Gene PCR Research Articles

Abstract B062: Transcriptomic analysis of whole blood samples from prostate cancer patients: A comparative study of patients from geographically diverse populations

Abstract Introduction: Prostate cancer is the most frequently diagnosed cancer across 112 countries. Among American men, an estimated 299, 010 new cases and 35,250 deaths are projected for 2024. In this study, the transcriptomic profile of prostate cancer patients from geographically and ethnically diverse cohorts (USA-Caucasian, RSA-Black) were compared to identify gene expression patterns associated with prostate cancer. Methods: Whole blood samples were collected from PCa and controls. Controls had the same age-range as PCa with no clinical indication of prostatic disease and serum PSA levels below 1.5 ng/ml. RNA was isolated, sequencing was carried out on Novaseq 6000, and data analyzed using PartekFlow to identify differential gene expression patterns (RSA-PCa: 6, RSA-Control: 6, USA-PCa: 7, USA-Control: 11). qRT-PCR was undertaken on a cohort of 27 biomarkers (PROSTest) related to PCa in whole blood. Results: Differential expression analysis identified distinct patterns in USA and RSA groups. In the USA PCa cohort, 2,193 genes were differentially expressed compared to the controls with fold change (FC) >1, p-value (p) <0.05, while in the RSA PCa cohort, 1,627 genes were differentially expressed compared to the controls. Pathway analyses identified enrichment for immune responsive genes in the RSA cohorts. We identified that 26 of 27 Wren’s PROSTest genes detectable in both groups. RNAseq and normalized PCR gene expression were well- correlated (r=0.44, p=0.0012, n=30 pairs). Conclusion: Based on this RNAseq study, there may be geographic differences related to ethnicity in overall blood RNA gene expression. We specifically identified alterations in immune function gene expression between the USA and RSA. Importantly, PROSTest genes were detected in both populations and exhibited a significant correlation. Development of gene signatures for biomarker discovery should, however, consider specific ethnicities and geographical regions. Citation Format: Srinivas V Koduru, Mark Kidd, Ané Pieters, S E Nagel, Robert P Millar, Abdel B Halim. Transcriptomic analysis of whole blood samples from prostate cancer patients: A comparative study of patients from geographically diverse populations [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr B062.

Large, but short-term, increase in fecal indicator bacteria following extreme flooding from Hurricane Harvey in Houston, TX

Hurricane Harvey caused widespread flooding along the Texas Gulf Coast in August 2017; some areas of Houston received >150 cm of rainfall within a few days. Due to concerns over fecal contamination of floodwaters, surface water samples were collected at six locations in the southeastern Houston area immediately before and after the hurricane and then every 1 to 2 weeks thereafter over a 2-month period. Total E. coli was enumerated using the IDEXX Quanti-Tray/2000 system. DNA extracted from water samples was analyzed via quantitative real-time PCR (qPCR) for general and source-specific total Bacteroidales and human Bacteroidales markers, and digital PCR (dPCR) for antibiotic resistance genes (ARG) and a plasmid (pBI143) associated with human waste. SourceTracker2 was used to determine human source contributions based on metagenomic analysis of PCR-amplified 16S rRNA gene fragments. Samples collected immediately after the hurricane had elevated levels of E. coli, ranging from 488 to 1,733 CFU 100 ml−1. After 1 week, E. coli levels decreased to <100 MPN 100 ml−1. Total Bacteroidales numbers were elevated immediately following the hurricane and remained high for 12 days. Human-source contributions, as assessed by PCR methods and metagenomic analysis, peaked within 12 days after the hurricane consistently across all sampling sites. Multiple regression analysis of environmental parameters, copies of ARG and pBI143, and metagenomic data confirmed that human waste caused the dramatic, short-term, high levels of fecal contamination of floodwaters generated by Hurricane Harvey. Fecal indicators approached normal background levels approximately 3 weeks after the rainfall ended.

