Mixed Genotype Research Articles

Hepatitis B genotyping in chronic hepatitis B patients not on treatment in a tertiary medical centre in Kuala Lumpur, Malaysia

Background: Distribution, clinical characteristics and clinical response to antiviral treatment vary among hepatitis B viral (HBV) genotypes. This study aimed to describe the prevalence, demography and clinical characteristics of the HBV genotypes among chronic hepatitis B patients who were not on treatment in Universiti Kebangsaan Malaysia Medical Centre (UKMMC). Methods & Materials: A total of 1200 plasma samples received by the Molecular Biology Laboratory from 2015 to 2017 for HBV viral load were screened for volume adequacy and viral load of at least 1000 IU/mL. Records were traced to include cases not on any antiviral therapy, cases without co-infection with other hepatitis virus or HIV, and cases without other causes of liver disease. Real-time polymerase chain reaction with 3 steps melting curve analysis was done. Sequencing was done in a private laboratory for 7 samples with undifferentiated genotypes using the platform of ABI 3730 XL (Applied Biosystems). The association among genotypes, demographic and clinical characteristic data (viral load, HBeAg, AntiHBe, ALT, AFP, cirrhosis/HCC) were analysed. Results: Eighty samples (n=80) were successfully genotyped. Of these, 73% (n=59) were genotype B, 20% genotype C (n=16) and 2.5% genotype A (n=2). One sample which constituted 1.3% (n=1) each for genotype D, mixed genotype A+B and mixed genotype B+C. Sixty percent were female (48/80) and mean age was 49 years. Chinese ethnic group recorded the highest cases (78.8%), followed by Malays (17.5%). Majority of the study population (85%) had no cirrhosis by ultrasonography. Normal level of AFP and ALT were predominant (median AFP: 2.46 μg/mL; median ALT: 26 U/L). Anti-Hbe reactivity was significantly higher in those with genotype B than C with normal ultrasound findings (p = 0.035). Individuals infected by genotype A and mixed genotype B+C showed more benign clinical characteristics while genotype B, C, and mixed genotype A+B were more aggressive although not statistically significant. Genotype B was also the most prevalent among the subjects with advanced liver disease in 4 out of 6 subjects (66.7%). Conclusion: Genotypes B and C were most prevalent among chronic hepatitis B patients who were not on treatment in UKMMC. Genotype B was also most prevalent among subjects with advanced liver disease.

Design of efficacious somatic cell genome editing strategies for recessive and polygenic diseases

Compound heterozygous recessive or polygenic diseases could be addressed through gene correction of multiple alleles. However, targeting of multiple alleles using genome editors could lead to mixed genotypes and adverse events that amplify during tissue morphogenesis. Here we demonstrate that Cas9-ribonucleoprotein-based genome editors can correct two distinct mutant alleles within a single human cell precisely. Gene-corrected cells in an induced pluripotent stem cell model of Pompe disease expressed the corrected transcript from both corrected alleles, leading to enzymatic cross-correction of diseased cells. Using a quantitative in silico model for the in vivo delivery of genome editors into the developing human infant liver, we identify progenitor targeting, delivery efficiencies, and suppression of imprecise editing outcomes at the on-target site as key design parameters that control the efficacy of various therapeutic strategies. This work establishes that precise gene editing to correct multiple distinct gene variants could be highly efficacious if designed appropriately.

Circulating HBV Genotypes among Animal and Non-Animal Handlers in Osun State, Nigeria

Aim: Hepatitis B virus (HBV) is not uncommon among animal and non-animal handlers. The brutality of HBV infection and the outcome of treatment is linked with exact HBV genotypes. No study on the circulation of HBV genotypes has been reported among animal and non-animal handlers in Nigeria. This study was intended to evaluate the genotypic distribution among animal and non-animal handlers in Osun State, Nigeria.
 Study Design: Cross-sectional study.
 Place and Duration of Study: Ladoke Akintola University of Technology (LAUTECH), Nigeria, between June 2015 and July 2019.
 Methods: Blood samples were obtained from HBsAg positive individuals and screened for HBV-DNA from cohorts of animal and non-animal handlers. HBV-DNA was extracted, amplified and genotyped using a multiplex PCR technique with primers specific for six genotypes of HBV (Genotype A, B, C, D, E and F).
 Results: Results showed that a total of 11 (6.1%) of the 180 animal and non-animal handlers evaluated were positive to HBsAg and 4.4% were positive for HBV-DNA by a semi-nested PCR using HBV specific primer pairs. The molecular analysis of the sera of 11 HBsAg positive animal and non-animal handlers showed that 72.7% of them had a true HBV infection. Results further show that genotype E (75.0%) was predominant over genotype A (12.5%) and mix genotypes (D and E) with 12.5% prevalence. Other genotypes were not detected. Of the 8 positive HBV-DNA samples, 7 (87.5%) were males and one (12.5%) was a female. All animal and non-animal handlers with true HBV infection were found to harbour HBV genotype E predominantly.
 Conclusion: The molecular analysis of HBV-DNA and genotypes circulating among animal and non-animal handlers shows that the majority of the subjects with true HBV infection were found to predominantly harbour HBV genotype E in Osun state, Nigeria. The study further highlights the predominance HBV genotype E in Nigeria.

