Novel Human Astroviruses: Prevalence and Association with Common Enteric Viruses in Undiagnosed Gastroenteritis Cases in Spain

Adenovirus; Children; Classic astrovirusAdenovirus; Nens; Astrovirus clàssicAdenovirus; Niños; Astrovirus clásicoA remarkable percentage of acute gastroenteritis cases remain etiologically undiagnosed. The aim of the study was to determine the prevalence of common and emerging enteric viruses, such as novel human astroviruses, among undiagnosed samples from children with acute gastroenteritis. Epidemiological studies for novel human astroviruses are still scarce. Stool samples collected over two consecutive winter seasons (2016-2017) from children with gastroenteritis in Spain, which were negative for bacteria, rotavirus, and adenovirus by routine diagnostics were screened by real-time RT-PCR assays for the presence of classical and novel astrovirus, rotavirus, norovirus GI and GII, sapovirus, and adenovirus. Overall, 220/384 stool samples (57.3%) were positive for at least one virus. Co-infections were identified in 21% of cases. Among a total of 315 viruses identified, adenovirus was the most prevalent (n = 103), followed by rotavirus (n = 51), sapovirus (n = 50), classical astrovirus (n = 43), novel astroviruses (n = 42), and norovirus (n = 26). Novel astroviruses were present in 13.3% of virus-positive cases. Most novel astroviruses were found in children <2-year-old (30/39 children, 77%, p = 0.01) and were found in co-infection (66%). Only classical astroviruses demonstrated significant differences in the Cq values during mono-infections compared to co-infections. In conclusion, common enteric viruses may be frequently found in children with undiagnosed gastroenteritis, indicating the need to implement more sensitive diagnostic methods. Novel astroviruses circulate in the community and could be the cause of gastroenteritis among young children.Supported in part by the Biotechnology Reference Network (XRB) program of the Generalitat de Catalunya. This work was also supported in part by the Certest Biotec Company. Diem-Lan Vu was recipient of a fellowship from the Geneva University Hospital

Human astrovirus MLB replication in vitro: persistence in extraintestinal cell lines

MLB astroviruses were identified 10 years ago in feces from children with gastroenteritis of unknown etiology and have been unexpectedly detected in severe cases of meningitis/encephalitis, febrile illness of unknown etiology, and respiratory syndromes. The aim of this study was to establish a cell culture system supporting MLB astrovirus replication. We used two clinical strains to infect several cell lines, an MLB1 strain from a gastroenteritis case, and an MLB2 strain associated with a neurologic infection. Efforts to propagate the viruses in the Caco-2 cell line were unsuccessful. In contrast, we identified two human nonintestinal cell lines, Huh-7 and A549, permissive for both genotypes. After serial passages in the Huh-7.5 cell line, the adapted strains were able to establish persistent infections in the Huh-7.5, Huh-7AI, and A549 cell lines, with high viral loads (up to 10 log10 genome copies/ml) detected by quantitative reverse transcription-PCR (RT-qPCR) in the culture supernatant. Immunofluorescence assays demonstrated infection in about 10% of cells in persistently infected cultures. Electron microscopy revealed particles of 32 to 33 nm in diameter after negative staining of cell supernatants and capsid arrays in ultrathin sections with a particularly high production in Huh-7.5 cells. Interferon (IFN) expression by infected cells and effect of exogenous IFN varied depending on the type of infection and the cell line. The availability of a cell culture system to propagate MLB astroviruses represents a key step to better understand their replicative cycle, as well as a source of viruses to conduct a wide variety of basic virologic studie

Vocational Orientation and the Need for Establishing Career Counselling Office in Vietnamese Schools

