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.

Purification and Characterization of a Small Thermostable Protease from Streptomyces sp. CNXK100.

Proteases derived from Streptomyces demonstrate numerous commendable properties, rendering it extensively applicable in biotechnology and various industrial sectors. This study focused on the purification and characterization of the thermostable protease obtained from Streptomyces sp. CNXK100. The purified protease exhibited an estimated molecular weight of 27 kDa, with optimal activity at 75°C and pH 8.0. Notably, the enzyme remained active even without any metal ions and fully active in the presence of Na+, K+, Mg2+, and Cu2+metal ions. The kinetic parameters were determined with a KM value of 3.13 mg/ml and a Vmax value of 3.28 × 106 U/mg. Furthermore, the protease has demonstrated notable stability when subjected to a treatment temperature of up to 65°C for 60 minutes, and across a broad pH range extending from 5.0 to 10.0. This protease also demonstrated resilience against a spectrum of harsh conditions, including exposure to organic solvents, surfactants, bleaching agents, and proteolytic enzymes. Additionally, the enzyme maintained its activity following treatment with commercial detergents, accomplishing complete thrombus lysis at a concentration of 2.50 mg/ml within 4 hours. Remarkably, the protease exhibited stability in terms of activity and protein concentration for 70 days at 4°C. These findings underscore the potential industrial applications of the thermostable protease from Streptomyces sp. CNXK100.

Lower Extremity Nerve Conduction Abnormalities in Vietnamese Patients with Type 2 Diabetes: A Cross-Sectional Study on Peripheral Neuropathy and Its Correlation with Glycemic Control and Renal Function.

Peripheral neuropathy is a common complication of type 2 diabetes mellitus (T2DM) that results in nerve conduction abnormalities. This study aimed to investigate the parameters of nerve conduction in lower extremities among T2DM patients in Vietnam. A cross-sectional study was conducted on 61 T2DM patients aged 18 years and older, diagnosed according to the American Diabetes Association's criteria. Data on demographic characteristics, duration of diabetes, hypertension, dyslipidemia, neuropathy symptoms, and biochemical parameters were collected. Nerve conduction parameters were measured in the tibial and peroneal nerves, including peripheral motor potential time, response amplitude M, and motor conduction speed, as well as sensory conduction in the shallow nerve. The study found a high rate of peripheral neuropathy among T2DM patients in Vietnam, with decreased conduction rate, motor response amplitude, and nerve sensation. The incidence of nerve damage was highest in the right peroneal nerve and left peroneal nerve (86.7% for both), followed by the right tibial nerve and left tibial nerve (67.2% and 68.9%, respectively). No significant differences were found in the rate of nerve defects between different age groups, body mass index (BMI) groups, or groups with hypertension or dyslipidemia. However, a statistically significant association was found between the rate of clinical neurological abnormalities and the duration of diabetes (p < 0.05). Patients with poor glucose control and/or decreased renal function also had a higher incidence of nerve defects. The study highlights the high incidence of peripheral neuropathy among T2DM patients in Vietnam and the association between nerve conduction abnormalities and poor glucose control and/or decreased renal function. The findings underscore the importance of early diagnosis and management of neuropathy in T2DM patients to prevent serious complications.

Extracellular Polysaccharide Extraction from Streptococcus thermophilus in Fermented Milk.

Lactic acid bacteria such as Streptococcus thermophilus are known to produce extracellular polysaccharide (EPS) in fermented foods that enhance the creaminess and mouthfeel of the product, such as yogurt. Strains producing larger amounts of EPS are highly sought-after, and therefore, robust and accurate quantification methodologies are important. This study found that two commonly used methodologies significantly underestimated the amount of EPS produced as measured using a milk matrix. To this end, a proteolytic step was implemented prior to EPS extraction (Method C). An initial proteolytic step using xanthan gum-spiked milk significantly increased recovery yield to 64%, compared to 27.8% for Method A and 34.3% for Method B. Method C showed no improvement when assessed using a chemically defined medium. Method C was further validated using three strains of S. thermophilus with varying EPS-production capabilities (STLOW, STMID, STHIGH). Overall, Method C demonstrated significant improvements in the EPS extraction yield for all three S. thermophilus strains in fermented milk. On average, Method C improved isolation yield by ∼3- to 6-fold compared with Method A and by ∼2- to 3-fold compared with method B. There were no significant differences between samples when they were grown in a chemically defined medium, highlighting the importance of a proteolytic step specifically for fermented milk samples. In commercial applications, accurate quantification of EPS-production is an important aspect when finding new strains. IMPORTANCE Extracellular polysaccharide (EPS) production by milk-fermenting microorganisms is a highly sought-after trait in improving the perceived thickness, creaminess, and mouthfeel of yogurt. Streptococcus thermophilus are commonly isolated and their EPS production is quantified in the search for higher-producing strains. In this study, we demonstrated that two commonly used methods for isolating EPS from milk samples significantly underestimated the true amount of EPS present. We demonstrated that the addition of a proteolytic step prior to EPS extraction isolated over 2-fold more EPS than identical samples processed using the traditional protocols. We further validated this method in fermented milk samples from three strains of S. thermophilus that included a low-, mid-, and high-EPS producing strain. Again, we showed significant improvements in EPS isolation using a proteolytic step. In the search for new S. thermophilus strains with enhanced EPS production, accurate quantification in an optimal medium is essential.

