Amplification Of Region Research Articles

Microbiome analysis in individuals with human papillomavirus oral infection

Microbiome gained attention as a cofactor in cancers originating from epithelial tissues. High-risk (hr)HPV infection causes oropharyngeal squamous cell carcinoma but only in a fraction of hrHPV+ individuals, suggesting that other factors play a role in cancer development. We investigated oral microbiome in cancer-free subjects harboring hrHPV oral infection (n = 33) and matched HPV− controls (n = 30). DNA purified from oral rinse-and-gargles of HIV-infected (HIV+) and HIV-uninfected (HIV−) individuals were used for 16S rRNA gene V3–V4 region amplification and sequencing. Analysis of differential microbial abundance and differential pathway abundance was performed, separately for HIV+ and HIV− individuals. Significant differences in alpha (Chao-1 and Shannon indices) and beta diversity (unweighted UniFrac distance) were observed between hrHPV+ and HPV-negative subjects, but only for the HIV− individuals. Infection by hrHPVs was associated with significant changes in the abundance of Saccharibacteria in HIV+ and Gracilibacteria in HIV− subjects. At the genus level, the greatest change in HIV+ individuals was observed for Bulleidia, which was significantly enriched in hrHPV+ subjects. In HIV− individuals, those hrHPV+ showed a significant enrichment of Parvimonas and depletion of Alloscardovia. Our data suggest a possible interplay between hrHPV infection and oral microbiome, which may vary with the HIV status.

True cancer stem cells exhibit relative degrees of dormancy and genomic stability.

Cancer stem cells in human tumors have been defined by stem cell markers, embryonal signaling pathways and characteristic biology, ie., namely the ability to repopulate the proliferating population. However, even if these properties can be demonstrated within a tumor cell subpopulation, it does not mean that they are truly hierarchical stem cells because they could have been derived from the proliferating population in a reversible manner. Using a human PDX, Mary-X, that overall expressed a strong cancer stem cell phenotype, the study conducted both GPP-labelled retroviral transfection and fluorescent microsphere uptake studies to distinguish proliferating from dormant cells and array CGH to identify regions of amplifications (gains) and deletions (losses) on the overall Mary-X population and then applied derived probes by FISH on individual cells to identify a genomically stable subpopulation. Whereas 97-99 % of the cells expressed retroviral GFP and not fluorescent particles and showed numerous gene amplifications and deletions, approximately 1-3 % of the cells showed the opposite. The subpopulation with the retained fluorescent microspheres and exhibiting genomic stability was significantly smaller in size than their GFP-expressing and genomically unstable counterparts. Sorting Mary-X spheroids on the basis of either CD133 or ALDH positivity further enriched for this subpopulation. These studies indicate that a truly biological cancer stem cell subpopulation exists that exhibits both dormancy and genomic stability. This subpopulation could not have been derived from the proliferating and resulting genomically unstable population and therefore represents a truly hierarchical stem cell subpopulation capable of only unidirectional differentiation.

Analysis of HIV transmission characteristics and intervention effects in Guangxi based on molecular networks.

This study evaluates changes in HIV transmission and the effectiveness of interventions after two rounds of the Guangxi AIDS Conquering Project (GACP) in Guangxi, China. Samples and epidemiological data from newly diagnosed people living with HIV (PLWH) between 2014-2020 were analyzed. Molecular networks were constructed using nested PCR amplification and sequencing of the pol region, and multivariable logistic regression identified factors associated with clustering and high-degree nodes. A total of 4,227 valid sequences (73.12% inclusion rate) were analyzed. Demographic changes included an increasing proportion of individuals aged ≥50 years (49.66%), with lower education (50.51%), peasants (76.82%), and heterosexual transmission (90.29%). The overall clustering rate was 86.89%, with higher clustering among individuals aged ≥50 (92.57%), those with primary school or below (89.09%), peasants (88.11%), and CRF08_BC infections (91.48%). Annual declines in cluster growth rate and clustering rates were observed, particularly among individuals aged <30, college graduates, men who have sex with men (MSM), and people who inject drugs (PWID). Key transmission hotspots were identified in Lingshan, particularly among older, less-educated individuals, and peasants. Factors associated with clustering included being male (aOR: 1.27), aged ≥50 (aOR: 3.84), and infected with CRF08_BC (aOR: 2.12). From 2017 to 2020, the risk of clustering and high-degree nodes was lower compared to 2014-2016, suggesting the effectiveness of interventions. Interventions in Guangxi effectively reduced HIV transmission among younger, high-degree populations. However, older, less-educated individuals remain at high risk, necessitating targeted strategies to address their specific needs and achieve better HIV control.

