Quantifying antimicrobial access and usage for paediatric diarrhoeal disease in an urban community setting in Asia.

Objectives: Antimicrobial-resistant infections are a major global health issue. Ease of antimicrobial access in developing countries is proposed to be a key driver of the antimicrobial resistance (AMR) epidemic despite a lack of community antimicrobial usage data. Methods: Using a mixed-methods approach (geospatial mapping, simulated clients, healthcare utilization, longitudinal cohort) we assessed antimicrobial access in the community and quantified antimicrobial usage for childhood diarrhoea in an urban Vietnamese setting. Results: The study area had a pharmacy density of 15.7 pharmacies/km2 (a pharmacy for every 1316 people). Using a simulated client method at pharmacies within the area, we found that 8% (3/37) and 22% (8/37) of outlets sold antimicrobials for paediatric watery and mucoid diarrhoea, respectively. However, despite ease of pharmacy access, the majority of caregivers would choose to take their child to a healthcare facility, with 81% (319/396) and 88% (347/396) of responders selecting a specialized hospital as one of their top three preferences when seeking treatment for watery and mucoid diarrhoea, respectively. We calculated that at least 19% (2688/14427) of diarrhoea episodes in those aged 1 to <5 years would receive an antimicrobial annually; however, antimicrobial usage was almost 10 times greater in hospitals than in the community. Conclusions: Our data question the impact of community antimicrobial usage on AMR and highlight the need for better education and guidelines for all professionals with the authority to prescribe antimicrobials.

Yeast-derived vacuoles as a novel carrier with enhanced hCMEC/D3 cell monolayer penetration.

The blood-brain barrier (BBB) is a brain protection structure that restricts drug delivery from the blood to the central nervous system. Thus, we developed a novel drug carrier using yeast vacuoles to overcome this problem. The purpose of this study was to assess the drug transportability of yeast vacuoles using a human cerebral microvascular endothelial cell line (hCMEC/D3) cell monolayer. Here, we used daunorubicin (DNR) as a microtubule-targeting agent with the ability to disaggregate pre-formed fibrils and prevent Tau fibrillization. An in vitro model was developed by culturing hCMEC/D3 cells on Transwell inserts in EBM-2 endothelial basal medium until the cells formed a monolayer. Next, nano-sized yeast vacuoles were loaded with DNR, and the signals inside and outside the hMEC/D3 cell monolayer were detected using the GloMax® Explorer fluorometer. DNR penetrated the cell monolayer and was regulated by endocytosis via receptor-mediated macropinocytosis on the surface of the cell. Confocal imaging showed a significant increase in intracellular DNR fluorescence when the cells were treated with the vacuole-encapsulated drug. These results indicate that the drug penetrated the hCMEC/D3 cell monolayer via encapsulation into the vacuoles. Overall, yeast-derived vacuoles are promising candidates as drug carriers to the brain.

Inhibition of tau phosphorylation and Aß accumulation by S. cerevisiae-derived vacuoles in LPS-induced SH-SY5Y cells.

Neurodegenerative diseases, such as Alzheimer's disease (AD), are characterized by the accumulation of intracellular tau and amyloid beta (Aß) proteins, which lead to neuroinflammation and neuronal apoptosis. In this study, we investigated the potential of a bioengineered vacuoles derived from Saccharomyces cerevisiae-derived vacuoles to treat neuroinflammation and protein accumulation in AD. The yeast-derived vacuole is a small organelle that achieves efficient degradation by utilizing a diverse array of hydrolytic enzymes. These hydrolytic enzymes break down and process proteins into smaller fragments. We found that vacuoles treatment significantly reduced LPS-primed cell apoptosis and diminished Aß42 secretion in vitro, potentially through the inhibition of the NF-kB p65 signaling pathway. Additionally, vacuole pre-treatment down-regulated NF-κB translocation and reduced phosphorylated tau levels in LPS-induced SH-SY5Y cells. Our results suggest that the vacuoles have potential as a therapeutic agent for neurodegenerative diseases. The vacuole's small size and diverse hydrolytic enzymes make it a promising drug delivery system for targeting intracellular proteins. Future studies may explore the use of vacuoles in animal models of AD to determine their therapeutic potential.

Alginate oligosaccharides can maintain activities of lysosomes under low pH condition.

The objective of this study was to report that lysosome extracted from egg white could be used as a drug through oral administration for treating diseases by using pH sensitive alginate oligosaccharides. Lysosome-alginate oligosaccharides composite were formulated for oral administration of lysosomes. The dissolution test confirmed the availability of the oral dosage form. When lysosome were used as an independent drug, the activity of protein was lost due to influence of low pH. Its antibacterial activity was also remarkably reduced. However, when lysosome-alginate oligosaccharides composite form was used, antimicrobial activity of lysozyme was maintained. At low pH, a gel-like matrix was formed by alginate oligosaccharides to protect the lysosome. When the pH was increased, alginate oligosaccharides were dissolved and the lysosome was released. SDS-polyacrylamide gel electrophoresis analysis of released lysosomes revealed that alginate oligosaccharide could effectively protect the lysosome from degradation or hydrolysis under acidic conditions for at least 2 h. The results of this study are important for application of lysosomes as therapeutic agents, and also it was confirmed that alginate oligosaccharides have potential as direct delivery system for the oral application of protein derived therapies.

Lysosomes Alteration in HeLa Cell Exposed with 1-Aminocyclopropane-1-Carboxylic Acid, the Ethylene Precursor of Plant Hormones.

