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Background and Objectives: Cervical cancer global burden is highly skewed towards poor countries primarily due to lack of awareness, poor screening, and low uptake of prophylactic vaccines. The purpose of our study is to educate and raise awareness among young girls and women about the importance of cervical screening and HPV vaccination. Materials and Methods: The present study, conducted from January 2023 to December 2023, focused on students, teachers, housewives, and healthcare professionals in the Jammu region to assess their awareness of cervical cancer and the HPV vaccine. HPV DNA testing was carried out using the Truenat Real-Time PCR method at Swastik Diagnostic Laboratory, Jammu. Results: Knowledge of cervical cancer, awareness of the HPV virus, and the vaccination status of women were assessed in survey. In the HPV screening test, out of 2,400 women, 106 tested positive for HPV. Among these 106 women, 19% had a high viral load (Ct < 20), 11% had a low viral load (25 = Ct < 30), indicating a low relative concentration of HPV viruses, 40% had a medium viral load (20 = Ct < 25), and 30% had very low viral loads (Ct = 30). Conclusion: These findings highlight the importance of routine cervical screenings, such as Pap smears and HPV tests, for the early detection of cervical cancer. There is an urgent need to implement cervical cancer screening and vaccination programs in the Jammu region. [ABSTRACT FROM AUTHOR]

Sexual abuse and rape are among the crimes that are increasing day by day in our country. According to 2019 TUIK data, 13.1% of children brought to security units are victimized by sexual crimes. According to data published by the World Population Review website in 2020, 1071 people were raped in Türkiye that year. The provability of crimes is a very important factor for the perpetrator to be punished. The field of forensic sciences tries to serve justice by examining the evidence found at the crime scene. In cases where the possible body fluids of the perpetrator and the victim are mixed, it is very difficult to make an identification because there is no method for the separation of these mixed samples. In this study, we aimed to differentiate mixed body fluids (blood and saliva) by microRNA method. The samples collected from volunteers for this study were prepared as two-donor mixtures in four different groups in five different dilutions. A total of 36 mixtures were prepared from fluids collected from 4 female and 4 male individuals. The miR-451A biomarker was used to indicate the presence of blood and the miR-203A-3p marker was used to indicate the presence of saliva in the mixture samples. The samples were first centrifuged and then RNA isolation was performed using miRNeasy Mini Kit. The concentration and purity of the obtained RNAs were determined by spectrophotometer. Then cDNA (complementary DNA) was synthesized from RNAs with Taqman microRNA Reverse Transcription Kit.U6 gene region was preferred as the reference gene and analyzes were performed by RT-PCR. The presence of blood samples in the mixture was shown in all dilutions, the best level was seen in 1x1 ratio. When all groups were analyzed, it was seen that the mixtures with male donors were expressed at a higher rate than the other mixtures. [ABSTRACT FROM AUTHOR]

Abstract Given the limitation of current routine approaches for pancreatic cancer screening and detection, the mortality rate of pancreatic cancer cases is still critical. The development of blood-based molecular biomarkers for pancreatic cancer screening and early detection which provide less-invasive, high-sensitivity, and cost-effective, is urgently needed. The goal of this study is to identify and validate the potential molecular biomarkers in white blood cells (WBCs) of pancreatic cancer patients. Gene expression profiles of pancreatic cancer patients from NCBI GEO database were analyzed by CU-DREAM. Then, mRNA expression levels of three candidate genes were determined by quantitative RT-PCR in WBCs of pancreatic cancer patients (N = 27) and healthy controls (N = 51). ROC analysis was performed to assess the performance of each candidate gene. A total of 29 upregulated genes were identified and three selected genes were performed gene expression analysis. Our results revealed high mRNA expression levels in WBCs of pancreatic cancer patients in all selected genes, including FKBP1A (p

Background and Objectives: Cervical cancer global burden is highly skewed towards poor countries primarily due to lack of awareness, poor screening, and low uptake of prophylactic vaccines. The purpose of our study is to educate and raise awareness among young girls and women about the importance of cervical screening and HPV vaccination. Materials and Methods: The present study, conducted from January 2023 to December 2023, focused on students, teachers, housewives, and healthcare professionals in the Jammu region to assess their awareness of cervical cancer and the HPV vaccine. HPV DNA testing was carried out using the Truenat Real-Time PCR method at Swastik Diagnostic Laboratory, Jammu. Results: Knowledge of cervical cancer, awareness of the HPV virus, and the vaccination status of women were assessed in survey. In the HPV screening test, out of 2,400 women, 106 tested positive for HPV. Among these 106 women, 19% had a high viral load (Ct < 20), 11% had a low viral load (25 ≤ Ct < 30), indicating a low relative concentration of HPV viruses, 40% had a medium viral load (20 ≤ Ct < 25), and 30% had very low viral loads (Ct ≥ 30). Conclusion: These findings highlight the importance of routine cervical screenings, such as Pap smears and HPV tests, for the early detection of cervical cancer. There is an urgent need to implement cervical cancer screening and vaccination programs in the Jammu region.

Abstract Vancomycin resistant enterococci (VRE) are a leading cause of ICU-acquired bloodstream infections in Europe. The bacterial load in enteral colonization may be associated with a higher probability of transmission. Here, we aimed to establish a quantitative vanA/vanB DNA real-time PCR assay on a high-throughput system. Limits of detection (LOD), linear range and precision were determined using serial bacterial dilutions. LOD was 46.9 digital copies (dcp)/ml for vanA and 60.8 dcp/ml for vanB. The assay showed excellent linearity between 4.7 × 101 and 3.5 × 105 dcp/ml (vanA) and 6.7 × 102 and 6.7 × 105 dcp/ml (vanB). Sensitivity was 100% for vanA and vanB, with high positive predictive value (PPV) for vanA (100%), but lower PPV for vanB (34.6%) likely due to the presence of vanB DNA positive anerobic bacteria in rectal swabs. Using the assay on enriched VRE broth vanB PPV increased to 87.2%. Quantification revealed median 2.0 × 104 dcp/ml in PCR positive but VRE culture negative samples and median 9.1 × 104 dcp/ml in VRE culture positive patients (maximum: 107 dcp/ml). The automated vanA/B_UTC assay can be used for vanA/vanB detection and quantification in different diagnostic settings and may support future clinical studies assessing the impact of bacterial load on risk of infection and transmission.

Vancomycin resistant enterococci (VRE) are a leading cause of ICU-acquired bloodstream infections in Europe. The bacterial load in enteral colonization may be associated with a higher probability of transmission. Here, we aimed to establish a quantitative vanA/vanB DNA real-time PCR assay on a high-throughput system. Limits of detection (LOD), linear range and precision were determined using serial bacterial dilutions. LOD was 46.9 digital copies (dcp)/ml for vanA and 60.8 dcp/ml for vanB. The assay showed excellent linearity between 4.7 × 101 and 3.5 × 105 dcp/ml (vanA) and 6.7 × 102 and 6.7 × 105 dcp/ml (vanB). Sensitivity was 100% for vanA and vanB, with high positive predictive value (PPV) for vanA (100%), but lower PPV for vanB (34.6%) likely due to the presence of vanB DNA positive anerobic bacteria in rectal swabs. Using the assay on enriched VRE broth vanB PPV increased to 87.2%. Quantification revealed median 2.0 × 104 dcp/ml in PCR positive but VRE culture negative samples and median 9.1 × 104 dcp/ml in VRE culture positive patients (maximum: 107 dcp/ml). The automated vanA/B_UTC assay can be used for vanA/vanB detection and quantification in different diagnostic settings and may support future clinical studies assessing the impact of bacterial load on risk of infection and transmission. [ABSTRACT FROM AUTHOR]