Occurrence of Pestalotiopsis sp. on Ornamental Plants Camellia japonica L. in Romanian Public Gardens

Camellias are evergreen shrubs native to the eastern and southern regions of Asia. In Romania, camellias are popular ornamental plants grown in pots. Pestalotiopsis species have a worldwide distribution and are pathogenic to a wide range of hosts. They also represent an important group of endophytes capable of producing a variety of bioactive secondary metabolites. This paper reports the occurrence of grey blight and twig or branch dieback caused by Pestalotiopsis sp. on Camellia japonica L. grown in pots and used in the landscaping of a public garden located in Iasi city, Romania (GPS coordinates: 47°09′24.0″ N 27°35′15.6″ E). Pestalotiopsis species were identified based on morphological characteristics and Sanger sequencing. Genomic analysis of the fungal isolate coded P_CJ_24, obtained from living plants, was based on ribosomal internal transcribed spacer region amplification using the LSU primers. This first report of Pestalotiopsis species infection of ornamental plants in Romania suggests that new plant pathogens can be introduced in new environmental conditions through plant pods and their incidence increases with the widespread of plant material among gardens.

The first assessment of Angiotensin-Converting Enzyme 1 (ACE1) D/I polymorphism and demographic factors in association with COVID-19 outcomes in the Moroccan Population.

SARS-CoV-2 responsible for the COVID-19 pandemic, infiltrates the human body by binding to the ACE2 receptor in the respiratory system cell membranes, leading to severe lung tissue damage. An analog of ACE2, ACE1, has gained attention due to its well-known Deletion/Insertion (D/I) polymorphism, which seems to be associated with COVID-19 outcomes. This study aims to reveal the allelic and genotypic frequencies of the rs4646994 polymorphism in the Moroccan population and investigate the association between COVID-19 outcomes and both genotypic and demographic data. We screened 162 Moroccan COVID-19 patients for the ACE1 gene D/I polymorphism using PCR amplification of the ACE1 polymorphic region within intron 16. Statistical analysis of the relationship between COVID-19 outcomes and each of the genetic and demographic data was performed using R software. The D allele was present in 74% of subjects. Homozygous (II) and heterozygous (DI) genotypes for the Insertion allele were present in 41.4% and 5.6% of patients, respectively. The median age in the COVID-19 'critical symptoms' category was significantly higher and gradually decreased with less severe symptoms. Similarly, males were significantly overrepresented in the 'critical symptoms' category, while females predominated in the 'mild symptoms' category. The present study reports the prevalence of ACE1 D/I alleles for the first time in the Moroccan population and confirms the strong association of severe COVID-19 outcomes with male sex and older age. Moreover, this work is the first to explore the relationship between ACE1 D/I polymorphism and COVID-19 clinical outcomes in North African adults. The lack of a significant association may be due to cohort size or population-specific factors. A comprehensive investigation in a larger North African cohort is highly recommended.

