Compensatory Modulation of Seed Storage Protein Synthesis and Alteration of Starch Accumulation by Selective Editing of 13 kDa Prolamin Genes by CRISPR-Cas9 in Rice.

Rice prolamins are categorized into three groups by molecular size (10, 13, or 16 kDa), while the 13 kDa prolamins are assigned to four subgroups (Pro13a-I, Pro13a-II, Pro13b-I, and Pro13b-II) based on cysteine residue content. Since lowering prolamin content in rice is essential to minimize indigestion and allergy risks, we generated four knockout lines using CRISPR-Cas9, which selectively reduced the expression of a specific subgroup of the 13 kDa prolamins. These four mutant rice lines also showed the compensatory expression of glutelins and non-targeted prolamins and were accompanied by low grain weight, altered starch content, and atypically-shaped starch granules and protein bodies. Transcriptome analysis identified 746 differentially expressed genes associated with 13 kDa prolamins during development. Correlation analysis revealed negative associations between genes in Pro13a-I and those in Pro13a-II and Pro13b-I/II subgroups. Furthermore, alterations in the transcription levels of 9 ER stress and 17 transcription factor genes were also observed in mutant rice lines with suppressed expression of 13 kDa prolamin. Our results provide profound insight into the functional role of 13 kDa rice prolamins in the regulatory mechanisms underlying rice seed development, suggesting their promising potential application to improve nutritional and immunological value.

Down-Regulation of Rice Glutelin by CRISPR-Cas9 Gene Editing Decreases Carbohydrate Content and Grain Weight and Modulates Synthesis of Seed Storage Proteins during Seed Maturation.

The glutelins are a family of abundant plant proteins comprised of four glutelin subfamilies (GluA, GluB, GluC, and GluD) encoded by 15 genes. In this study, expression of subsets of rice glutelins were suppressed using CRISPR-Cas9 gene-editing technology to generate three transgenic rice variant lines, GluA1, GluB2, and GluC1. Suppression of the targeted glutelin genes was confirmed by SDS-PAGE, Western blot, and q-RT-PCR. Transgenic rice variants GluA1, GluB2, and GluC1 showed reduced amylose and starch content, increased prolamine content, reduced grain weight, and irregularly shaped protein aggregates/protein bodies in mature seeds. Targeted transcriptional profiling of immature seeds was performed with a focus on genes associated with grain quality, starch content, and grain weight, and the results were analyzed using the Pearson correlation test (requiring correlation coefficient absolute value ≥ 0.7 for significance). Significantly up- or down-regulated genes were associated with gene ontology (GO) and KEGG pathway functional annotations related to RNA processing (spliceosomal RNAs, group II catalytic introns, small nucleolar RNAs, microRNAs), as well as protein translation (transfer RNA, ribosomal RNA and other ribosome and translation factors). These results suggest that rice glutelin genes may interact during seed development with genes that regulate synthesis of starch and seed storage proteins and modulate their expression via post-transcriptional and translational mechanisms.

Exploring the Sub-nanoscale Structure of Cobalt Molybdenum Sulfide and the Role of a Cobalt Promoter in Catalytic Hydrogen Evolution.

Cobalt-promoted molybdenum sulfide (CoMoS) is known as a promising catalyst for H2 evolution reaction and hydrogen desulfurization reaction. This material exhibits superior catalytic activity as compared to its pristine molybdenum sulfide counterpart. However, revealing the actual structure of cobalt-promoted molybdenum sulfide as well as the plausible contribution of a cobalt promoter is still challenging, especially when the material has an amorphous nature. Herein, we report, for the first time, on the use of positron annihilation spectroscopy (PAS), being a nondestructive nuclear radiation-based method, to visualize the position of a Co promoter within the structure of MoS at the atomic scale, which is inaccessible by conventional characterization tools. It is found that at low concentrations, a Co atom occupies preferably the Mo-vacancies, thus generating the ternary phase CoMoS whose structure is composed of a Co-S-Mo building block. Increasing the Co concentration, e.g., a Co/Mo molar ratio of higher than 1.12/1, leads to the occupation of both Mo-vacancies and S-vacancies by Co. In this case, secondary phases such as MoS and CoS are also produced together with the CoMoS one. Combining the PAS and electrochemical analyses, we highlight the important contribution of a Co promoter to enhancing the catalytic H2 evolution activity. Having more Co promoter in the Mo-vacancies promotes the H2 evolution rate, whereas having Co in the S-vacancies causes a drop in H2 evolution ability. Furthermore, the occupation of Co to the S-vacancies leads also to the destabilization of the CoMoS catalyst, resulting in a rapid degradation of catalytic activity.

