIs) are one of the main causes of hospitalization and mortality causing substantial economic burden to healthcare systems globally. As opposed to the previous belief that respiratory infections are caused by a single pathogen, studies have shown that most RTIs are a result of a combination of bacterial and/or viral pathogens infecting the host. The clinical manifestation of RTIs is very similar (i.e., syndrome), often showing Influenza-like illness (ILI) symptoms. While COVID-19 has dominated ILI over the past few years, there are many other pathogens that are responsible for ILI. In addition, it is not uncommon to have coinfections with multiple pathogens in patients presenting with ILI and that such coinfections can even exacerbate the disease severity of RTIs. Therefore, an insight into coinfections can help with accurate disease prognosis, patient care management and outcomes. The goal of this study was to identify the different organisms in symptomatic patients presenting with ILI.

Accurate and timely testing has become an essential measure in combatting the COVID-19 global pandemic. Currently, polymerase chain reaction (PCR) based assays are the most relied on methods for SARS-CoV-2 detection. This traditional workflow involves a viral RNA extraction from the viral transport media storing nasopharyngeal swabs collected from patients, followed by PCR based detection. While accurate, this methodology is time consuming and resource heavy, causing for delays in receiving results or limited access to testing. Herein, we demonstrate a validated method for SARS-CoV-2 detection from viral transport media using a two-step, direct-to-PCR workflow revolving around shaker-mill homogenization. This method completely bypasses the extraction steps of the traditional workflow, replacing it with 30 seconds of mechanical disruption sufficient to allow for COVID-19 detection with a 96.43% sensitivity and 100% specificity when compared to traditional extraction to PCR based methods.

Mosquitoes genus Aedes are the most important arthropod disease vector. Dengue Virus (DENV) and Chikungunya Virus (CHIKV) are the main arboviruses distributed in the world. Based on entomo-virological surveillance, appropriating public health strategies can be addressed to contain cases and control outbreaks. This study aims to show the potential performance of two new molecular methods for detecting DENV serotypes and CHIKV in mosquitoes. Mosquitoes were collected in urban areas and sylvatic areas between July to August 2023, DENV and CHIKV were screened using new multiplex RT-PCR and RT-q PCR. A total of 2150 mosquitoes were trapped including 976 Aedes (959 Ae. egypti, 6 Ae. furcifer and 11 Ae. Vittatus) and 1174 Culex from 39 pools of a maximum of 30 mosquitoes each, tested by molecular screening, 7.7% (3/39) were positive for DENV, DENV-1 was detected in one pool (1/3), and DENV-3 in two pools (2/3). All the pools were negative for CHIKV. The overall minimum infection rate (MIR) of DENV in this study was 3.07 (95% CI: 2.24 —19.86). This study shows the usefulness of our new molecular tools in DENV serotypes and CHIKV surveillance.

In this study, a novel method is proposed to estimate surface-spectral reflectance from camera responses that combines model-based and training-based approaches. An imaging system is modeled using the spectral sensitivity functions of an RGB camera, spectral power distributions of multiple light sources, unknown surface-spectral reflectance, additive noise, and a gain parameter. The estimation procedure comprises two main stages: (1) selecting the local optimal reflectance dataset from a reflectance database and (2) determining the best estimate by applying a neural network to the local optimal dataset only. In stage (1), the camera responses are predicted for the respective reflectances in the database, and the optimal candidates are selected in the order of lowest prediction error. In stage (2), most reflectance training data are obtained by a convex linear combination of local optimal data using weighting coefficients based on random numbers. A feed-forward neural network with one hidden layer is used to map the observation space onto the spectral reflectance space. In addition, the reflectance estimation is repeated by generating multiple sets of random numbers, and the median of a set of estimated reflectances is determined as the final estimate of the reflectance. Experimental results show that the estimation accuracies exceed those of other methods.

