Growth and development of transgenic peanut (Arachis hypogaea) lines containing chitinase 42 kDa gene from Trichoderma asperellum SH16

Peanut (Arachis hypogaea L.) is vulnerable to many diseases. Vietnam and other regions where peanut is widely cultivated have a high threat of fungal and other plant diseases. Various fungicides are available to control the fungal disease but these have various harmful effects on the natural flora, fauna, and environment. Transgenic peanut lines which possess antifungal activity provide a possible solution in managing fungal diseases apart from the traditional resistance and fungicide usage. Therefore, this study evaluated the probable growth and development of chitinase transgenic peanut lines against Sclerotium rolfsii, a pathogen that causes “southern blight” in plants, under greenhouse conditions. This study provided evidence that through Agrobacterium itumefaciens mediated transformation, 42 kDa chitinase genes from Trichoderma asperellum, which is under the regulation of 35S promoter, were successfully incorporated into the peanut’s (A. hypogaea L.) genome and expressed in their plants. This evidence also demonstrated that transgenic peanut lines were suitable for growing and developing in the greenhouse. Further, it was reported that transgenic peanut lines took approximately 133 to 145 days from planting to maturity. These results also revealed that various growth characteristics of transgenic peanut lines having two synthetic genes (syncod Chi42-2 i.e. S2-2, S2-4, S2-6, and syncod Chi42-1 i.e. S1-1, S1-2, S1-3) were greater than that from the wild-type Chi42 (WT-1, WT-2, and WT-3). In addition, yield-related parameters including the number of mature pods, 100 pods weight and 100 seeds weight for all the transgenic peanut lines were higher than that of the non-transformed plant. Among the transgenic lines, line S2-4 exhibited significantly higher growth and yield than the other transgenic lines. These results demonstrated that 42 kDa chitinase genes overexpressing peanut lines could be a candidate for improvement against plants to phytopathogenic fungus S. rolfsii and high yield.

TẠO DÒNG CÁC GEN MÃ HÓA CHITINASE 42 kDa CỦA Trichoderma asperellum VÀO VECTOR BIỂU HIỆN THỰC VẬT pMYV719 ĐỂ PHỤC VỤ CHUYỂN GEN

In this study, chitinase genes containing a signal peptide sequence, such as Chi42, syncodChi42-1 and syncodChi42-2, were cloned in the plant expression vector pMYV719 and successfully transferred into Agrobacterium tumefaciens LBA 4404. Among them, Chi42 is a wild-type gene of Trichoderma asperellum SH16. Both genes syncodChi42-1 and syncodChi42-2 are derived from Chi42, which was optimized for codon usage for plant expression. Agrobacterium bacteria-harbouring pMYV719/chitinase vector was used for genetic transformation into peanuts (Arachis hypogaea L.) to enhance resistance to phytopathogenic fungi in further studies.Trong nghiên cứu này, các gen chitinase mang trình tự peptide tín hiệu như Chi42, syncodChi42-1 và syncodChi42-2 đã được tạo dòng trong vector biểu hiện thực vật pMYV719 và biến nạp thành công vào vi khuẩn Agrobacterium tumefaciens LBA 4404. Trong đó, gen Chi42 là kiểu gen hoang dại từ chủng nấm Trichoderma asperellum SH16. Hai gen syncodChi42-1 và syncodChi42-2 có nguồn gốc từ gen Chi42 đã được tối ưu hóa bộ ba sử dụng để biểu hiện thực vật. Vi khuẩn A. tumefaciens mang các gen chitinase được sử dụng để chuyển gen vào cây lạc (Arachis hypogaea L.) trong các nghiên cứu tiếp theo để cải thiện khả năng kháng nấm bệnh của chúng

Multiple Wolbachia strains provide comparative levels of protection against dengue virus infection in Aedes aegypti.

The insect bacterium Wolbachia pipientis is being introgressed into Aedes aegypti populations as an intervention against the transmission of medically important arboviruses. Here we compare Ae. aegypti mosquitoes infected with wMelCS or wAlbB to the widely used wMel Wolbachia strain on an Australian nuclear genetic background for their susceptibility to infection by dengue virus (DENV) genotypes spanning all four serotypes. All Wolbachia-infected mosquitoes were more resistant to intrathoracic DENV challenge than their wildtype counterparts. Blocking of DENV replication was greatest by wMelCS. Conversely, wAlbB-infected mosquitoes were more susceptible to whole body infection than wMel and wMelCS. We extended these findings via mosquito oral feeding experiments, using viremic blood from 36 acute, hospitalised dengue cases in Vietnam, additionally including wMel and wildtype mosquitoes on a Vietnamese nuclear genetic background. As above, wAlbB was less effective at blocking DENV replication in the abdomen compared to wMel and wMelCS. The transmission potential of all Wolbachia-infected mosquito lines (measured by the presence/absence of infectious DENV in mosquito saliva) after 14 days, was significantly reduced compared to their wildtype counterparts, and lowest for wMelCS and wAlbB. These data support the use of wAlbB and wMelCS strains for introgression field trials and the biocontrol of DENV transmission. Furthermore, despite observing significant differences in transmission potential between wildtype mosquitoes from Australia and Vietnam, no difference was observed between wMel-infected mosquitoes from each background suggesting that Wolbachia may override any underlying variation in DENV transmission potential

