Medical Research Archives

Human diversity did not only remain restricted to their socio-cultural and linguistic domains but also have penetrated deep inside their genetic root. Africa harbors more genetic diversity than any other part of the world. Diversification of the African lineages were complex, involving long-distance gene flow. Data from Africansis needed to better understand the origin and evolution of modern humans, the genetic basis local adaptation, and the evolution of complex traits and related diseases. This analysis formed the basis for this study of determining the origin and migration of the Ameru community in Kenya. Blood samples was collected from 132 male adults of 65 year and above. DNA was extracted and analyzed for the Hyper variable region 1and 2. The sequences were sequenced using Sanger sequence alignment and analyzed using Geneious. Phylogenetic analysis was done using Mega-X while haplotype analysis was done using DNASP software. L1 haplogroup (2.9%) was found among Igembe (7%), Tharaka (6%), and Chuka (7%) and is common in West, Central, and parts of East Africa. L2 haplogroup (6.7%) was present in all subgroups except Imenti and Tigania, indicating West and Central African maternal ancestry. L1 and L2 haplotypes indicate that most Ameru subgroups share partial maternal ancestry from West and Central Africa, while Imenti and Tigania have different maternal lineages. L0-L4 haplogroups indicate predominant East, Central, and West African maternal origins, with subgroups showing variation in haplotype frequencies (e.g., L1 and L2 in Igembe, Tharaka, Chuka; L3 in Tharaka, Mwimbi, Chuka; L4 across all subgroups). Subgroup differences suggest that certain communities, particularly Imenti, have distinct maternal lineages, with less contribution from L1, L2, and L3 but potential links to Afro-Asiatic groups via L4 (found in the Middle East). Non-African haplogroups (N and R) point to historical interactions or shared ancestry with populations in Eurasia and the Horn of Africa, primarily in Tigania and Imenti. Overally, the Ameru maternal gene pool is heterogeneous, shaped by multiple migration routes and interactions across East Africa and beyond, with subgroup-specific maternal histories.

IntroductionIndividuals with COPD can be classified according to their levels of physical activity (PA) and physical capacity (PC). The relationship between nutrition and body composition within these classifications remains unclear. ObjectivesTo compare the body composition and food intake of people with COPD and verify the associations. MethodsCross-sectional exploratory analysis study in which body composition and food intake were assessed in individuals with COPD. Classification was based on six-minute walk test (PC) and accelerometry(PA): Quadrant "can do, dont do" (I-preserved PC, low PA); quadrant "can do, do do" (II-preserved PC, preserved PA). Results72 individuals with COPD, 39 in quadrant I and 33 in quadrant II, with mean ages of (69 {+/-} 6) (67 {+/-} 7), respectively. Group I had a higher proportion of males, whereas group II had a higher proportion of females. A positive trend in skeletal muscle mass (p=0.011) (B= 2.883) and a negative trend in basal metabolic rate (p=0.010) (B=-0.092) for group I. ConclusionBrazilians with COPD classified in quadrants I and II showed similar results in terms of body composition and food intake. A positive trend in skeletal muscle mass was observed for the group I. These findings align with the pathophysiological model of COPD, in which the preservation of muscle mass and adequate protein intake support functional capacity and the maintenance of higher physical activity levels.