Study of the Effect of Different Hepatitis C Virus Genotypes on Splenomegaly.

BackgroundSeveral recent studies have shown that the hepatitis C virus (HCV) and its different genotypes are a predominant and leading cause of cirrhosis and splenomegaly in different regions of the world. Advanced stage of cirrhosis leads to portal hypertension that causes splenomegaly. This complication may have many other manifestations such as anemia, infections, and bleeding disorders in severe stages. This study aimed to determine the effect of different HCV genotypes on the development of splenomegaly and to assess which HCV genotypes are more associated with it.Materials and methodsA total of 483 conveniently sampled HCV patients were included in this descriptive cross-sectional study. Six genotypes (1, 2a, 2b, 3a, 3b, and mixed) were studied, and 80 patients for each of these genotypes were included. Data were collected from patient medical records regarding patient demographic details, HCV serology and genotyping, and sonographic size of the spleen.ResultsIn total, splenomegaly was present in 14.1% (n=68) patients. The development of splenomegaly was significantly associated with old age, as 25.2% (n=26) of patients above 60 years of age (n=103) developed splenomegaly (P=0.005). Our study determined that splenomegaly was significantly related to HCV genotypes 3a, 3b, and 1 (P<0.001, P=0.017, and P=0.019, respectively). By taking mixed genotype as a reference, the odds of developing splenomegaly with genotype 3a were the highest (OR = 9.481; CI=95%).ConclusionsOur study concludes that HCV genotype 3a, 3b, and 1, and age above 60 years have a significant association with splenomegaly. Genotype 3a has the highest risk of developing splenomegaly. Therefore, our study demands screening, early diagnosis, and prompt treatment of these particular HCV genotypes to prevent complications and risk of mortality.

HPV-EM: an accurate HPV detection and genotyping EM algorithm

Accurate HPV genotyping is crucial in facilitating epidemiology studies, vaccine trials, and HPV-related cancer research. Contemporary HPV genotyping assays only detect < 25% of all known HPV genotypes and are not accurate for low-risk or mixed HPV genotypes. Current genomic HPV genotyping algorithms use a simple read-alignment and filtering strategy that has difficulty handling repeats and homology sequences. Therefore, we have developed an optimized expectation–maximization algorithm, designated HPV-EM, to address the ambiguities caused by repetitive sequencing reads. HPV-EM achieved 97–100% accuracy when benchmarked using cell line data and TCGA cervical cancer data. We also validated HPV-EM using DNA tiling data on an institutional cervical cancer cohort (96.5% accuracy). Using HPV-EM, we demonstrated HPV genotypic differences in recurrence and patient outcomes in cervical and head and neck cancers.

Promoting mixed genotype infections in CpGV: analysis on field and laboratory sprayed apple leaves

ABSTRACT The success of biological control of the codling moth, Cydia pomonella, with commercial products based on C. pomonella granulovirus (CpGV) isolates relies both on the ingestion of a sufficient amount of occlusion bodies (OBs) between their hatching from the egg and the penetration inside the fruits, and the correct replication of the virus in the insect larvae. Larvae resistant to one isolate, CpGV-M are frequently found in European orchards. In recent years, efforts have been devoted to obtaining virus isolates able to control resistant insects, among them, CpGV-R5. Ingestion of CpGV-M by a resistant larva does not allow replication of CpGV-M. Ingestion of CpGV-R5 results in virus replication and larval death. Ingestion of a mixture of CpGV-M and CpGV-R5 allows replication of both genotypes and better insect control. Both CpGV-M and CpGV-R5 are present in the commercial product Carpovirusine® Evo 2 (NPP, Arysta LifeSciences). No recent data have been collected concerning the actual amount of virus ingested by the neonate larvae. As mixed infection CpGV-M and CpGV-R5 provide better protection, this is an interesting property to explore to improve the strategy of treatment in orchards. We analysed the distribution of virus droplets on leaves for two orchards using two different dispersion techniques and leaves sprayed in laboratory conditions. We observed the mortality of larvae allowed to feed on these contaminated leaves for selected amounts of time. We then used High resolution melting (HRM) to estimate the occurrence of each genotype alone and mixed genotypes in individual larvae.