The study of senescence preservative on cut flowers helps boost the commercial value of flowers. Senescence in cut flower is associated with an increase of ethylene production, and is significantly influenced by ethylene pathway. This study was conducted to investigate whether S-adenosyl-L-methionine (SAM) and aminocyclopropane-1-carboxylic acid (ACC) involved in the ethylene synthesis process are correlated with the lysosome. The alterations of lysosome which was treated with the ethylene precursors ACC and SAM in HeLa cell using the confocal laser scanning microscope were investigated. According to the experimental results, the activity of lysosomes increased concentration dependently by ACC treatment, however, no change was observed by SAM treatment. In addition, Liquid chromatography-mass spectrometry (LC/MS) analysis was performed to confirm the effect of lysosomal enzyme (LE) extracted from egg white on ACC reduction, but no change was observed. On the contrary, to confirm the effect of ACC on lysosomes, lysosomes were extracted from HeLa cells treated with 5 mM ACC and confirmed by FE-SEM. The results showed that the size of lysosomes treated with ACC is larger than that of the control, which was treated with distilled water. The lysosomes in the control group were distributed in various ranges from 0 to 800 nm, but those treated with 5 mM ACC were in the range of 400 nm to 800 nm or more. Therefore, lysosomes had no effect on ACC, the precursor of ethylene, the aging hormone of cut flowers, however, ACC had effect on lysosomes.

Rapamycin persistently improves cardiac function in aged, male and female mice, even following cessation of treatment.

Even in healthy aging, cardiac morbidity and mortality increase with age in both mice and humans. These effects include a decline in diastolic function, left ventricular hypertrophy, metabolic substrate shifts, and alterations in the cardiac proteome. Previous work from our laboratory indicated that short-term (10-week) treatment with rapamycin, an mTORC1 inhibitor, improved measures of these age-related changes. In this report, we demonstrate that the rapamycin-dependent improvement of diastolic function is highly persistent, while decreases in both cardiac hypertrophy and passive stiffness are substantially persistent 8 weeks after cessation of an 8-week treatment of rapamycin in both male and female 22- to 24-month-old C57BL/6NIA mice. The proteomic and metabolomic abundance changes that occur after 8 weeks of rapamycin treatment have varying persistence after 8 further weeks without the drug. However, rapamycin did lead to a persistent increase in abundance of electron transport chain (ETC) complex components, most of which belonged to Complex I. Although ETC protein abundance and Complex I activity were each differentially affected in males and females, the ratio of Complex I activity to Complex I protein abundance was equally and persistently reduced after rapamycin treatment in both sexes. Thus, rapamycin treatment in the aged mice persistently improved diastolic function and myocardial stiffness, persistently altered the cardiac proteome in the absence of persistent metabolic changes, and led to persistent alterations in mitochondrial respiratory chain activity. These observations suggest that an optimal translational regimen for rapamycin therapy that promotes enhancement of healthspan may involve intermittent short-term treatments.

Late-life restoration of mitochondrial function reverses cardiac dysfunction in old mice.

Diastolic dysfunction is a prominent feature of cardiac aging in both mice and humans. We show here that 8-week treatment of old mice with the mitochondrial targeted peptide SS-31 (elamipretide) can substantially reverse this deficit. SS-31 normalized the increase in proton leak and reduced mitochondrial ROS in cardiomyocytes from old mice, accompanied by reduced protein oxidation and a shift towards a more reduced protein thiol redox state in old hearts. Improved diastolic function was concordant with increased phosphorylation of cMyBP-C Ser282 but was independent of titin isoform shift. Late-life viral expression of mitochondrial-targeted catalase (mCAT) produced similar functional benefits in old mice and SS-31 did not improve cardiac function of old mCAT mice, implicating normalizing mitochondrial oxidative stress as an overlapping mechanism. These results demonstrate that pre-existing cardiac aging phenotypes can be reversed by targeting mitochondrial dysfunction and implicate mitochondrial energetics and redox signaling as therapeutic targets for cardiac aging.

Malnutrition among 6-59-Month-Old Children at District 2 Hospital, Ho Chi Minh City, Vietnam: Prevalence and Associated Factors.

OBJECTIVES: Childhood malnutrition is major health concern in many low- and middle-income countries, including Vietnam. It was a major risk factor for child mortality and adult ill-health. Malnutrition could increase the risk of serious infections; conversely current diseases also had a negative impact on the growth of child. This study, therefore, examines the prevalence of stunting and underweight among 6-59-month-old outpatient children in District 2 Hospital, Ho Chi Minh City, Vietnam. METHODS: A cross-sectional study involved a sample of 225 children aged 6-59 months who were randomly selected from the Outpatient Department in District 2 Hospital. Anthropometric measurements and blood test of children were taken to assess the nutritional status and anaemia. A structured questionnaire was also used to collect mothers' and children's characteristics to examine associated risk factors. RESULTS: The prevalence of stunting, underweight, overweight, and anaemia among children aged 6-59 months was 9.8%, 8.4%, 25.8%, and 30.7%, respectively. Underweight significantly correlated only to having breastfeeding in the first hour after birth (RR: 0.02; 95% CI: 0.01-0.17; p<0.001), while stunting was related to age of starting complementary foods from equal to/more than 6 months (RR=0.70, 95%CI=0.50-0.99, p<0.05) and normal birth weight (RR = 0.29, 95%CI = 0.15-0.56, p<0.001). CONCLUSIONS: This study emphasized the importance of measuring the overall nutritional status for children, who have coexisting infectious diseases and anaemia. The high prevalence of malnutrition and anaemia underlined the need for routine screening as well as treatment of children. Additionally, health information strategies should be focused on young children feeding practices to minimize stunting and underweight.