The domestic ducks (Anas platyrhynchos domesticus) are resistant to most of the highly pathogenic avian influenza virus (HPAIV) infections. In this study, we characterized the lung proteome and phosphoproteome of ducks infected with HPAI H5N1 virus (A/duck/India/02CA10/2011/Agartala) at 12 hr, 48 hr and 5 days post infection condition. Total 2082 proteins were differentially expressed and 320 phosphorylation sites mapping to 199 phosphopeptides corresponding to 129 proteins were identified. Functional annotation of proteome data analysis reveals activation of RIG-I-like receptor and Jak-STAT signaling pathways, that lead to induction of interferon stimulated genes (ISGs) expression. The pathway analysis of phosphoproteome datasets also confirms the activation of RIG I, Jak-STAT signaling, NF-kappa B signaling and MAPK signaling pathways in the lung tissues. The induction of ISGs proteins (STAT1, STAT3, STAT5B, STAT6, IFIT5 and PKR) establishes protective antiviral immune response in duck lung tissue. Further, the protein-protein interaction network analysis identified proteins like AKT1, STAT3, JAK2, RAC1, STAT1, PTPN11, RPS27A, NFKB1 and MAPK1 as main hub proteins that might play important roles in disease progression in ducks. Together, functional annotation of proteome and phosphoproteome datasets reveals the molecular basis of the disease progression and disease resistance mechanism in ducks infected with HPAI H5N1 virus.

Oncolytic virotherapy is a promising immunotherapy approach for cancer treatment, utilizing viruses to preferentially infect and eliminate cancer cells while stimulating immune responses. In this review we synthesize current literature on the molecular circuits of immune sensing and response to oncolytic virotherapy; focusing on viral DNA or RNA sensing by infected cells, cytokine and danger-associated signal sensing by neighboring cells, and subsequent downstream activation of immune pathways. The sequential sense-and-response mechanisms involve the triggering of molecular sensors by viruses or infected cells to activate transcription factors and related genes for a breadth of immune responses. We describe how molecular signals induced in the tumor upon virotherapy can trigger diverse immune signaling pathways, activating both antigen-presenting cell-based innate and T cell-based adaptive immune responses. Insights into these complex mechanisms provide valuable knowledge for enhancing oncolytic virotherapy strategies.

Cleomaceae is an important group that is medicinally and economically important. However, its systematic position has been controversial. In this study, we have obtained three chloroplast genomes sequencing results of Cleomaceae species. The genome sizes ranged from 157,073 bp (Sieruela rutidosperma) to 158,152 bp (Gynandropsis gynandra). These chloroplast genomes have a typical quadripartite formation with a large single-copy (LSC) region (86,422-87,242bp), a small single-copy (SSC) region (18,485-18,548bp), and a pair of inverted repeat (IR) regions (26,083-26,184bp). The chloroplast genomes encoded 131-132 genes, comprising 8 rRNA genes, 37 tRNA genes, and 79-84 protein-coding genes. The overall GC contents of the three chloroplast genomes are almost identical (35.81%-36.02%). The phylogenetic trees concluding 8 Cleomaceae plants were constructed based on ML and BI methods. The results showed that Gynandropsis gynandra and Sieruela rutidosperma were in the same branch, with Corynandra viscosa as their sister. Six hypervariable areas were identified, containing clpP, matK, ndhF, atpH-atpI, atpF-atpH and petN-psbM which could be used in species identification and population genetics studies. This study provides valuable information for further species identification, evolutionary and genetic studies of Cleomaceae in the future.

The present study provides a detailed analysis of the chloroplast genome of Microula sikkimensis. The genome was consisted of altogether 149,428bp and distinct regions, including large single-copy (81329bp), small single-copy (17261bp), and inverted repeat (25419bp). The genome contained 112 genes, including 78 protein-coding genes, 30 tRNA genes, and 4 rRNA genes, some exhibited duplication in the IR region. The chloroplast genome displayed different GC content across regions, with the IR region exhibiting the highest. Codon usage analysis and the identification of simple sequence repeats (SSRs) offer valuable genetic markers. Comparative analysis with other Boraginaceae species highlighted conservation and diversity in coding and noncoding regions. Phylogenetic analysis placed M. sikkimensis within the Boraginaceae family, revealing its distinct relationship with specific species.

In the first part of this article, we discuss and generalize the complete convergence introduced by Hsu and Robbins (1947) to the r-complete convergence introduced by Tartakovsky (1998). We also establish its relation to the r-quick convergence first introduced by Strassen (1967) and extensively studied by Lai (1976). Our work is motivated by various statistical problems, mostly in sequential analysis. As we show in the second part, generalizing and studying these convergence modes is important not only in probability theory but also to solve challenging statistical problems in hypothesis testing and changepoint detection for general stochastic non-i.i.d. models.