Background: Vinegar-quenched Testudinis Carapax et Plastrum (TCP) and vinegar-quenched Trionycis Carapax (TC) are made from the shells of Chinemys reevesii and Trionyx sinensis. However, some cheap substitutes from non-medicinal species such as Trachemys scripta, Mauremys sinensis, Chelydra serpentina , and Apalone ferox are often seen on the market, which exposes public health to high risk. Conventional methods are not applicable for the identification of these two products since their intrinsic constituents are largely destroyed after being highly processed. Objectives: A new quantitative polymerase chain reaction (qPCR) method was developed to detect DNA components of Vinegar-quenched TCP and vinegar-quenched TC based on simple sequence repeat differential genes of animal-derived mitochondria. Materials and Methods: Six species-specific primer reactions were designed and further validated for specificity, sensitivity, and repeatability. Finally, the developed method was used to assess the authenticity of their commercial products. Results: The assay exhibited good specificity, and the limit of detection was above 1 × 102 copies/µL. There was good linearity in the concentration, ranging from 1 × 102 to 1 × 107 copies/µL. In addition, the method was validated through repeatability assessment (coefficient variation <1%). While qPCR was applied for the analysis of 36 batches of commercial products of these two tonics, it could effectively identify the authenticity of the products, and consequently, seven batches were fake or adulterated. Conclusion: This newly proposed method is promising for the quality evaluation of highly processed animal-derived Chinese medicines, which will assist in ensuring safety and efficacy in clinical practice and protect fair trade in the market. [ABSTRACT FROM AUTHOR]

Introduction: Tuberculosis (TB) is a major global health problem caused by Mycobacterium tuberculosis (MTB). The National Tuberculosis Elimination Programme (NTEP) emphasises early diagnosis and treatment of TB cases. The Cartridge-Based Nucleic Acid Amplification Test (CBNAAT) is a semi-automated molecular assay that rapidly detects MTB Deoxyribonucleic Acid (DNA) and Rifampicin (RIF) resistance-associated mutations using Real-Time Polymerase Chain Reaction (RT-PCR). Aim: This cross-sectional study aims to assess the association between the Cycle Threshold (Ct) value of CBNAAT and the clinical severity of Pulmonary Tuberculosis (PTB). Materials and Methods: The study was conducted at the Department of Respiratory Medicine, Navodaya Medical College Hospital and Research Centre-Raichur, from January 2020 to July 2021. A sample size of 106 participants, aged ≥18 years and meeting the NTEP guidelines for presumptive PTB cases, were enrolled. Participants underwent chest radiography, sputum smear microscopy for Acid-Fast Bacilli (AFB), and CBNAAT testing. Clinical severity of PTB was categorised as mild, moderate, or severe using the Bandim TB score. Data was entered into Microsoft Excel 2017, and statistical analysis was performed using SPSS version 23. Descriptive statistics, frequencies, percentages, means, and standard deviations were calculated, and the chi-square test was utilised to assess associations between qualitative variables. Results: Among the participants, the majority (55.7%, n=59) had moderate clinical severity. Of these, 12.3% (n=13) had high CBNAAT Ct values, 20.8% (n=22) had medium Ct values, 11.3% (n=12) had low Ct values, and 11.3% (n=12) had very low Ct values. A total of 31.1% (n=33) had severe clinical severity, with 13.2% (n=6) having high Ct values, 12.3% (n=13) having medium Ct values, and 6.6% (n=7) each having low and very low Ct values. Additionally, 13.2% (n=14) had mild clinical severity, with 0.9% (n=1) having high Ct values, 3.8% (n=4) having medium Ct values, 5.7% (n=6) having low Ct values, and 2.8% (n=3) having very low Ct values. The chi-square value was 4.697, with a p-value of 0.58. Conclusion: There is no association between the Ct value of CBNAAT and the clinical severity of PTB.

Introduction: Tuberculosis (TB) is a major global health problem caused by Mycobacterium tuberculosis (MTB). The National Tuberculosis Elimination Programme (NTEP) emphasises early diagnosis and treatment of TB cases. The Cartridge-Based Nucleic Acid Amplification Test (CBNAAT) is a semi-automated molecular assay that rapidly detects MTB Deoxyribonucleic Acid (DNA) and Rifampicin (RIF) resistance-associated mutations using Real-Time Polymerase Chain Reaction (RT-PCR). Aim: This cross-sectional study aims to assess the association between the Cycle Threshold (Ct) value of CBNAAT and the clinical severity of Pulmonary Tuberculosis (PTB). Materials and Methods: The study was conducted at the Department of Respiratory Medicine, Navodaya Medical College Hospital and Research Centre-Raichur, from January 2020 to July 2021. A sample size of 106 participants, aged ≥18 years and meeting the NTEP guidelines for presumptive PTB cases, were enrolled. Participants underwent chest radiography, sputum smear microscopy for Acid-Fast Bacilli (AFB), and CBNAAT testing. Clinical severity of PTB was categorised as mild, moderate, or severe using the Bandim TB score. Data was entered into Microsoft Excel 2017, and statistical analysis was performed using SPSS version 23. Descriptive statistics, frequencies, percentages, means, and standard deviations were calculated, and the chi-square test was utilised to assess associations between qualitative variables. Results: Among the participants, the majority (55.7%, n=59) had moderate clinical severity. Of these, 12.3% (n=13) had high CBNAAT Ct values, 20.8% (n=22) had medium Ct values, 11.3% (n=12) had low Ct values, and 11.3% (n=12) had very low Ct values. A total of 31.1% (n=33) had severe clinical severity, with 13.2% (n=6) having high Ct values, 12.3% (n=13) having medium Ct values, and 6.6% (n=7) each having low and very low Ct values. Additionally, 13.2% (n=14) had mild clinical severity, with 0.9% (n=1) having high Ct values, 3.8% (n=4) having medium Ct values, 5.7% (n=6) having low Ct values, and 2.8% (n=3) having very low Ct values. The chi-square value was 4.697, with a p-value of 0.58. Conclusion: There is no association between the Ct value of CBNAAT and the clinical severity of PTB. [ABSTRACT FROM AUTHOR]

Urinary system stones have a complex pathophysiology affected by environmental and genetic factors. To confirm whether ALPL gene polymorphisms are an effective universal risk factor for the development of urolithiasis, we aimed to investigate ALPL gene polymorphism in Turkish population. Our study was carried out with 187 patients with urolithiasis and 92 healthy volunteers (control group) who were not diagnosed with urolithiasis either in themselves or in their families, applied to Emergency Medicine Clinic of Health Sciences University Haydarpasa Numune Training and Research Hospital in Istanbul, Turkey between November 2021 and February 2022, prospectively. In order to evaluate the relationship between ALP gene (rs1256328) polymorphism and urolithiasis, blood samples were analyzed by quantitative Real Time PCR (qPCR) method. Male gender (OR:3.785; 95% CI:2.118-6.763; p<0.001), increased BUN level (OR:1.082; 95% CI:1.013-1.156; p=0.019), increased NLR level (OR:1.149; 95 %CI:1.033-1.277; p=0.011) and heterozygous genotype (OR:2.353; 95% CI:1.31-4.225; p=0.004) were determined as independent risk factors for the development of urolithiasis. The presence of heterozygous (CT) genotype in the ALPL rs1256328 gene region in the Turkish population is associated with an increased risk of urolithiasis. This suggests that ALPL rs1256328 gene polymorphism can be used as a genetic marker in the Turkish population. [ABSTRACT FROM AUTHOR]

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Introduction: The Coronavirus Disease-2019 (COVID-19) pandemic has emerged from Wuhan, China in late 2019 and spread across the world in few months. This pandemic has affected people all over the world in terms of not only health but also psychologically, economically and socially. Early diagnosis by Real Time Polymerase Chain Reaction (RT-PCR) test helped the health system to isolate the patient and prevent the spread of the disease in community. Aim: To know the prevalence of asymptomatic COVID-19 infection among healthcare workers. Materials and Methods: This study was a cross-sectional study and focused on early detection of asymptomatic COVID19 infection among Healthcare Workers (HCW) by RT-PCR test. Total of 356 samples were collected of all HCW working in Nootan Medical College and Research Centre Visnagar, Gujarat, India, in month of May 2021. Data were analysed on basis of job profile, working area and vaccination status. Chi-square test was used for statistical analysis. Results: Out of total 356 samples, six were positive showing prevalence of 1.68%. Out of these six positive workers two had exposure to COVID-19 positive patients at home in last 10 days. No significance was found in positivity rate in relation to job profile, working area in hospital and vaccination status. Conclusion: This study reveals low prevalence of asymptomatic infection among HCWs engaged in COVID-19 related duties. The HCWs are at highest risk of contracting infection, hence it is necessary to protect their safety and health as infection among them can spread to other co-workers and patients.