The effect of different media and temperature conditions for Salmonella bacteriophage preservation

This research aimed to determine the optimal media and temperature conditions for the long-term storage of bacteriophages. In this study, the viability of Salmonella phages in 50% glycerol, 10% sodium chloride-magnesium sulfate (SM) buffer, and 5% dimethyl sulfoxide (DMSO) media at room temperature, 4oC, -20oC, and -80oC for 12 months was determined. In 50% glycerol, at the end of the experiment, no significant difference was found between four temperature conditions on phage density, ranging from 6.20-6.23 log10 PFU/mL (P>0.05). Under 10% SM medium, phage preservation at room temperature provided the optimum density at 6.31 log10 PFU/mL. In addition, phages preserved in a 5% DMSO medium were of similar density values across all temperature treatments. Still, their availability after 12 month-storage (88.0-88.5%) was significantly lower (P<0.05) than that of 50% glycerol and 10% SM. Moreover, for phage lysis capacity, low temperatures (4oC, -20oC, and -80oC) were superior to room temperature used for preservation. Considering the density, lysis capacity, and practical convenience, storing phages at 4°C in a 50% Glycerol medium is recommended

Genome evolution of dengue virus serotype 1 under selection by <i>Wolbachia pipientis</i> in <i>Aedes aegypti</i> mosquitoes.

The introgression of antiviral strains of Wolbachia into Aedes aegypti mosquito populations is a public health intervention for the control of dengue. Plausibly, dengue virus (DENV) could evolve to bypass the antiviral effects of Wolbachia and undermine this approach. Here, we established a serial-passage system to investigate the evolution of DENV in Ae. aegypti mosquitoes infected with the wMel strain of Wolbachia. Using this system, we report on virus genetic outcomes after twenty passages of serotype 1 of DENV (DENV-1). An amino acid substitution, E203K, in the DENV-1 envelope protein was more frequently detected in the consensus sequence of virus populations passaged in wMel-infected Ae. aegypti than wild-type counterparts. Positive selection at residue 203 was reproducible; it occurred in passaged virus populations from independent DENV-1-infected patients and also in a second, independent experimental system. In wild-type mosquitoes and human cells, the 203K variant was rapidly replaced by the progenitor sequence. These findings provide proof of concept that wMel-associated selection of virus populations can occur in experimental conditions. Field-based studies are needed to explore whether wMel imparts selective pressure on DENV evolution in locations where wMel is established

Chemical Components of Essential Oils From the Leaves of Seven Species Belonging to Rutaceae Family from Binh Chau-Phuoc Buu Nature Reserve, Vietnam

Several plant species of the Rutaceae family are medicinal plants, oil bearing and food crops. To provide more information for utilization of some species of this family in Binh Chau-Phuoc Buu Nature Reserve, we extracted essential oils from the leaves of seven species of the Rutaceae family: Acronychia pedunculata (L.) Miq., Atalantia citroides Pierre ex Guillaumin, Clausena excavata Burm.f., Glycosmis pentaphylla (Retz.) DC., Luvunga scandens (Roxb.) Buch.-Ham. ex Wight & Arn, Melicope pteleifolia (Champ. ex Benth.) T.G. Hartley, and Micromelum sp., via hydrodistillation, and identified their components using GC/MS analysis. A total of 60 compounds were identified from essential oils of seven species. The main components of the essential oils isolated from five species, including A. pedunculata, C. excavata, M. pteleifolia, G. pentaphylla, and Micromelum sp., were caryophyllene (57.63% and 55.41% in A. pedunculata and C. excavata, respectively), 1,9-decadiyne (32.59%, M. pteleifolia), β-ocimene (23.10%, G. pentaphylla), and 3-carene (58.03%, Micromelum sp.). Additionally, this study revealed the chemical composition of essential oils of L. scandens and A. citroides for the first time. The main constituent of A. citroides was 7-oxabicyclo[4.1.0] heptane, 3-oxiranyl- (53.91%) and that of L. scandens was caryophyllene (34.66%). These findings provide the basis for further application of these species in medicine

Successful Psoriasis Treatment Using NB-UVB with Methotrexate: The Vietnamese Experience

AIM: To compare the effectiveness of narrowband ultraviolet B (NBUVB) and oral methotrexate (MTX) to oral MTX alone in Vietnamese psoriasis patients, from May 2016 to May 2018. METHODS: We conducted a non-randomized trial on 70 patients with plaque-type psoriasis of moderate to severe. Thirty-five patients apply NBUVB once/day in 5 days/week for 4 weeks plus oral MTX 7.5 mg/week and 35 patients oral MTX 7.5 mg/week and both two groups treatment for 3 months. The extent of the lesion was assessed by the Psoriasis Area and Severity Index (PASI). RESULTS: The proportion of decreasing PASI was comparable (68.49% in NBUVB and MTX versus 57.62% in MTX alone); p &lt; 0.05. Inside, good 28.58%, moderate 68.57% and poor 2.85% in NBUVB and MTX better than good 2.85%, moderate 71.4% and poor 25.72% in MTX alone; p &lt; 0.05. The recurrence rate after 24 months of the NBUVB and MTX group (42.9%) was lower than the MTX alone group (71.4%); p &lt; 0.05. CONCLUSION: NBUVB and oral MTX have affected treatment with chronic plaque psoriasis better than oral MTX alone

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