Introduction Obstructive lung diseases (OLDs) are responsible for high rates of illness and death worldwide. Inflammation, chronic airflow limitation, and bronchial remodeling occur in OLD and eventually result in the unique respiratory sounds. Despite its subjective and having low reproducibility, still traditional auscultation using a manual stethoscope is the main method used to identify the lung sounds. Nevertheless, the combination of recent advancements in digital stethoscopes and AI (Artificial Intelligence) has permitted the objective measurement of lung sounds. Nevertheless, there is a lack of standardized, region-specific databases for AI training and validation. Even though lung sound classification is an emerging aspect in research and telerehabilitation the lobar wise acoustic pattern is still novel due to lack of prevailing database to train AI models. Identifying this gap this study aims to develop an acoustic repository and analyze the data using segmental lung sounds from patients with OLDs and healthy controls through an electronic stethoscope. Methods and analysis This is a cross sectional observational study involving 120 participants (60 OLD patients and 60 healthy controls). Lobar wise acoustic signals will be captured using an electronic stethoscope in healthy and diseases population. The data will be analyzed using Audacity software for annotations and then it will be used for feature extraction and statistical analysis. The acoustic features extracted through Audacity, will include frequency, intensity, pitch, and root mean square (RMS) energy. Repeated measures ANOVA will be applied to compare mean sound intensities across lung segments while Pearson correlation will be used to assess associations with body composition parameters. The data will then be standardized for AI-based diagnostic applications. Ethics and dissemination The study is being reviewed from the Ethics Review Committee, Faculty of Medicine, University of Peradeniya (2025/EC/87) will be sought. Informed consent will be obtained in writing. The dissemination of results will take place through peer-reviewed publications and the creation of a public database containing lung sounds from the region.

Dengue virus (DENV) infection generates a significant health burden throughout the world, and there are no clinically approved antiviral drugs, as of now. The virus also depends on lipid metabolism in the host to conduct effective replication and this makes lipid-directed compounds promising as therapeutic options. We assessed the antiviral effect of lauric acid, a 12-carbon medium-chain fatty acid, against DENV serotype 2 (DV2) in the presence of a stable cell line, DV2-replicon, expressing all the non-structural proteins (NS1-NS5) and a luciferase reporter. Active viral replication in replicon cells was established by morphological examination and immunofluorescence of cells. The MTT assay was used to determine the cytotoxicity of lauric acid revealing the LD50 of 2.52 uM, so higher concentrations were toxic as the effect of the drug is dose-related. The antiviral effect was tested through replicon inhibition (luciferase) assay which showed an incredible inhibition of viral RNA replication with a IC50 of 1.70 uM and this is equivalent to antiviral mycophenolphycic acid. The cytopathic effects, as well as a decrease in the activity of luciferase, proved the presence of viral translation and replication inhibition within the process of the treatment of the lauric acid. These results propose that lauric acid has cytotoxic and antiviral dual effect and can be a possible inhibitor of DENV replication. The toxicity needs to be reduced and future research is necessary to explain its molecular pathway and also to come up with the best delivery methods. IMPORTANCEDengue virus (DENV) remains a significant health challenge to the world since there are no effective antiviral agents. This work will recognize lauric acid as a possible dengue virus replication inhibitor in a model of a DV2 replicon, exhibiting antiviral action that is similar to that of mycophenolic acid. These results support lipid-directed compounds as potential dengue antiviral targets, but more research is needed to minimize toxicity and better understand the molecular mechanism of action.

Background: Chronic Obstructive Pulmonary Disease (COPD) is the third leading cause of global mortality, with persistent lung inflammation contributing to disease progression. This inflammation is partly associated with reduced levels of histone deacetylase 2 (HDAC2). Previous studies suggest that Vitamin D may modulate HDAC2 levels. This study aimed to evaluate the effect of Vitamin D supplementation on HDAC2 expression in stable COPD patients. This experimental study aimed to evaluate the effect of vitamin D supplementation on HDAC2 expression in stable COPD patients at Jemursari Islamic Hospital. Methods: Five COPD patients received a daily dose of 5000 IU of Vitamin D for three months. Serum levels of 25(OH)D3 and HDAC2 were measured before and after the intervention. Results: Vitamin D supplementation resulted in a significant increase in both 25(OH)D and HDAC2 levels. Pulmonary function parameters showed an increasing trend, however, no statistically significant differences were observed. Conclusion: Vitamin D supplementation was associated with increased HDAC2 levels, suggesting a potential anti-inflammatory effect. However, no significant improvement in pulmonary function was observed. Further studies are needed to determine its clinical impact.