Astaxanthin mediated repair of tBHP-Induced cellular injury in chondrocytes

ABSTRACT Objective This study investigates how astaxanthin (AST) counters tert-butyl hydroperoxide (tBHP)-induced cellular damage in C28/I2 chondrocytes, focusing on the circ-HP1BP3/miR-139-5p/SOD1 signaling pathway and its use in sustained-release microspheres for osteoarthritis treatment. Methods We employed a variety of techniques including real-time quantitative PCR, Western blot, ELISA, and dual-luciferase reporter gene assays to explore AST's molecular effects. Additionally, the efficacy of AST-loaded sustained-release microspheres was evaluated in vitro and in a mouse model of osteoarthritis. Results AST significantly enhanced SOD1 expression, reducing apoptosis and inflammation in damaged cells. The AST-loaded microspheres showed promising in vitro drug release, improved cell viability, and reduced oxidative stress. In the osteoarthritis mouse model, they effectively decreased joint inflammation and increased the expression of chondrocyte markers. Conclusion Astaxanthin effectively mitigates oxidative stress and inflammation in chondrocytes via the circ-HP1BP3/miR-139-5p/SOD1 pathway. The development of AST-loaded microspheres offers a novel and promising approach for osteoarthritis therapy, potentially extending to osteoarthritis treatment.

Does Fusobacterium in Colorectal Cancer Sites Originate From the Oral Cavity? A Pilot Study.

Fusobacterium can contribute to oral diseases, but also pose as a systemic risk factor. This genus, and especially F. nucleatum, can be found in colorectal cancer (CRC) tissue and is involved in multiple aspects of this type of cancer. Previous studies indicated a possible oral origin of these bacteria; however, stronger evidence is needed to reach a definitive conclusion. This pilot study aimed to establish a method to successfully compare, at the strain level, fusobacteria from the oral cavity and CRC resection material for future cohort studies of CRC patients. In a first cohort of eight periodontitis patients, gingival crevicular fluid and saliva were collected. Fusobacterium was isolated on two different media. In a second cohort, saliva and CRC resection material were collected from ten CRC patients. These samples were used for screening of Fusobacterium with culturing, 16S rRNA gene profiling and a PCR-based approach. In the first cohort, different Fusobacterium species were identified in GCF and saliva samples. However, as the total yield of Fusobacterium seemed slightly higher in saliva samples, it was therefore preferred for subsequent sample collection. Thus, in the second cohort, patient-matched saliva and CRC resection material were screened for Fusobacterium and this showed that nine patients were culture-positive in the saliva samples; however, no Fusobacterium could be isolated from the resection material. On the other hand, 16S rRNA gene profiling of the resection material indicated that eight CRC patients were positive for Fusobacterium. All eight of these patients carried Fusobacterium in their saliva, indicated by both marker gene PCR and culture-based screening. These pilot results are compatible with data from previous studies, indicating a possible link between oral and CRC-associated Fusobacterium, and a more in-depth analysis of specific strains and their characteristics in a larger cohort is justified. The protocol was registered at clinicaltrials.gov (NCT05945082).

The effect of next-generation, dual-active-ingredient, long-lasting insecticidal net deployment on insecticide resistance in malaria vectors in Benin: results of a 3-year, three-arm, cluster-randomised, controlled trial

