Three new minor steroidal glycosides from the whole plants of Hoya parasitica (Wall. ex Hornem.) Wight.

Three new minor steroidal glycosides were isolated from the whole plants of Hoya parasitica (Wall. ex Hornem.) Wight. Their structures were further elucidated as 3ß,4α-dihydroxy-5ß-spirost-(25)27-en-1ß-yl O-α-L-arabinopyranoside (1), 3ß-hydroxy-5ß-spirost-25(27)-en-1ß-yl O-α-L-rhamnopyranosyl-(1→2)-ß-D-xylopyranoside (2), and (23S,24S,25S)-3ß,4α,23,24-tetrahydroxy-5ß-spirostan-1ß-yl O-α-L-rhamnopyranosyl-(1→2)-ß-D-fucopyranoside (3) through interpretation of the spectroscopic data (one-dimensional and two-dimensional) and mass spectrometry (HR-ESI-MS). To the best of our knowledge, this is the first report of the isolation of spirostane-type steroidal saponins from the Hoya genus.

Happiness in Old Age: The Daughter Connection.

Family and intergenerational relationships are becoming increasingly important as sources of support and care for the elderly population in rapidly ageing Asian societies. However, this has also raised concerns over reinforcement of cultural preference for sons as a source of old-age security. This paper therefore revisits the question-what determines happiness in old age-by investigating the role of adult children's gender in the context of Thailand, an ageing Asian country with no legacy of sex-preference in fertility. We employ nationally representative data to examine the association between old-age happiness and presence of a co-residing child. Compared to living alone, living with at least one child is found to positively associate with older persons' happiness. However, this result is specific to daughters. Moreover, compared to older men, women systematically benefit from a "daughter effect". Co-residing daughters with university education and those who maintain a good relationship with their parents help explain the positive happiness effect on older persons. Co-residing daughters are also positively linked to  reduced loneliness; improved self-rated health; and improved economic conditions of older parents. Our findings suggest that policies that increase human capital of the girl child and enhance family solidarity are likely to have long term intergenerational wellbeing benefits.

Diagnostic accuracy of two-dimensional and three-dimensional transesophageal echocardiography in detecting pannus and thrombus in left mechanical valve obstruction.

The aim of this study was to determine the accuracy of two-dimensional and three-dimensional transesophageal echocardiography (TEE) in the detection of pannus and thrombus in left mechanical valve obstruction (LMVO) compared with surgical and histopathology findings. Materials and methods: Patients with suspected LMVO on transthoracic echocardiography were enrolled consecutively. All patients underwent two-dimensional and three-dimensional TEE, and open-heart surgery to replace obstructed valves. Macroscopic and microscopic analysis of the excised masses was used as the gold standard for the diagnosis of thrombus and/or pannus. Results: Forty-eight patients [34 women (70.8%), age 49±13 years, New York Heart Association II: 68.8%, New York Heart Association III: 31.2%] were enrolled. In the diagnostic of thrombus, the sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of three-dimensional TEE were 89.2, 72.7, 85.4, 91.7, and 66.7%, respectively, compared with those of two-dimensional TEE (42.2, 66.7, 43.8, 95, and 7.1%, respectively). In the diagnosis of pannus, the sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of three-dimensional TEE were 53.3, 100, 85.4, 100, and 82.5%, respectively; compared with those of two-dimensional TEE (7.4, 90.5, 43.8, 50, and 43.2%, respectively). Receiver operating characteristic curves depict that the area under the curves of three-dimensional TEE was higher than the area under the curves of two-dimensional TEE in both diagnoses of thrombus and pannus (0.8560 vs. 0.7330, P=0.0427 and 0.8077 vs. 0.5484, P=0.005, respectively). Conclusions: This study indicated that three-dimensional TEE had a higher diagnostic value than two-dimensional TEE in the detection of thrombus and pannus in patients with LMVO, and can be a reliable imaging modality to identify the causes of LMVO.

An Alternative Splicing Variant of the Mixed-Lineage Leukemia 5 Protein Is a Cellular Adhesion Receptor for ScaA of Orientia tsutsugamushi.