Real-world effectiveness of glecaprevir/pibrentasvir and ledipasvir/sofosbuvir for mixed genotype hepatitis C infection: A multicenter pooled analysis in Taiwan.

Data on direct-acting antiviral agent (DAA) treatment for mixed genotype hepatitis C virus (HCV) infection are scant. This study examined the effectiveness of glecaprevir/pibrentasvir (GLE/PIB) and ledipasvir/sofosbuvir (LDV/SOF) for mixed HCV genotype infection in a real-world setting in Taiwan. We analysed the data from all patients with mixed HCV genotype infections treated with GLE/PIB or LDV/SOF from 2017 to 2019 in three Chang Gung Memorial Hospitals in Taiwan. The primary treatment outcome was sustained virologic response 12weeks after treatment cessation (SVR12). Adverse events (AEs) were also evaluated. A total of 5190 HCV patients received DAA treatment during this time period. Among them, 116 patients (2.2%) had mixed infections of any 2 or 3 genotypes of 1a, 1b, 2, 3 and 6. Fifty-four patients received GLE/PIB and 62 received LDV/SOF. SVR12 rates for LDV/SOF vs GLE/PIB therapy were 96.6% (56/58) vs 100% (51/51) by the per-protocol analysis and 90.3% (56/62) vs 94.4% (51/54) by the evaluable population analysis. Two patients with 1b+6 and 1b+2 genotype infections in the LDV/SOF group had relapse. Evaluating the GLE/PIB vs LDV/SOF groups for the most common AEs revealed pruritus (16.7% vs 4.8%), abdominal discomfort (5.6% vs 8%) and fatigue (5.6% vs 4.8%). One patient with AE-related treatment discontinuation presented with liver decompensation after 4-week GLE/PIB therapy. DAA-related significant laboratory abnormalities occurred in two patients with >3× elevated bilirubin level in the GLE/PIB group. GLE/PIB and LDV/SOF are well tolerated and achieve high SVR12 rates for patients with mixed HCV genotype infection.

Molecular surveillance of the Pfmdr1 N86Y allele among Congolese pregnant women with asymptomatic malaria

BackgroundMalaria in pregnancy is associated with considerable morbidity and mortality. Regular surveillance of artemisinin-based combination therapy tolerance, or molecular makers of resistance, is vital for effective malaria treatment, control and eradication programmes. Plasmodium falciparum multiple drug resistance-1 gene (Pfmdr1) N86Y mutation is associated with reduced susceptibility to lumefantrine. This study assessed the prevalence of Pfmdr1 N86Y in Brazzaville, Republic of Congo.MethodsA total 1001 of P. falciparum-infected blood samples obtained from asymptomatic malaria pregnant women having a normal child delivery at the Madibou Integrated Health Centre were analysed. Pfmdr1 N86Y genotyping was conducted using PCR-restriction fragment length polymorphism.ResultsThe wild type Pfmdr1 N86 allele was predominant (> 68%) in this study, whereas a few isolates carrying the either the mutant allele (Pfmdr1 86Y) alone or both alleles (mixed genotype). The dominance of the wildtype allele (pfmdr1 N86) indicates the plausible decline P. falciparum susceptibility to lumefantrine.ConclusionThis study gives an update on the prevalence of Pfmdr1 N86Y alleles in Brazzaville, Republic of Congo. It also raises concern on the imminent emergence of resistance against artemether–lumefantrine in this setting. This study underscores the importance to regular artemether–lumefantrine efficacy monitoring to inform the malaria control programme of the Republic of Congo.