ASSOCIATION ANALYSIS OF MULTIDRUG RESISTANCE GENE ABCB1 IN BREAST CANCER PATIENTS OF PUNJAB

Aims: This study aims to determine the frequency distribution of genetic polymorphisms of ABCB1 (rs1128503 and rs1045642) gene in breast cancer patients and the healthy controls and the association of these genotypes, if any, with susceptibility to breast cancer. Methods: The present case control study was conducted on the population of Punjab (North India). The cases included 300 breast cancer confirmed patients and the control group included (n=300) healthy female volunteers from general public after taking clearance from Institutional Ethical Committee (IEC) of Punjabi University, Patiala. Genomic DNA was isolated from whole blood collected in EDTA coated tubes by employing the standard inorganic (Salting Out) extraction method (Miller et al, 1988). PCR-RFLP technique was used for the amplification of the target region of the gene by using a single pair of primers. Genotyping of the PCR products and digested fragments obtained after RFLP was done by agarose gel electrophoresis. For PCR products, 1% gel was made and for the restriction digestion products, 3.5% gel was made. After separation, the bands representing products/ fragments were visualized under UV light for genotyping. Results:The frequency distribution of CC, CT, and TT genotypes of rs1045642 (C3435T) among breast cancer patients were 34%, 46.67% and 19.33% and among controls were 38.67%, 45.67% and 15.67% respectively. The frequencies of genotypes CC, CT and TT of rs1128503 (C1236T) among cases were 28%, 45.67% and 26.33% and among controls were 28.33%, 42% and 29.67%, respectively. The frequency of the wild allele (C) and mutant allele (T) of rs1045642 among cases was 57.33% and 42.67% and among controls, 61.5% and 38.5% respectively. The frequencies of the wild allele (CC) and mutant allele (T) of rs1128503 among cases were 50.83% and 49.17% and among controls were 49.33% and 50.67%, respectively.For association analysis, the contingent chi square test did not show any statistically significant differences between cases and controls for rs1045642 and rs1128503 gene polymorphisms neither at genotypic level and nor at allelic level. Conclusion: We found no evidence that reveals any statistically significant association of C3435T and C1236T polymorphisms with susceptibility to breast cancer in Punjab, India.

Optical refractive inverse weak value tiltmeter

We present an expanded mathematical model of the optical weak value amplification device introduced by Duck, Stevenson, and Sudarshan in 1989 [Phys. Rev. D 40, 2112 (1989)PRVDAQ0556-282110.1103/PhysRevD.40.2112]. We model the pointer behavior in and around the weak value and inverse weak value amplification regions with respect to the incidence angle of the birefringent crystal. We present laboratory measurements that support the validity of this model. We measure a tilt sensitivity of ∼.58m/rad, which represents an amplification of 390 times over nonamplified refraction. This corresponds to an increase in SNR of ∼110 times with respect to shot noise. This paper demonstrates the novel utility of a birefringence-based WVA device as a sensitive tiltmeter, to our knowledge.

The relationship between mitochondrial DNA haplotype and its copy number on body weight and morphological traits of Wuliangshan black-bone chickens.

Mitochondria play a pivotal role as carriers of genetic information through their circular DNA molecules. The rapid evolution of the D-loop region in mitochondria makes it an ideal molecular marker for exploring genetic differentiation among individuals within species and populations with close kinship. However, the influence of mtDNA D-loop region haplotypes and mtDNA copy numbers on phenotypic traits, particularly production traits in chickens, remains poorly understood. In this comprehensive study, we conducted D-loop region amplification and sequencing in the blood mitochondria of 232 female Wuliangshan black-bone chickens. Our investigation identified a total of 38 haplotypes, with a focus on 10 haplotypes that included more than five individuals. We meticulously analyzed the correlations between these haplotypes and a range of traits, encompassing body weight, tibial length, tibial circumference, body oblique length, chest width, and chest depth. The results unveiled significant disparities in specific tested traits across different haplotypes, indicating a tangible association between mtDNA haplotypes and traits in chickens. These findings underscore the potential impact of mitochondrial DNA variations on energy metabolism, ultimately leading to divergent chicken phenotypes. Furthermore, our examination revealed positive correlations between mtDNA copy numbers and tested traits for select haplotypes, while other haplotypes exhibited non-uniform relationships between traits and mtDNA copy numbers. In addition, phylogenetic analysis disclosed the involvement of two subspecies of red jungle chicken in the origin of Wuliangshan black-bone chickens. Consequently, our research contributes novel insights into mitochondrial genomic selection, augments comprehension of the roles played by haplotypes and mtDNA copy numbers in chicken population genetics and phylogenetic analysis, and furnishes fundamental data crucial for the preservation and provenance determination of black-bone chickens.