Scrub typhus is a mite-borne disease caused by the obligately intracellular bacterium Orientia tsutsugamushi. We previously demonstrated that ScaA, an autotransporter membrane protein of O. tsutsugamushi, is commonly shared in various genotypes and involved in adherence to host cells. Here, we identified a mixed-lineage leukemia 5 (MLL5) mammalian trithorax group protein as a host receptor that interacts with ScaA. MLL5, identified by yeast two-hybrid screening, is an alternative splicing variant of MLL5 (vMLL5) which contains 13 exons with additional intron sequences encoding a tentative transmembrane domain. Indeed, vMLL5 is expressed on the plasma membrane as well as in intracellular compartments in eukaryotic cells and colocalized with adherent O. tsutsugamushi. In addition, ScaA-expressing Escherichia coli showed significantly increased adherence to vMLL5-overexpressing cells compared with vector control cells. We mapped the C-terminal region of the passenger domain of ScaA as a ligand for vMLL5 and determined that the Su(var)3-9, Enhancer of zeste, Trithorax (SET) domain of MLL5 is an essential and sufficient motif for ScaA binding. We observed significant and specific inhibition of bacterial adhesion to host cells in competitive inhibition assays using the C-terminal fragment of ScaA or the SET domain of vMLL5. Moreover, immunization with the C-terminal fragment of ScaA provided neutralizing activity and protective immunity against lethal challenge with O. tsutsugamushi as efficiently as vaccination with the whole passenger domain of ScaA. These results indicate that vMLL5 is a novel cellular receptor for ScaA-mediated adhesion of O. tsutsugamushi and facilitates bacterial adhesion to host cells, thereby enhancing bacterial infection. IMPORTANCE O. tsutsugamushi is a mite-borne pathogen that causes scrub typhus. As an obligately intracellular pathogen, its adhesion to and invasion of host cells are critical steps for bacterial growth. However, the molecular basis of the bacterial ligand and host receptor interaction is poorly defined. Here, we identified a splicing variant of MLL5 (vMLL5) as a cellular adhesion receptor of ScaA, an outer membrane autotransporter protein of O. tsutsugamushi. We mapped the interacting domains in the bacterial ligand and host receptor and confirmed their functional interaction. In addition, immunization with the C-terminal region of ScaA, which involves an interaction with the SET domain of vMLL5, not only induces enhanced neutralizing antibodies but also provides protective immunity against lethal challenge with O. tsutsugamushi.

Antibiotic Resistance of Helicobacter pylori in Children with Gastritis and Peptic Ulcers in Mekong Delta, Vietnam.

BACKGROUND: Helicobacter pylori (H. pylori) infection causes gastritis, duodenal and gastric ulcers, and gastric cancer. H. pylori eradication efficacy is low worldwide, and antibiotic resistance is the leading cause of therapy failure; therefore, this study was performed to determine the characteristics of antibiotic resistance of H. pylori in children with gastritis, duodenal and gastric ulcer. METHODS: A cross-sectional study was conducted on 237 pediatric patients diagnosed with gastroduodenal inflammation and ulcer at two hospitals in Vietnam from March 2019 to April 2022. Pediatric patients with positive H. pylori tests continued to do E-tests to measure the minimum inhibitory concentration of the antibiotic so that we could prescribe effective antibiotics based on the sensitivity. RESULTS: In 237 pediatric patients (51.1% males) with a median age of 10.3 years (range 5-16 years), endoscopic images showed that inflammatory lesions and peptic ulcers accounted for 69.2% and 30.8%, respectively. Resistance rates of H. pylori were 80.6% to clarithromycin (CLR), 71.7% to amoxicillin (AMX), 49.4% to metronidazole (MTZ), 45.1% to levofloxacin (LEV), and 11.4% to tetracycline (TET); dual resistance to AMX + CLR was 64.2%, AMX + LEV 35%, AMX + MTZ 33.3%, CLR + MTZ 32.5%, and TET + MTZ 7.2%. The frequency of clarithromycin resistance was significantly increased, particularly in pediatric patients who had received prior H. pylori treatment. The percentage of amoxicillin resistance increased with age; amoxicillin resistance of H. pylori was more prevalent among pediatric patients with peptic ulcers than those with gastroduodenal inflammation and higher in males than females. CONCLUSIONS: The proportions of resistance to CLR, AMX, MTZ, and LEV were extremely high, in contrast to TET, which was lower in pediatric patients. Our study suggests that the standard triple therapy with CLR should be limited as the empiric therapy for pediatric patients, and we should consider using eradication regimens with TET for children over 8 years of age if the medical facility is not qualified to perform antibiotic susceptibility tests of H. pylori in the Mekong Delta.

Women's education and desire for additional children in Vietnam: regional differences and the role of son preference.

This paper investigates the relationship between women's education and desire for additional children across the six economic regions of Vietnam. The study employed data from the nationally representative Vietnam Multiple Indicator Cluster Survey (MICS) 2014. Probit regression results showed that for women with one child, higher levels of education were associated with higher fertility desire in two out of six regions. Similar results were found for women with two or more children. Children's sex composition played a role in the desire for additional children, reflecting both son preference and mixed-gender preference. In Vietnam overall, among women with at least one boy, those with lower levels of education were more likely not to want another child. The results, however, differed by region. The findings suggest that the social and economic context of each region, particularly sex ratio at birth and total fertility rate, should be taken into account when designing population policies in Vietnam.