The use of apitherapy and natural herbal medicines for combating toxoplasmosis has garnered major attention from many researchers. However, there is no available information regarding the potential use of a combination of propolis and wheat germ oil (WGO) in the treatment of toxoplasmosis. In the present study, the potential effects of propolis, WGO, and their combination in the treatment of chronic toxoplasmosis in Swiss albino mice were investigated. Following induction of chronic toxoplasmosis, the potential antiparasitic effects of these substances were evaluated by parasitological assessment and by counting of Toxoplasma cysts. Additionally, the effects of the treatments on parasite loads were analyzed using TaqMan real-time quantitative PCR targeting the Toxoplasma P29 gene followed by investigation of the major histopathological changes in the brain, uterus, and kidney. Interestingly, the combination of propolis and WGO significantly (P ≤ 0.05) decreased the parasite burden in experimental animals compared with burdens seen in groups treated with propolis or WGO alone. Furthermore, the quantification of the DNA concentrations of Toxoplasma P29 gene after the treatment with propolis and WGO revealed a reduction in parasite load in treated groups versus the control group (infected untreated animals). Importantly, the severity of histopathological lesions was significantly (P ≤ 0.05) improved following treatment with propolis and WGO. Collectively, the present study indicated a potential novel role for propolis and WGO as an active apitherapy and natural herbal medication for treating chronic toxoplasmosis, combat the disease, and which could also help overcome the side effects of chemical drugs.

Introduction: Members of Enterobacterales are very common pathogens, which continue to show resistance to many antibiotics. Carbapenem performed well for some time. Colistin was the final hope for the carbapenem-resistant Enterobacterales, but resistance against it has virtually tied the clinician's hands, especially when it comes to treating critically ill patients. Purpose: Detection of colistin resistance by the agar method as well as by the polymerase chain reaction (mobilized colistin resistance-1 gene) in carbapenem-resistant Enterobacterales. Materials and Methods: A cross-sectional study from Dec 2019 to Dec 2020 was conducted at the Department of Microbiology, Army Medical College, National University of Medical Sciences Rawalpindi Pakistan. Antimicrobial susceptibility of Enterobacterales was determined according to the Kirby–Bauer disc diffusion method except for colistin. Colistin agar was used, in concentrations of 2 μg/mL and 4 μg/mL. Results were interpreted according to Clinical and Laboratory Standards Institute guidelines 2020. Mobilized colistin-resistant-1 gene in the carbapenem resistant Enterobacterales was detected by performing real-time polymerase chain reaction assay. Results: Among the 172 carbapenem-resistant Enterobacterales 18 isolates were resistant using the colistin agar test. Whereas by molecular method colistin resistance was detected among 10 isolates that carried mobilized colistin resistance 1 gene, making the frequency of the MCR-1 gene 5.81%. Seventy percent of isolates were from paired blood samples. Eight patients, from whom the colistin resistant gene was isolated expired. Conclusion: Colistin resistance is a very serious issue and should not be missed in a clinical microbiology laboratory. The phenotypic agar test method is an excellent option for routine use, as it combines ease of performance with affordable cost. However, molecular methods are essential for the detection of mobilized colistin resistance gene (1– 9) for epidemiological purposes. [ABSTRACT FROM AUTHOR]

Maternal‐fetal medicine (FM) is currently a highly demanding branch and is gaining importance as increasing number of genetic disorders rise in incidence. Prenatal testing helps to detect such abnormalities that could affect the health status of the developing fetus like birth defects or genetic disorders. Considering the rising trend of genetic disorders, there is a need for a highly sensitive way of noninvasive prenatal testing (NIPT) that may reduce the incidence of unnecessary invasive procedures and iatrogenic fetal loss. The concept of NIPT for screening of genetic disorders is continuously evolving over the last two decades and multiple techniques have come up to utilize this in the field of FM. The crucial factor which decides the accuracy of NIPS is cell free fetal DNA (cffDNA) that is present in extremely low fraction (10%–15%) in the maternal plasma. Among the available methods, the next generation sequencing (NGS) is considered as the gold standard. However, the higher cost diminishes its utility in low‐resource settings. Droplet digital Polymerase chain reaction (ddPCR), a type of digital PCR is a novel technique that is frugal, equally sensitive, less labor intensive, less time‐consuming and plain algorithm dependent method for detecting cffDNA fraction. Considering these impressive attributes of ddPCR, we decided to critically review the existing literature on ddPCR for NIPT whilst highlighting the clinical utility, challenges and its advantages over NGS. [ABSTRACT FROM AUTHOR]

Introduction: Hepatitis C virus (HCV) has posed a major public health problem globally. Since majority of HCV infected patients are asymptomatic, diagnosis of HCV infection is mainly based on the detection of anti-HCV antibodies by the Enzyme Linked Immunosorbent Assay (ELISA) or Rapid Diagnostic Tests (RDTs) and HCV Ribonucleic Acid (RNA) by real time Polymerase Chain Reaction (PCR) of serum or plasma samples. Aim: To assess the performance of RDT and 4th generation ELISA against real time reverse transcriptase PCR for the detection of HCV. Materials and Methods: A hospital-based cross-sectional study was carried out in the Virology Section, Department of Microbiology, Jawaharlal Nehru Institute of Medical Sciences (JNIMS), Imphal, Manipur, India, for a period of two years from June 2019 to May 2021. The study included 3,254 plasma samples from suspected cases of HCV monoinfection, and HCV/HIV co-infection. The plasma samples were subjected to anti-HCV antibodies by RDT (SD BIOLINE, South Korea) and 4th generation ELISA (Monolisa™ HCV Ag-Ab ULTRA, Bio-Rad, France), and HCV RNA by real time PCR. Data analysis was done using descriptive statistics, and performance of the assays was evaluated by using Cohen kappa test (1) and Receiver Operating Characteristic (ROC) curve. Results: PCR is considered as the gold standard test. HCV was detected by RDT in 453 (13.92%), ELISA in 413 (12.69%) and RT-PCR in 367 (11.28%) samples. The present study demonstrated sensitivity of 97.55% and specificity of 96.71% with Positive Predictive Value (PPV) of 79.03% by HCV-RDT. The 4th generation ELISA showed high sensitivity of 99.46% and specificity of 98.34%. Using ROC curve, the area under the curve was 81% for ELISA with diagnostic accuracy of 98.46%. Conclusion: 4th generation ELISA is more sensitive and specific than RDT for the detection of HCV infection. Confirmatory HCV-RNA assay could be performed to clear doubts related to false-positive and false-negative findings of the primary screening assays. [ABSTRACT FROM AUTHOR]

All four dengue serotypes cause infection, with one of them predominantly reported from a particular geographical region. Coinfection by more than one serotype is reported from hyperendemic regions. These coinfections are clinically more severe than infection with a single serotype. This study was carried out to detect the predominant dengue serotype and presence of coinfections. Acute-phase serum samples of patients suffering from dengue infection were collected. They were screened for the presence of IgM, IgG and NS1Ag by a rapid test. Conventional multiplex reverse transcriptase polymerase chain reaction (RT-PCR) and multiplex real-time RT-PCR assays were carried out for detection and serotyping of the dengue virus. A total of 196 samples were positive by the rapid card test. Of these, 139 were NS1Ag positive, 40 were positive only for IgM, 5 were positive only for IgG and 12 samples were positive for different combinations of antigen and antibodies. All four serotypes were detected in these samples by PCR. DENV-3 was found to be most common circulating serotype. A total of 22 cases were found to have coinfection with more than one dengue serotypes. Samples having only antibodies and no antigen on rapid card test were also positive for virus by PCR. Prevalence of dengue co-infections is increasing. Moreover, it is important to screen for dengue virus in those samples also which do not show NS1Ag on rapid tests and have either one or both the antibodies. Real-time multiplex RT-PCR is found to be more sensitive in detecting coinfection than conventional multiplex RT-PCR. [ABSTRACT FROM AUTHOR]