BackgroundBronchiectasis in adults often goes undiagnosed following the routine assessment. Cystic Fibrosis (CF) is usually diagnosed during childhood, but some cases are identified in adulthood when disease is mild. High-resolution computed tomography (HRCT) of chest may offer structural information that can indicate CF as an underlying etiology. ObjectiveTo compare the HRCT features of adult patients with CF and non-CF bronchiectasis and to determine the radiologic features that may be suggestive of CF. MethodsThis retrospective, analytical, cross-sectional study was carried out in Bangladesh Medical University after IRB clearance. Total 130 adults (12 with CF and 118 with non-CF bronchiectasis) of both sexes, whose bronchiectasis was confirmed by chest HRCT were included. Imaging findings were assessed based on Reid morphological classification, anatomical distribution and extent of spread within the lungs, and their association was tested using chi-square test with statistical significance of p<0.05. ResultsCystic bronchiectasis was more common in CF than non-CF patients (83.3% vs 29.7; p<0.001). Mixed central-peripheral extension had been found a considerable associated with CF (66.7% vs. 42.4; p=0.034). There was no statistically significant difference in right lung lobar distribution (p=0.540) but combined upper and lower lobe involvement on the left side was more common in CF patients (54.5%) than non-CF patients (21.3) (p=0.054). ConclusionAdult CF had unique chest HRCT imaging characteristics when compared to non-CF bronchiectasis, especially cystic morphology and mixed extension. Identification of such features could help physician in the early diagnosis and selection of treatment strategy.

O_LIPlants and fungi are major sources of natural products beneficial to society, making the study of distinct species essential for discovering new drugs and bioactive compounds. The medicinal mushroom "Lingzhi" or "Reishi" (Ganoderma lingzhi) is widely used in traditional medicine and extensively studied for its bioactive triterpenoids, yet it is commonly identified as Ganoderma lucidum, the type species of the genus, which lacks a type specimen. C_LIO_LIWe sequenced a G. lucidum specimen preserved in the Kew fungarium, which agreed with the original description and was collected from wood of Corylus avellana in southern England. Using this reference specimen, we compiled genomic and ITS barcoding datasets to explore the genetic and geographic variation within this species. C_LIO_LIWe showed that G. lingzhi and G. lucidum diverged more than 12 million years ago and that all seven "G. lucidum" genomes deposited in public databases belong to other species. More than 1000 barcoding sequences showed that the widely used homology-based ITS barcoding is not working in this group, which can be mitigated by a phylogenetic placement approach. The 149 sequences assigned to G. lucidum with high confidence showed a Eurasian distribution and introductions to North and South America and Africa. C_LIO_LIOur study underscores the importance of accurate species identification and provides guidance for a group of pharmaceutical and socially significant species. To further support future studies and the wider public in differentiating between G. lingzhi and G. lucidum, we propose using "False Lingzhi" as the English name for G. lucidum. C_LI Societal Impact StatementTraditional Chinese Medicine has expanded far beyond Asia, with growing markets in North America and Europe for supplements and functional foods. Lingzhi or Reishi (Ganoderma lingzhi), a well-known medicinal mushroom, is valued for its anti-inflammatory and anticancer properties. However, it is often misidentified with species that may not provide the same health benefits. This confusion poses risks to consumer safety, product regulation, and research. Here, we establish a reference using morphological and molecular tools for the most commonly misidentified species (Ganoderma lucidum) and propose the name "False Lingzhi" to distinguish it, supporting accurate identification, safer product development, and reliable research.

