Diversity of Rotavirus Strains in Children; Results From a Community-Based Study in Nepal

Objective: The objectives of this study were to describe the incidence and genetic diversity of Rotavirus (RV) infection among children up to 3 years of age in a community in Nepal.Methods: We investigated community-acquired cases of asymptomatic and symptomatic RV infections in children from birth to 36 months of age in a community-based birth cohort in Bhaktapur, Nepal. Monthly surveillance and diarrheal stool samples were collected from 240 children enrolled at birth, of which 238 completed the 3 years of follow-up. Samples were screened for rotavirus by Enzyme Immuno Assay (EIA). All RV screened positives were further genotyped by reverse transcription-polymerase chain reaction for the capsid genes VP7 and VP4.Results: In total, 5,224 stool samples were collected from 238 children, followed from birth to 36 months of age. Diarrhea occurred in 92.4% (230/238) of all children in the cohort. During the 3 years study period, RV was more frequently seen in children with symptoms (7.6%) than in non-symptomatic children (0.8%). The highest RV detection rate was found in younger children between 3 and 21 months of age. Although rotavirus is known as winter diarrhea, it was detected throughout the year except in August. The highest positivity rate was observed in the months between December and March, with a peak in January. Four common G types were seen: G2 (30%), G1 (29%), G12 (19%), and G9 (16%). The most predominant genotypes seen were G2P[4] (30%), followed by G1P[8] (27.0%), G12P[6] (14.0%), G9P[8] (10%), and remaining were mixed, partial, and untyped.Conclusion: Our study confirms that rotavirus is a common cause of gastroenteritis in young children in the community. The prevalence and pathogenicity of rotavirus infection differed by age. There was substantial variability in circulating strains in the community samples compared to samples collected from hospitals. This shows the importance of including community-based surveillance systems to monitor the diversity of circulating rotavirus strains in Nepal.

Increased Serum Level of 8-Hydroxy-2’-Deoxyguanosine may be an Indication of Pesticides Induced DNA Damage among Indian Farmers

The aim of the present hospital based study is to assess the serum levels of 8-hydroxy-2’-deoxyguanosine (8-OHdG) among 152 subjects. A total of 152 subjects, categorized into 3 groups: (i) pesticides exposed group (N=60), (ii) pesticide un-exposed group (N=42) and (iii) healthy controls group (N=50) were recruited for the study following the inclusion and exclusion criteria. The blood was drawn from the eligible subjects and Enzyme Immuno Assay (EIA) was performed to assess the 8-OHdG levels in serum. Appropriate statistical methods were used to analyse the study data. Assessment showed that pesticides exposed group has higher levels of serum 8-OHdG as compared to un-exposed group and healthy controls. Age and duration of exposure had an impact on the levels of serum 8-OHdG.The higher serum levels of 8-OHdG may be a marker for pesticides-induced oxidative DNA damage. Key words: pesticides; exposure; oxidative stress; 8-hydroxy-2’-deoxyguanosine (8-OHdG).

Serologic and molecular characterization of rotavirus from children with acute gastroenteritis in Chennai, Tamil Nadu, India

Background: Rotavirus gastroenteritis is the leading cause of diarrhea in infants and young children worldwide. Although, Rotavirus vaccine has been introduced in 2017 in states like Tamil Nadu, there are reports of the role of Rotavirus as one of high disease burden agents with genetic variants arising, especially from low-income countries like India.Methods: Authors evaluated stool samples from 507 children with acute gastroenteritis Rotavirus A among the hospitalized children (>5 years) to provide baseline information on changing profile in this state. The stool samples were collected and screened for Rotaviral Antigen by Enzyme Immuno Assay and use of semi-multiplex RT PCR technique was conceded out in order to conclude the P and G genotypes of human rotavirus in rotavirus-positive samples from January 2014 to December 2016 in and around Chennai, India.Results: Of 507 samples collected 213 (42.01%) were positive for rotavirus antigen by Enzyme Immuno Assay (EIA). The maximum positivity (75%) was in the age group of one to two years. Rotavirus positives were subjected to further VP7 and VP4 molecular characterization and the predominant genotypes identified were G9P[4] followed by G9P[8], G1P[8], G3P[8], G2P[4] and mixed types of G2G9 with P[4] and G4P[6][11] with few untypable strains.Conclusions: This study had demonstrated the Rota Virus Gastro Enteritis (RVGE) is a common disease affecting the pediatric population and G9P[4], G9P[8] circulating types among the gastroenteritis cases reported in the city and its suburban area. This study in comparison to previous ones shows that the dominant serotypes and circulating genotypes changes from time to time within country. The results have reemphasized the need of rotavirus vaccines with broad serotype coverage which may help in decreasing the disease burden in this region of the country.