Introduction: The coronavirus disease 2019 (COVID-19) pandemic has required changes in the management of pediatric cardiac surgery. We would like to share the patient treatment and surgical management strategies employed in our Pediatric Cardiovascular Surgery Clinic during the COVID-19 pandemic. Methods: A total of 112 patients were followed up in our clinic between 11.03.2020 and 02.07.2020. Their mean age was 1,118 (4–5,740) days. Management and treatment were performed by our pediatric heart team (pediatric cardiac anesthetists, general pediatricians, pediatric cardiologists, pediatric cardiac surgeons, and an infectious diseases specialist). We prepared new protocols and a surveillance system specific to the pandemic to prevent in-hospital transmission and reduce postoperative mortality and morbidity; our operations were performed according to these protocols. All decisions pertaining to the operation timing and treatment strategy of our COVID-19-positive patients were made by the same team Results: During the study period, a total of 112 patients, 69 boys and 43 girls, were hospitalized in our clinic. A total of 333 COVID-19 real-time polymerase chain reaction tests were performed on patients and accompanying persons; positive results were found in three patients and two accompanying individuals. Conclusion: By employing new protocols and a surveillance system throughout the healthcare system, we think that early diagnosis and treatment of the pediatric congenital heart disease population, which is susceptible to infections, can continue unperturbed. This and similar approaches can increase postoperative success and prevent transmission in the pediatric population – which are frequently COVID-19 asymptomatic [ABSTRACT FROM AUTHOR]

Introduction: Hepatitis C virus (HCV) has posed a major public health problem globally. Since majority of HCV infected patients are asymptomatic, diagnosis of HCV infection is mainly based on the detection of anti-HCV antibodies by the Enzyme Linked Immunosorbent Assay (ELISA) or Rapid Diagnostic Tests (RDTs) and HCV Ribonucleic Acid (RNA) by real time Polymerase Chain Reaction (PCR) of serum or plasma samples. Aim: To assess the performance of RDTs and fourth generation ELISA against real time reverse transcriptase PCR for the detection of HCV. Materials and Methods: A hospital-based cross-sectional study was carried out in the Virology Section, Department of Microbiology, Jawaharlal Nehru Institute of Medical Sciences (JNIMS), Imphal, Manipur, India, for a period of two years from June 2019 to May 2021. The study included 3,254 plasma samples from suspected cases of HCV monoinfection, and HCV/HIV co-infection. The plasma samples were subjected to anti-HCV antibodies by RDTs (SD BIOLINE, South Korea) and fourth generation ELISA (Monolisa™ HCV Ag-Ab ULTRA, Bio-Rad, France), and HCV RNA by real time PCR. Data analysis was done using descriptive statistics, and performance of the assays was evaluated by using Cohen kappa test (κ) and Receiver Operating Characteristic (ROC) curve. Results: PCR is considered as the gold standard test. HCV was detected by RDTs in 453 (13.92%), ELISA in 413 (12.69%) and RT-PCR in 367 (11.28%) samples. The present study demonstrated sensitivity of 97.55% and specificity of 96.71% with Positive Predictive Value (PPV) of 79.03% by HCV-RDT. The fourth generation ELISA showed high sensitivity of 99.46% and specificity of 98.34%. Using ROC curve, the area under the curve was 81% for ELISA with diagnostic accuracy of 98.46%. Conclusion: Fourth generation ELISA is more sensitive and specific than RDTs for the detection of HCV infection. Confirmatory HCV-RNA assay could be performed to clear doubts related to false-positive and false-negative findings of the primary screening assays.

ABSTRACT Introduction: The coronavirus disease 2019 (COVID-19) pandemic has required changes in the management of pediatric cardiac surgery. We would like to share the patient treatment and surgical management strategies employed in our Pediatric Cardiovascular Surgery Clinic during the COVID-19 pandemic. Methods: A total of 112 patients were followed up in our clinic between 11.03.2020 and 02.07.2020. Their mean age was 1,118 (4-5,740) days. Management and treatment were performed by our pediatric heart team (pediatric cardiac anesthetists, general pediatricians, pediatric cardiologists, pediatric cardiac surgeons, and an infectious diseases specialist). We prepared new protocols and a surveillance system specific to the pandemic to prevent in-hospital transmission and reduce postoperative mortality and morbidity; our operations were performed according to these protocols. All decisions pertaining to the operation timing and treatment strategy of our COVID-19-positive patients were made by the same team. Results: During the study period, a total of 112 patients, 69 boys and 43 girls, were hospitalized in our clinic. A total of 333 COVID-19 real-time polymerase chain reaction tests were performed on patients and accompanying persons; positive results were found in three patients and two accompanying individuals. Conclusion: By employing new protocols and a surveillance system throughout the healthcare system, we think that early diagnosis and treatment of the pediatric congenital heart disease population, which is susceptible to infections, can continue unperturbed. This and similar approaches can increase postoperative success and prevent transmission in the pediatric population - which are frequently COVID-19 asymptomatic.

Background and Objectives: Acinetobacter baumannii is recognized as an important pathogen responsible for serious infections causing episodes of hospital infection. Carbon nanotubes (CNTs) have recently emerged as superior materials against antibiotic-resistant bacteria. In this study, a new chemical compound was designed in order to combat A. baumannii infections. Subsequently, the effect of this novel carbon nanotube coated with an antibacterial compound on Extensively Drug-Resistant (XDR), Multidrug-Resistant (MDR) and Pan-Drug-Resistance (PDR) strains of A. baumannii was investigated. Materials and Methods: A total of 122 clinical isolates of A. baumannii were cultured from burn patients and their susceptibility to antibiotics were checked using disk diffusion method and Minimum inhibitory concentration. Antimicrobial effects of the coated carbon nanotube were evaluated on XDR, MDR and PDR isolates of A. baumannii. Cell viability was determined using tetrazolium reduction assay (MTT) on human fibroblast cell line (HDFa). Wound healing processes were assessed by quantitative polymerase chain reaction. Results: Of the 50 A. baumannii isolates, 38 (76%) were found to be MDR and 12 (24%) were XDR. No PDR strains were detected. Results indicated that the carbon nanotube combined with mercury had antibacterial effect against different A. baumannii species and it also was able to increase the expression of epidermal growth factor, platelet-derived growth factor and vascular endothelial growth factor A mRNA levels which are involved in wound healing. Conclusion: The engineered carbon nanotube compound can potentially be used for treatment of burn related infections. This can potentially give clinicians a new tool for treating A. baumannii infections.

The SARS-CoV-2 pandemic is the most devastating health crisis our generation has seen. The present study tries to gather more epidemiological data and improve the knowledge of the demographic factors responsible for a higher incidence of COVID-19. We analyzed the real time polymerase chain reaction (RT-PCR) results obtained in one of the biggest tertiary hospitals in Bulgaria during the first year of the COVID-19 pandemic (May 2020–April 2021). For this period, almost 40% of all tested samples of hospitalized patients and health care workers in University Hospital “St. Marina,” Varna, were SARS-CoV-2 positive. The most affected individuals were 60–79-year-old people. Male sex is a significant risk factor only for the active ages (20–59 years), while both men and women of advanced age have the same risk to be infected with SARS-CoV-2. Interestingly, girls under 19 years were more susceptible to the infection than boys in the same age group.