Metabolite profiling of PGPR Bacillus subtilis BGKMR1: A potential strategy for managing Fusarium equiseti causing wilt in bitter gourd (Momordica charantia L.)

Wilt caused by Fusarium equiseti is one of the most destructive diseases that leads to substantial yield loss in bitter gourd. The F. equiseti strain CBEFE1((PQ111513.1) which was identified morphologically with falcate shaped three septate macroconidia and oval shaped microconidia and molecularly confirmed through amplification of the ITS region at 560bp was used for this study. Plant growth promoting rhizobacteria (PGPR) act as a sustainable biocontrol agent against major plant pathogens through multiple mode of actions. PGPR were isolated from native rhizopshere region in bitter gourd. The isolated PGPR Bacillus subtilis BGKMR 1 shows maximum mycelial inhibition of 68.73 % against F. equiseti in dual plate assay. The presence of metabolites produced by B. subtilis and F. equiseti along with its interaction were identified through the GCMS profiling. Primarily, pathogenic compound squalene (8.88 %) was identified in F. equiseti and antifungal compound 1,4-benzenedicarboxylic acid (6.1 %) was identified in B. subtilis. Further, B. subtilis BGKMR 1 during its interaction shows propanoic acid (14.69 %) which was found to have effectiveness against F. equiseti. KEGG analysis revealed pyrimidine metabolism in F. equiseti and nitrogen, alanine, aspartate and glutamate metabolism in B. subtilis and propanoate metabolism pathways were detected during its interaction. B. subtilis BGKMR 1 promote the seed germination rate upto 96 % and vigor index to 1919.04 when compared to control. The seed treatment with soil application of B. subtilis BGKMR1 reduced disease incidence upto 25.95 % under glass house condition. Moreover, B. subtilis BGKMR1 treated plants shows enhanced defense enzymes activity such as peroxidase, polyphenol oxidase and phenylalanine ammonia-lyase indicating induction of systemic resistance. Therefore, B. subtilis BGKMR 1 contains novel antifungal metabolites which were identified via GC-MS and induced defense enzyme activity highlights its practical application as a sustainable and eco-friendly solution for managing wilt disease in bitter gourd.

Fungal endophytes diversity and influencing factors in Liquidambar orientalis Mill. in Türkiye

Fungal endophytes were isolated from the leaves and petioles of Liquidambar orientalis Mill., an endangered species in Türkiye. Plant material was collected from 10 sites in September 2023, yielding 499 fungal isolates, classified into 38 morphological groups. DNA extraction and polymerase chain reaction (PCR) amplification of ITS and Beta-tubulin regions were conducted on representative isolates. All fungi belonged to the Ascomycota phylum, comprising 11 genera and 26 species across 15 families, with one group unidentified. The most prevalent families were Diaporthaceae (34.9%), Pleosporaceae (23.4%), and Botryosphaeriaceae (22.2%), with Diaporthe eres (15.0%) and Phomopsis sp. (12.4%) being dominant species. Fungal diversity was assessed using Shannon, Simpson, and Chao1 indices, revealing tissue type as the strongest factor influencing species diversity, followed by media and spatial factors. The presence of pathogenic families, such as Botryosphaeriaceae, highlights potential threats to the species. This is the first study to report fungal endophytes in L. orientalis, as well as the first records in Türkiye for several species, including Alternaria destruens, Alternaria alstroemeriae, Stemphylium majusculum, Diaporthe cynaroidis, Pseudopithomyces rosae, Nothophoma variabilis, Cladosporium endophyticum, Cladosporium colombiae, Muyocopron sp., Sphaerulina rhododendricola, Constantinomyces macerans, and Aequabiliella effusa.