School psychology is a field that has been receiving a lot of attention recently in Vietnam as it prepares to restructure the general education curriculum. In particular, vocational orientation and career counseling are most concerned due to the emergence of new subjects in the educational curriculum - Experimental and Vocational Orientation Activities. The objective of this study is to investigate the current situation of vocational orientation problems and the need for career counseling of Vietnamese high school students to provide evidence to develop the school counseling activities and the School Psychology in Vietnam. This is a quantitative study used a questionnaire to examine the current situation of vocational orientation problems and the need for career counseling of 1200 high school students in Vietnam to illustrate the importance of vocational orientation work. The results showed that Vietnamese high school students faced many vocational orientation problems and wanted support from school counselors. But the reality did not meet the needs of students: Vietnamese schools lacked a team of school counselors both in quality and quantity, also lack of school counseling offices in almost high schools. This result is expected to contribute to the development of the School Psychology in Vietnam, but firstly, to promote the establishment of counseling office at least one office per school

In vitro growth and content of vincristine and vinblastine of Catharanthus roseus L. hairy roots in response to precursors and elicitors

Catharanthus roseus L. is a medicinal plant that produces numerous indole terpenoid alkaloids, including vincristine and vinblastine, which are used for cancer treatment. The effect of specified precursors (L-phenylalanine, L-tyrosine) and elicitors (chitosan, methyl jasmonate) on C. roseus hairy roots (CHR) growth has been examined in order to increase the content of vincristine and vinblastine. Our results showed that CHR generated by an Agrobacterium rhizogenes strain isolated in Vietnam was capable of producing both vincristine and vinblastine when subjected to precursors, but only vinblastine when exposed to elicitors. However, both precursors and elicitors were evaluated to have an effect on increasing the accumulation of TIAs in CHR. In particular, the use of elicitors required more time to find the appropriate induction conditions, while the use of precursors gave outstanding efficiency in the treatment with 1 µM phenylalanine. The greatest yields of vincristine (51.99 µg g-1 DW) and vinblastine (699.92 µg g-1 DW) were obtained in the 7th week (with 0.306 g DW biomass). This result is the first time we might boost the levels of vincristine and vinblastine in our CHR clone generated by the Vietnam strain of A. rhizogenes

Novel Human Astroviruses: Prevalence and Association with Common Enteric Viruses in Undiagnosed Gastroenteritis Cases in Spain

A remarkable percentage of acute gastroenteritis cases remain etiologically undiagnosed. The aim of the study was to determine the prevalence of common and emerging enteric viruses, such as novel human astroviruses, among undiagnosed samples from children with acute gastroenteritis. Epidemiological studies for novel human astroviruses are still scarce. Stool samples collected over two consecutive winter seasons (2016-2017) from children with gastroenteritis in Spain, which were negative for bacteria, rotavirus, and adenovirus by routine diagnostics were screened by real-time RT-PCR assays for the presence of classical and novel astrovirus, rotavirus, norovirus GI and GII, sapovirus, and adenovirus. Overall, 220/384 stool samples (57.3%) were positive for at least one virus. Co-infections were identified in 21% of cases. Among a total of 315 viruses identified, adenovirus was the most prevalent (n = 103), followed by rotavirus (n = 51), sapovirus (n = 50), classical astrovirus (n = 43), novel astroviruses (n = 42), and norovirus (n = 26). Novel astroviruses were present in 13.3% of virus-positive cases. Most novel astroviruses were found in children <2-year-old (30/39 children, 77%, p = 0.01) and were found in co-infection (66%). Only classical astroviruses demonstrated significant differences in the Cq values during mono-infections compared to co-infections. In conclusion, common enteric viruses may be frequently found in children with undiagnosed gastroenteritis, indicating the need to implement more sensitive diagnostic methods. Novel astroviruses circulate in the community and could be the cause of gastroenteritis among young children

Adsorption of Co(II) from the simulated solution by zeolite NaX derived from rice husk ash

The adsorption of Co(II) from the simulated solution was investigated using zeolite NaX derived from rice husk ash as an alternative adsorbent. The adsorption behavior of Co(II) depended strongly on the equilibrium pH, Co (II) concentration, zeolite NaX dosage, and reaction time. The high adsorption efficiency of Co(II) by zeolite NaX was obtained under the conditions: pH 3.0, 100 mg/L of Co(II), 5 g/L of zeolite NaX, and a reaction time of 75 min. The loading behavior of Co(II) onto the zeolite NaX was well-fitted to the Freundlich adsorption isotherm and the Co(II) loading capacity by zeolite NaX was around 38 mg/g. The obtained results indicate that synthesized zeolite NaX from rice husk ash is a potential adsorbent to remove cobalt from waste solutions due to its high adsorption

