Mycology laboratory diagnostic capacity for invasive fungal diseases in public hospitals in Vietnam.

This was a cross-sectional study on the availability of laboratory infrastructure and capacity for the diagnosis of invasive fungal diseases in 24 public hospitals in Vietnam in 2023. Among the hospitals surveyed, 66.7% (14/21) had specialised personnel assigned for mycology testing, and 95.8% (23/24) had a separate microbiology laboratory space. Microscopy and culture methods are available in nearly all laboratories for isolate identification. Antifungal susceptibility testing is only performed for yeasts in 16/24 (66.7%) laboratories. Non-culture methods are hardly used in laboratories. Strengthening local laboratory capacities is essential to meeting health needs in these endemic regions.

There was a need for investment in fungal diagnostics to improve health services in the settings with burden of endemic fungal infections.

Direct Quantitative Analysis of Fluorine in Solid Samples by Cryogenic Laser Ablation and Ionization Time-of-Flight Mass Spectrometry.

The determination of fluorine, the lightest element in halogens, suffers from high ionization potential and spectral interference from water molecules in mass spectrometry. Herein, we introduced a liquid nitrogen cooling unit into the laser ablation and ionization source for the first time to construct a cryogenic laser ablation and ionization time-of-flight mass spectrometry (Cryo-LAI-TOFMS) system. With this system, the interference of water-related species at m/z 19 was effectively eliminated, and fluorine atomization and ionization efficiency could reach 6.3%. A direct quantitative analysis method was developed to determine fluorine contents in phosphate rock, copper ore, industrial byproduct gypsum, stream sediment, and soil. Considering the simplicity, high sensitivity, and low spectral interference of this technique, it can be extended to the determination of fluorine content as low as µg/g in complex solid samples.

Nanoscale Three-Dimensional Imaging of Drug Distributions in Single Cells via Laser Desorption Post-Ionization Mass Spectrometry.

Exploring the three-dimensional (3D) drug distribution within a single cell at nanoscale resolution with mass spectrometry imaging (MSI) techniques is crucial in cellular biology, yet it remains a great challenge due to limited lateral resolution, detection sensitivities, and reconstruction problems. Herein, a microlensed fiber laser desorption post-ionization time-of-flight mass spectrometer (MLF-LDPI-TOFMS) was developed for the 3D imaging of two anticancer drugs within single cells at a 500 × 500 × 500 nm3 voxel resolution. Nanoscale desorption was obtained with a microlensed fiber (MLF), and a 157 nm post-ionization laser was introduced to enhance the ionization yield. Furthermore, a new type of alignment method for 3D reconstruction was developed on the basis of our embedded uniform circular polystyrene microspheres (PMs). Our findings demonstrate that this 3D imaging technique has the potential to provide information about the 3D distributions of specific molecules at the nanoscale level.

Standardized xeno- and serum-free culture platform enables large-scale expansion of high-quality mesenchymal stem/stromal cells from perinatal and adult tissue sources.

BACKGROUND AIMS: Mesenchymal stem/stromal cells (MSCs) are of interest for the treatment of graft-versus-host disease, autoimmune diseases, osteoarthritis and neurological and cardiovascular diseases. Increasing numbers of clinical trials emphasize the need for standardized manufacturing of these cells. However, many challenges related to diverse isolation and expansion protocols and differences in cell tissue sources exist. As a result, the cell products used in numerous trials vary greatly in characteristics and potency. METHODS: The authors have established a standardized culture platform using xeno- and serum-free commercial media for expansion of MSCs derived from umbilical cord (UC), bone marrow and adipose-derived (AD) and examined their functional characteristics. RESULTS: MSCs from the tested sources stably expanded in vitro and retained their biomarker expression and normal karyotype at early and later passages and after cryopreservation. MSCs were capable of colony formation and successfully differentiated into osteogenic, adipogenic and chondrogenic lineages. Pilot expansion of UC-MSCs and AD-MSCs to clinical scale revealed that the cells met the required quality standard for therapeutic applications. CONCLUSIONS: The authors' data suggest that xeno- and serum-free culture conditions are suitable for large-scale expansion and enable comparative study of MSCs of different origins. This is of importance for therapeutic purposes, especially because of the numerous variations in pre-clinical and clinical protocols for MSC-based products.