Background and Objectives: The entire globe is undergoing an unprecedented challenge of COVID-19. Considering the need of rapid and accurate diagnostic tests for SARS-CoV-2, this study was planned to evaluate the cost effective extraction free RT-PCR technique in comparison to the standard VTM based RT-qPCR method. Materials and Methods: Paired swabs from nasopharynx and oropharynx were collected for SARS-CoV-2 testing, from 211 adult patients (≥18 years) in VTM and plain sterile tubes (dry swabs). These samples were processed and RT-qPCR was carried out as per standard protocols. Results: 54.5% of the patients were females and 45.5% were males with sex ratio 1:1.19 (M: F). 38.86% were symptomatic, of which fever (86.59%), cough (79.23%) and breathlessness (46.34%) were the most common symptoms. The positivity by VTM based method and index method was 31.27% and 13.27% respectively. Of the 27 inconclusive results from index method, 37.04% were positive, 48.15% were negative by VTM based method. However, in 40 inconclusive results by VTM based method, 90% were negative and rest remained inconclusive by index method. The sensitivity and specificity of the index method were 39.39% and 85.71% respectively. The overall agreement between VTM based method and index method was 49.59% with estimated Kappa value of 0.19. Conclusion: VTM based method showed higher sensitivity compared to the index method. The higher positivity by VTM based method, suggests that VTM based method could plausibly be a better detection method of SARS-CoV-2. Still, the index method might add value in a resource limited setups for detection of SARS-CoV-2. [ABSTRACT FROM AUTHOR]

Background and Objectives: The entire globe is undergoing an unprecedented challenge of COVID-19. Considering the need of rapid and accurate diagnostic tests for SARS-CoV-2, this study was planned to evaluate the cost effective extraction free RT-PCR technique in comparison to the standard VTM based RT-qPCR method. Materials and Methods: Paired swabs from nasopharynx and oropharynx were collected for SARS-CoV-2 testing, from 211 adult patients (≥18 years) in VTM and plain sterile tubes (dry swabs). These samples were processed and RT-qPCR was carried out as per standard protocols. Results: 54.5% of the patients were females and 45.5% were males with sex ratio 1:1.19 (M: F). 38.86% were symptomatic, of which fever (86.59%), cough (79.23%) and breathlessness (46.34%) were the most common symptoms. The positivity by VTM based method and index method was 31.27% and 13.27% respectively. Of the 27 inconclusive results from index method, 37.04% were positive, 48.15% were negative by VTM based method. However, in 40 inconclusive results by VTM based method, 90% were negative and rest remained inconclusive by index method. The sensitivity and specificity of the index method were 39.39% and 85.71% respectively. The overall agreement between VTM based method and index method was 49.59% with estimated Kappa value of 0.19. Conclusion: VTM based method showed higher sensitivity compared to the index method. The higher positivity by VTM based method, suggests that VTM based method could plausibly be a better detection method of SARS-CoV-2. Still, the index method might add value in a resource limited setups for detection of SARS-CoV-2.

Background: Obesity and diabetes prevalence are increasing worldwide. We aimed to detect the possible association of osteoprotegerin (OPG) gene expression with visceral adiposity indices and cardiometabolic risk factors among obese women. Methods and results: The study enrolled 150 controls and 150 obese cases subdivided into two subgroups non-diabetic (n = 70) and 80 patients with type 2 diabetes mellitus (T2DM). Circulating OPG gene expression levels were figured out by real time PCR (Polymerase Chain Reaction). Serum OPG levels were assessed by Enzyme Linked Immunosorbent Assay. Our results explored that OPG serum levels were lower in the obese women compared to control group (p < 0.001) and obese diabetics had higher serum levels of OPG in comparison to obese non-diabetic patients (p < 0.001). Expression levels of OPG were higher in obese women than controls (p < 0.001). Moreover, the blood expression levels of OPG gene were higher in diabetic obese patients than non-diabetics. We found positive correlations between parameters of metabolic syndrome and obesity indices. After adjustment of the traditional risk factors, stepwise linear regression analysis test revealed that OPG expression levels were independently correlated with glycated hemoglobin, high-density lipoprotein-cholesterol, and waist-to-hip ratio. Conclusions: OPG mRNA levels were associated with surrogate markers of insulin resistance in Egyptian obese women. [ABSTRACT FROM AUTHOR]

A nanoparticle-assembled photonic crystal (PC) array was used to detect single nucleotide polymorphism (SNP). The assay platform with PC nanostructure enhanced the fluorescent signal from nanoparticle-hybridized DNA complexes due to phase matching of excitation and emission. Nanoparticles coupled with probe DNA were trapped into nanowells in an array by using an electrophoretic particle entrapment system. The PC/DNA assay platform was able to identify a 1 base pair (bp) difference in synthesized nucleotide sequences that mimicked the mutation seen in a feline model of human autosomal dominant polycystic kidney disease (PKD) with a sensitivity of 0.9 fg/mL (50 aM)-sensitivity, which corresponds to 30 oligos/array. The reliability of the PC/DNA assay platform to detect SNP in a real sample was demonstrated by using genomic DNA (gDNA) extracted from the urine and blood of two PKD- wild type and three PKD positive cats. The standard curves for PKD positive (PKD+) and negative (PKD-) DNA were created using two feline-urine samples. An additional three urine samples were analyzed in a similar fashion and showed satisfactory agreement with the standard curve, confirming the presence of the mutation in affected urine. The limit of detection (LOD) was 0.005 ng/mL which corresponds to 6 fg per array for gDNA in urine and blood. The PC system demonstrated the ability to detect a number of genome equivalents for the PKD SNP that was very similar to the results reported with real time polymerase chain reaction (PCR). The favorable comparison with quantitative PCR suggests that the PC technology may find application well beyond the detection of the PKD SNP, into areas where a simple, cheap and portable nucleic acid analysis is desirable.

The incidence of melanoma is one of the fastest growing of all tumor types in the United States and the number of cases worldwide has doubled in the past 30 years. Melanoma, which arises from melanocytes, is an extremely aggressive tumor that invades the vascular and lymphatic systems to establish tumors elsewhere in the body. Melanoma is a particularly resilient cancer and systemic therapy approaches have achieved minimal success against metastatic melanoma resulting in only a few FDA-approved treatments with limited benefit. Leading treatments offer minimal efficacy with response rates generally under 15% in the long term with no clear effect on melanoma-related mortality. Even the recent success of the specific BRAF mutant inhibitor vemurafenib has been tempered somewhat since acquired resistance is rapidly observed. Thus, understanding the mechanism(s) of melanoma carcinogenesis is paramount to combating this deadly disease. Not only for the treatment of melanoma but, ultimately, for prevention. In this report, we will summarize our work to date regarding the characterization of ultraviolet radiation (UVR)-mediated melanomagenesis and highlight several promising avenues of ongoing research. © 2014 Elsevier B.V. All rights reserved.

The study objective is to compare the results of semen analysis using the standard cell culture method and real time polymerase chain reaction (RT-PCR).Materials and methods. Between 2016 and 2018, a prospective study including 143 males who applied to the Urology Clinic of the Ivanovo State Medical Academy due to infertility and for periconceptional preparation was performed. Semen was examined using cell culture method and RT-PCR. The group 1 included 54patients with leukospermia and bacteriospermia; the group 2 included 58patients without leukospermia but with bacteriospermia; the control group consisted of 31 males without leukospermia and bacteriospermia based on RT-PCR.Results. Per RT-PCR, the most frequent microorganisms in the groups 1 and 2 were Corynebacterium spp. (51.9 and 60.3 %) and Enterobacteriaceae spp./Enterococcus spp. (51.9 and 62.1 %), but there weren’t any significant differences between the groups. The range of detected microorganisms per cell culture method and RT-PCR was the same in most cases, but in 22 (15.4 %) patients RT-PCR detected Enterobacteriaceae/Enterococcus spp., which weren’t observed in cell culture. These males were prescribed bovhyaluronidase azoximer (Longidase®) 2 times a week for 1 month for anti-inflammatory therapy and to decrease leukospermia. Post-therapy control semen cultures detected microorganisms that were previously detected by RT-PCR.Conclusion. Our study demonstrates the advantages of RT-PCR compared to the standard cell culture semen analysis. RT-PCR can be recommended as a screening method. To increase the accuracy of microorganism detection, Longidase® is recommended because it promotes prostatic acini drainage.