Clinicoepidemiological and Genotyping Correlation of Pediatric Scrub Typhus from Chandigarh, India

We studied the clinical phenotypes and prevalent genotypes of Orientia tsutsugamushi in our area using indirect immunofluorescence assay (IFA). We prospectively screened all febrile children presenting to our hospital over three years. From among children who were scrub typhus positive by ELISA we selected a sample of convenience for IFA testing to determine the genotypes of O. tsutsugamushi using four strains namely Boryong, Gilliam, Karp and Kato. Of all scrub positive patients (n=77), we tested 14 samples using IFA and all 14 samples were IFA positive. Karp genotype (n=7) was most prevalent followed by Kato (3), Boryong (1) and Gilliam (1) genotypes; 2 patients were positive for mixed genotype. There was high prevalence of organ dysfunction among IFA positive children. Three most common organ dysfunctions included hematological derangement in all, liver involvement in 10 (71%), and encephalopathy and shock in 4 each. Karp was the most prevalent genotype of O. tsutsugamushi in our area.

BREEDING PRACTICES AND TRAITS OF PREFERENCE FOR SELECTION OF PIGS BY MALE AND FEMALE SMALLHOLDER FARMERS IN NIGERIA

&lt;p&gt;&lt;strong&gt;Background.&lt;/strong&gt; Pig farming is one of the most important livestock raised in Nigeria by smallholder farmers. Objective. To determine the breeding practices and breeding traits of preference by pig farmers in Nasarawa State, Nigeria. &lt;strong&gt;Methodology.&lt;/strong&gt; A total of 120 pig rearers (62 males and 58 females) were randomly sampled. Primary data were collected through individual structured questionnaire administration. Chi square (χ2) statistics was used to compare categorical variables. Arithmetic means and their standard errors of continuous variables between gender were tested using T- Test. Ranking of the eight traits of preference (body size, body conformation, mothering ability, survival, heat tolerance, disease resistance, fertility and temperament) was carried out using the non-parametric Friedman Test (Wilcoxon Signed-rank test for post hoc analysis) as well as Kruskal–Wallis H Test (with Mann– Whitney U Test for means separation). Multivariate Clustering Analysis was also explored to group the farmers on gender basis. &lt;strong&gt;Results.&lt;/strong&gt; The source as well as the number of foundation stock varied between the sexes (P= 0.048 and 0.028, respectively). Average flock size was higher (20.31±2.21 versus 14.78±1.53; P&amp;lt;0.05) where the pigs were under the management of male farmers. This reflected more in pigs of mixed genotypes compared to the indigenous ones. The number of piglets/sow/annum (14.77±0.77 versus 12.29±0.54; 16.36±0.76 versus 14.02±0.58; P ≤ 0.05) was also higher in flocks of male farmers for both indigenous pigs and those of mixed blood. Between-gender, all the preference traits were ranked similarly (P&amp;gt;0.05) apart from temperament which was more highly rated by the males. However, the multivariate analysis revealed that body size and body conformation were more associated with the female farmers while survival, disease resistance, heat tolerance, mothering ability and temperament were more preferred by their male counterparts. &lt;strong&gt;Implication.&lt;/strong&gt; The traits of economic importance obtained in this study may guide future community-based pig genetic improvement programmes in a tropical environment. &lt;strong&gt;Conclusion.&lt;/strong&gt; The present study revealed the importance of body size, body conformation, mothering ability, survival, heat tolerance, disease resistance and fertility in the selection of breeding pigs. However, these traits were differentially rated by male and female farmers.&lt;/p&gt;

Landmarks of the Knowledge and Trypanosoma cruzi Biology in the Wild Environment.