Insecticide resistance among malaria vector species now occurs in 84 malaria-endemic countries and territories worldwide. Novel vector-control interventions, including long-lasting insecticidal nets (LLINs) that incorporate new active ingredients with distinct modes of action, are urgently needed to delay the evolution and spread of resistance and to alleviate reversals in malaria-control gains. We aimed to assess the longitudinal effect of two dual-active-ingredient LLINs on insecticide resistance during a cluster-randomised, controlled trial in Benin. This 3-year, three-arm, cluster-randomised, controlled trial was conducted between Oct 17, 2019, and Oct 24, 2022, in three districts in southern Benin, to compare the effects of LLINs containing chlorfenapyr-pyrethroid or pyriproxyfen-pyrethroid with LLINs containing pyrethroid only. In 19 292 mosquitoes (Anopheles gambiae sensu lato) collected over 36 months-3 months of baseline followed by 3 years post-intervention-we measured longitudinal phenotypic insecticide resistance profiles using bioassays and genotypic resistance profiles using quantitative, real-time, reverse transcriptase PCR of metabolic resistance genes in two clusters per trial group. The trial was registered with ClinicalTrials.gov, NCT03931473. In all three trial groups, a significant effect of LLINs on insecticide resistance selection was evident, with the median lethal dose (LD50) of α-cypermethrin approximately halving between baseline and 12 months post-LLIN distribution (pyrethroid-only LLIN cluster 21: LD50 78·78 μg/ml [95% CI 65·75-94·48] vs 35·93 [29·41-43.86] and cluster 31: 79·26 [65·40-96·44] vs 38·71 [30·88-48·53]; chlorfenapyr-pyrethroid LLIN cluster 43: 104·30 [82·97-133·58] vs 43·99 [35·30-54·86]; and pyriproxyfen-pyrethroid LLIN cluster 36: 63·76 [52·14-77·75] vs 37·96 [30·88-46·69] and cluster 53: 77·67 [57·63-104·56] vs 39·72 [29·26-53·97]). Over the subsequent 2 years, the LD50 of α-cypermethrin increased past baseline values in all three trial groups (year 3 pyrethroid-only LLIN cluster 21: 141·01 [111·70-181·90] and cluster 31: 115·15 [93·90-143·09]; chlorfenapyr-pyrethroid LLIN cluster 43: 97·00 [77·24-123·54] and cluster 55: 126·99 [102·34-161·26]; and pyriproxyfen-pyrethroid LLIN cluster 36: 142·29 [112·32-184·84] and cluster 53: 109·88 [79·31-157·70]). We observed minimal reductions in chlorfenapyr susceptibility and variable but significant reductions in fertility after pyriproxyfen exposure, with an overall trend of increasing susceptibility across trial years. Several metabolic genes were implicated in resistance selection, including CYP6P4 in the pyriproxyfen-pyrethroid LLIN group, which encodes an enzyme known to metabolise pyriproxyfen in vitro, and CYP6P3 and CYP9K1 in the chlorfenapyr-pyrethroid LLIN group, both of which encode enzymes that are involved in pro-insecticide activation. After 24 months of use, chlorfenapyr-pyrethroid LLINs no longer mitigated pyrethroid resistance selection in this area of southern Benin, which has high malaria transmission dominated by highly resistant A gambiae sensu lato. This finding raises issues for current net-procurement schedules, which are based on an operational net lifespan of 3 years. Knowledge of the effects of next-generation LLINs on insecticide-resistance selection is crucial for the pragmatic design of prospective resistance-management strategies. UNITAID and The Global Fund to Fight AIDS, Tuberculosis and Malaria.

DNA Barcoding Using 18S rRNA Gene Fragments for Identification of Tick-Borne Protists in Ticks in the Republic of Korea

The objective of this study was to evaluate the diversity and prevalence of tick-borne protists in the Republic of Korea via DNA barcoding using 18S rRNA gene fragments and PCR. Between 2021 and 2022, questing ticks were collected using the flagging method, with a total of 13,375 ticks collected and pooled into 1003 samples. Of these, 50 tick pools were selected for DNA barcoding targeting the V4 and V9 regions of 18S rRNA using the MiSeq platform. A taxonomic analysis of the amplicon sequence variants identified three genera of protozoa, namely Hepatozoon canis, Theileria luwenshuni, and Gregarine sp. However, the number and abundance of protists detected were different depending on the primer sets, and T. gondii was not identified in DNA barcoding. Furthermore, conventional PCR confirmed the presence of H. canis, Toxoplasma gondii, T. luwenshuni, and Theileria sp. in the collected ticks. This study identified H. canis and T. gondii in Ixodes nipponensis for the first time. It demonstrated that the results of DNA barcoding using 18S rRNA gene fragments can vary depending on the primer sets and further optimization is required for library construction to identify tick-borne protists in ticks. Despite these limitations, the findings highlight the potential of DNA barcoding using 18S rRNA gene fragments for screening the diversity of tick-borne protists in ticks.

Use of modified human hemangioma tissue cultures and human umbilical vein endothelial cell cultures to gain mechanistic insights into imiquimod treatment for infantile hemangioma.