Introduction Recent respiratory illness, especially influenza, may trigger acute cardiac events via elevated inflammatory mediators. During the 2018 influenza season in Bangladesh, this study examined whether recent acute clinical respiratory illness (CRI) or laboratory-confirmed influenza was associated with elevated hs-CRP and IL-6, linked to acute cardiac events. Methods A total of 139 participants aged [&ge;]40 were recruited from a Dhaka cardiac hospital: 70 with acute myocardial infarction (AMI), 30 with other acute cardiac events, and 39 healthy individuals. CRI was defined as fever with cough and/or respiratory symptoms within seven days. Respiratory swabs were tested for influenza, and blood was analyzed for hs-CRP and IL-6. Results Median hs-CRP and IL-6 were higher in participants with CRI or influenza but not significantly. Cardiac patients had elevated hs-CRP (9.98 mg/L in other cardiac; 4.86 mg/L in AMI vs. 1.73 mg/L in healthy) and IL-6 (0.1 pg/mL in other cardiac; 0.145 pg/mL in AMI vs. 0.08 pg/mL in healthy) (p<0.001). CRI was not significantly associated with elevated hs-CRP or IL-6, though influenza in healthy participants was linked to higher IL-6. Cardiac patients had a higher risk of hs-CRP [&ge;]3 mg/L and elevated IL-6. Conclusion Cardiac patients showed significantly increased inflammatory markers, but CRI was not clearly linked to inflammation. Further research should assess biomarker utility for early cardiac risk.

BackgroundThe primary cause of death associated with opioids is opioid-induced respiratory depression (OIRD). Naloxone is used to reverse OIRD, but this drug is a competitive antagonist of {micro}-opioid receptor (MOR) and reverses analgesia, which limits its therapeutic use. Alternative non-opioid receptor antagonist-based approaches to OIRD treatment and prevention are needed. The aim of this study was to evaluate if setmelanotide (SET) is capable of reversing OIRD in a mouse model. MethodsC57BL/6J male and female mice and Sprague-Dawley rats were given IP morphine or fentanyl and then treated 15 min later with either SET or vehicle VEH (IP) in a random order. Breathing was recorded by barometric plethysmography, and pain sensitivity was measured by the tail-flick test. ResultsIn mice with OIRD, SET induced a 3-fold reduction of the apnea index, and decreased apnea duration as compared to the VEH treatment. SET increased respiratory rate and did not affect opioid-induced analgesia. Photostimulation of MC4R+ ChR2-expressing fibers in the parafacial region of MC4R-Cre mice elicited short-latency excitatory postsynaptic current in rostral ventral respiratory group (rVRG) pre-motoneurons projecting to the phrenic nucleus in the C3-C4 ventral horns of the spinal cord. Fentanyl inhibited the activity of rVRG neurons and SET reversed this effect. ConclusionsSET effectively treated OIRD by increasing respiratory rate and inducing a significant decrease in the number of apneas without decreasing analgesia.

Acidification of the oral environment has been implicated in the initiation and progression of oral pathologies including oral cancer, but how acidic environments modulate normal oral epithelial cell (OEC) responses to microbial ligands is not understood. This study examined the impact of acidic stress on OEC morphological, molecular, and functional responses to toll-like-receptor ligand engagement in vitro. OEC cultures were exposed to either normal (pH:=:8.0) or acidified growth media (pH:=:3.0) for 24 hours prior to machine-learning-guided morphological analysis and exposure to either toll-like receptor (TLR)5 (flagellin) or TLR2/TLR1 (Pam3CSK4) agonists. Multiplex gene expression technology was used to quantify the transcriptional responses of metabolic-and immune-related genes at 6 hours post-TLR agonist exposure. OEC-mediated production of transforming growth factor-beta (TGF-{beta}) was assessed by enzyme-linked immunosorbent assay at 2-, 6-, and 24-hours post-agonist exposure. Results showed that acid exposure induced significant changes to OEC morphology resembling epithelial-mesenchymal transition, the differential expression of n=197 metabolic-and n=43 immune-related genes and significantly increased OEC TGF-{beta}1 production. The results demonstrate that acid stress skews normal OECs towards pro-inflammatory and pro-oncogenic phenotypes when faced with concomitant microbial ligand challenge and provide key molecular clues to OEC survival strategies with potential implications for elucidating the early molecular events in the development of epithelial dysplasia. Article HighlightsO_LIAcute acid exposure reduces survival of OECs C_LIO_LIA subpopulation of OECs is resistant to acid-mediated cell loss and undergo morphometric changes consistent with epithelial-mesenchymal transition C_LIO_LIConcurrent acid stress and TLR stimulation modulates transcription of immune and metabolic genes in OECs C_LIO_LIAcid stress increases TGF-{beta}1 protein production of OECs following TLR agonist stimulation C_LI