Mutations on the A and C genomes of Brassica napus confer herbicide resistance to acetohydroxyacid synthase inhibitors

ABSTRACT The point mutations conferring resistance to the sulfonylurea herbicide, chlorsulfuron, are reported for two Brassica napus lines (19c and 30a) developed by seed mutagenesis using ethyl methanesulfonate. Crosses with the herbicide-susceptible wild-type line generated F2 and backcrossed progeny and confirmed that chlorsulfuron resistance was inherited as a dominant mutation at a single locus in both lines. F2 progeny from crossing 19c and 30a segregated in a 15 resistant: 1 susceptible ratio, establishing that the two herbicide-resistant loci are independently inherited and that the mutations for resistance occur at different loci. PCR amplification and DNA sequencing of regions from the acetohydroxyacid synthase gene revealed two separate point mutations at different nucleotides in the same codon. This results in the substitution of proline-197 for leucine in 19c and proline-197 for serine in 30a. The different amino acid substitutions in the acetohydroxyacid synthase enzyme explain the differing responses of 19c and 30a to sulfonylurea and imidazolinone herbicides. Based on attempts at introgression of chlorsulfuron resistance to a wide range of forage brassicas, we conclude that the 19c mutation resides in the Brassica C genome, whereas the 30a mutation is in the Brassica A genome.

PoxtA amplification and mutations in 23S rRNA confer enhanced linezolid resistance in Enterococcus faecalis.

This study aimed to explore the evolutionary patterns and resistance mechanisms of an Enterococcus faecalis strain harbouring poxtA under linezolid exposure. A poxtA-carrying E. faecalis electrotransformant DJH702 with a linezolid minimum inhibitory concentration of 4 mg/L was exposed to increasing concentrations of linezolid (8-64 mg/L). The derived strains growing at 8, 16, 32 and 64 mg/L, designed DJH702_8, DJH702_16, DJH702_32 and DJH702_64, were obtained. The amplification and overexpression of poxtA were measured using sequencing and RT-PCR, the fitness cost by competition assays and the stability of the repeat units by serial passage. In all derived strains, high-level linezolid resistance develops through poxtA amplification. The relative copy numbers and transcription levels of poxtA were significantly increased. However, in the presence of higher linezolid concentrations, DJH702_32 and DJH702_64 showed reduced poxtA copy numbers and transcription levels compared with DJH702_8 and DJH702_16, but additional mutations in the 23S rRNA (G2505A). IS1216E-mediated formation of translocatable units with subsequent tandem amplification of these translocatable units supported the gain of poxtA segments. However, these amplicons were not stable and were lost frequently in the absence of a linezolid selection pressure. The amplification of the poxtA region did not result in a fitness cost, but mutations in 23S rRNA did. poxtA-carrying E. faecalis electrotransformants used two distinct mechanisms to resist linezolid selection pressure: at lower concentrations, strains prioritized increasing poxtA expression levels, while at higher concentrations, a combination of increased poxtA expression and mutations in 23S rRNA was observed.

Integrated analyses of multi-omic data derived from paired primary lung cancer and brain metastasis reveal the metabolic vulnerability as a novel therapeutic target