Introduction: First spotted in Wuhan, China, World Health Organisation declared the deadly outbreak caused by novel Severe Acute Respiratory Syndrome Coronavirus-2 (SARS CoV-2) as global pandemic on March 11, 2020. With over 72 million cases globally till December 2020, countries need to gear up to detect, isolate and treat Corona Virus Disease-2019 (COVID-19) cases. Laboratories play a crucial role in diagnosis thereby instituting early contact tracing and quarantine measures. The laboratory based COVID-19 diagnostic testing data may help in formulating strategies to contain the spread of infection. Aim: To describe key patient variables of the respiratory samples processed for SARS-CoV-2 by Real Time Polymerase Chain Reaction (RT-PCR) at a diagnostic laboratory. Materials and Methods: In this descriptive, single centre study carried out at ICMR approved COVID-19 diagnostic laboratory, nasopharyngeal swabs received in Viral Transport Medium (VTM) were tested for SARS-CoV-2 infection by RTPCR. Key patient variables, like age, gender, symptoms and sample positivity rate were tabulated. The demographic and clinical data of samples tested were summarised by medians and Interquartile Range (IQR) for continuous variables and by proportions for categorical variables. Results: A total of 12,187 samples were received between 21st March to 8th July 2020. The data from 11,196 individuals were complete and were included in the analysis. Overall, 2,053 samples were tested positive for SARS-CoV-2 indicating positivity rate of 18.33%. Sample positivity was highest (63.91%) among high-risk contacts of a laboratory confirmed case. The maximum number of samples tested belonged to age group of 21-40 years and male predominance was observed. Conclusion: Although social distancing, mask usage, hand hygiene and respiratory etiquettes are important measures for containment of COVID-19, strengthening and capacity building of laboratory network is crucial for mitigating the pandemic.

Introduction: First spotted in Wuhan, China, World Health Organisation declared the deadly outbreak caused by novel Severe Acute Respiratory Syndrome Coronavirus-2 (SARS CoV-2) as global pandemic on March 11, 2020. With over 72 million cases globally till December 2020, countries need to gear up to detect, isolate and treat Corona Virus Disease-2019 (COVID-19) cases. Laboratories play a crucial role in diagnosis thereby instituting early contact tracing and quarantine measures. The laboratory based COVID-19 diagnostic testing data may help in formulating strategies to contain the spread of infection. Aim: To describe key patient variables of the respiratory samples processed for SARS-CoV-2 by Real Time Polymerase Chain Reaction (RT-PCR) at a diagnostic laboratory. Materials and Methods: In this descriptive, single centre study carried out at ICMR approved COVID-19 diagnostic laboratory, nasopharyngeal swabs received in Viral Transport Medium (VTM) were tested for SARS-CoV-2 infection by RTPCR. Key patient variables, like age, gender, symptoms and sample positivity rate were tabulated. The demographic and clinical data of samples tested were summarised by medians and Interquartile Range (IQR) for continuous variables and by proportions for categorical variables. Results: A total of 12,187 samples were received between 21st March to 8th July 2020. The data from 11,196 individuals were complete and were included in the analysis. Overall, 2,053 samples were tested positive for SARS-CoV-2 indicating positivity rate of 18.33%. Sample positivity was highest (63.91%) among high-risk contacts of a laboratory confirmed case. The maximum number of samples tested belonged to age group of 21-40 years and male predominance was observed. Conclusion: Although social distancing, mask usage, hand hygiene and respiratory etiquettes are important measures for containment of COVID-19, strengthening and capacity building of laboratory network is crucial for mitigating the pandemic. [ABSTRACT FROM AUTHOR]

Detection of pathogenic bacteria, such as Listeria monocytogenes, in food is crucial for safeguarding public health in Iraq. Forty five samples of frozen meat (15 samples of each of minced red meat, chicken, and fish) were collected from different markets in Baghdad city. Molecular (RT-PCR) and culturing (conventional microbiological examination) methods were used to determine the level of contamination of L. monocytogenes in these types of meat. For the culturing method, TSYEB broth was used as an enrichment medium, whereas BALCAM medium (HiMedia) with the listeria selective supplement FD061 was used as a selective medium, for the isolation and identification of this bacterium. The isolates were confirmed microscopically and biochemically. The results of the culturing method showed that the total number of the isolates of L. monocytogenes was 14/45 (31.1%). The incidence of this bacterium was high in fish (11/15, 73.3%), while it was low in the other two types of meat. 2/15 (13.3%) in red meat and 1/15 (6.7 %) in chicken. Molecular detection of each sample of the bacteria was performed using RTPCR technique after preparing the Genomic DNA extraction of these samples according to the protocol provided by ReliaPrep™ Blood gDNA Miniprep System kit (Promega, USA). The PCR primers and the hybridization probe ((Macrogen, Korea) were used to target the inlA gene sequence (specific for L. monocytogenes). The results of the RT-PCR assay showed that 10/45 (22.2%) of the samples were positive for L. monocytogenes, which was detected only in fish samples ((10/15, 66.7%), while not found in minced red meat and chicken. However, our results showed differences when compared to other previous works because there were many studies found that the highest contamination rate was in red meat products. We conclude that the PCR kit used for the detection of L monocytogenes appears to give accurate results in the diagnoses of this bacterium in meat products and in comparison with the other routine diagnosis methods in the laboratory, which included culturing and doing biochemical tests which last for approximately 7 days, the RT-PCR technique was able to confirm the findings within 48 hours. [ABSTRACT FROM AUTHOR]

Background and Objectives: Acinetobacter baumannii is recognized as an important pathogen responsible for serious infections causing episodes of hospital infection. Carbon nanotubes (CNTs) have recently emerged as superior materials against antibiotic-resistant bacteria. In this study, a new chemical compound was designed in order to combat A. baumannii infections. Subsequently, the effect of this novel carbon nanotube coated with an antibacterial compound on Extensively Drug-Resistant (XDR), Multidrug-Resistant (MDR) and Pan-Drug-Resistance (PDR) strains of A. baumannii was investigated. Materials and Methods: A total of 122 clinical isolates of A. baumannii were cultured from burn patients and their susceptibility to antibiotics were checked using disk diffusion method and Minimum inhibitory concentration. Antimicrobial effects of the coated carbon nanotube were evaluated on XDR, MDR and PDR isolates of A. baumannii. Cell viability was determined using tetrazolium reduction assay (MTT) on human fibroblast cell line (HDFa). Wound healing processes were assessed by quantitative polymerase chain reaction. Results: Of the 50 A. baumannii isolates, 38 (76%) were found to be MDR and 12 (24%) were XDR. No PDR strains were detected. Results indicated that the carbon nanotube combined with mercury had antibacterial effect against different A. baumannii species and it also was able to increase the expression of epidermal growth factor, platelet-derived growth factor and vascular endothelial growth factor A mRNA levels which are involved in wound healing. Conclusion: The engineered carbon nanotube compound can potentially be used for treatment of burn related infections. This can potentially give clinicians a new tool for treating A. baumannii infections. [ABSTRACT FROM AUTHOR]

The SARS-CoV-2 pandemic is the most devastating health crisis our generation has seen. The present study tries to gather more epidemiological data and improve the knowledge of the demographic factors responsible for a higher incidence of COVID-19. We analyzed the real time polymerase chain reaction (RT-PCR) results obtained in one of the biggest tertiary hospitals in Bulgaria during the first year of the COVID-19 pandemic (May 2020–April 2021). For this period, almost 40% of all tested samples of hospitalized patients and health care workers in University Hospital "St. Marina," Varna, were SARS-CoV-2 positive. The most affected individuals were 60–79-year-old people. Male sex is a significant risk factor only for the active ages (20–59 years), while both men and women of advanced age have the same risk to be infected with SARS-CoV-2. Interestingly, girls under 19 years were more susceptible to the infection than boys in the same age group. [ABSTRACT FROM AUTHOR]