A Spanish case-control study in <5 year-old children reveals the lack of association between MLB and VA astrovirus and diarrhea

Novel human astroviruses (HAstV) were discovered 10 years ago and have been associated with fatal cases of central nervous system infections. Their role in gastroenteritis is controversial, as they have been identified in symptomatic and asymptomatic subjects. The aim of the study was to investigate novel HAstV in a gastroenteritis case-control study including a pediatric population in Spain over a one-year period. We included stool samples from patients with gastroenteritis and negative results for viruses screened by routine diagnostics, and stool samples of control subjects who sought for a routine medical consultation. All samples were screened by real-time RT-PCR assays for novel HAstV. An additional screening for rotavirus, norovirus GI, GII, sapovirus, classic HAstV and adenovirus was also performed for the control group. Overall, 23/363 stool samples from case patients (6.3%) and 8/199 stool samples from control patients (4%) were positive for ≥1 novel HAstV. MLB1 was predominant (64.5% of positives). Seasonality was observed for the case group (p = 0.015), but not the control group (p = 0.95). No difference was observed in the prevalence of novel HAstV between the case and control groups (OR 1.78, 95% CI 0.68-5.45; p = 0.30). Nevertheless, MLB genome copy numbers/ml of fecal suspension was significantly higher in the control group than in the case group (p = 0.008). In our study, we identified a lack of association between novel HAstV and gastroenteritis in the studied population, which could indicate a potential role of reservoir for children, especially given the higher viral load observed in the asymptomatic group for some of them

ẢNH HƯỞNG CỦA MÔI TRƯỜNG NUÔI CẤY LÊN SINH TRƯỞNG CỦA CALLUS CÂY GIẢO CỔ LAM (Gynostemma pentaphyllum (Thunb.) Makino) – MỘT CÂY DƯỢC LIỆU CÓ GIÁ TRỊ

Gynostemma pentaphyllum (Thunb.) Makino (Jiaogulan) is a traditional medicinal herb belonging to the Cucurbitaceae family. G. pentaphyllum grows widely in Southern China, Japan, Korea, and Vietnam. The essential bioactive components of Jiaogulan are saponin glycosides (gypenosides) and antioxidants. Jiaogulan exhibits bioactive activities such as anticancer, antioxidant, cholesterol-reducing agent, immunopotentiation, and others. In this study, the primary callus of Jiaogulan was used as a material to evaluate the influence of the culture medium on the induction and growth of secondary calli. The results reveal that the callus cultured on the MS medium supplemented with 2.0 mg/L kinetin and 0.5 mg/L indole-3-butyric acid has the best growth ability, high rate of secondary callus induction (100%), and good callus quality for suspension culture. The concentration of gypenoside and Rb1 in callus is 36.298 and 0.009 mg/g dry weight. The gypenoside concentration of callus is lower than that of leaves (65.58%) and almost similar to that of stems (92.38%) from natural samples. The suitable solvent for the extraction of gypenoside is methanol. The obtained callus will be used as material for cell suspension culture in further studies.Giảo cổ lam (Gynostemma pentaphyllum (Thunb.) Makino) là một cây thuốc dân gian thuộc họ Bầu bí. Cây phân bố rộng ở miền Nam Trung Quốc, Nhật Bản, Hàn Quốc và Việt Nam. Các thành phần có hoạt tính sinh học quan trọng của Giảo cổ làm là saponin glycoside (gypenoside) và các chất chống oxy hóa. Giảo cổ lam được sử dụng hỗ trợ chống ung thư, chống oxy hóa, giảm cholesterol, tăng cường miễn dịch và các tác dụng khác. Trong nghiên cứu này, callus sơ cấp của cây Giảo cổ lam được sử dụng làm nguyên liệu để đánh giá ảnh hưởng của môi trường nuôi cấy lên khả năng phát sinh và sinh trưởng của callus thứ cấp. Kết quả cho thấy, callus được nuôi cấy trên môi trường MS có bổ sung 2,0 mg/L kinetin và 0,5 mg/L indole-3-butyric acid sinh trưởng tốt nhất; tỷ lệ tạo callus thứ cấp cao (100%); callus đáp ứng được tiêu chuẩn để nuôi cấy huyền phù. Hàm lượng gypenoside và Rb1 trong callus là 36,298 và 0,009 mg/g chất khô; gypenoside thấp hơn trong lá (65,58%) và gần tương đương với mẫu thân (92,38%) của sản phẩm thu mua từ thị trường. Dung môi thích hợp để tách chiết gypenoside là methanol. Callus thu được sẽ được sử dụng làm nguyên liệu cho nuôi cấy tế bào huyền phù trong các nghiên cứu tiếp theo