Infantile hemangiomas (IH) are a common entity encountered by dermatologists, otolaryngologists, and other surgeons. Oral propranolol is a mainstay of treatment for IH and is well-tolerated, though propranolol-refractory IH and other drug-related adverse events are documented and can limit its usage. There are few in vitro testing systems for putative treatment agents. To address this, we modified a tissue culture system for human hemangioma treatment testing to evaluate the treatment impact of the immune modifier, imiquimod. Human umbilical vein endothelial cells (HUVEC) and hemangioma cultures were treated with several concentrations of imiquimod followed by MTT assays, reporter gene assays, PCR, ELISA, and Western blotting for IL-8, VEGF, Cyclin D1, and IFNα and immunohistochemistry for Cyclin D1 and Ki-67. HUVEC showed acute decreases in IL-8, VEGF, and Cyclin D1 promoter activity and increases in IFNα mRNA after imiquimod treatment. Hemangioma samples showed no change in Ki-67 or Cyclin D1 staining after treatment with imiquimod after 27 d, with significantly increased IL-8 and VEGF. From this preliminary analysis, we discerned that hemangioma tissues can be grown in tissue culture and used for drug treatment studies. We also conclude acute and chronic modulation of cell cycle, angiogenesis factors, and immunostimulatory conditions may be associated with imiquimod mechanisms of action in hemangioma involution.

Exploring Fresh Lettuce (Lactuca sativa) as a Dairy-Free Probiotic Source

Probiotics are living microorganisms that, when given in appropriate amounts, have optimistic effects on body. This study explores the potential of fresh lettuce (Lactuca sativa) leaves as a probiotic source for individuals with dairy allergies. Gram staining and the catalase test were used to identify the bacteria that were recovered from lettuce leaves as gram-positive, catalase-negative strains. The resistance to antibiotics and their capacity to tolerate low pH and bile salt concentrations both essential for surviving the digestive tract were evaluated. The bacteria showed a high tolerance to bile salts and low pH, but they were susceptible to ampicillin, streptomycin, gentamycin, chloramphenicol, and neomycin. Tests for gas production in the presence of glucose revealed no gas production. Molecular techniques such as PCR and 16S rRNA gene sequencing revealed the presence of Enterococcus lactis, Enterococcus durans, Lactobacillus paracasei, and Lactobacillus casei

Diversity of the Chicken Growth Hormone Gene and Effects on Growth Desi and Fayoumi Chicken Traits at District Kashmore, Sindh-Pakistan

Chicken growth hormone (cGH) gene, one of the candidate genes for economic traits that control body weight and fat deposition, is associated with regulating both growth-axon patterning and metabolism. The growth hormone gene located on the chromosome 2 it enhances and encodes growth hormones as well as protein which are responsible for regulating of growth and development of body tissues. Growth hormone gene as economically very important because of growth rate, efficiency of feeds, body weight, and also for egg production. It also used for the Genetic Engineering for production of desirable GH- related genes. Genomic DNA from four different chicken breeds were screened for single nucleotide polymorphisms (SNPs) of cGH gene by denaturing high-performance liquid chromatography, as well as sequencing. SNPs and Nucleotide Diversity of the cGH Gene PCR amplification of 700 bp region from each individual from two chicken breeds was performed, which covered partial exons/introns encoding fragments corresponding to the investigated translated peptide. An average of one SNP was found every 86 bp, with a total of eight SNPs discovered. Nine SNPs were in introns, while 4 and one each was in the 5'UTR and 3'UTR respectively of these, five of them (01 from GWAS linkage signals) had significant associations.

Relationships between fecal indicator abundance in water and sand and the presence of pathogenic genes in sand of recreational beaches.