Objective of the investigation was to study the infection of ethnic Tatars with the Epstein–Barr virus (EBV) and to analyze the genetic structure of the oncogene of the virus, the latent membrane protein 1 (LMP1), in the virus strains of Tatar origin. Materials and methods. The materials for the study were samples of boucle flushes of 60 students from the Kazan State Medical University who are ethnic Tatars (Tatars no less than in the 3rd generation). Amplified from DNA of boucle flushes the nucleotide sequences of the LMP1 samples translated into DNA amino acid sequences, have undergone classification based on the well-known and widely used in literature the R.H. Edwards et al. classification. Results. The analysis of nucleotide and deductive amino acid sequences of the 41 LMP1 amplicons revealed their homology with only three gene variants from the R.H. Edwards et al. classification (1999): 95.8/A (29.3 %; 12/41), Med– (14.6 %; 6/41) and China1 (7.3 %, 3/41). Such variants of LMP1 as Alaskan, Med+, Chinа2, China3 and NC, were not found. Among the LMP1 samples of Tatar origin in 20 cases (48.8 %), the composition of the mutations found did not allow them to be assigned to any of the oncogene variants listed above. Out of this number, in 7 (17.1 %) cases a mono group of LMP1 samples was found, differing not only from representatives of the Slavs, inhabitants of the European part of Russia, but also from other Kazan samples, and was designated as LMP1-TatK. The remaining 13 samples of LMP1 (31.7 %), not belonging to any of the known classifications, formed the group designated by us as an LMP1 group beside the classification (LMP1BC). Conclusion. Continuation of the study of the molecular-biological and functional properties of LMP1 in TatK and BC groups, which constitute 48.8 % of the number of gene samples studied, and an analysis of the peculiarities of the ethnic Tatar genotype, will probably help to clarify whether certain EBV strains influence morbidity and mortality in Tatar population with malignant neoplasms, which include EBVassociated cases.

Purpose: The aim of this study was to report the results of the use of real-time polymerase chain reaction (PCR) for the diagnosis of hepatitis B virus (HBV) infection in cornea donors at the Central Eye Bank of Iran. Methods: Between 2014 and 2016, all cornea donors that had negative screening serologic results for hepatitis B (HB) surface antigen, HB surface antibody (Ab), hepatitis C virus Ab, human immune deficiency virus Ab, human T-cell leukemia virus Ab, and syphilis, and positive serology for HB core Ab were subjected to real-time PCR with a detection limit of 400 IU/mL to identify HBV DNA. Positive results for HBV DNA were considered occult HBV infections in these donors. Results: Over the 3-year period, 122 out of 10448 cornea donors had negative screening serologic tests outside of HB core Ab. Of which, 90 cases were subjected to real-time PCR. Occult HBV was detected in 11 cases (12.2%), resulting in the rejection of the corresponding corneas. The remaining 79 cases (87.8%) had negative results for HBV DNA and the corresponding corneas were used for transplantation. Conclusion: Implementation of PCR for the detection of occult HBV in cornea donors is necessary to not only increase the security level of cornea donation but also minimize the rejection rate of donors that have isolated HB core Ab reactivity.

Objective: To explore the changes in microbial composition and the corresponding impact after lactitol treatment in constipated patients. Methods: Altogether 29 consecutive outpatients diagnosed with chronic constipation from three centers were recruited and stratified based on their history of diabetes mellitus. All patients were administered with oral lactitol for 2 weeks, and a symptoms diary of constipation was recorded. Fecal samples were collected before and after lactitol treatment, and were analyzed by 16S rRNA sequencing and real‐time polymerase chain reaction (PCR) to detect gut microbiota. Results: Twenty patients with diabetes mellitus and nine without, all with chronic constipation, were enrolled in this study. After 2‐week administration of lactitol, their subscale scores and constipation symptoms significantly decreased (P < 0.05). An analysis of fecal flora using 16S rRNA sequencing found an increasing trend of abundance of Bifidobacterium in the post‐lactitol group (P = 0.08). Actinobacteria, Actinobacteria, Bifidobacteriales, Bifidobacteriaceae and Bifidobacterium were significantly more abundant after lactitol administration. Real‐time PCR showed significantly high DNA copy numbers of Bifidobacterium after lactitol treatment (1.39 × 1010 vs 2.74 × 109 copies/μL, P = 0.01). The results of 16S rRNA sequencing and real‐time PCR illustrated an increasing trend of Bifidobacterium in both patients with and without diabetes. In addition, Bifidobacterium was negatively correlated with constipation subscale scores. Conclusions: Alterations in fecal flora composition after lactitol supplementation, especially in terms of an increasing trend of Bifidobacterium, alleviated constipation symptoms. Lactitol may be a promising prebiotic candidate for patients with constipation, regardless of diabetes mellitus. [ABSTRACT FROM AUTHOR]

Lately, there was an outbreak of Coronavirus disease (COVID-19) in Wuhan, a city of province Hubei, China. The infectious and transferrable virus originated in bats, seafood and animals. COVID-19 belongs to Coronviradae family. Severe Acute Respiratory Syndrome (SARS) and Middle East Respiratory Syndrome (MERS) also belong to this family and their outbreak occurred in China and Saudi Arabia back in 2003 and 2012. The mortality rate of COVID-19 is much greater than that of SARS and MERS. The clinical symptoms of infected individuals from virus are almost the same but diagnostically they are different from one another. Reverse transcription polymerase chain reaction (RT-PCR) laboratory technique is a standard test for evaluation and confirmation of COVID-19 disease. In early stages, imaging modalities can be used for screening purposes. X-Ray is used to identify the gross changes within the chest region while Computed Tomography scan chest is used for detailed modification which occurs due to coronavirus. CT scan evaluates the different characteristics in the individuals infected from COVID-19 such as presence of bilateral ground glass opacities, presence of diffuse consolidation, presence of nodules with halo sign, enlarge thoracic lymph nodes, presence of thicken inter-lobar septa, presence of bronchiectasis and pleural effusion. Each lung lobe was assigned different score on the basis of lobe area involvement. A lobe having no involvement is of score 0, minimal involvement lobe score is 1, mild involvement lobe score is 2, moderate involvement lobe score is 3, and severe involvement lobe score is 4. The lobes are assessed for different degree of involvement and then are divided as none (0%), minimal (1% to 25%), mild (26% to 50%), moderate (51% to 75%) and severe 76% to above level. These all techniques are very effective and responsible for the exact location of the area involved and also show the extension of pathology that can only be possible on CT scan. CT scan not only evaluates the disease but it also provides the assessment of the disease severity. [ABSTRACT FROM AUTHOR]

Purpose. Sturgeon is the common name for the 27 species of fish belonging to the family Acipenseridae. The family is grouped into four genera: Acipenser, Huso, Scaphirhynchus and Pseudoscaphirhynchus. Four species may now be extinct. Two closely related species, Polyodon spathula (paddlefish) and Psephurus gladius (Chinese paddlefish, possibly extinct) are of the same order, Acipenseriformes, but belong to the family Polyodontidae and are not considered as "true" sturgeons. The aim of our paper is to develop and carry out a new DNA identification methodology for the sturgeons species with the use of real time polymerase chain reaction. Methodology. A PCR with the detection of results in real time was used for DNA-identification of sturgeon species. Findings. The new real-time PCR was developed and tested for rapid and accurate identification of DNA extracted from eggs and meat of four sturgeon species such as beluga (Huso huso, Linnaeus), Russian sturgeon (Acipenser gueldenstaedtii, Brandt), sterlet (Acipenser ruthenus, Linnaeus) and stellate sturgeon (Acipenser stellatus, Pallas). Based on the results of analytical specificity and comparison of the amplification parameters, we detected specific amplification and absence of cross reactions for fin fragments and eggs of the studied fish species. The proposed DNA-identification method can be used for the identification of sturgeon species that will allow controlling the production of competitive aquaculture products, which will meet CITES requirements. Originality. For the first time, the development and approbation of the methodology for species identification of sturgeon fish by using real-time PCR was carried out. Practical value. Species identification of sturgeon fish will allow controlling the production of competitive and legalized aquaculture products that meet CITES requirements as well as will ensure the implementation of the country strategic plan for food security in Ukraine.