Spinal cord injury (SCI) is associated with cardiovascular deficits that affect cerebral blood flow, cerebral perfusion, and cerebrovascular control. While several studies use neuroimaging techniques such as functional magnetic resonance imaging (fMRI) to understand neuroplasticity following SCI, more work needs to be done to evaluate the cerebrovascular changes following SCI. Understanding these effects using neuroimaging is essential as these deficits also affect neurovascular coupling and how we interpret neuroplasticity measured based on neuroimaging. Hence, we conducted a pilot study in twelve healthy males and thirteen males with thoracolumbar SCI using functional near-infrared spectroscopy (fNIRS) to understand the effects of breath-holding induced hypercapnia on the hemodynamics of the sensorimotor cortex and prefrontal cortex (PFC) after SCI. Participants performed 30 seconds of regular breathing alternated by 15 seconds of breath-holding for 5 minutes. Compared to controls, the SCI group presented with a greater initial decrease in oxy-hemoglobin concentration change and a delayed subsequent increase in oxy-hemoglobin concentration change in response to hypercapnia at p<. Additionally, the net increase in oxy-hemoglobin concentration change following BH in the PFC was negatively correlated with the level of injury at p=0.005, where higher levels of injury were associated with a smaller increase in oxy-hemoglobin concentration following hypercapnia. These findings confirm that a) SCI, including lower levels of injury (below T6) are associated with cerebrovascular changes that are quantifiable using fNIRS, and b) fNIRS could be a robust tool to understand the neuroplastic and cerebrovascular changes in people with SCI.

Since the initial distribution of the SARS-CoV-19 vaccine, its widespread use has been hypothesized to act as a selective pressure that drives the COVID-19 virus to mutate. This study aims to investigate the correlation between global vaccination rates and the mutation rate of the SARS-CoV-2 Beta variant (B.1.351). From January to July 2021, nucleotide diversity increased in tandem with vaccination rates, demonstrating that the virus evolved more rapidly in response to selective pressure from mass vaccination. Statistical analysis revealed statistically significant positive correlations between both vaccination rates and vaccine doses administered with nucleotide diversity. Thus, our findings indicate a positive correlation between rising vaccination rates and nucleotide diversity, suggesting that increased vaccination coverage acted as a selective pressure that accelerated viral evolution of SARS CoV2.

Dental enamel, the hardest mineralised tissue in the human body, has proven to be an excellent source of ancient proteins, which have been found to survive within dental enamel for at least twenty million years. In archaeological and palaeontological contexts, the enamel proteome is generally considered to be rather small, consisting of about twelve proteins, most of which are unique to enamel. During amelogenesis these proteins undergo in vivo digestion by matrix metalloproteinase 20 (MMP20) and kallikrein 4 (KLK4) as well as serine phosphorylation by family with sequence similarity member 20-C (FAM20C) that alter their characteristics. Gaining knowledge of the previously understudied influence of amelogenesis on the archaeological human dental enamel proteome could benefit various palaeoproteomic analysis, especially in an human evolutionary context. Here we present archaeological dental enamel proteomes and explore protein cleavage patterns and sequence coverage to estimate the effects of in vivo digestion, as well as explore phosphorylation patterns. Additionally, we present a new marker based on phosphorylation to estimate genetic sex.

Primary human bronchial epithelial cells (pHBECs) of the airways of smokers are chronically exposed to cigarette smoke, which may induce chronic obstructive pulmonary disease (COPD) ranked fourth among the most common global causes of death. Using an established protocol for differentiation of pHBECs to a pseudostratified epithelium at an air liquid interface (ALI), we analyzed functional expression of transient receptor potential vanilloid 4 (TRPV4) proteins after application of cigarette smoke extract (CSE), which upregulated seven smoke exposure regulated genes (SERGs). TRPV4 protein expression in the plasma membrane and localization next to the cilia of ciliated cells was reduced, while cell barrier function was not altered after chronic exposure to CSE for 28 days compared to untreated control cells. Accordingly, TRPV4-mediated Ca2+ influx was blocked in pHBECs after CSE exposure. Moreover, Os-9 protein, which after binding mediates protection from degradation of TRPV4 protein by polyubiquitination, was significantly less expressed in pHBECs upon CSE exposure. Most interestingly, overexpression of OS-9 in pHBECs rescued reduced TRPV4 protein levels induced by CSE. Our study identifies a novel molecular mechanism of toxicity by CSE interfering with TRPV4 and OS-9 expression in pHBECs, which may blaze the trail for new therapeutic options in COPD.