For decades, the risk of exposure to infectious diseases in recreational beaches has been evaluated through the quantification of fecal indicator bacteria in water samples using culture methods. The analyses of sand samples have recently been developed as a complement to the monitoring of recreational waters in beach quality assessments. The growing use of molecular techniques for environmental monitoring allows for the rapid detection of pathogenic genes, thus providing more accurate information regarding the health risk of exposure to contaminated sand. The aim of this work was to determine the relationship between the fecal indicators abundance in water and sand and the presence of Shiga toxin-producer Escherichia coli (STEC) in sand by analyzing samples from touristic beaches using culture-dependent (fecal coliforms assay) and culture-independent (real-time PCR of stx1, stx2, and eae genes) techniques. We found a high concentration of coliform bacteria in water and sand in several beaches in eastern Uruguay, with different levels of sanitation networks and levels of urbanization. The presence of STEC virulence genes (mainly stx1) was confirmed in 8 out of 20 sand samples. The recreational use of sandy beaches may imply a risk to the health of its users, especially near streams and creek outflows, thus highlighting the need of monitoring sand bacteriological quality and pathogens using molecular tools.

Predictors of COVID-19 severity among a cohort of Egyptian patients

BackgroundAs the outbreak of COVID-19 progresses, prognostic markers for the identification of high-risk individuals are urgently needed. The angiotensin system is implicated in the pathogenesis of COVID-19 as ACE2 is the cellular receptor for SARS-COV-2 virus, and expression of the ACE2 gene could regulate an individual’s susceptibility to infection. In addition, the balance between ACE and ACE2 activity may play a role in the severity of COVID-19.Aim of workThe aim of the work is to explore the role of ACE1 I/D and ACE2 G8790A gene variants and serum ACE l/ACE2 ratio as risk factors for severity of COVID-19 infection.MethodsOne hundred and eighty COVID-19 patients were divided into three groups: mild (60 patients), moderate (60 patients), and severe (60 patients). The enzyme levels of ACE and ACE2 were measured by ELISA. ACE I/D (rs4646994) was assayed using PCR and ACE2 (rs2285666) gene variant was determined using real-time PCR.ResultsACE/ACE2 ratio was significantly lower in the mild group than in the moderate-to-severe group (P < 0.001). GG (reference) genotype and G allele of ACE2 were more frequent in mild group, AA (variant) genotype, and A allele were more frequent in severe group (P value < 0.001). In the multiple logistic regression, COVID-19 severity was associated with older age (> 50y) (OR 10.4, 95% CI 3.8–28.4, P < 0.001), comorbidities (OR 8.2, 95% CI 1.6–42.1, P 0.012), and higher ACE/ACE2 ratio (OR 8.3, 95% CI 3.7–18.6 P < 0.001) were independent significant predictors of severity. Haplotype analysis revealed that patients with D allele of the ACE gene combined with the A allele of the ACE2 gene had nearly double the risk of having severe COVID infection (OR = 1.9, 95% CI 1.1–3.5, P = 0.024).ConclusionOld age (> 50 years), presence of comorbidities, and a high ACE/ACE2 ratio are recognized as pivotal predictors of COVID-19 severity.

Intra-abdominal nocardiosis and scedosporiosis in a dog: case report and literature review.

A 2-y-old, intact female, mixed-breed dog was presented to the veterinary hospital with abdominal distension, anemia, and lethargy following a chronic history of nonspecific gastrointestinal signs. CBC and serum biochemistry revealed moderate nonregenerative anemia with neutrophilia, hypoalbuminemia, hyperglobulinemia, hypoglycemia, decreased urea and creatinine, and hypercholesterolemia. Abdominal radiographs and ultrasound revealed a large heterogeneous mesenteric mass and ascites. Abdominocentesis confirmed septic peritonitis with filamentous bacteria. Fine-needle aspiration of the mass yielded pyogranulomatous inflammation and hyphae. An exploratory laparotomy revealed a large cranial abdominal mass with granulomas present throughout the abdominal cavity. Due to the poor prognosis and disseminated disease, the owner elected euthanasia. Postmortem and histologic examinations detected intralesional mycetomas and bacterial colonies within the mesenteric masses. 16S ribosomal RNA gene PCR and sequencing using formalin-fixed, paraffin-embedded sections identified Nocardia yamanashiensis, Nocardioides cavernae, and Nocardioides zeicaulis. Fungal culture, PCR, and sequencing confirmed Scedosporium apiospermum. Our report highlights the importance of molecular methods in conjunction with culture and histologic findings for diagnosing coinfections caused by infrequent etiologic agents. Additionally, we provide a comprehensive literature review of Scedosporium apiospermum infections in dogs.

