OPTIMIZING THE PRODUCTION OF A FUNCTIONAL TYPE A RECOMBINANT ENDOCHITINASE FROM Trichoderma asperellum IN Escherichia coli

Chitinases from the genus Trichoderma fungi are mainly responsible for their anti-fungal activities, which allow them to become the most widely used fungal biocontrol. Therefore, several Trichoderma chitinases have been cloned and expressed to facilitate their production and applications. A previous study of the same authors has characterized an endochitinase from a relatively novel Trichoderma spp., Trichoderma asperellum. To produce this enzyme more economically and efficiently, we reported the synthesis and expression of its synthetic encoding gene in the Escherichia coli M15 strain and established the optimal conditions for preparative scale production of the enzyme in its functional form. By lowering the induction temperatures, we observed substantial improvement in the expression levels of the active enzyme.  At 30 oC and 0.5 mM IPTG induction, 1 L of cells yielded approximately 80 - 100 mg of soluble protein, accounting for about 9-11 % of total soluble protein. This figure may be an underestimation of the actual yield, as deduced from the SDS-PAGE data. The recombinant enzyme can be retrieved by simple repeated freezing and thawing cycles and purified to near homogeneity using Ni-NTA chromatography. The purified enzyme showed in vitro colloidal chitin hydrolysis activity. These results could be scaled up to produce soluble 42 kDa chitinase in E. coli. The study demonstrated an economical method to produce chitinases for various agricultural and environmental applications

High Proportion of Genome-Wide Homology and Increased Pretreatment pvcrt Levels in Plasmodium vivax Late Recurrences: a Chloroquine Therapeutic Efficacy Study.

Chloroquine (CQ) is the first-line treatment for Plasmodium vivax malaria in most countries where malaria is endemic. Monitoring P. vivax CQ resistance (CQR) is critical but remains challenged by the difficulty to distinguish real treatment failure from reinfection or liver relapse. The therapeutic efficacy of CQ against uncomplicated P. vivax malaria was evaluated in Gia Lai Province, Vietnam. Sixty-seven patients were enrolled and followed for 42 days using microscopy and quantitative PCR. Adequate clinical and parasitological response (ACPR) was 100% (66/66) on day 28 but 75.4% (49/65) on day 42. Eighteen recurrences (27.7%) were detected, with a median time to recurrence of 42 days (interquartile range [IQR], 35 to 42) and blood CQ concentration of 98% identity by descent to paired day 0 samples. This study shows that CQ remained largely efficacious to treat P. vivax in Gia Lai; i.e., recurrences occurred late (>day 28) and in the presence of low blood CQ concentrations. However, the combination of both WGS and gene expression analysis (pvcrt) data with clinical data (PCT) allowed us to identify potential emergence of low-grade CQR, which should be closely monitored. (This study has been registered at ClinicalTrials.gov under identifier NCT02610686.)

Awareness and preparedness of healthcare workers against the first wave of the COVID-19 pandemic: A cross-sectional survey across 57 countries.

BACKGROUND: Since the COVID-19 pandemic began, there have been concerns related to the preparedness of healthcare workers (HCWs). This study aimed to describe the level of awareness and preparedness of hospital HCWs at the time of the first wave. METHODS: This multinational, multicenter, cross-sectional survey was conducted among hospital HCWs from February to May 2020. We used a hierarchical logistic regression multivariate analysis to adjust the influence of variables based on awareness and preparedness. We then used association rule mining to identify relationships between HCW confidence in handling suspected COVID-19 patients and prior COVID-19 case-management training. RESULTS: We surveyed 24,653 HCWs from 371 hospitals across 57 countries and received 17,302 responses from 70.2% HCWs overall. The median COVID-19 preparedness score was 11.0 (interquartile range [IQR] = 6.0-14.0) and the median awareness score was 29.6 (IQR = 26.6-32.6). HCWs at COVID-19 designated facilities with previous outbreak experience, or HCWs who were trained for dealing with the SARS-CoV-2 outbreak, had significantly higher levels of preparedness and awareness (p<0.001). Association rule mining suggests that nurses and doctors who had a 'great-extent-of-confidence' in handling suspected COVID-19 patients had participated in COVID-19 training courses. Male participants (mean difference = 0.34; 95% CI = 0.22, 0.46; p<0.001) and nurses (mean difference = 0.67; 95% CI = 0.53, 0.81; p<0.001) had higher preparedness scores compared to women participants and doctors. INTERPRETATION: There was an unsurprising high level of awareness and preparedness among HCWs who participated in COVID-19 training courses. However, disparity existed along the lines of gender and type of HCW. It is unknown whether the difference in COVID-19 preparedness that we detected early in the pandemic may have translated into disproportionate SARS-CoV-2 burden of disease by gender or HCW type