BackgroundIn the conflict-ridden Gaza Strip, burn injuries pose significant challenges for affected individuals. Often stemming from accidents involving generators and substandard gas cylinders, these burns can have profound physical and psychological impacts. Access to comprehensive medical care is limited in this context, making specialized burn treatment centers critical lifelines for those in need. This study seeks to explore the outcomes of rehabilitation and physiotherapy for post-surgical skin grafts in specialized burn treatment centers in Gaza, recognizing the crucial role such interventions play in improving the quality of life for burn survivors in this challenging environment. MethodsWe carried out a retrospective descriptive study of patients with post-surgical skin graft (SSG) scars subsequent to burns injury, and enrolled in the MSF post-op outpatient clinics in the Gaza Strip (Gaza City, Beit Lahia, and Khan Younis), between January 2018 and December 2020. Our analysis used five outcome measures to assess the effects of physiotherapy and post-surgical treatment: the reducibility score, which indicates the extent to which scars can be reduced in size or prominence; the Functional Activity for Burn (FAB) score, which evaluates the functional capabilities of patients following burn injury; the Vancouver Scar Scale (VSS), which assesses various aspects of scar appearance such as pigmentation, vascularity, pliability, and height; the Visual Analgesic Scale (VAS), which quantifies pain levels experienced by patients; and the itching score, which measures the severity of itching associated with scar formation ResultsA total of 177 patient records were examined, revealing that the majority of burn victims in Gaza were children under the age of 18 (n=136, 76.8%), with scalding from liquid burns being the primary cause (n=119, 67.6%). The outcomes of the physiotherapy program varied depending on the type of pressure therapy and insert material utilized. However, significant improvements were observed in various outcome measures following enrollment in the program. These improvements included reductions in pain scores (mean initial score: 5.3, SD: 2.5; mean final score: 1.4, SD: 1.8), itching (mean initial score: 3.7, SD: 2.7; mean final score: 2.7, SD: 2.2), scar pigmentation, vascularity, pliability, and height (mean initial VSS: 7.1, SD: 1.8; mean final VSS: 5.7, SD: 1.7). Moreover, treatment correlated with enhancements in overall function (mean initial score: 25.6, SD: 7.1; mean final score: 34.6, SD: 2.3) and a reduction in skin contracture (mean initial score: 2.3, SD: 1.4; mean final score: 0.8, SD: 0.9). ConclusionsOur findings underscore the important role of rehabilitation and physiotherapy in optimizing outcomes for post-surgical skin graft patients in conflict-affected regions like the Gaza Strip. Despite the challenging environment, MSFs clinic in Gaza has demonstrated the feasibility and effectiveness of delivering comprehensive care to burn survivors. Moving forward, further research is needed to refine and validate best practices in rehabilitation interventions tailored to the specific needs of patients in conflict zones, ensuring continued progress in enhancing the quality of care and quality of life for those affected by burn injuries.

