Expression, purification and evaluation of recombinant L-asparaginase inmehthylotrophic yeast Pichia pastoris: Expression, purification and evaluation of recombinant L-asparaginase in mehthylotrophic yeast Pichia pastoris: Research article

L-asparaginase (EC 3.5.1.1), a therapeutic enzyme used in the treatment of childhood acute lymphoblastic leukemia (ALL). Hence, the goal of this work is study the expression and evaluation of hydrolysis activity of native sequence (X12746) encoding for L-asparaginase from Erwinia chrysanthemi NCPBB1125 in the popular expression system Pichia pastoris. The sequence of asn encoded for mature protein was expressed in P. pastoris SMD1168 and X33. SDS-PAGE analysis showed recombinant L-asparaginase was secreted efficiently. Stable and high hydrolysis activity of extracellular L-asparaginase in P. pastoris SMD1168 making it a potential candidate to produce recombinant protein. After purification, a specific band whose appearance approximately 45 kDa indicating the glycosylated protein with specific activity by 6.251 Umg-1 and about 3 folds purifications.L-asparaginase (EC 3.5.1.1), một loại enzyme được sử dụng trong điều trị bệng ung thư bạch cầu mãn tính ở trẻ em. Mục tiêu của nghiên cứu này là biểu hiện và đánh giá hoạt tính thủy phân của L-asparaginase mã hóa bởi đoạn gene (X12746) tương ứng từ Erwinia chrysanthemi NCPBB1125 được biểu hiện trong nấm men Pichia pastoris. Gene đã được cắt signal peptide và biểu hiện trong P. pastoris SMD1168 and X33. Qua phân tích kết quả điện di SDS-PAGE của môi trường sau lên men, L-asparaginase tái tổ hợp được tìm thấy trong dịch ngoại bào của P. pastoris. Với khả năng sản xuất protein có hoạt tính cao hơn so với chủng P. pastoris X33, SMD1168 được lựa chọn để biểu hiện L-asparaginase tái tổ hợp. Sau khi tinh sạch, sự xuất hiện của một băng có kích khối lượng phân tử xấp xỉ 45 kDa trên điện di SDS-PAGE cho thấy protein tái tổ hợp đã bị glycosyl hóa với hoạt tính riêng 6.251 Umg-1 và đạt độ sạch 3.471 lần

TỔNG HỢP VẬT LIỆU Ni/UiO-66 THEO PHƯƠNG PHÁP TRỰC TIẾP VÀ GIÁN TIẾP

An archetypal metal-organic framework, UiO-6,6, synthesized with the solvo-thermal method, has a large specific surface area of 1071 m2·g–1. This material was modified with NiCl2 in the direct (Nix(tt)/UiO-66) and indirect (Nix(gt)/UiO-66) methods. The two modified materials were characterized by using powder X-ray diffraction, scanning electron microscopy, and infrared spectroscopy. Their crystalline and chemical structure is not much different. However, the thermal and porous properties differ from each other.UiO-66 là họ vật liệu khung hữu cơ kim loại tổng hợp bằng phương pháp nhiệt dung môi có diện tích bề mặt riêng lớn (1071 m2·g–1). Vật liệu UiO-66 này được biến tính bằng NiCl2 theo phương pháp trực tiếp (Nix(tt)/UiO-66) và gián tiếp (Niy(gt)/UiO-66). Kết quả đặc trưng hai vật liệu biến tính này bằng nhiễu xạ tia X, hiển vi điện tử quét và phổ hồng ngoại khác nhau không đáng kể. Tuy nhiên, phân tích nhiệt trọng lượng và đẳng nhiệt hấp phụ – khử hấp phụ N2 cho thấy có sự khác nhau giữa các phương pháp tổng hợp

Isolation and identification of phenolic compounds from the leaf extract of Cassia alata L.

Cassia alata is one of the most important species of the genus Cassia which is rich in anthraquinones and polyphenols. This plant is used as a medicinal material of which the leaves are known to have laxative and antibiotic properties. In our study, the methanol leaf extract of C. alata showed a significant antibacterial activity against human pathogenic bacteria strains Staphylococcus aureus and Bacillus cereus. The organic layers such as n-hexane, ethyl acetate, and aqueous layers, were prepared by partitioning the methanol extract with n-hexane and ethyl acetate successively.  We successfully isolated and identified the structures of five compounds from C. alata leaves. Their structures were elucidated by MS and NMR spectroscopic methods as well as comparison with literature data. These compounds were determined to be methyl 2,4,6-trihydroxybenzoate (1), kaempferol (2), (-)epiafzelechin (3), kaempferol-3-O-glucoside (4) and kaempferol-3-O-gentiobioside (5). Keywords. Cassia alata L., epiafzelechin, kaempferol, kaempferol-3-O-glucoside, kaempferol-3-O-gentiobioside