Longevity of seropositivity and neutralizing antibodies in recovered MERS patients: a 5-year follow-up study.

OBJECTIVES: We aimed to assess the longevity of spike-specific antibody responses and neutralizing activity in the plasma of recovered Middle East respiratory syndrome (MERS) patients. METHODS: We traced the antibody responses and neutralizing activity against MERS coronavirus (MERS-CoV) in peripheral blood samples collected from 70 recovered MERS patients for 5 years after the 2015 MERS outbreak in South Korea. We also measured the half-life of neutralizing antibody titres in the longitudinal specimens. RESULTS: The seropositivity rate persisted for up to 4 years (50.7-56.1%), especially in MERS patients who suffered from severe pneumonia, and then decreased (35.9%) in the fifth year. Although the spike-specific antibody responses decreased gradually, the neutralizing antibody titres decreased more rapidly (half-life: 20 months) in 19 participants without showing negative seroconversion during the study period. Only five (26.3%) participants had neutralizing antibody titres greater than 1/1000 of PRNT50, and a high neutralizing antibody titre over 1/5000 was not detected in the participants at five years after infection. DISCUSSION: The seropositivity rate of the recovered MERS patients persisted up to 4 years after infection and significantly dropped in the fifth year, whereas the neutralizing antibody titres against MERS-CoV decreased more rapidly and were significantly reduced at 4 years after infection.

GmDREB6, a soybean transcription factor, notably affects the transcription of the NtP5CS and NtCLC genes in transgenic tobacco under salt stress conditions.

Soil is contaminated with salinity, which inhibits plant growth and development and reduces crop yields. The DREB (dehydration responsive element binding protein) gene responds to salt stresses through enhanced transcriptional expression and activation of genes involved in plant salinity resistance. In this study, we present the results of the analysis of the expression of the GmDREB6 transgene, a gene that encodes the soybean DREB6 transcription factor, regulating the transcription of the NtP5CS and NtCLC genes in transgenic tobacco under salt stress conditions. The transcription of GmDREB6, NtP5CS, and NtCLC in transgenic tobacco lines was confirmed by qRT-PCR. Under salt stress conditions, the GmDREB6 gene transcription levels in the transgenic tobacco lines L1 and L9 had increased from 2.40- to 3.22- fold compared with the condition without salinity treatment. Two transgenic lines, L1 and L9, had transcription levels of the P5CS gene that had increased from 1.24- to 3.60- fold compared with WT plants. For the NtCLC gene, under salt stress conditions, the transgenic lines had transcription levels that had increased by 3.65-4.54 (fold) compared with WT plants (P < 0.05). The L1-transgenic tobacco line showed simultaneous expression of both the GmDREB6 transgene and two intrinsic genes, the NtP5CS and NtCLC genes. This study demonstrated that expression of the GmDREB6 gene from soybean increases the transcription levels of the NtP5CS and NtCLC genes in transgenic tobacco plants under salt stress conditions. The analysis results have suggested that the GmDREB6 gene is a potential candidate for improving the salt tolerance of plants, opening up research and development opportunities for salt stress-tolerant crops to respond to climate change and the rise in sea levels.

Recycled Concrete Aggregate for Medium-Quality Structural Concrete.

This paper reports an evaluation of the properties of medium-quality concrete incorporating recycled coarse aggregate (RCA). Concrete specimens were prepared with various percentages of the RCA (25%, 50%, 75%, and 100%). The workability, mechanical properties, and durability in terms of abrasion of cured concrete were examined at different ages. The results reveal insignificant differences between the recycled concrete (RC) and reference concrete in terms of the mechanical and durability-related measurements. Meanwhile, the workability of the RC reduced vastly since the replacement of the RCA reached 75% and 100%. The ultrasound pulse velocity (UPV) results greatly depend on the porosity of concrete and the RC exhibited higher porosity than that of the reference concrete, particularly at the transition zone between the RCA and the new paste. Therefore, the sound transmission in the RC required longer times than that in the reference concrete. Moreover, a predictive equation relating the compressive strength to the UPV was developed.

The Orientia tsutsugamushi ScaB Autotransporter Protein Is Required for Adhesion and Invasion of Mammalian Cells.