Genetic structure and population connection of two Bouyei populations in northern Vietnam based on short tandem repeat analysis.

OBJECTIVES: Genetic characteristics were investigated based on short tandem repeat (STR) data to assess the relationship between two Vietnamese Bouyei populations in Vietnam. METHODS: We collected hair and buccal swab samples from two separate Bouyei populations in the mountainous region of Northern Vietnam, which are the Bo Y in Ha Giang Province and the Tu Di in Lao Cai Province. The study included data of 23 autosomal and 27 Y-chromosome STRs loci of 96 unrelated participants from a total Vietnamese Bouyei population of under 3300 individuals. RESULTS: The results showed that these STR markers are valuable for differentiation of individuals and human genetic studies in Vietnamese Bouyei populations. Genetic analysis indicated that Tu Di and Bo Y people were from the same Bouyei population in China. CONCLUSIONS: The results supported the official historical records of the region and the classification of the Vietnamese government. Furthermore, the genetic data provided in this study will be helpful in investigating the genetic genealogy evolution and settlement or migration patterns of the Bouyei populations in Vietnam.

Effect of organic matter on nitrogen removal through the anammox process.

In the anaerobic ammonium oxidation (anammox) process, the anammox bacterial activity is inhibited by high chemical oxygen demand (COD) contents. In this study, the effect of the COD/total nitrogen (TN) ratio and hydraulic retention time (HRT) on the efficiency of the anammox process in a fixed-bed reactor was assessed via experiments with increasing COD/TN ratios from 0 to 7.0 and different HRTs of 6, 9, and 12 hr. Felibendy cubes were used as the biomass carrier. The presence of organic compounds affects the performance of the treatment process, and the level of influence increases with the increase in influent COD. With COD greater than 300 mg/L, the anammox process was inhibited, TN efficiency was the lowest, and effluent water quality did not yield effluent standards. The kinetic parameters Umax (the maximum substrate removal rate constant) and KB (the saturation constant) of the Stover-Kincannon model corresponding to the different COD/TN ratios were determined in this study. The kinetic parameter values of the ammonium removal process decreased gradually from Umax  = 0.685 g/L/day and KB  = 0.846 g/L/day, for COD/TN = 0, to Umax  = 0.314 g/L/day and KB  = 0.498 g/L/day, for COD/TN = 6. The kinetic parameter of the TN removal process also decreased gradually as the COD/TN ratio in wastewater increased gradually. This means that the anammox process was inhibited as much as high organic compounds. PRACTITIONER POINTS: The higher COD/TN ratios the more anammox activity is affected, especially COD/TN = 7.0 (COD = 350 mg/L). The tolerance threshold of anammox process about organic matter concentration is 300 mg/L as COD (COD/TN ratio <6). With the COD/TN ratios from 1.0 to 6.0, the values of Umax and KB constants of Stover-Kincannon model for ammonium removal by Anammox process decrease gradually from 0.685 to 0.314 and from 0.846 to 0.498, respectively.

Postnatal depressive symptoms display marked similarities across continents.