Astrovirus replication in human intestinal enteroids reveals multi-cellular tropism and an intricate host innate immune landscape.

Human astroviruses (HAstV) are understudied positive-strand RNA viruses that cause gastroenteritis mostly in children and the elderly. Three clades of astroviruses, classic, MLB-type and VA-type have been reported in humans. One limitation towards a better understanding of these viruses has been the lack of a physiologically relevant cell culture model that supports growth of all clades of HAstV. Herein, we demonstrate infection of HAstV strains belonging to all three clades in epithelium-only human intestinal enteroids (HIE) isolated from biopsy-derived intestinal crypts. A detailed investigation of infection of VA1, a member of the non-canonical HAstV-VA/HMO clade, showed robust replication in HIE derived from different patients and from different intestinal regions independent of the cellular differentiation status. Flow cytometry and immunofluorescence analysis revealed that VA1 infects several cell types, including intestinal progenitor cells and mature enterocytes, in HIE cultures. RNA profiling of VA1-infected HIE uncovered that the host response to infection is dominated by interferon (IFN)-mediated innate immune responses. A comparison of the antiviral host response in non-transformed HIE and transformed human colon carcinoma Caco-2 cells highlighted significant differences between these cells, including an increased magnitude of the response in HIE. Additional studies confirmed the sensitivity of VA1 to exogenous IFNs, and indicated that the endogenous IFN response of HIE to curtail the growth of strains from all three clades. Genotypic variation in the permissiveness of different HIE lines to HAstV could be overcome by pharmacologic inhibition of JAK/STAT signaling. Collectively, our data identify HIE as a universal infection model for HAstV and an improved model of the intestinal epithelium to investigate enteric virus-host interactions

Quantifying antimicrobial access and usage for paediatric diarrhoeal disease in an urban community setting in Asia.

OBJECTIVES: Antimicrobial-resistant infections are a major global health issue. Ease of antimicrobial access in developing countries is proposed to be a key driver of the antimicrobial resistance (AMR) epidemic despite a lack of community antimicrobial usage data. METHODS: Using a mixed-methods approach (geospatial mapping, simulated clients, healthcare utilization, longitudinal cohort) we assessed antimicrobial access in the community and quantified antimicrobial usage for childhood diarrhoea in an urban Vietnamese setting. RESULTS: The study area had a pharmacy density of 15.7 pharmacies/km2 (a pharmacy for every 1316 people). Using a simulated client method at pharmacies within the area, we found that 8% (3/37) and 22% (8/37) of outlets sold antimicrobials for paediatric watery and mucoid diarrhoea, respectively. However, despite ease of pharmacy access, the majority of caregivers would choose to take their child to a healthcare facility, with 81% (319/396) and 88% (347/396) of responders selecting a specialized hospital as one of their top three preferences when seeking treatment for watery and mucoid diarrhoea, respectively. We calculated that at least 19% (2688/14427) of diarrhoea episodes in those aged 1 to <5 years would receive an antimicrobial annually; however, antimicrobial usage was almost 10 times greater in hospitals than in the community. CONCLUSIONS: Our data question the impact of community antimicrobial usage on AMR and highlight the need for better education and guidelines for all professionals with the authority to prescribe antimicrobials