Genetic diversity and evolution of porcine hemagglutinating encephalomyelitis virus in Guangxi province of China during 2021-2024.

Porcine hemagglutinating encephalomyelitis virus (PHEV) is the only known porcine neurotropic coronavirus, which is prevalent worldwide at present. It is of great significance to understand the genetic and evolutionary characteristics of PHEV in order to perform effective measures for prevention and control of this disease. In this study, a total of 6,986 tissue samples and nasopharyngeal swabs were collected from different regions of Guangxi province in southern China during 2021-2024, and were tested for PHEV using a quadruplex RT-qPCR. The positivity rate of PHEV was 2.81% (196/6,986), of which tissue samples and nasopharyngeal swabs had 2.05% (87/4,246) and 3.98% (109/2,740) positivity rates, respectively. Fifty PHEV positive samples were selected for PCR amplification and gene sequencing. Sequence analysis revealed that the nucleotide homology and amino acid similarities of S, M, and N genes were 94.3%-99.3% and 92.3%-99.2%, 95.0%-99.7% and 94.7%-100.0%, 94.0%-99.5% and 93.5%-99.3%, respectively, indicating M and N genes were more conservative than S gene. Phylogenetic trees based on these three genes revealed that PHEV strains from different countries could be divided into two groups G1 and G2, and the PHEV strains from Guangxi province obtained in this study distributed in subgroups G1c and G2b. Bayesian analysis revealed that the population size of PHEV has been in a relatively stable state since its discovery until it expanded sharply around 2015, and still on the slow rise thereafter. S gene sequences analysis indicated that PHEV strains existed variation of mutation, and recombination. The results indicated that the prevalent PHEV strains in Guangxi province had complex evolutionary trajectories and high genetic diversity. To the best of our knowledge, this is the first report on the genetic and evolutionary characteristics of PHEV in southern China.

Phylogenetic tree and Molecular detection of Theileria velifera in sheep southern Iraq

Background: The purpose of this research was to ascertain T. velifera's evolutionary state in the Iraqi province of Al-Diwaniyah. In this investigation, the clinical assessment of fifty infected Blood samples (2.5 ml per animal) were collected from sheep, and a PCR technique was used to target the samples. Aim: This suggests that there may be intimate ties between certain global strains and our local ones. The vacuum tubes were coated with EDTA. Materials And Methods: The PCR and partial gene sequencing were conducted with the 18S rRNA gene serving as the molecular target. All of the examined samples had the targeted segments amplified at 400 bp according to the PCR, which used the 18S rRNA gene as genetic targets. Local T. velifera isolates' 18S rRNA gene sequences were compared to global reference strains of the species that were listed in the GenBank. Results: The local strains matched Chinese isolates 100% of the time and Chinese isolates Theileria luwenshuni 99% of the time. High fever (40.3–41.5°C), decreased appetite or appetence, asymmetrical enlargement of superficial lymph nodes, especially the pre-scapular ones, diarrhea in some cases, pale or icteric eye mucus membrane, lacrimation, sclera bleeding, abnormal movement, depression, dullness, and salivation were among the clinical signs displayed by diseased sheep. Conclusion: The information gathered from the current inspection effort may show how the local strains differ genetically from other strains listed in the GeneBank.

Alternating between even and odd ploidy levels switches on and off the recombination control, even near the centromeres.

Meiotic recombination is a key biological process in plant evolution and breeding, as it generates genetic diversity in each generation through the formation of crossovers (COs). However, due to their importance in genome stability, COs are highly regulated in frequency and distribution. We previously demonstrated that this strict regulation of COs can be modified, both in terms of CO frequency and distribution, in allotriploid Brassica hybrids (2n = 3x = 29; AAC) resulting from a cross between Brassica napus (2n = 4x = 38; AACC) and Brassica rapa (2n = 2x = 20; AA). Using the recently updated B. napus genome now including pericentromeres, we demonstrated that COs occur in these cold regions in allotriploids, as close as 375 kb from the centromere. Reverse transcription quantitative PCR (RT-qPCR) of various meiotic genes indicated that Class I COs are likely involved in the increased recombination frequency observed in allotriploids. We also demonstrated that this modified recombination landscape can be maintained via successive generations of allotriploidy (odd ploidy level). This deregulated meiotic behavior reverts to strict regulation in allotetraploid (even ploidy level) progeny in the second generation. Overall, we provide an easy way to manipulate tight recombination control in a polyploid crop.