BACKGROUND: Postnatal depressive symptoms measured by the Edinburgh Postnatal Depression Scale (EPDS) are reported to display measurement variance regarding factor structure and the frequency of specific depressive symptoms. However, postnatal depressive symptoms measured by EPDS have not been compared between women representing three continents. METHODS: A cross-sectional study including birth cohort samples from Denmark, Vietnam and Tanzania. Women were included during pregnancy at routine care sites. Depressive symptoms were self-reported 40-90 days postpartum using the EPDS. Exploratory and confirmatory factor analyses and generalized additive regression models were performed. RESULTS: A total of N = 4,516 participated in the study (Denmark N = 2,069, Vietnam N = 1,278, Tanzania N = 1,169). Factor analyses identified three factors (anhedonia, anxiety and depression) that were almost identical in the three study populations. The only variation between countries was that the item 'self-harm' loaded differently. Women from Tanzania and Denmark were more likely to have an EPDS total score above cut-off 12 (12.6% and 6.4%), compared to women from Vietnam (1.9%) (p<0.001). A low level of education was associated with significantly more depressive symptoms after adjusting for country (p<0.001). LIMITATIONS: EPDS data was collected at a later time point in the Danish sample. CONCLUSIONS: Postnatal depressive symptoms constitute a three-factor model across cultures including the factors anhedonia, anxiety and depression. The frequency of postnatal depressive symptoms differs between high-, medium-, and low-income countries. However, clinicians should bear in mind that low-educated women worldwide are more likely to experience postnatal depressive symptoms.

Genetic characteristics of 23 Y-chromosomal STRs in the Kinh population in Northern Vietnam.

We collected and analysed the genetic characteristics of 23 Y-STR data of 200 unrelated Kinh individuals living in the North of Vietnam. Haplotype frequencies and forensic parameters were calculated, showing high discrimination value. Population comparison analysis was performed to determine the genetic relationship with neighbouring ethnicities, in particular with Thai and Han populations.

Emotional violence exerted by intimate partners and postnatal depressive symptoms among women in Vietnam: A prospective cohort study.

BACKGROUND: Previous studies have shown a relation between intimate partner violence (IPV) and postpartum depression (PPD). However, these studies have primarily focused on physical and sexual violence as predictors for postpartum depression and little attention has been given to emotional violence (EV), despite emotional violence having been well reported as the most common type of violence experienced by women. This present study aimed to investigate the association between various types of emotional experience during life with present partner and postnatal depressive symptoms among women in Vietnam. METHODS: A total of 1,274 pregnant women were recruited from 24 communities in the Dong Anh District, Hanoi, Vietnam. They were interviewed four times: (a) at enrolment (before week 24 of pregnancy); (b) at a gestational age of 30-34 weeks; (c) 24-48 hours after delivery; and (d) 4-12 weeks after delivery. Emotional violence and postnatal depressive symptoms were measured using a questionnaire developed by the World Health Organization (WHO) and the Edinburgh Postpartum Depression Scale (EPDS), respectively. RESULTS: A total of 639 (50.4%) women experienced at least one type of emotional violence with their present partner, whereas 104 women (8.2%) experienced postnatal depressive symptoms. Women exposed to emotional violence were more likely to experience postnatal depressive symptoms (OR = 3.15; 95%CI: 1.17-8.51). Other statistically significant predictors of increased postnatal depressive symptoms included type of employment, lack of family support after delivery, lower level of education, husband's preference for a specific sex of child, presence of mental disorder, and depression during pregnancy. CONCLUSIONS: Among Vietnamese women, there was a statistically significant association between exposure to emotional violence with their present partner and postpartum depression. The findings indicate an urgent need for screening for all acts of emotional violence as risk factors for postnatal depressive symptoms.

Intimate partner violence (IPV): The validity of an IPV screening instrument utilized among pregnant women in Tanzania and Vietnam.

BACKGROUND: Intimate partner violence (IPV) is a global problem that affects one-third of all women. The present study aims to develop and determine the validity of a screening instrument for the detection of IPV in pregnant women in Tanzania and Vietnam and to determine the minimum number of questions needed to identify IPV. METHOD: An IPV screening instrument based on eight questions was tested on 1,116 Tanzanian and 1,309 Vietnamese women who attended antenatal care before 24 gestational weeks. The women were re-interviewed during their 30th-34th gestational week where the World Health Organization (WHO) IPV questionnaire was used as the gold standard. In all, 255 combinations of eight different questions were first tested on the Tanzanian study population where sensitivity, specificity, positive predictive value, negative predictive value and accuracy were calculated. In the evaluation of the performance of the question combinations, different IPV types and the frequency of abusive acts were considered. The question combinations that performed best in Tanzania were subsequently evaluated in the Vietnamese study population. RESULTS: In Tanzania, a combination of three selected questions including one question on emotional IPV, one on physical IPV and one on sexual IPV was found to be most effective in identifying women who are exposed to at least one type of IPV during pregnancy (sensitivity = .80; specificity = .74). The performance of the identified combination was slightly less effective in Vietnam (sensitivity = .74; specificity = .68). Focusing on different IPV types, the best performance was found for exposure to physical IPV in both Tanzania (sensitivity = .93; specificity = .70) and Vietnam (sensitivity = .96; specificity = .55). In both countries, the sensitivity increased with the frequency of abuse whereas the specificity decreased. CONCLUSION: By asking pregnant women three simple questions we were able to identify women who were exposed to IPV during pregnancy in two different countries. The question combination performed best in assessing physical IPV where it identified 93% and 96% of Vietnamese and Tanzanian women, respectively, who were exposed to physical IPV.