BackgroundLung cancer brain metastases (LC-BrMs) are frequently associated with dismal mortality rates in patients with lung cancer; however, standard of care therapies for LC-BrMs are still limited in their efficacy. A deep understanding of molecular mechanisms and tumor microenvironment of LC-BrMs will provide us with new insights into developing novel therapeutics for treating patients with LC-BrMs.MethodsHere, we performed integrated analyses of genomic, transcriptomic, proteomic, metabolomic, and single-cell RNA sequencing data which were derived from a total number of 154 patients with paired and unpaired primary lung cancer and LC-BrM, spanning four published and two newly generated patient cohorts on both bulk and single cell levels.ResultsWe uncovered that LC-BrMs exhibited a significantly greater intra-tumor heterogeneity. We also observed that mutations in a subset of genes were almost always shared by both primary lung cancers and LC-BrM lesions, including TTN, TP53, MUC16, LRP1B, RYR2, and EGFR. In addition, the genome-wide landscape of somatic copy number alterations was similar between primary lung cancers and LC-BrM lesions. Nevertheless, several regions of focal amplification were significantly enriched in LC-BrMs, including 5p15.33 and 20q13.33. Intriguingly, integrated analyses of transcriptomic, proteomic, and metabolomic data revealed mitochondrial-specific metabolism was activated but tumor immune microenvironment was suppressed in LC-BrMs. Subsequently, we validated our results by conducting real-time quantitative reverse transcription PCR experiments, immunohistochemistry, and multiplexed immunofluorescence staining of patients’ paired tumor specimens. Therapeutically, targeting oxidative phosphorylation with gamitrinib in patient-derived organoids of LC-BrMs induced apoptosis and inhibited cell proliferation. The combination of gamitrinib plus anti-PD-1 immunotherapy significantly improved survival of mice bearing LC-BrMs. Patients with a higher expression of mitochondrial metabolism genes but a lower expression of immune genes in their LC-BrM lesions tended to have a worse survival outcome.ConclusionsIn conclusion, our findings not only provide comprehensive and integrated perspectives of molecular underpinnings of LC-BrMs but also contribute to the development of a potential, rationale-based combinatorial therapeutic strategy with the goal of translating it into clinical trials for patients with LC-BrMs.

Electrochemical Detection of DNA in Agriculture Using Lamp-Based Amplification

The variety and complexity of genetically modified organisms (GMOs) are increasing, which makes their detection harder than ever before. This impacts the traceability, safety, and monitoring of food. Current research indicates that GM crop technology may pose risks to human health and the environment. The aim of this study is the development of a LAMP-based GMO electrochemical detection device. LAMP amplification of P-35S, P-FMV, and T-NOS regions (three elements that are most often present in genetically modified plants as part of the transgenic construct) is done using gBlocks, synthetic dsDNA fragments that have an identical DNA sequence to that of the targeted part of a transgenic construct.Using single-stranded DNA probes containing methylene blue as a redox probe, the electrochemical DNA sensor detects LAMP products that are complementary to the target DNA. The probes were attached to the gold electrodes functionalized by gold nanoparticles, and 2D nanomaterials (MoS2, Mxenes, and reduced graphene oxide). The electrochemical methods, such as alternating current voltammetry, square wave voltammetry, and differential pulse voltammetry were employed in testing the electrochemical detection response of the DNA probes immobilized on 2D nanomaterials.The research has shown that the obtained DNA detection signal depends significantly on the methods (and setup parameters) used in electrochemical detection, and the nanomaterials used on electrodes. The electrochemical detection resulted in a signal decrease consequential to the conformational changes of the DNA probes upon attachment of the target DNA. The methylene blue molecules attached at the top of the DNA probes get higher resistance in the electron transfer to the electrode materials, which makes the target DNA detection quantitatively and qualitatively visible. These results are promising for the development of a device that will enable a specific, sensitive, fast, cost-effective, and precise in-field detection system for the routine detection of GMOs.**Acknowledgments:*This project is supported by The Science Fund of the Republic of Serbia DEVELOPMENT Program - Green program of cooperation between science and industry - LABOUR project (grant agreement No. 6710), and The Ministry of Science, Technological Development and Innovation of the Republic of Serbia (451-03-66/2024-03/200358).*This work is supported through the ANTARES project that has received funding from the European Union's Horizon 2020 research and innovation program under grant agreement SGA-CSA. No. 739570 under FPA No. 664387. https://doi.org/10.3030/739570 Figure 1

The Impact of Green Tea Kombucha on the Intestinal Health, Gut Microbiota, and Serum Metabolome of Individuals with Excess Body Weight in a Weight Loss Intervention: A Randomized Controlled Trial.