Trypanosomatids are ancient parasitic eukaryotes that still maintain prokaryotic characteristics. Trypanosoma cruzi, a primarily wild mammal parasite, infected humans already long before European colonization of the Americas. T. cruzi heterogeneity remains an unsolved question, and until now, it has still not been possible to associate T. cruzi genotypes with any biological or epidemiological feature. One of the first biochemical attempts to cluster the T. cruzi subpopulations recognized three main subpopulations (zymodemes) that have been associated with the transmission cycles in the wild (Z1; Z3) and in the domestic environment (Z2). The description of wild mammal species harboring Z2 two decades later challenged this assemblage attempt. Currently, the genotypes of T. cruzi are assembled in seven discrete typing units (DTUs). The biology of T. cruzi still shows novelties such as the description of epimastigotes multiplying and differentiating to metacyclic trypomastigotes in the lumen of the scent glands of Didelphis spp. and the capacity of the true meiosis in parallel to clonal reproduction. The study of the transmission cycle among wild animals has broken paradigms and raised new questions: (i) the interaction of the T. cruzi DTUs with each of its mammalian host species displays peculiarities; (ii) the impact of mixed genotypes and species on the transmissibility of one or another species or on pathogenesis is still unknown; (iii) independent T. cruzi transmission cycles may occur in the same forest fragment; (iv) the capacity to act as a reservoir depends on the peculiarities of the host species and the parasite genotype; and (v) faunistic composition is a defining trait of the T. cruzi transmission cycle profile. The development of models of environmental variables that determine the spatial distribution of the elements that make up T. cruzi transmission by spatial analysis, followed by map algebra and networking, are the next steps toward interpreting and dealing with the new profile of Chagas disease with its many peculiarities. There is no way to solve this neglected disease once and for all if not through a multidisciplinary look that takes into account all kinds of human and animal activities in parallel to environmental variations.

Genotyping and Quantification of In Situ Hybridization Staining in Zebrafish.

In situ hybridization (ISH) is an important technique that enables researchers to study mRNA distribution in situ and has been a critical technique in developmental biology for decades. Traditionally, most gene expression studies relied on visual evaluation of the ISH signal, a method that is prone to bias, particularly in cases where sample identities are known a priori. We have previously reported on a method to circumvent this bias and provide a more accurate quantification of ISH signals. Here, we present a simple guide to apply this method to quantify the expression levels of genes of interest in ISH-stained embryos and correlate that with their corresponding genotypes. The method is particularly useful to quantify spatially restricted gene expression signals in samples of mixed genotypes and it provides an unbiased and accurate alternative to the traditional visual scoring methods.

The Mixed AII and BIII Genotypes of Human Giardia lamblia Isolate Circulating in Mazandaran Province, Northern Iran

Giardiasis is one of the human parasitic diseases by a flagellate protozoan named Giardia lamblia. This parasite is one of the most common organisms causing diarrhea in humans and a common gastrointestinal parasite in vertebrates. The purpose of this investigation was to determine the molecular epidemiology of Giardia lamblia (G. lamblia) by PCR-RFLP method in the Mazandaran Province and to investigate the possible relationship between parasite genotype and clinical symptoms. In this study, 50 stool samples were collected from human cases of G. lamblia. referred to health centers in Sari, Babol, Nowshahr cities. The samples were examined by formalin-ether concentration procedure. DNA extraction was carried out on samples containing adequate Giardia cysts, and then PCR-RFLP was done on glutamate dehydrogenase (gdh) marker. In these patients, the most common signs were abdominal pain (42.9%), and diarrhea (47.6%); and least marked common sign was malaise (4.7%). Of all 21 isolates, 9 samples (42.8%) were found as genotype mixed AII and BIII, 6 samples (28.6%) belonged to genotype BIII and 6 (28.6%) were of genotype AII. Additionally, there was a significant correlation between AII genotype and clinical symptoms (P <0.05). According to the statistical tests, there was a significant relationship between abdominal pain and the intensity of infection (P<0.05). However, genotype AII and mixed AII and BIII genotypes were more common, respectively. The relatively high prevalence of mixed infection of these genotypes indicated the potential of zoonotic source of giardiasis among patients in Mazandaran Province.

Evaluation towards stud bulls with different mixed genotypes relating to somatotropin cascade genes by origin

A preliminary evaluation towards a breeding value of the Holstein breeding bulls with different genotypes of somatotropin cascade genes is presented: pituitary transcription factor or growth hormone factor (PIT1), prolactin (PRL), somatotropin (GH), somatotropin releasing hormone (GHRH), insulin-like (IGF-1) in the conditions of the Republic of Tatarstan. The research studies the signs (indices) of milk productivity (milk yield and fat in milk) of the closest female ancestors of bulls with different genotypes of somatotropin cascade genes. Studies and analysis have shown that the bulls with the mixed genotypes ABAALLBBAA (10269 kg), AAAALLBBAB (4.13 %) and BBAALLABBB (9402 kg and 4.06 %) had the highest and optimum evaluation by origin, judging by the bulls’ pedigree (geneological) index. The frequency to meet bulls with the desired mixed genotypes was low and amounted to 4.28–5.71 %.

HCV Genotype Distribution of Patients with Chronic Hepatitis C in Istanbul.