Enhanced lipid productivity of Chlamydomonas reinhardtii with combination of NaCl and CaCl2 stresses.

Salinity (NaCl) stress treatment is a strategy to induce lipid accumulation in microalgae. This study aimed to investigate the effect of a combination of two salts (NaCl/CaCl2) on lipid productivity of Chlamydomonas reinhardtii. C. reinhardtii was cultured in a two-stage culture comprising 9-day active growth in C medium followed by 3-day salt stress in C medium with various concentrations of NaCl (50‒200 mM)/CaCl2 (100 mM). In salt stress stage, NaCl (200 mM), CaCl2 (100 mM), and the NaCl/CaCl2 mixture inhibited growth but increased the lipid content in C. reinhardtii in comparison with NaCl (0, 50, and 100 mM) conditions. Combinatorial treatment with 100 mM NaCl/100 mM CaCl2 resulted in the highest lipid content (73.4%) and lipid productivity (10.9 mg/L/days), being 3.5- and 2.1-fold, respectively, in salt-free control conditions, and 1.8- and 1.5-folds, respectively, with 200 mM NaCl. Furthermore, 100 mM NaCl/100 mM CaCl2 treatment markedly upregulated glycerol-3-phosphate dehydrogenase (GPDH), lysophosphatidic acid acyltransferase (LPAAT), and diacylglycerol acyltransferase (DAGAT), which are involved in lipid accumulation in C. reinhardtii. The upregulation of these genes with 100 mM NaCl/100 mM CaCl2 resulted in the highest lipid content in C. reinhardtii. Therefore, stress treatment using two salts, 100 mM NaCl/100 mM CaCl2, is a potentially promising strategy to enhance lipid productivity in microalgae.

Micro-Lensed Fiber Laser Desorption Mass Spectrometry Imaging Reveals Subcellular Distribution of Drugs within Single Cells.

The visualization of temporal and spatial changes in the intracellular environment has great significance for chemistry and bioscience research. Mass spectrometry imaging (MSI) plays an important role because of its unique advantages, such as being label-free and high throughput, yet it is a challenge for laser-based techniques due to limited lateral resolution. Here, we develop a simple, reliable, and economic nanoscale MSI approach by introducing desorption laser with a micro-lensed fiber. Using this integrated platform, we achieved 300 nm resolution MSI and successfully visualized the distribution of various small-molecule drugs in subcellular locations. Exhaustive dynamic processes of anticancer drugs, including releasing from nanoparticle carriers entering nucleus of cells, can be readily acquired on an organelle scale. Considering the simplicity and universality of this nanoscale desorption device, it could be easily adapted to most of laser-based mass spectrometry applications.

Chemical and Topographical Single-Cell Imaging by Near-Field Desorption Mass Spectrometry.

Simultaneously acquiring chemical and topographical information within a single cell at nanoscale resolutions is vital to cellular biology, yet it remains a great challenge due to limited lateral resolutions and detection sensitivities. Herein, the development of near-field desorption mass spectrometry for correlated chemical and topographical imaging is reported, thereby bridging the gap between laser-based mass spectrometry (MS) methods and multimodal single-cell imaging. Using this integrated platform, an imaging resolution of 250 nm and 3D topographically reconstructed chemical single-cell imaging were achieved. This technique offers more in-depth cellular information than micrometer-range laser-based MS imaging methods. Considering the simplicity and compact size of the near-field device, this technique can be introduced to MALDI-MS, expanding the multimodal abilities of MS at nanoscale resolutions.

Approaching Standardless Quantitative Elemental Analysis of Solids: Microsecond Pulsed Glow Discharge and Buffer-Gas-Assisted Laser Ionization Time-of-Flight Mass Spectrometry.