Autotransporter proteins are widely present in Gram-negative bacteria. They play a pivotal role in processes related to bacterial pathogenesis, including adhesion, invasion, colonization, biofilm formation, and cellular toxicity. Bioinformatics analysis revealed that Orientia tsutsugamushi, the causative agent of scrub typhus, encodes six different autotransporter genes (scaA-scaF). Although four of these genes (scaA, scaC, scaD, and scaE) are present in diverse strains, scaB and scaF have been detected in only a limited number of strains. Previous studies have demonstrated that ScaA and ScaC are involved in the adherence of host cells. However, the putative function of other O. tsutsugamushi Sca proteins has not been studied yet. In this study, we show that scaB is transcribed and expressed on the surface of O. tsutsugamushi Boryong strain. Using a heterologous Escherichia coli expression system, we demonstrated that ScaB-expressing E. coli can successfully mediate adherence to and invasion into non-phagocytic cells, including epithelial and endothelial cells. In addition, pretreatment with a recombinant ScaB polypeptide inhibits the entry of O. tsutsugamushi into cultured mammalian cells. Finally, we also identified the scaB gene in the Kuroki and TA686 strains and observed high levels of sequence variation in the passenger domains. Here, we propose that the ScaB protein of O. tsutsugamushi can mediate both adhesion to and invasion into host cells in the absence of other O. tsutsugamushi genes and may play important roles in bacterial pathogenesis.

Genotypic Heterogeneity of Orientia tsutsugamushi in Scrub Typhus Patients and Thrombocytopenia Syndrome Co-infection, Myanmar.

Serologic and molecular surveillance of serum collected from 152 suspected scrub typhus patients in Myanmar revealed Orientia tsutsugamushi of genotypic heterogeneity. In addition, potential co-infection with severe fever with thrombocytopenia syndrome virus was observed in 5 (3.3%) patients. Both scrub typhus and severe fever with thrombocytopenia syndrome are endemic in Myanmar.

Overexpression of the GmDREB6 gene enhances proline accumulation and salt tolerance in genetically modified soybean plants.

Soybean plants are sensitive to the effects of abiotic stress and belong to the group of crops that are less drought and salt tolerant. The identification of genes involved in mechanisms targeted to cope with water shortage is an essential and indispensable task for improving the drought and salt tolerance of soybean. One of the approaches for obtaining lines with increased tolerance is genetic modification. The dehydration-responsive element binding proteins (DREBs), belonging to the AP2 family, are trans-active transcription factors that bind to the cis-sequences of the promoter for activating the expression of the target genes that mediate drought and salt tolerant responses. In this study, the GmDREB6 transgene was introduced into DT84 cultivar soybean plants, using Agrobacterium-mediated transformation. The efficacy of GmDREB6 overexpression in enhancing the transcriptional level of GmP5CS and proline accumulation in genetically modified (GM) soybean plants was also assayed. The results demonstrated that ten GM soybean plants (T0 generation) were successfully generated from the transformed explants after selecting with kanamycin. Among these plantlets, the presence of the GmDREB6 transgene was confirmed in nine plants by Polymerase Chain Reaction (PCR), and eight plants showed positive results in Southern blot. In the T1 generation, four GM lines, labelled T1-2, T1-4, T1-7, and T1-10, expressed the recombinant GmDREB6 protein. In the T2 generation, the transcriptional levels of the GmP5CS gene were higher in the GM lines than in the non-transgenic plants, under normal conditions and also under conditions of salt stress and drought, ranging from 1.36 to 2.01 folds and 1.58 to 3.16 folds that of the non-transgenic plants, respectively. The proline content was higher in the four GM soybean lines, T2-2, T2-4, T2-7, and T2-10 than in the non-transgenic plants, ranging from 0.82 µmol/g to 4.03 µmol/g. The proline content was the highest in the GM T2-7 line (7.77 µmol/g). In GM soybean lines, T2-2, T2-4, T2-7, and T2-10 proline content increased after plants were subjected to salt stress for seven days, in comparison to that under normal conditions, and ranged from 247.83% to 300%, while that of the non-GM plants was 238.22%. These results suggested that GmDREB6 could act as a potential candidate for genetic engineering for improving tolerance to salt stresses.

Photostability of cosmetic UV filters on mammalian skin under UV exposure.

Previous studies showed that the common UV filter substances benzophenone-3 (BP-3), butyl methoxydibenzoylmethane (BM-DBM), octocrylene (OCR), ethylhexyl methoxycinnamate (EHMC), ethylhexyl salicylate (EHS) and ethylhexyl triazone (EHT) were able to react with amino side chains of different proteins in vitro. To transfer the results to mammalian skin conditions, sunscreen products were applied on both prepared fresh porcine skin and glass plates, followed by UV irradiation and the determination of depletion of the respective UV filters. Significantly lower recoveries of the UV filters extracted from skin samples than from glass plates indicated the additional reaction of the UV filters with skin constituents, when proteins will be the most important reactants. Among the products tested, BP-3 showed the greatest differences in recoveries between glass and skin samples of about 13% and 24% after 2 and 4 h of irradiation, respectively, followed by EHS > BM-DBM > OCR > EHMC > EHT. The obtained results raise the question, whether the common in vitro evaluations of sunscreens, using inert substrate materials like roughened quartz or polymethyl methacrylate (PMMA) plates are really suitable to fully replace in vivo methods, as they cannot include skin-typical reactions.