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

A Physically Based Model for the Streaming Potential Coupling Coefficient in Partially Saturated Porous Media

International audienceThe electrokinetics methods have great potential to characterize hydrogeological processes in porous media, especially in complex partially saturated hydrosystems (e.g., the vadose zone). The dependence of the streaming coupling coefficient on water saturation remains highly debated in both theoretical and experimental works. In this work, we propose a physically based model for the streaming potential coupling coefficient in porous media during the flow of water and air under partially saturated conditions. The proposed model is linked to fluid electrical conductivity, water saturation, irreducible water saturation, and microstructural parameters of porous materials. In particular, the surface conductivity of porous media has been taken into account in the model. In addition, we also obtain an expression for the characteristic length scale at full saturation in this work. The proposed model is successfully validated using experimental data from literature. A relationship between the streaming potential coupling coefficient and the effective excess charge density is also obtained in this work and the result is the same as those proposed in literature using different approaches. The model proposes a simple and efficient way to model the streaming potential generation for partially saturated porous media and can be useful for hydrogeophysical studies in the critical zone

Synthesis and characterization of magnetic properties of nanocrystalline perovskite-type La1-xEuxFeO3 (0 ≤ x ≤ 0.5)

Perovskite ABO3 are kinds of materials caring many interesting electric and magnetic properties; specially their properties can change dramatically by doping with rare-earth metals. In this paper, nanocrystalline La1-xEuxFeO3 (0 ≤ x ≤ 0.5) was prepared by sol-gel method. The research on the structure by X-ray diffraction (XRD) analysis and Scanning Electron Microscope (SEM) shows that the structure of crystal changes from cubic to orthorhombic when x &gt; 0.3. Average size of particles equals 19nm and decreases with the increasing of Eu doped. The study on magnetic properties shows that these materials are weak ferromagnetism closing to superparamagnetism. The ratio of magnetic remanence and saturation Mr/Ms is nearly equal to zero. Magnetic properties of materials varying with the increasing of unit volume proves that magnetic properties depend on crystal distortion of material

Electroosmotic Coupling in Porous Media, a New Model Based on a Fractal Upscaling Procedure

International audienceElectrokinetic and electroosmotic couplings can play important roles in water and ions transport in charged porous media. Electroosmosis is the phenomena explaining the water movement in a porous medium subjected to an electrical field. In this work, a new model is obtained through a new up-scaling procedure, considering the porous medium as a bundle of tortuous capillaries of fractal nature. From the model, the expressions for the electroosmosis pressure coefficient, the relative electroosmosis pressure coefficient, the maximum back pressure, the maximum flow rate, the flow rate-applied back pressure relation and the product of the permeability and formation factor of porous media are also obtained. The sensitivity of the relative electroosmosis pressure coefficient is then analyzed and explained. The model predictions are then successfully compared with published datasets. Additionally, we deduce an expression for the relative streaming potential coefficient and then compare it with a previously published model and experimental data from a dolomite rock sample. We find a good agreement between those models and experimental data, opening up new perspectives to model electroosmotic phenomena in porous media saturated with various fluids

Knowledge, attitude and practice on fall risk factors and prevention among rural older community-dwellers in Vietnam.

Falls among the elderly are an important global health problem. This study assesses knowledge regarding risk factors of falls, as well as attitudes and practices towards fall prevention among older adults in the rural community. A cross-sectional study was performed in four rural communes in Thai Binh province, Vietnam. A total of 3038 older people were recruited. Knowledge was assessed by using Falls Risk Awareness Questionnaire. Questions about attitudes were based on the Health Belief Model. Other questions regarding attitudes and practices regarding fall prevention were also asked. Multivariate regression was performed to identify associated factors with knowledge, attitudes and practices. Results showed that the mean score of knowledge regarding risk factors of falls was low at 11.37/32. The highest scores were observed in terms of drug aspects, followed by medical condition and behavioural aspects. Older people mostly agreed with perceived severity, susceptibility, benefits and barriers, but their attitudes about cues to action, health motivation and actions were most neutral. Meanwhile, more than half of the participants practised recommended fall preventive measures. To conclude, health education interventions and fall prevention services that enhance community-based fall prevention knowledge, attitudes and practices for older adults should be performed to reduce the burden of falls in this population