Green tea kombucha (GTK) has emerged as a promising probiotic fermented beverage. Few studies have investigated its effect on human health, mainly focusing on intestinal health, microbiota composition, and metabolomics. The present study is a pioneer in investigating the effect of GTK consumption in individuals with excess body weight. This is a randomized controlled trial, lasting ten weeks, with two groups placed under an energy-restricted diet: control (CG, n = 29), kombucha (KG, n = 30; 200 mL/d). Biological samples and questionnaires were collected before and after the intervention. Microbiota analysis used an amplification of the V4 region of 16S rRNA. Serum untargeted metabolomics used HPLC-TOF mass spectrometry. Intestinal permeability considered the urine excretion of lactulose and mannitol, plasma zonulin, and LPS-binding protein. After the intervention, no differences related to intestinal permeability and microbiota were found between groups, but only the CG had increased fecal pH, lactulose/mannitol ratio, and zonulin. In addition to this, the KG reported lower gastrointestinal symptoms related to motility compared to the CG, and discriminant metabolites (e.g., diethyl malonate) were found strictly in the KG. GTK did not significantly improve gut microbiota and intestinal permeability. However, GTK ameliorated gastrointestinal symptoms and positively influenced the serum metabolome, which may contribute to enhancing the metabolic health of individuals with excess body weight.

Tick Infestation and Molecular Detection of Tick-Borne Pathogens from Indian Long-Eared Hedgehogs (Hemiechinus collaris) in Pakistan.

Hedgehogs can act as reservoirs for the transmission of tick-borne pathogens (TBPs) to domestic livestock, wild animals, and humans. Understanding host-tick dynamics is essential to evaluate the impact of TBPs. This study was conducted in Pakistan and aimed to determine the prevalence and species of TBPs in the blood and ticks of Indian long-eared hedgehogs captured from various environments. A total of 64 hedgehogs were captured to check for tick infestation. Tick species were identified morphologically and molecularly including ITS-2 region amplification by PCR and subsequent Sanger sequencing. Moreover, TBPs were identified in both ticks and the blood of hedgehogs through conventional PCR and sequencing, targeting the regions msp1b, 18S rRNA, and cytb for Anaplasma spp., Babesia spp., and Theileria spp., respectively. Out of 64 hedgehogs, 16 (25%) were found to be infested with ticks. Morphological and molecular analysis identified all 109 collected ticks as Rhipicephalus turanicus. Only one hedgehog (6.2%) was infected with A. marginale. From the tick samples, 3.7% tested positive for Theileria lestoquardi, 2.8% for Anaplasma marginale, and another 2.8% for Babesia bigemina. This study provides critical insights into circulating TBPs in this region and what possible role hedgehogs might play in disease maintenance for Anaplasma marginale while identifying multiple pathogens that are of concern to human and animal health.

Fault Detection selection method for distribution network based on regional amplification of grounding current and double closed-loop feedback control

Abstract The susceptibility of high-resistivity ground faults in power systems, coupled with systemic imbalances, can result in the distortion of fault characteristics, leading to erroneous fault line selection. In addition, the full compensation of fault arc suppression conceals the fault characteristics, which brings difficulties to fault detection. In order to take into account the needs between fault arc suppression and fault detection, and tackle the issue of high-impedance fault line discrimination with a comprehensive and innovative approach, this paper studies the approach to line selection based on double closed-loop feedback control and amplification of the grounding current region. It enables adaptable grounding of the neutral point when a transient ground fault arises, and priority is given to ensuring fast and reliable arc suppression. In the event of a persistent grounding fault, the flexible grounding of the neutral point through the current source serves to mitigate the deleterious effects of fault deterioration, and the fault current is flexibly modulated within a relatively safe range until the requirements of fault arc suppression and fault detection are met.