Clostridioides difficile infections: Update and therapeutic guidelines

Clostridioides difficile infection (CDI) represents a significant challenge due to its increasing incidence, severity, and treatment difficulty. Effective management requires a multifactorial approach that includes preventive strategies, prudent antibiotic use, and adapted therapeutic options. Ongoing research and innovation offer promising prospects for improving ICD management, making vigilance and informed practices essential among healthcare professionals. Two main complications of ICD are pseudomembranous colitis (PMC) and toxic megacolon. PMC involves severe colonic inflammation due to C.difficile toxins, leading to pseudomembrane formation. Diagnosis relies on clinical criteria, microbiological tests, and endoscopy. Toxic Megacolon is characterized by severe colonic dilation and systemic toxicity, requiring immediate medical intervention. ICD diagnosis combines clinical signs and microbiological tests. These tests include toxin tests, GDH antigen detection, PCR for toxin genes, and stool culture. Imaging techniques assess colonic inflammation and complications. Combined diagnostic criteria from the American Gastroenterological Association (AGA) and European guidelines emphasize integrating clinical and laboratory findings for accurate diagnosis. ICD treatment involves stopping the implicated antibiotics and starting specific antimicrobial therapy. Common treatments include mainly fidaxomicin and oral vancomycin. Fecal microbiota transplantation (TMF) is recommended for recurrent cases unresponsive to standard treatments. Bezlotoxumab, an antibody targeting C.difficile toxin B, is used to prevent recurrence in high-risk adults. ICD poses a major challenge due to its increasing incidence, severity, and difficulty in treatment. A multifactorial approach involving rigorous preventive strategies, prudent antibiotic management, and adapted therapeutic options is essential for controlling the infection. Ongoing research and innovations in treatment offer promising prospects for improving patient management. Healthcare professionals must remain vigilant and informed to ensure effective practices in combating this infection and utilizing available resources optimally.

DNA barcoding, pollen and seed characteristics for documentation of four monocot Mediterranean endemic plant species in Egypt

Endemic taxa are critical constituents in the flora of most parts of the world. Recently, most of these taxa have become exposed to extinction. This study aims to increase understanding of four monocot Mediterranean threatened endemic taxa in Egypt (Allium mareoticum Bornm. & Gauba, Bromus aegyptiacus Tausch, Muscari salah-eidii (Täckh. & Boulos) Hosni, Pancratium arabicum Sickenb.) by description of their pollen and seed and authentication of DNA barcoding using rbcL and matK genes. Twenty field visits were conducted from March 2017 till April 2023 including different localities through Mediterranean area for covering the presence of these species. Characterization of pollen grains and seeds were examined using a light (LM) and scanning electron microscopes (SEM). DNA barcoding was carried out for documentation of the four species. Maturase K (matK) and ribulose-bisphosphate carboxylase (rbcL) were used as primers for PCR amplification and gene sequencing. The two genes were successful in identifying Allium mareoticum, Bromus aegyptiacus, and Muscari salah-eidii at the genus level and Pancratium arabicum at both species and genus levels for rbcL marker, and the genus level for matK marker. This study is considered a leading study for new barcodes of (A) mareoticum, (B) aegyptiacus, and M. salah-eidii which were initially made available to the NCBI.