H2_{2}O2_{2} production in Lactobacillus strains isolated from the intestinal microbiome of healthy people

Lactobacillus sp. in the digestive tract are capable of producing H2O2 to inhibit the growth of harmful bacteria and balance the gut microflora. In this study, we have isolated 115 strains of Lactobacillus spp. from stool samples of healthy people in Ha Noi. Of the 50 tested Lactobacillus strains, 9 strains were capable of producing H2O2, of which the Lac.VFE-14 strain produced highly H2O2 with a concentration of 2.183 mM, followed by Lac.VFE-08 strains (2.081 mM) and Lac.VFE-04 (2.067 mM). All three strains grew well in MRS medium supplemented with bile salts or adjusted to low pH value. With 0.3% of bile salt, the survival rates of these 3 strains were 99%, 95% and 97%, respectively. At pH 3.0, after 3 hours of cultivation, the survival rates of the three strains were 98.54%, 94.15% and 95.27%, respectively. In addition, each of the cell-free culture supernatants of these three strains that inhibit the growth of S. aureus ATCC-23235. The inhibition zone diameters of the three strains were 19.0±1.0 mm, 14.0±1.0 mm and 11.7±1.3 mm, respectively. The results of 16S rRNA gene analyses showed that Lac.VFE-14, Lac.VFE-08 and Lac.VFE-04 had high similarity scores with L. plantarum ZZU 23 (100%), L. rhamnosus JCM 1136 (99%) and L. plantarum S7 (98.65%), respectively. This study indicates that all three strains have the potential to be used as probiotics in the future. 

SIMULATION AS A TEACHING METHOD FOR NURSING EDUCATION IN HEALTHCARE-ASSOCIATED INFECTION PREVENTION AND CONTROL IN ASIAN COUNTRIES: A QUALITATIVE STUDY

BACKGROUND: Applying simulation for nursing education, especially in healthcare-associated infection prevention and control (HAI-PC) in developing countries has limited evidence. The study was conducted to explore educators’ perceptions of simulation as a teaching method for nursing education in HAI-PC in two Vietnamese and two Cambodian universities. METHODS: An exploratory qualitative design was applied. A focus group of 37 educators from four universities was conducted for data collection. Inductive and deductive qualitative content analysis was applied in analysing the data. RESULTS: The core category was constructed to reflect educators’ perception of scenario-based simulation (SBS) as a teaching method for nursing education in HAI prevention and control. This main category included three subcategories: (i) enhancing nursing competence; (ii) preparing students for simulation; and [1] promoting simulation pedagogy competence. CONCLUSIONS: The findings identified the importance and benefits of applying simulation as a teaching method in nursing education. Additionally, it emphasized the necessity of enhancing knowledge associated with HAIs and providing additional training on simulation for educators to improve the quality of conducting simulations

EXPERIENCES OF THE NURSING STUDENT IN LEARNING HAI PREVENTION AND CONTROL IN ASIAN COUNTRIES THROUGH THE USE OF SCENARIO-BASED SIMULATION: AN EXPLORATIVE QUALITATIVE STUDY

BACKGROUND: Healthcare-associated infections (HAIs) have posed a major threat to both patients and to the safety healthcare personnel worldwide. According to the World Health Organization, 10% of hospitalized patients are affected by HAIs worldwide. OBJECTIVE: The objective of this study was to explore the experiences of nursing students in learning HAIs prevention and control by the application of the scenario-based simulation pedagogy now in use in two Vietnamese and two Cambodian universities. METHODS: A qualitative study was conducted among 160 nursing students from 2 Cambodian universities and 2 Vietnamese universities, and by using the purposive-sampling method. The data were collected through a focus group discussion and analyzed by the Graneheim and Lundman method (Graneheim & Lundman, 2004). RESULTS: Two themes and six categories were generated. 1) First theme: factors for enhancing student learning on the prevention and control of HAIs by use of scenario-based simulation; and 2) Second theme: factors hindering students learning on HAI prevention and control by use of scenario-based simulation. CONCLUSION: The findings showed that SBS is an effective learning method for nursing students that can be applied to enhance the quality of nursing education in the Asian countries as SBS not only improves the clinical skills, but also the soft skills of nursing students. However, the effective outcomes and impacts can only be achieved in the context with the appropriate learning materials and equipment, simulation facilities and the instructors with pedagogical skills

The global response: How cities and provinces around the globe tackled Covid-19 outbreaks in 2021