The Willingness of Parents to Vaccinate Their Children Aged from Five to under Twelve Years with COVID-19 Vaccines between February and March 2022 in Vietnam.

The current study used data surveyed with 5357 parents/guardians (parents would be used to represent both 'parents' and 'guardians' hereafter) between February and March 2022 in Vietnam to examine their willingness to vaccinate their children with current COVID-19 vaccines. It applied the multinomial logistic regression model to examine the association between the willingness of parents and selected influential factors. In addition, the reasons that made parent hesitant or unwilling to vaccinate their children were investigated. Moreover, it identified parents' preferences for vaccine origins. Approximately, 75.4% of the parents were willing, 21.3% were hesitant and 3.3% were unwilling to vaccinate their children. The most common reasons that made the parents hesitant or unwilling to vaccinate their children were their concerns about the vaccine safety, efficacy and immunity. The most and the second most preferred vaccines were those developed/originated in the US and EU, respectively. Parents who were more likely to vaccinate their children included those whose children were insured, who regularly vaccinated their children, who belonged to the vaccine priority groups, who possessed sufficient knowledge about the ways to prevent the virus or about the herd immunity, and who perceived that their children might be infected with the virus and whose children were afraid of needles. Parents who were less likely to vaccinate their children included those who were the family main income source, who had savings, and who had tertiary education or higher.

Electronic Device Use before Bedtime and Sleep Quality among University Students.

Using electronic devices before bedtime impacts sleep quality and has become a major public health issue. This study aims to investigate the associations between electronic devices (EDs) use before bedtime and sleep quality in Vietnamese university students. A total of 369 university students from three departments were recruited. Participants completed self-report surveys, including demographic characteristics, lifestyle, ED-use behaviors, the Pittsburgh Sleep Quality Index, and the Center for Epidemiologic Studies Depression Scale. A total of 48.8% of the students experienced poor sleep quality, and 98.1% reported using at least one type of ED every day within two hours before bedtime. Smartphones are the most used devices (92.3%). ED usage within two hours before bedtime (p = 0.031), lack of exercise (p = 0.006), alcohol consumption (p = 0.025), and coffee intake after 4 pm (p = 0.018) were associated with poor sleep quality. ED use near bedtime for a duration longer than 30 min (p = 0.001) and depression (p < 0.001) were associated with poorer sleep quality among university students. ED use near bedtime more than 30 min was significantly associated with poorer sleep quality after adjusting depression status, exercise, and caffeine/alcohol intake in the latter part of the day. This study emphasizes the importance of adequate sleep and restriction of ED use near bedtime, which are necessary for better sleep in university students.

Clinical performance of a semi-quantitative assay for SARS-CoV2 IgG and SARS-CoV2 IgM antibodies.

BACKGROUND: While the diagnosis of SARS-CoV-2 infection is primarily based on detection of viral RNA, the detection of SARS-CoV-2 antibodies is useful for assessing past prevalence of the disease, and in corroborating a current infection in challenging cases. Sensitive and specific immunoassays provide the ability to identify exposure to SARS-CoV-2, to determine seroconversion, to confirm eligibility for donation of convalescent plasma as well as play an essential part in epidemiological studies. We report on the validation of the Ansh Laboratories SARS-CoV-2 IgG and SARS-CoV-2 IgM ELISA immunoassays. These assays were evaluated for detection of anti-SARS-CoV-2 IgG and IgM antibodies for clinical use in our hospital as part of an orthogonal testing algorithm recommended by the CDC. METHODS: Diagnostic specificity and sensitivity of the IgG and IgM ELISA assays were tested using samples confirmed to be negative or positive for COVID-19 by RT-PCR. We also evaluated precision, analytical interference, and cross-reactivity with known cases of infection with other viruses. Additionally, we validated concordance with molecular and other serological testing and evaluated seroconversion in our patient population. RESULTS: The IgG and IgM ELISA assays showed acceptable precision, were robust to analytical interference and did not exhibit cross reactivity with specimens positive for common respiratory viruses. Both assays exhibited 95% agreement with a primary screening serological assay utilized at our institution as well as with a reference laboratory semi-quantitative method. Concordance with RT-PCR was excellent > 6 days after symptom onset (100%). CONCLUSIONS: The Ansh SARS-CoV-2 ELISA assays have good analytical performance suitable for clinical use.

Study on evaluation of bioavailability of nifedipine tablets

The HPLC method described herein has sufficient sensitivity to determine the pharmacokinetic parameters of nifedipine in plasma. A single dose cross study was carried out in volunteers comparing the test sustained release nifedipine formulation to Adalat retard tablets. Pharmacokenetic variables were calculated from the nifedipine plasma concentration data and evaluated statistically. The results showed the test formulation to be bioequivalent to the reference tablets.