Objectives:Hepatitis C virus (HCV), which has no protective vaccine, is a common cause of chronic hepatitis, which is a severe public health threat. There are differences in nucleotide and amino acid sequences in different regions of the HCV genome. As a result of these differences, HCV has been shown to have at least seven major genotypes and many subtypes. In Turkey, the prevalence of genotype 1 is between 51.7% and 97.1%, the highest rate among all genotypes, while subtype 1b is the genotype with the highest rate. It is important to detect mixed genotype infection reliably as it causes treatment failure. This study aims to reveal the distribution of the HCV genotypes in our hospital in Istanbul over the years and to contribute to the epidemiological data of Turkey.Methods:For this purpose, 385 patient samples sent to Sisli Hamidiye Etfal Training and Research Hospital, Clinical Microbiology Laboratory for HCV genotype determination between January 2016 and June 2019 were evaluated retrospectively. Anti-HCV was screened by enzyme immunoassay and confirmation was performed by Line immunoassay. HCV genotyping assays targeting highly conserved 5’UTR and most variable region NS5B regions were used.Results:The most common genotype was genotype 1 (81.3%) with 313 cases and subtypes 1a and 1b were detected at the rates of 10.9% and 67.8%, respectively. In addition, genotype 3, 2, 4, 5 were detected at the rates of 8.8%, 3.4%, 2.9%, 0.8%, respectively and mixed genotype was found in 2.9% of cases. Although genotype 5 is seen in South Africa, it is found in the Middle East region, albeit at a low rate. In our study, it was observed that genotype 5 was detected in different years from patients of Syrian origin.Conclusion:In this study, genotype 1 was the most common genotype with a rate of 81.3% and subtype 1b was 67.8%, in accordance with the literature. However, genotypes 3, 2, 4 and 5 were also present at low rates. It is important to monitor these rare genotypes since some of them are dominant in surrounding countries. In addition, 2.9% of HCV mixed genotype was detected and this should be considered concerning management of HCV infection.

Evaluation of an ensemble-based distance statistic for clustering MLST datasets using epidemiologically defined clusters of cyclosporiasis.

Outbreaks of cyclosporiasis, a food-borne illness caused by the coccidian parasite Cyclospora cayetanensis have increased in the USA in recent years, with approximately 2300 laboratory-confirmed cases reported in 2018. Genotyping tools are needed to inform epidemiological investigations, yet genotyping Cyclospora has proven challenging due to its sexual reproductive cycle which produces complex infections characterized by high genetic heterogeneity. We used targeted amplicon deep sequencing and a recently described ensemble-based distance statistic that accommodates heterogeneous (mixed) genotypes and specimens with partial genotyping data, to genotype and cluster 648 C. cayetanensis samples submitted to CDC in 2018. The performance of the ensemble was assessed by comparing ensemble-identified genetic clusters to analogous clusters identified independently based on common food exposures. Using these epidemiologic clusters as a gold standard, the ensemble facilitated genetic clustering with 93.8% sensitivity and 99.7% specificity. Hence, we anticipate that this procedure will greatly complement epidemiologic investigations of cyclosporiasis.

The glucocorticoid receptor in the nucleus accumbens plays a crucial role in social rank attainment in rodents

Social hierarchy in social species is usually established through competitive encounters with conspecifics. It determines the access to limited resources and, thus, leads to reduced fights among individuals within a group. Despite the known importance of social rank for health and well-being, the knowledge about the processes underlying rank attainment remains limited. Previous studies have highlighted the nucleus accumbens (NAc) as a key brain region in the attainment of social hierarchies in rodents. In addition, glucocorticoids and the glucocorticoid receptor (GR) have been implicated in the establishment of social hierarchies and social aversion. However, whether GR in the NAc is involved in social dominance is not yet known. To address this question, we first established that expression levels of GR in the NAc of high anxious, submissive-prone rats are lower than that of their low anxious, dominant-prone counterparts. Furthermore, virally-induced downregulation of GR expression in the NAc in rats led to an improvement of social dominance rank. We found a similar result in a cell-specific mouse model lacking GR in dopaminoceptive neurons (i.e., neurons containing dopamine receptors). Indeed, when cohabitating in dyads of mixed genotypes, mice deficient for GR in dopaminoceptive neurons had a higher probability to become dominant than wild-type mice. Overall, our results highlight GR in the NAc and in dopaminoceptive neurons as an important regulator of social rank attainment.