Background: Tackling the spread of COVID-19 remains a crucial part of ending the pandemic. Its highly contagious nature and constant evolution coupled with a relative lack of immunity make the virus difficult to control. For this, various strategies have been proposed and adopted including limiting contact, social isolation, vaccination, contact tracing, etc. However, given the heterogeneity in the enforcement of these strategies and constant fluctuations in the strictness levels of these strategies, it becomes challenging to assess the true impact of these strategies in controlling the spread of COVID-19.Methods: In the present study, we evaluated various transmission control measures that were imposed in 10 global urban cities and provinces in 2021 Bangkok, Gauteng, Ho Chi Minh City, Jakarta, London, Manila City, New Delhi, New York City, Singapore, and Tokyo.Findings: Based on our analysis, we herein propose the population-level Swiss cheese model for the failures and pit-falls in various strategies that each of these cities and provinces had. Furthermore, whilst all the evaluated cities and provinces took a different personalized approach to managing the pandemic, what remained common was dynamic enforcement and monitoring of breaches of each barrier of protection. The measures taken to reinforce the barriers were adjusted continuously based on the evolving epidemiological situation.Interpretation: How an individual city or province handled the pandemic profoundly affected and determined how the entire country handled the pandemic since the chain of transmission needs to be broken at the very grassroot level to achieve nationwide control

Safety and efficacy of fluoxetine on functional outcome after acute stroke (AFFINITY): a randomised, double-blind, placebo-controlled trial

Background Trials of fluoxetine for recovery after stroke report conflicting results. The Assessment oF FluoxetINe In sTroke recoverY (AFFINITY) trial aimed to show if daily oral fluoxetine for 6 months after stroke improves functional outcome in an ethnically diverse population. Methods AFFINITY was a randomised, parallel-group, double-blind, placebo-controlled trial done in 43 hospital stroke units in Australia (n=29), New Zealand (four), and Vietnam (ten). Eligible patients were adults (aged ≥18 years) with a clinical diagnosis of acute stroke in the previous 2–15 days, brain imaging consistent with ischaemic or haemorrhagic stroke, and a persisting neurological deficit that produced a modified Rankin Scale (mRS) score of 1 or more. Patients were randomly assigned 1:1 via a web-based system using a minimisation algorithm to once daily, oral fluoxetine 20 mg capsules or matching placebo for 6 months. Patients, carers, investigators, and outcome assessors were masked to the treatment allocation. The primary outcome was functional status, measured by the mRS, at 6 months. The primary analysis was an ordinal logistic regression of the mRS at 6 months, adjusted for minimisation variables. Primary and safety analyses were done according to the patient's treatment allocation. The trial is registered with the Australian New Zealand Clinical Trials Registry, ACTRN12611000774921. Findings Between Jan 11, 2013, and June 30, 2019, 1280 patients were recruited in Australia (n=532), New Zealand (n=42), and Vietnam (n=706), of whom 642 were randomly assigned to fluoxetine and 638 were randomly assigned to placebo. Mean duration of trial treatment was 167 days (SD 48·1). At 6 months, mRS data were available in 624 (97%) patients in the fluoxetine group and 632 (99%) in the placebo group. The distribution of mRS categories was similar in the fluoxetine and placebo groups (adjusted common odds ratio 0·94, 95% CI 0·76–1·15; p=0·53). Compared with patients in the placebo group, patients in the fluoxetine group had more falls (20 [3%] vs seven [1%]; p=0·018), bone fractures (19 [3%] vs six [1%]; p=0·014), and epileptic seizures (ten [2%] vs two [<1%]; p=0·038) at 6 months. Interpretation Oral fluoxetine 20 mg daily for 6 months after acute stroke did not improve functional outcome and increased the risk of falls, bone fractures, and epileptic seizures. These results do not support the use of fluoxetine to improve functional outcome after stroke

Mapping 123 million neonatal, infant and child deaths between 2000 and 2017

Since 2000, many countries have achieved considerable success in improving child survival, but localized progress remains unclear. To inform efforts towards United Nations Sustainable Development Goal 3.2—to end preventable child deaths by 2030—we need consistently estimated data at the subnational level regarding child mortality rates and trends. Here we quantified, for the period 2000–2017, the subnational variation in mortality rates and number of deaths of neonates, infants and children under 5 years of age within 99 low- and middle-income countries using a geostatistical survival model. We estimated that 32% of children under 5 in these countries lived in districts that had attained rates of 25 or fewer child deaths per 1,000 live births by 2017, and that 58% of child deaths between 2000 and 2017 in these countries could have been averted in the absence of geographical inequality. This study enables the identification of high-mortality clusters, patterns of progress and geographical inequalities to inform appropriate investments and implementations that will help to improve the health of all populations