Molecular survey of pfmdr-1, pfcrt, and pfk13 gene mutations among patients returning from Plasmodium falciparum endemic areas to Turkey

BackgroundIn recent years, there has been an increasing trend in the number of imported Plasmodium falciparum cases in Turkey. To improve treatment success and to better understand malaria epidemiology among imported cases, it is necessary to determine anti-malarial drug resistance. This study aimed to survey polymorphisms of resistance genes in imported P. falciparum patients using archived thin smear preparations and EDTA blood samples.MethodsA total of 100 imported P. falciparum patients admitted to Bakırköy Dr. Sadi Konuk Research and Training Hospital between 2017 and 2022 were included in this study. DNA extraction was performed using an archived slide and EDTA blood samples that were microscopically diagnosed. After confirming the samples by real-time PCR, the pfmdr1, pfcrt, and pfk13 genes were amplified and sequenced. Single nucleotide polymorphisms (SNPs) were screened using Geneious R9 software, with the reference P. falciparum clone 3D7 isolate.ResultsAll studied samples were confirmed to be P. falciparum using real-time PCR. Nested PCR was conducted and the pfcrt (92 samples), pfmdr1 (91 samples), and pfk13 (93 samples) genes were successfully amplified. Sequence analysis revealed the highest mutation rate in the pfmdr1 (74.5%) gene, with the identification of five haplotypes: NYSND (wild-type, 23%), NFSND (56%), NYSDD (2.2%), NFSDD (15.4%), and YFSND (3.4%)]. The pfcrt mutation was identified in 11 samples (12.2%), whereas the pfk13 mutation was found in only two samples.ConclusionThis study is the first molecular survey of anti-malarial drug resistance genes in Turkey. With the increasing number of imported Plasmodium malaria cases and recent reports of sporadic indigenous P. falciparum cases, malaria is becoming a growing concern in Turkey. Although molecular screening for resistance markers in P. falciparum malaria is not routinely conducted, the data from this study will enhance treatment success rates and contribute to global malaria elimination.

Bartonella spp. in bats from the Brazilian Amazon Forest.

Despite the great diversity of bats (64 species) in the State of Acre, northwestern Brazil, there are no studies on occurrence and diversity of Bartonella spp. in bats in this region. The present study investigated the occurrence and molecular identity of Bartonella spp. in spleen samples (n = 271) from bats of 30 different species from this region, within the Amazon biome. Twenty-one out of 208 (10.1%) samples positive in the PCR for the mammalian gapdh endogenous genes were positive in the qPCR for Bartonella spp. based on the nuoG gene. The two gltA Bartonella genotypes detected grouped with those previously identified in bats from other locations, expanding the diversity of genotypes associated with bats. This study provided the first molecular evidence of the presence of Bartonella spp. in bats in the state of Acre and in bats of the species Lophostoma silvicolum, Vampyressa thyone, Tonatia saurophila and Phyllostomus elongatus.

IGF 1, BDNF, and NGF mediate the neuro-modulatory role of stem cells in acrylamide-induced hippocampal toxic changes in rats.

Acrylamide (ACR), a common industrial chemical, is a strong neurotoxic material. The hippocampus is a brain area of interest mostly affected by Alzheimer's disease. Mesenchymal stem cells (MSCs) usefulness in various neurological diseases including Alzheimer's is being debated. In this work, the authors aim to explore the role of MSCs in ACR-induced hippocampal neurodegeneration and elucidate the mediating mechanism. For this purpose, ten rats served as control, another ten were injected ACR (i.p. 50 mg/kg/day for 2 weeks), and the last ten rats were injected ACR in addition to MSCs (i.p. 1 × 10⁷ MSCs single injection). ACR induced neurodegenerative histopathological hippocampal changes and adversely altered hippocampal oxidative stress markers SOD, MDA, and GSH. ACR had induced hippocampal demyelination as detected by silver staining. ACR significantly (P < 0.05) up-regulated the ELISA hippocampal TNF-alpha and IL-6 and produced microglial & astrocyte activation (as tracked by Iba1 & GFAP immunohistochemistry respectively). ACR significantly reduced hippocampal PCR gene expression of IGF 1 (insulin growth factor-1), BDNF (brain-derived neurotrophic factor), and NGF (nerve growth factor). MSCs administration had mitigated all the previous deleterious changes. Acrylamide caused detrimental effects on the hippocampus and demonstrably altered the hippocampal architecture. Bone marrow mesenchymal stem cells offered a promising therapeutic role against these neurotoxic effects of acrylamide, presumably through modulation of IGF 1, BDNF, and NGF gene expressions.