Among various ionization sources for mass spectrometry, microsecond pulsed glow discharge (MP-GD) and buffer-gas-assisted laser ionization (BGA-LI) sources have the potential for direct quantitative elemental analysis of solids without the requirement of standard reference materials. The analytical potential of these two ionization sources has been evaluated by coupling them to orthogonal time-of-flight mass spectrometry (MS). A straightforward method was proposed to achieve the quantitative result: if a spectrum contains little interference and elemental peak currents are proportional to their concentrations, then the molar concentration of each element is equal to its ion current proportion in the total ion current. Two series of metal standards were applied for the evaluation. Explicit spectra with little interference can be acquired by both techniques. The interferences contribute only a very small portion to the total ion current for MP-GD-MS and BGA-LI-MS; therefore, their influence on the quantitative result can be ignored. All metal elements can be determined quite accurately by the proposed quantitation method, while gaps exist for nonmetal elements due to the high ionization potentials or gas species interference. Between the two techniques, BGA-LI-MS offers a more accurate quantitative result, primarily due to its higher plasma temperature.

Mutational screening of germline RB1 gene in Vietnamese patients with retinoblastoma reveals three novel mutations.

Purpose: Retinoblastoma (Rb) is a rare and unique eye cancer that usually develops in the retinas of children less than 5 years old due to mutations in the RB1 gene. About 40% of affected individuals have the heritable form making genetics testing of the RB1 gene important for disease management. This study aims to identify germline mutations in RB1 in a cohort of patients with Rb from northern Vietnam. Methods: Genomic DNA was extracted from peripheral blood of 34 patients with Rb (nine unilateral and 25 bilateral cases) and their available parents. Twenty-seven exons, flanking sequences, and the promoter region of RB1 gene were screened for mutations with direct PCR sequencing. Multiplex ligation-dependent probe amplification (MLPA) was applied for patients with negative sequencing results. In the mutation-positive patients, their available parental DNA was analyzed to determine the parental origin of the mutation. Results: Germline mutations in RB1 were identified in 25 (73.53%) of 34 patients (four unilateral and 21 bilateral cases). Of these mutations, 19 were detected, including seven nonsense, six frameshift, four splice-site (one was identified in two siblings), and one missense, with Sanger sequencing. Three novel frameshift mutations were discovered in one unilateral and two bilateral patients. MLPA detected mutations in the RB1 gene in six bilateral cases, of whom five had a whole gene deletion (three familial cases) and one had a partial gene deletion (from exon 4 to exon 27) in one allele of the RB1 gene. Parental testing showed five mutations originated from the fathers and one was inherited from a mother who was mosaic for the mutation. Conclusions: This study provides a data set of germline mutations in the RB1 gene in Vietnamese patients with retinoblastoma. Screening of mutations in the RB1 gene can help to identify heritable Rb and contribute to clinical management and genetic counseling for affected families.

Two new guaiane sesquiterpene lactones from the aerial parts of Artemisia vulgaris.

Two new guaiane sesquiterpene lactones, vulgarolides A and B (1 and 2), were isolated from Artemisia vulgaris aerial parts using various chromatographic separations. The structure elucidation was performed by combination of spectroscopic experiments including 1D and 2D NMR, HR ESI MS, and CD. Their in vitro cytotoxic activities against five human cancer cell lines were also evaluated using SRB method.

Dietary free L-glutamate contributes to maintaining a low sodium intake among Vietnamese.