BackgroundTuberculosis (TB) is the second-leading cause of deaths from infectious agents and remains a global health threat. Ethionamide (ETH) is a prodrug used in regimens for multidrug-resistant TB, and, partly due to side effects that can lead to low treatment adhesion, resistance arises. Changes in EthA, the monooxygenase that activates ETH, are the main mechanism of resistance. Yet, of hundreds of EthA substitutions found in resistant isolates, only a handful have been annotated as resistance determinants. ResultsAn in silico analysis was carried out on a previously described panel of Mycobacterium tuberculosis clinical isolates for which genomes and ETH susceptibility testing results were available. EthA substitutions were mapped, revealing the existence of hotspots in its sequence. Visualization of the hotspots in the EthA structural model shows that they cluster in three regions, including ligand binding pockets. Models were built of twenty-three variants found in resistant isolates and changes in local configuration was mapped to identify investigate impact on ETH activation. Information from these models contributed to establishing five criteria for scoring whether substitutions are most likely to lead to resistance. Using these criteria, EthA D58G was selected and its expression is shown to increase growth in high ETH concentrations. ConclusionFunctionally relevant regions of EthA are revealed and point out priority substitutions for functional studies, enhancing identification and detection of substitutions not been previously associated with resistance.

Pentosan polysulfate (PPS) is a semisynthetic sulfated polysaccharide that was approved by the United States Food and Drug Administration (FDA) for treatment of interstitial cystitis (IC). A 2018 study by our group described a vision-threatening macular toxicity associated with long-term use of PPS. However, given the relatively recent characterization of PPS maculopathy, we have limited knowledge of its pathophysiology. The present study therefore investigated the pathophysiology of PPS maculopathy in a cell culture model, assessing impacts of PPS exposure on morphology and mitochondrial function. We treated ARPE-19 cells with increasing doses of PPS and investigated both mitoprotective and cytoprotective mechanisms, mitochondrial reactive oxygen species production (ROS) and respiration, cellular structure, and retinal pigment epithelium (RPE) dysfunction through phagocytosis assays. We found that PPS increased mitochondrial superoxide accumulation and that increased doses of PPS impaired basal and maximal respiration in a Seahorse assay without the expected response of increases in the cellular energy sensor pAMPK. PPS exposure disrupted mitochondrial and cell protective mechanisms against ROS accumulation as assessed through examination of mitochondrial biogenesis markers PGC-1 and SIRT1 and autophagy markers LC3 and p62. PINK1 expression increased with increasing duration of exposure to PPS. Further, we found that PPS led to functional and structural changes to RPE cells, which exhibited an increase in cell aspect ratio and impaired phagocytosis with higher doses of PPS. Lastly, we found an increase in cell death in response to higher doses of PPS, evident through ethidium homodimer cell viability assays. Taken together, our study shows PPS exposure has profound effects on RPE viability and function through impairment of mitochondrial respiration and mito- and cyto-protective mechanisms and highlights mitochondrial insult as a potential focus of future PPS research.

Breastfeeding provides health benefits in childhood, reducing the frequency of gastrointestinal and respiratory infections. Breastmilk (BM) is a rich source of bioactive molecules including extracellular vesicles (EVs), which exert immunomodulatory signalling in recipient cells, with cargo that is affected by maternal characteristics. Here we investigated the biophysical characteristics of BM-EVs from mothers with (asthmatic BM-EVs) or without asthma (control BM-EVs) and their effect on the release of cytokines from primary human hTERT-immortalized airway smooth muscle cells (hASMs) from asthmatic or non-asthmatic (control) donors. BM-EVs were isolated using size exclusion chromatography (N=5/group), characterized biophysically and by EV-specific protein markers. In addition, BM-EV were co-cultured (48h) with primary hASM cells from both non-asthmatic (control) and asthmatic donors to determine the effect on cytokine release. All participants were Caucasian and the BM was collected 12-15 weeks postpartum. BM-EVs showed the presence of intact and small-EVs ([~]100 nm). Asthmatic BM-EVs appeared to have a smaller average EV size (135.6 nm) vs. controls (148.3 nm, p=0.0613), but [~]5-fold higher concentration of both total (p=0.0014) and small EVs (p=0.0016). The expression of EV subtype protein expression was reduced in asthmatic BM-EVs vs. control BM-EVs: CD63 by 86% (p=0.0224), flotillin-1 by 40% (p=0.0196), CD9 by 24% (p=0.0646) and HSP70 by 69% (p=0.0873). Asthmatic BM-EVs co-cultured with hASMs from control donors decreased pro-inflammatory cytokine release: MCP-1 by 55% (p=0.0286), IL-6 by 45% (p=0.0801) and IL-2 by 32% (p=0.0970) vs. control-BM-EVs. Conversely, asthmatic BM-EVs co-cultured with hASMs from asthmatic donors increased secretion of anti-inflammatory cytokine IL-10 by 32% (p=0.0660), and IL-1Ra by 75% (p=0.0875), and pro-inflammatory IL-2 by 57% (p=0.0688) vs. control-BM-EVs. Internalization of control and asthmatic BM-EVs was confirmed by labelled EV uptake experiments. No detrimental effects on cell viability with BM-EV treatment were observed. In summary, asthmatic BM-EVs are smaller and enriched in BM, and exert differential effects on cytokine release in a BM-donor and recipient-cell specific manner. Given that BM can enter infant airways, the immunomodulatory effects of BM-EVs on hASMs warrants further investigation to delineate the under underlying mechanisms.