Intimate partner violence during pregnancy in Vietnam: prevalence, risk factors and the role of social support.

Background: Violence against women is a global public health problem. A better understanding of risk factors for intimate partner violence (IPV) exposure during pregnancy is important to develop interventions for supporting women being exposed to IPV. Objective: The purpose of this study was to measure the prevalence of IPV during pregnancy and analyse how social support and various risk factors are associated with IPV. Methods: A cross-sectional study conducted among 1309 pregnant women in Dong Anh district, Vietnam. Information about socio-economic conditions and previous exposure to IPV was collected when women attended antenatal care before the 24th gestational week. Information about social support information and exposure to IPV during pregnancy was collected in the 30th-34th gestational week. Multivariable regression was used to identify associations between IPV, social support and other potential risk factors. Results: The prevalence of IPV exposure during pregnancy was 35.2% (Emotional violence: 32.2%; physical violence: 3.5% and sexual violence: 9.9%). There was a statistically significant association between previous IPV exposure, lack of social support and IPV exposure during pregnancy. After adjustment for socioeconomic characteristics, pregnant women who had previously been exposed to IPV were more likely to be exposed IPV at least one time (AOR = 6.3; 95% CI: 4.9-8.2) as well as multiple times (AOR = 6.0; 95% CI: 4.5-8.0). Similarly, pregnant women having a lack of social support had a higher likelihood of being exposed to IPV at least one time (AOR = 3.1; 95% CI: 2.4-3.9) or multiple times (AOR = 2.9; 95% CI: 2.2-3.8). Conclusion: IPV is relatively high during pregnancy in Vietnam. Previous exposure to IPV and lack of social support is associated with increased risk of violence exposure among pregnant women in Vietnam.

The prime questions in authentic patient's consultations: a call for additional research on current and new paradigms.

Although the 3 prime questions ("What did your doctor tell you the medication is for?" "How did your doctor tell you to take the medication?," and "What did your doctor tell you to expect?") have been recommended as a way to implement an interactive approach to patient's counseling in pharmacy, research examining how these questions are actually used in practice is relatively sparse. Qualitative approaches might assist to inform pertinent questions that might challenge prevailing paradigms. This commentary calls for a close look at how novice pharmacists in training manage these questions in real-life patient's consultations. These examples are aimed to provide preliminary observations about (1) how the prime questions in their original and modified forms are treated by pharmacists in training and patients, and (2) the interactional functions that the prime questions and similar questions may serve. Preliminary observations based on a conversation analysis of these examples show that the open-ended nature of the original prime questions sometimes leads to interactional problems such as delays in patients' responses and pharmacists' revision of the questions. Modified question formats that involve the use of specific knowledge expected to be possessed by a pharmacist, such as declarative questions and Q-word questions with concrete information, may lead to smoother interaction. Finally, questions about the purpose of the therapy may also be used to create opportunities to express empathy toward the patient or to shift the zone of expertise to the doctor. These initial findings suggest a more context sensitive and adaptive approach to communication in pharmacy.

An efficient colorimetric assay for RNA synthesis by viral RNA-dependent RNA polymerases, using thermostable pyrophosphatase.

RNA-dependent RNA polymerase (RdRp) is essential for the replication of RNA genome-containing positive-strand RNA viruses. We developed a simple colorimetric assay to quantify the RNA synthesis activity of RdRp by measuring the pyrophosphates released during nascent RNA synthesis. RNA polymerase reaction was quenched by heating at 70 °C for 5 min, during which thermostable inorganic pyrophosphatase converted the accumulated pyrophosphates into inorganic phosphates. Subsequently, the amount of inorganic phosphate was measured using a color-developing reagent. Using RdRp's from hepatitis C virus and foot-and-mouth disease virus, we demonstrate that this colorimetric assay facilitates the measurement of RNA polymerase activity.

Characterization of bacteriophage φPto-bp6g, a novel phage that lyses Pseudomonas tolaasii causing brown blotch disease in mushrooms.