Background: The World Health Organization (WHO) and the Food and Agriculture Organization (FAO) recommend the consumption of less than 2,000 mg of sodium/day to reduce blood pressure and the risk of conditions such as cardiovascular disease and coronary heart disease. The sodium intake among Vietnamese was reported to be 7,200 mg/d or more. Free L-glutamate enhances flavor when it is added to food and improves the taste of sodium-reduced foods. Objective: This study aims to investigate whether the intake of free L-glutamate-rich seasonings contributes to maintaining a low sodium intake in a cross-over study. Methods: From a total of 145 subjects, 42 participants were screened for participation in the cross-over design study. Subjects were randomly allocated to the Low free L-glutamate group (Low free L-Gl) and the Normal free L-glutamate group (Normal free L-Gl). Both received a direct educational guideline to reduce sodium intake. The Low free L-Gl group started with a restriction in the variety of free L-glutamate-rich seasonings, and the Normal free L-Gl group had no restriction in the variety of seasonings. Blood pressure was measured at week 0 (baseline), week 2, week 4, and week 6, while body weight, height, urine sodium and potassium excretion, chromogranin-A (CgA pmol/mg protein) from saliva, and free L-glutamate from food were measured at week 0, week 3, and week 6. Results: In Low free L-Gl, the amount of free L-glutamate in food decreased significantly from baseline to week 6 (p < 0.00), while it did not change in the Normal free L-Gl (p > 0.05). However, the reduction of sodium excretion at week 6 was 22% in Low free L-Gl (5,875 mg/d vs. 4,603 mg/d, p < 0.01) and 46% in Normal free L-Gl (6,107 mg/d vs. 3,277 mg/d, p < 0.00), both lower than the baseline. CgA (pmol/mg protein) did not show any difference between the two groups. Conclusion: The group with Normal free L-Gl intake showed a 46% reduction in sodium excretion by week 6 compared to the baseline. This suggests that the consumption of L-glutamate-rich seasonings when complemented with direct educational guidelines, can contribute to maintaining a low sodium intake.

Avian H6 Influenza Viruses in Vietnamese Live Bird Markets during 2018-2021.

Avian influenza viruses of the H6 subtype are prevalent in wild ducks and likely play an important role in the ecology of influenza viruses through reassortment with other avian influenza viruses. Yet, only 152 Vietnamese H6 virus sequences were available in GISAID (Global Initiative on Sharing All Influenza Data) prior to this study with the most recent sequences being from 2018. Through surveillance in Vietnamese live bird markets from 2018 to 2021, we identified 287 samples containing one or several H6 viruses and other influenza A virus subtypes, demonstrating a high rate of co-infections among birds in Vietnamese live bird markets. For the 132 H6 samples with unique influenza virus sequences, we conducted phylogenetic and genetic analyses. Most of the H6 viruses were similar to each other and closely related to other H6 viruses; however, signs of reassortment with other avian influenza viruses were evident. At the genetic level, the Vietnamese H6 viruses characterized in our study encode a single basic amino acid at the HA cleavage site, consistent with low pathogenicity in poultry. The Vietnamese H6 viruses analyzed here possess an amino acid motif in HA that confers binding to both avian- and human-type receptors on host cells, consistent with their ability to infect mammals. The frequent detection of H6 viruses in Vietnamese live bird markets, the high rate of co-infections of birds with different influenza viruses, and the dual receptor-binding specificity of these viruses warrant their close monitoring for potential infection and spread among mammals.

Solvothermal fabrication of rubidium tungsten bronze for the absorption of near infrared light.

For fabricating the crystal of Rb(x)WO3, the solvothermal process was performed using a mixed medium of water and ethanol as solvent. It has been found that the morphological evolutions of samples were greatly changed as disordered nanoparticles, bundle of nanorods and microfibers with increasing content of water in the starting solution. Among them, sample synthesized with assistance of 3 ml water consisted of the nanorods with a diameter in the range of 80-150 nm, showing suitable size and morphology as near-infrared shielding material. On the merits of this nanostructure, the thin film of the Rb(x)WO3 nanorods realized excellent shielding of near-infrared light, meanwhile, it selectively transmitted the major part of visible light.

Dysregulation of T Cell Differentiation and the IL17A(+)Foxp3(+)Treg Subset in Chronic Hepatitis B Patients with Hepatitis Flare.