Pemberian resveratrol oral mencegah peningkatan F2-Isoprostan urin tikus Wistar (Rattus norvegicus) jantan yang dipapar tartrazine

This study was aimed to prove that oral administration of resveratrol could prevent urinary F2-isoprostane elevation in tartrazine-induced male Wistar rats (Rattus norvegicus). This was an experimental study using the pretest-posttest control group design. Subjects were 24 rats (Rattus norvegicus), Wistar strain, healthy, 2-3 months old, weighing 200-220 g, divided into 2 groups with 12 rats each. The first group (P0), the control group, was given a placebo of 2 ml distilled water 2 hours prior to the administration of tartrazine 75 mg/kg body weight. The second group (P1), the treatment group, was given resveratrol of 20 mg/kg 2 hour prior to the administration of tartrazine 75 mg/kg. Rats’ urine was collected before and after treatment for 4 weeks. Level of F2-isoprostane was examined by using an 8-iso-PGF2α enzyme immuno assay kit. The comparative analysis of the pretest groups showed that there was no difference between the average levels of F2-isoprostane in both groups (5.45±0.62 ng/mL in P0 group vs 5.42±0.64 ng/mL in P1 group) (P > 0.05). Meanwhile, after treatment for 4 weeks, the average level of F2-isoprostane in the P0 group was significantly different from the P1 group (6.61±0.93 ng/mL vs 3.79±0.48 ng/mL) (P < 0.01). Analysis of the treatment effect showed a significant increase of F2-isoprostane level in the P0 group, and a significant decrease in the P1 group (P < 0.01). Conclusion: Oral administration of resveratrol could prevent urinary F2-isoprostane elevation in tartrazine-induced male Wistar rats (Rattus norvegicus).Keywords: tartrazine, resveratrol, F2-isoprostaneAbstrak: Tujuan penelitian untuk membuktikan pemberian resveratrol oral dapat mencegah peningkatan F2-isoprostan dalam urin tikus (Rattus norvegicus) Wistar jantan yang dipapar tartrazine. Jenis penelitian eksperimental dengan menggunakan pretest-posttest control group design. Subyek penelitian ialah 24 ekor tikus (Rattus norvegicus), galur Wistar, sehat, berumur 2-3 bulan, dengan berat badan 200-220 gr, dibagi menjadi 2 kelompok, masing-masing berjumlah 12 ekor tikus. Kelompok pertama (P0) ialah kelompok kontrol, diberikan tartrazine 75 mg/kg BB dan 2 jam setelahnya diberikan plasebo berupa aquadest 2 ml. Kelompok kedua (P1) ialah kelompok perlakuan, diberikan tartrazine 75 mg/kg BB dan 2 jam setelahnya diberikan resveratrol 20 mg/kg BB. Saat sebelum dan sesudah perlakuan selama 4 minggu, urin tikus dikoleksi untuk pemeriksaan kadar F2-isoprostan menggunakan 8-iso-PGF2α enzyme immuno assay kit. Analisis komparasi sebelum perlakuan (pretest) menunjukkan rerata kadar F2-isoprostan pada kedua kelompok tidak berbeda nyata (5,45±0,62 ng/mL vs 5,42±0,64 ng/mL) (P > 0,05). Setelah perlakuan selama 4 minggu, rerata kadar F2-isoprostan pada kelompok P0 berbeda nyata dibandingkan kelompok P1 (6,61±0,93 ng/mL vs 3,79±0,48 ng/mL) (P < 0,01). Analisis efek perlakuan menunjukkan terjadi peningkatan kadar F2-isoprostan pada kelompok P0 dan penurunan bermakna pada kelompok P1 (P < 0,01). Simpulan: Pemberian resveratrol oral dapat mencegah peningkatan F2-isoprostan urin tikus (Rattus norvegicus) Wistar jantan yang dipapar tartrazine.Kata kunci: tartrazine, resveratrol, F2-isoprostan, urin

Comparitive performance of in-house RT-PCR for NSP4 gene with a commercial enzyme immunoassay for the detection of rotavirus in stool samples of children with diarrhoea

Rotaviruses are the most significant pathogen in the etiology of watery diarrhoea in infants and young children all over the world. Laboratory diagnosis is important both for clinical management and estimation of the disease bur-den. We collected faecal samples from 260 children under the age of five years hospitalized with acute diarrhoea and tested them for rotavirus anti-gen by Enzyme immuno assay (EIA) kit [RIDASCREEN]. RNA was also extract-ed from supernatant of 10% stool suspensions in Phosphate Buffered Saline (PBS) and amplified by Reverse transcriptase polymerase chain reaction (RT-PCR) for Non Structural Protein 4 (NSP4) gene. EIA was positive in 58(22.3%) samples while RT-PCR was positive in 45(17.3%) samples. Compared to EIA, RT-PCR was 77.5% sensitive and 100% specific. The Positive Predictive Value (PPV) and Negative Predictive Value (NPV) were 100% and 93.9% respective-ly. We infer that RT-PCR is a moderately sensitive but highly specific diagnos-tic test for rotavirus infection in this study.