Cryptosporidium parvum is a protozoan parasite responsible for cryptosporidiosis, significantly threatening immunocompromised individuals, particularly HIV/AIDS patients, by causing severe diarrhea and potential mortality. Current treatments are largely ineffective, prompting investigations into new therapeutic options. This study evaluated two antiparasitic drugs: Mebendazole, used for helminth infections, and Artemisinin, used for malaria. The SKSR gene family encodes virulence factors in C. parvum, and Calcium-dependent protein kinase1 (CpCDPK1) regulates the life cycle of C. parvum; targeting these proteins may reduce growth and infection in hosts. In the current study, molecular docking was conducted taking Mebendazole and Artemisinin drugs as ligands, SKSR gene family and CpCDPK1 proteins as drug targets. Results with SKSR showed binding energy of -4.9 kcal/mol, -6.72 kcal/mol for Mebendazole and Artemisinin, respectively. Whereas, with CpCDPK1, the binding energies were -6.44 kcal/mol, -9.18 kcal/mol for Mebendazole and Artemisinin, respectively. Docking of Nitazoxanide (an in-use drug for C. parvum) with SKSR and CpCDPK1 revealed binding energies -4.2 kcal/mol, -4.81 kcal/mol, respectively. The stability of the proteins (targets) upon binding to the ligands was assessed by performing all-atom MD simulations for 100ns using the GROMACS package. No major variations were observed upon binding of Artemisinin and Mebendazole to SKSR and CpCDPK1. The findings of MD simulations imply that both proteins maintain their stability upon binding of Artemisinin and Mebendazole. Molecular Docking and MD simulation studies suggest that Artemisinin and Mebendazole are potential candidates for repurposing in the treatment of C. parvum infections, with recommendations for in vitro studies to validate these findings.

The origin of eukaryotes is a key event in the evolution of cellular life hypothesized to involve a symbiotic integration between a member of the Asgard archaea and the Alphaproteobacteria. Recent work has provided evidence for additional genetic input from other prokaryotes to the eukaryotic proteome yet the extent and sources of these contributions remain debated. Here we aimed to further resolve the prokaryotic origins of eukaryotic genes to inform our understanding of eukaryogenesis. Specifically, we developed a phylogenetic framework to investigate the origins of eukaryotic gene families associated with metabolism and informational processing for comparison. We found that informational processing genes were predominantly derived by archaea whereas eukaryotic metabolism is highly chimeric in its origin. In contrast to previous studies, we report a substantial number of archaeal origins of diverse metabolic enzymes including key metabolic regulators. This highlights an overlooked participation of archaeal metabolism and pinpoints potential metabolic integrations during eukaryogenesis. Apart from the alphaproteobacterial contributions to the eukaryotic metabolism, we found an additional dominant phylogenetic signal of genes potentially derived from Myxococcota, especially for gene families associated with lipid metabolism. By systematically analysing the origins of eukaryotic metabolism, this research offers novel insights into the origin of eukaryotic membranes and refine our current models for the origin of the eukaryotic cell.