The regulatory T (Treg) and T helper 17 (Th17) cells modulate the immune response in chronic hepatitis B virus (HBV) infection by promoting immune tolerance and restricting liver damage or stimulating inflammatory response and rendering hepatocyte injury. These cells act through signaling transcription factors and secreting cytokines. We aimed to observe the percentages of Treg, Th17 cells, and their messenger RNA (mRNA) level of forkhead box protein 3 (Foxp3) and retinoid orphan receptor γt (RORγt) in the chronic hepatitis B (CHB)-infected group and CHB patients with hepatitis flare (HF). We recruited 103 participants, including 88 CHB-infected cases and 15 healthy controls (HCs) in Ho Chi Minh City. CHB cases were enrolled into two groups: HBeAg+ CHB infection (e+CHBI; n = 42) and HF (including 20 mild HF and 26 severe HF [sHF]). The Foxp3(+)Treg and Th17 cells were measured by flow cytometry. The mRNA levels of Foxp3 and RORγt were analyzed by real-time polymerase chain reaction. The percentages of Foxp3(+)Treg, of Th17, and the Foxp3(+)Treg/Th17 ratio were significantly higher in the sHF compared to the e+CHBI group. The sHF and e+CHBI groups had significantly higher mRNA levels of Foxp3 and RORγt compared to the HC group. Furthermore, a special subset, interleukin 17A(+)Foxp3(+)Treg cells, were observed with a significantly higher percentage in the sHF compared to the e+CHBI group. This finding revealed the contributions of this new subset on the severe flare cases. Our results explained the diversity of T cells and their subsets in the immune response in CHB. This subset should be further investigated as a specific tool in HBV immune response.

The anxiety and depression disorder in adults with atopic dermatitis: experience of a dermatology hospital.

The objectives of this study are to identify the proportion of atopic dermatitis adult patients having anxiety and depression disorder and to measure the relationship between anxiety and depression disorder and characteristics of atopic dermatitis. A cross-sectional study with convenience sampling was conducted. Diagnostic criteria for atopic dermatitis were based on modified Hanifin and Raijka criteria and the severity of anxiety-depression disorder was evaluated using the hospital anxiety and depression scale. In this study, 208 patients were enrolled. The percentage of patients with anxiety and subthreshold anxiety were 11.1% and 34.1%, respectively. 5.3% of patients had depression and 39.4% of patients suffered from subthreshold depression. The proportion of patients with mixed anxiety-depressive disorder was 1.44%. Patients with severe atopic dermatitis were more likely to endure anxiety but not depression. Allergies or autoimmune diseases and scoring atopic dermatitis C were two independent risk factors of depression whereas edema and excoriation were two independent risk factors related to anxiety in atopic dermatitis patients. These findings suggest that atopic dermatitis is associated with anxiety and depression. Allergies, autoimmune diseases, pruritus, and insomnia had a correlation with anxiety and depression disorder.

Cassava Witches' Broom Disease in Southeast Asia: A Review of Its Distribution and Associated Symptoms.

Cassava witches' broom disease (CWBD) is one of the main diseases of cassava in Southeast Asia (SEA). Affected cassava plants show reduced internodal length and proliferation of leaves (phyllody) in the middle and top part of the plant, which results in reduced root yields of 50% or more. It is thought to be caused by phytoplasma; however, despite its widespread distribution in SEA still little is known about CWBD pathology. The overarching goal of this study was to review and corroborate published information on CWBD biology and epidemiology considering recent field observations. We report the following: (1) CWBD symptoms are conserved and persistent in SEA and are distinct from what has been reported as witches' broom in Argentina and Brazil. (2) In comparison with cassava mosaic disease, another major disease of cassava in SEA, symptoms of CWBD develop later. (3) Phytoplasma detected in CWBD-affected plants belong to different ribosomal groups and there is no association study available indicating phytoplasma as the causing agent of CWBD. These findings are essential clues for designing surveillance and management strategies and for future studies to better understand the biology, tissue localization and spatial spread of CWBD in SEA and other potential risk areas.

Mechanical Properties of Ternary Composite from Waste Leather Fibers and Waste Polyamide Fibers with Acrylonitrile-Butadiene Rubber.