Methodology comparison of environmental sediment fungal community analysis

Fungi play important roles in ecosystems. Analyzing fungal communities in environments has long been a challenge due to the large difference in compositions retrieved using different methods or sequencing regions, obscuring the true abundance and species information. Our study aimed to compare and determine more accurate approach for evaluating fungal populations in river sediment. To achieve this, different primer sets in the internal transcribed spacer (ITS) (ITS5/ITS1R, ITS1F/ITS2), 18S rRNA gene (0817F/1196R) for High-throughput sequencing (HTS), metagenomic shotgun sequencing (MS) directly from environmental samples, and HTS using ITS primers for the fungal samples collected from plate cultivation were used to characterize the fungal communities. We calculated diversity index and used FungalTraits to analyze methods preferences for fungal species. The study revealed that when analyzing the fungal species directly from environmental samples, amplification and sequencing of ITS region demonstrated more accuracy than MS and 18S rRNA gene sequencing methods, but displayed significant primer preference. Over 30 % fungal species from HTS after plate cultivation were not present in HTS from the environmental samples. NMDS analysis demonstrated significant disparities in species diversity among different methods, suggesting potential complementarity between them. Over 85% species identified by HTS using ITS primers belonged to filamentous fungi, while the MS mostly identified yeast (62%). Therefore, to get more accurate fungal community information in sediment, multiple methods were recommended by using cultivation, molecular biological methods dependent on PCR techniques like ITS1F/ITS2 primer for HTS and PCR independent method such as metagenomic shotgun sequencing techniques.

Non-destructive and rapid identification of yeasts by nearinfrared spectroscopy and machine learning

This study aimed to apply near-infrared (NIR) spectroscopy combined with machine learning techniques to identify yeast strains rapidly and practically, comparing the results with traditional molecular identification methods. Yeasts were isolated from the digestive tracts of aquatic mining insects collected in the extreme north of the Western Amazon (Roraima), Brazil, and preserved through cryopreservation and mineral oil methods. Molecular identification involved PCR amplification and sequencing of ribosomal DNA regions. NIR spectroscopy, coupled with multivariate analysis and machine learning algorithms such as principal component analysis (PCA), hierarchical cluster analysis (HCA), k-nearest neighbor (KNN), and soft independent modeling by class analogy (SIMCA), was used to analyze and classify the yeast samples, accurately identified yeast strains at the genus and species levels, achieving 100% accuracy in both the calibration and validation sets. The results indicate that this method provides a rapid, non-destructive, and environmentally friendly alternative to traditional molecular techniques, making it suitable for real-time, in situ analysis with minimal sample manipulation.

Spatiotemporal variations of global precipitation concentration and potential links to flood-drought events in past 70 years

The evaluation of precipitation events is crucial for predicting severe droughts and floods, particularly in the context of global warming. This study presents a comprehensive spatiotemporal analysis of the global precipitation concentration index (CI) from 1950 to 2020, comparing CI with nine extreme precipitation indices to assess its applicability. The world map was divided into three distinct regions related to flood-drought events using the standardized precipitation index (SPI). Key findings include: 1) CI exhibited significant spatial heterogeneity, with elevated values in seasonally warm coastlines and arid regions. Seasonal changes in CI were not synchronized, particularly lower in Northern Hemisphere winters, while the Southern Hemisphere displayed minor seasonal variation, alongside a long-term growth trend in approximately 36.5 % of the global area. 2) CI's spatiotemporal variations helped delineating regional precipitation patterns, revealing a strong correlation (R2 = 0.87) with the extreme precipitation index R95c, but a negligible relationship with total precipitation (P) (R2 = −0.05). This facilitated the global partitioning into three regions: Humidity Amplification Region (I), Dryness Intensification Region (II), and Climate Fluctuation Region (III). 3) Flood-drought events were potentially linked to variations in CI and P, where In Region I experienced increased flood risk due to rising CI and P, Region II faced heightened drought risk from rising CI and declining P, and Region III showed opposing effects on floods due to reduced CI and increased P, with P variability significantly influencing flood frequency. 4) The main atmospheric circulation factors varied by region, typically including the Antarctic Oscillation (AAO), Atlantic Multi-decadal Oscillation (AMO) and Arctic Oscillation (AO), with AAO being most significant. This research underscores the intricate relationships between climatic indices and hydrological extremes, emphasizing the challenges posed by global warming, atmospheric circulation changes, and data uncertainties in assessing drought and flood disasters.