Hepatitis B virus genotypes among chronic hepatitis B patients from Baghdad, Iraq and their impact on liver function

Hepatitis B virus (HBV) infection is a significant global health problem. Populations of different ethnicities show great heterogeneity in HBV genotype frequency distributions. A cross-sectional study was conducted during June–October 2018 to determine frequency of HBV genotypes among chronic HBV patients from Baghdad, Iraq. The method of detection was nested polymerase chain reaction system. Further, the study assessed the impact of HBV genotypes on serum level of liver-function tests: total serum bilirubin, alkaline phosphatase, alanine aminotransferase and aspartate aminotransferase. Eighty chronic HBV patients were enrolled in the study. Six HBV genotypes were identified (A, B, C, D, E and F). The most frequently encountered genotypes were B and F (72.5% each), while E genotype was the least observed with frequency of 12.5%. In addition, the majority of patients (77.5%) showed mixed HBV genotypes (two, three, four or five) with different genotypic combinations. Single HBV genotype patients represented only 17.5%. When patients positive for mixed HBV genotypes were inspected for serum level of liver-function parameters, only alkaline phosphatase showed a significant difference. It exhibited a gradual increase median that positively paralleled the number of positive genotypes; patients positive for one genotype showed the lowest median (87.0 U/L), while patients positive for five genotypes demonstrated the highest median (130 U/L). In conclusion, the HBV genotypes B and F were the most frequent among the investigated sample of Iraqi chronic HBV patients from Baghdad.

Rotavirus Burden, Genetic Diversity and Impact of Vaccine in Children under Five in Tanzania.

In Tanzania, rotavirus infections are responsible for 72% of diarrhea deaths in children under five. The Rotarix vaccine was introduced in early 2013 to mitigate rotavirus infections. Understanding the disease burden and virus genotype trends over time is important for assessing the impact of rotavirus vaccine in Tanzania. When assessing the data for this review, we found that deaths of children under five declined after vaccine introduction, from 8171/11,391 (72% of diarrhea deaths) in 2008 to 2552/7087 (36% of diarrhea deaths) in 2013. Prior to vaccination, the prevalence of rotavirus infections in children under five was 18.1–43.4%, 9.8–51%, and 29–41% in Dar es Salaam, Mwanza and Tanga, respectively, and after the introduction of vaccines, these percentages declined to 17.4–23.5%, 16–19%, and 10–29%, respectively. Rotaviruses in Tanzania are highly diverse, and include genotypes of animal origin in children under five. Of the genotypes, 10%, 28%, and 7% of the strains are untypable in Dar es Salaam, Tanga, and Zanzibar, respectively. Mixed rotavirus genotype infection accounts for 31%, 29%, and 12% of genotypes in Mwanza, Tanga and Zanzibar, respectively. The vaccine effectiveness ranges between 53% and 75% in Mwanza, Manyara and Zanzibar. Rotavirus vaccination has successfully reduced the rotavirus burden in Tanzania; however, further studies are needed to better understand the relationship between the wildtype strain and the vaccine strain as well as the zoonotic potential of rotavirus in the post-vaccine era.

A Fluorescence Sensing Method with Reduced DNA Typing and Low-Cost Instrumentation for Detection of Sample Tampering Cases in Urinalysis.

This work presents a method to unequivocally detect urine sample tampering in cases where integrity of the sample needs to be verified prior to urinalysis. The technique involves the detection of distinct patterns of a triplex short tandem repeats system in DNA extracted from human urine. The analysis is realized with single-dye fluorescence detection and using a regular smartphone camera. The experimental results had demonstrated the efficacy of the analytical approach to obtaining distinct profiles of amplicons in urine from different sample providers. Reproducibility tests with fresh and stored urine have revealed a maximum variation in the profiles within an interval of 5 to 9%. Cases of urine sample tampering via mixture were simulated in the study, and the experiments have identified patterns of mixed genotypes from dual mixtures of urine samples. Moreover, sample adulteration by mixing a non-human fluid with urine in a volume ratio over 25% can be detected. The low cost of the approach is accompanied by the compatibility of the technique to use with different DNA sample preparation protocols and PCR instrumentation. Furthermore, the possibility of realizing the method in an integrated microchip system open great perspectives to conducting sample integrity tests at the site of urine sample reception and/or at resource-limited settings.