This study aimed to improve the mechanical properties of a composite material consisting of waste leather fibers (LF) and nitrile rubber (NBR) by partially replacing LF with waste polyamide fibers (PA). A ternary recycled composite NBR/LF/PA was produced by a simple mixing method and vulcanized by compression molding. The mechanical properties and dynamic mechanical properties of the composite were investigated in detail. The results showed that the mechanical properties of NBR/LF/PA increased with an increase in the PA ratio. The highest tensile strength value of NBR/LF/PA was found to have increased about 1.26 times, that is from 12.9 MPa of LF50 to 16.3 MPa of LF25PA25. Additionally, the ternary composite demonstrated high hysteresis loss, which was confirmed by dynamic mechanical analysis (DMA). The presence of PA formed a non-woven network that significantly enhanced the abrasion resistance of the composite compared to NBR/LF. The failure mechanism was also analyzed through the observation of the failure surface using scanning electron microscopy (SEM). These findings suggest that the utilization of both waste fiber products together is a sustainable approach to reducing fibrous waste while improving the qualities of recycled rubber composites.

Four new N-phenethylbenzamide derivatives from the stems of piper betle and their antimicrobial activity.

Four new N-phenethylbenzamide derivatives, named piperbetamides A-D (1-4), and six allylbenzene derivatives (5-10) were isolated from the stems of Piper betle L. Their structures were determined by HR-ESI-MS and NMR spectroscopic methods. Compounds 1-10 were evaluated for their inhibitory effects on the growth of nine microorganisms including five Gram-negative (Escherichia coli, Salmonella enterica serovar Typhimurium, Shigella flexneri, Pseudomonas aeruginosa, and Extended-spectrum beta-lactam resistant Klebsiella pneumoniae), three Gram-positive (Listeria monocytogenes, Methicilin-resistant Staphylococcus aureus, Vancomycin-resistant Enterococcus faecalis), and one yeast (Candida albicans) strains. Compounds 1, 3, 4, 6 and 10 exhibited potential antimicrobial activity against S. flexneri, L. monocytogenes, methicillin-resistant S. aureus and vancomycin-resistant E. faecalis with minimum inhibitory concentration (MIC) values in the range of 16-32 µg/mL.

Ceratobasidium sp. is associated with cassava witches' broom disease, a re-emerging threat to cassava cultivation in Southeast Asia.

Cassava witches' broom disease (CWBD) is a devastating disease of cassava in Southeast Asia (SEA), of unknown etiology. Affected plants show reduced internodal length, proliferation of leaves and weakening of stems. This results in poor germination of infected stem cuttings (i.e., planting material) and significant reductions in fresh root yields and starch content, causing economic losses for farmers and processors. Using a metagenomic approach, we identified a fungus belonging to the Ceratobasidium genus, sharing more than 98.3-99.7% nucleotide identity at the Internal Transcribed Spacer (ITS), with Ceratobasidium theobromae a pathogen causing similar symptoms in cacao. Microscopy analysis confirmed the identity of the fungus and specific designed PCR tests readily showed (1) Ceratobasidium sp. of cassava is strongly associated with CWBD symptoms, (2) the fungus is present in diseased samples collected since the first recorded CWBD outbreaks in SEA and (3) the fungus is transmissible by grafting. No phytoplasma sequences were detected in diseased plants. Current disease management efforts include adjustment of quarantine protocols and guarantee the production and distribution of Ceratobasidium-free planting material. Implications of related Ceratobasidium fungi, infecting cassava, and cacao in SEA and in other potential risk areas are discussed.

Continued Circulation of Highly Pathogenic H5 Influenza Viruses in Vietnamese Live Bird Markets in 2018-2021.

We isolated 77 highly pathogenic avian influenza viruses during routine surveillance in live poultry markets in northern provinces of Vietnam from 2018 to 2021. These viruses are of the H5N6 subtype and belong to HA clades 2.3.4.4g and 2.3.4.4h. Interestingly, we did not detect viruses of clade 2.3.4.4b, which in recent years have dominated in different parts of the world. The viruses isolated in this current study do not encode major determinants of mammalian adaptation (e.g., PB2-E627K or PB1-D701N) but possess amino acid substitutions that may affect viral receptor-binding, replication, or the responses to human antiviral factors. Several of the highly pathogenic H5N6 virus samples contained other influenza viruses, providing an opportunity for reassortment. Collectively, our study demonstrates that the highly pathogenic H5 viruses circulating in Vietnam in 2018-2021 were different from those in other parts of the world, and that the Vietnamese H5 viruses continue to evolve through mutations and reassortment.