The bacteriophage, φPto-bp6g, exhibited strong bactericidal activity against Pseudomonas tolaasii, the bacterium that causes brown blotch disease in cultivated mushrooms. Analysis of phage morphology with an electron microscope revealed that φPto-bp6g contains an icosahedral head and a long tail, which is classified as the family of Siphoviridae. The phage was observed to lyse P. tolaasii in the broth about 4h after inoculation, indicating a putative lytic pathway exists during bacterial growth. The whole genome of φPto-bp6g was completely sequenced, with a length of 26,499 bp and a G+C content of 42.7%. A total of 77 open reading frames (ORFs) as putative coding sequences were identified and annotated, whereas 43 ORFs possessed no homologs. Proteins of several ORFs showed similarity with proteins of a diverse group of phages, including Siphoviridae (5 ORFs), Myoviridae (11 ORFs), and Podoviridae (4 ORFs). Phage proteins were grouped into three categories based on their predicted functions: (i) DNA replication and nucleotide metabolism, (ii) phage particle formation, and (iii) host interaction. Since there is no identified gene encoding integrase and toxins in phage genome, phage φPto-bp6g could be potentially applicable as a safe biological control reagent against brown blotch disease in mushroom cultivation.

The commodification of obstetric ultrasound scanning in Hanoi, Viet Nam.

Growing numbers of pregnant women across the world now routinely have ultrasound scans as part of antenatal care, including in low-income countries. This article presents the findings of anthropological research on the use of obstetric ultrasonography in routine antenatal care in Hanoi, Viet Nam. The findings come from observation, a survey and interviews with women seeking ultrasound scans at a main maternity hospital and interviews with doctors providing ultrasound there. We found a dramatic overuse of ultrasound scanning; the 400 women surveyed had had an average of 6.6 scans and 8.3 antenatal visits during pregnancy, while one-fifth had had ten scans or more. Doctors considered obstetric ultrasound an indispensable part of modern antenatal care. For two-thirds of the women, the main reason for frequent scans was reassurance of normal fetal development. However, the women often also said their doctor had recommended the scans. This overuse must be seen in the context of growing commercialisation in the Vietnamese health care system, where ultrasound provides an important source of revenue for both private and public providers. There is an urgent need in Viet Nam for policy and practice guidelines on the appropriate use of ultrasonography in pregnancy and how best to combine it with essential antenatal care, and information dissemination to women.

Fetal conditions and fatal decisions: ethical dilemmas in ultrasound screening in Vietnam.

In the context of globalization, new technologies of pregnancy are spreading rapidly from affluent to low-income countries. Yet, to date, there is very little research on the application of prenatal diagnostic technology in developing country settings or the dilemmas that prenatal screening may give rise to in situations where health-care resources are scarce. In this article, we describe how obstetrical ultrasound scanning is used as the most important technology for prenatal diagnosis in Vietnam. We explore the social context that shapes moral sentiments and ethical deliberations within everyday medical interactions, drawing attention to the complex social processes through which ethical dilemmas are configured. The data include observations in the scanning room, and semi-structured interviews with patients and staff at a major maternity hospital in the country's capital Hanoi. We found that pregnant women received very limited information and support when a fetal anomaly was detected by ultrasound. Most women left the hospital feeling uncertain about what was wrong with the fetus and decision-making about abortion centred around the difficulties of parenting a disabled child in Vietnam. We conclude that the ethical problems surrounding prenatal screening are intensified in low-income settings such as Vietnam and point to the need for research that takes into account the wider social context that structures ethical dilemmas.

Constructing 'expertness': a novice pharmacist's development of interactional competence in patient consultations.

It has been argued that being able to utilize expertise in discourse is as important as having expertise (C. Candlin and S. Candlin 2002). From a constructivist perspective, knowledgeability does not exist outside of social interaction: being expert involves performing knowledge and skills effectively in social practices (cf. Butler 1990; Lave and Wenger 1991). It is, then, crucial that novice professionals develop the interactional competence to construct themselves as experts in interaction. This study brings together perspectives from research on expert-lay communication, social interaction, and situated learning to examine how a pharmacy intern performed his professional knowledge in the practice of patient consultation over time. Using conversation analysis complemented by the interactional competence framework (Hall 1995; Young 2002), I demonstrate that as the novice pharmacist gained more experience over the course of one internship, he was able to employ professional knowledge in ways that were sensitive to the sequential organization of talk, and to share the patients' lifeworld perspectives (Mishler 1984) more effectively, thus successfully managing the transactional and interpersonal functions of the consultations. This study contributes to research on expert-lay interaction, particularly in pharmacy, and to the under-studied area of interactional competence development in the professions.