Introduction: In the era of Direct Acting Antiviral (DAA), elimination of HCV is possible but persistence in extra hepatic sites is hindrance. Detection of HCV has been reported in platelets, however its impact on DAA therapy not been studied. Aim: Detection and quantification of HCV RNA from the platelets and to compare with parallel plasma levels. Materials and Methods: Whole blood collected from 62 chronic HCV infected patients. HCV RNA quantitation was done from paired plasma and platelet pool at baseline using commercial real time PCR assay (Abbott). Correlation calculated by Bland-Altman analysis. Bivariate association and p-value calculated using chi-square tests. Results: A total of 45 patients were viremic while 17 were nonviremic. Median plasma viral load was log105.7 (IQR: log103.55- 6.7) IU/mL, median platelet viral load was log104.13 (IQR: log100- 5.5) IU/mL. Genotype (GT) 3 (66.6%) was commonest followed by GT 1 (33.3%). Sustained Virologic Response (SVR) was attained in 39/45 patients (92.8%) patients while 3/45 patients (6.67%) were non-responders. Viral load at baseline in platelet pool was higher in non-responders when compared to those achieved SVR (median log104.46 vs. log103.27, p=0.456). Conclusion: HCV-RNA was efficiently detected in platelets. Persistence in platelets could be one of the factors associated with treatment failure.

Abstract The nucleotide alignment of all 11 genes of human Rotavirus A (RVA) strains revealed suitability of NSP2, NSP3 and VP6 genes for the development of real time PCR (qRT-PCR). Evaluation of qRT-PCR assays using known rotavirus ELISA positive and negative fecal specimens showed non-overlapping ranges of Mean ±3SD cycle threshold (Ct) values for NSP3 and VP6 based assays. Using serial dilutions of purified RVA, high sensitivity of VP6 qRT-PCR assay (1.95 × 10−5 pg/μL of RNA) was recorded as compared to NSP2 and NSP3 qRT-PCR assays (1.95 × 10-4 pg/μL of RNA). Further, evaluation of the VP6 qRT-PCR assay involving 266 fecal specimens and frequency polygon analysis of the data indicated cut-off value of 35 for Ct with high sensitivity (126/131, 96%) and specificity (12/12, 100%). This VP6 qRT-PCR assay will be a useful diagnostic tool to evaluate clinical presentations in rotaviral gastroenteritis under different conditions such as breast feeding and administration of rotavirus vaccines. [ABSTRACT FROM AUTHOR]

Abstract Visceral leishmaniasis (VL) is a zoonosis caused by the protozoa of the genus Leishmania. Among the species, L. infantum and/or L. infantum (chagasi) are the most important species affecting the Americas. Domestic dogs are the main reservoir of the parasite and participate effectively in the parasite' transmission cycle. The Canine Visceral Leishmaniasis Control Program (PCLV) adopted in Brazil present as strategies the vector control, health education and serological diagnosis of CVL in dogs followed by culling of the seropositive ones. The resolution to eliminate seropositive dogs by euthanasia, when necessary, are the most controversial and least accepted by society. The diagnostic methods for canine visceral leishmaniasis, currently indicated and approved in Brazil by the Ministry of Health from Brazil are the Dual Path Platform (DPP)® as a screening test and the Enzyme immunoassay test (ELISA®). This study aimed to verify the presence of Leishmania spp. DNA in peripheral blood samples of dogs presenting positive serological results byDPP® and ELISA® tests,throughreal-time polymerase chain reaction (rt-PCR), using the pair of primers 150–152 already described. For this purpose, were collected blood samples from 185 seropositive dogs among them, 41 (22%) exhibited some clinical signal of disease, whereas 144 (78%) was asymptomatic. The animals were also analyzed according to gender, race and hair size. According to the results of rt-PCR, it was observed that among the185 seropositive dogs analyzed, only 132 (71%) presented positive results for CVL and 53 (29%) presented negative results. From this, 41/41 symptomatic dogs were positive (100%), while among the asymptomatic dogs, 91/144 were positive (63, 2%) and 53/144 were negative (36, 8%). Concerning the hair size of seropositive dogs, we found that 41 (22%) had long hair, while 144 (78%) had short hair. No statistical significance occurred between the results of rt-PCR, ELISA and DPP tests and the profile of the animals (gender, size of the dogs and hair size), probably due to the small number of samples and the sampling differences of each profile. But statistical significance occurred between the results of rt-PCR and the clinical evaluation, since the rt-PCR was positive in all symptomatic dogs. Thus, through these results, we reached at the following question, which may contribute to an important current debate: the dogs presenting CVL seropositive diagnosis confirmed by tests distributed by the Ministry of Health were in reality ill or were they seropositive by living in an endemic area of the disease? Would these asymptomatic seropositive dogs spread the disease to the inhabitants even presenting a low parasite charge circulating in the blood. Highlights • In 185 seropositive dogs analyze rt-PCR specific for Leishmania´ DNA had positive results in 41 symptomatic dogs. • Concerning the hair size of seropositive dogs, were found that 41 (22%) had long hair, while 144 (78%) had short hair. • Serological tests for CVL should be supplemented by clinical findings and DNA detection of the parasite circulating. • The culling of seropositive dogs has an emotional impact in the owners. • Molecular techniques should be used to help CLV diagnosis supplementary to serology achieves. [ABSTRACT FROM AUTHOR]

Background: Research interest has grown around the potential therapeutic use of cannabidiol in mood-related disorders, due to its anxiolytic and antidepressant-like effects. These have been partially attributed to its action as an allosteric modulator of 5-HTR1A. However, the exact mechanism supporting cannabidiol properties remains unclear.Aims: To assess the effects of cannabidiol on different targets of the hypothalamus-pituitary-adrenal axis under baseline and stress conditions.Methods: We administered cannabidiol (5 mg/kg, 15 mg/kg or 30 mg/kg, intraperitoneally) or vehicle to male C57BL/6J mice 90 min before single restraint stress exposure (20 min). Using real-time polymerase chain reaction analysis, we measured alterations in the relative gene expression of corticotropin-releasing factor in the paraventricular nucleus, pro-opiomelanocortin in the arcuate nucleus of the hypothalamus, glucocorticoid receptor in the hippocampus, and serotonin 5-HTR1A receptor in the hippocampus and amygdala.Results: Under baseline conditions, cannabidiol did not modify any element of the hypothalamus-pituitary-adrenal axis. In contrast, all doses induced alterations in 5-HTR1A in the amygdala and hippocampus. Interestingly, cannabidiol at low (5 mg/kg) and intermediate doses (15 mg/kg) successfully blocked the effects induced by acute stress on corticotropin-releasing factor, pro-opiomelanocortin and glucocorticoid receptor gene expression. Also, restraint stress induced the opposite effects in 5-HTR1A gene expression in the hippocampus and amygdala, an effect not seen in mice treated with cannabidiol at low doses.Conclusions: Taken together, these data suggest the ability of cannabidiol to regulate acute stress hypothalamus-pituitary-adrenal axis activation might be explained, at least in part, by its action on 5-HTR1A receptors. [ABSTRACT FROM AUTHOR]

Helicobacter pylori (H. pylori) may enter a non-replicative, non-culturable, low metabolically active state, the so-called coccoid form, to survive in extreme environmental conditions. Since coccoid forms are not susceptible to antibiotics, they could represent a cause of therapy failure even in the absence of antibiotic resistance, i.e., relapse within one year. Furthermore, coccoid forms may colonize and infect the gastric mucosa in animal models and induce specific antibodies in animals and humans. Their detection is hard, since they are not culturable. Techniques, such as electron microscopy, polymerase chain reaction, loop-mediated isothermal amplification, flow cytometry and metagenomics, are promising even if current evidence is limited. Among the options for the treatment, some strategies have been suggested, such as a very high proton pump inhibitor dose, high-dose dual therapy, N-acetycysteine, linolenic acid and vonoprazan. These clinical, diagnostic and therapeutic uncertainties will represent fascinating challenges in the future.