Micropatterning in BioMEMS for Separation of Cells/Bioparticles

Biofluids remain a difficult issue in some drug delivery processes for separation of bioparticles through microchannels. This chapter reviews the techniques which have been substantiated and proven helpful for the separation of particles depending on mass and size with some constraints of high throughput. In this study, a key focus will be on separation criterion by patterning of a microchannel and utilize sieve type channels based on spherical bioparticles. The first part focuses on the designing of the pattern for issues of the network like clogging and theoretical experiments by both hydrodynamic and other passive methods for sorting/separation. The second part focuses on the simulations for separation for small and larger bio particles depending on mass and size, samples of blood and other Klebsiella infected fluidic samples for the experiment. The theme provided for mass and size-based separation is simple and can accomplish operations in microfluidics for several biological experiments, diagnosis approaches and zoological researches

Targeting neurodegenerative phenotypes and mitochondrial dysfunction in a Drosophila Melanogaster model of Parkinson’s Disease

Parkinson's disease is the most common neurodegenerative disease, characterised by the abnormal accumulation of protein aggregates. It involves gradual loss of dopaminergic neurons in the substantia nigra part of brain, which leads to motor symptoms such as tremor, rigidity, and bradykinesia, as well as non-motor symptoms includes mood disorders, sleep disturbances, and autonomic dysfunction. Mitochondrial dysfunction, particularly involving Complex I of the electron transport system and mitophagy pathways, is significantly affected in the pathogenesis of PD. This study investigates the roles of NUBPL and PINK1 mutations in mitochondrial dysfunction, focusing on ageing as a risk factor, using Drosophila melanogaster and human fibroblast cells as models for PD. The multidisciplinary approach investigated the mutations in Drosophila genotypes, including W1118, PINK1B9, NUBPL, and the double mutant PINK1B9, NUBPL. Mitochondrial function was evaluated using Drosophila's survival assays, qRT-PCR, mitochondrial respiration assays, ATP measurements, protein expression analysis, and RNA sequencing. In parallel, human fibroblast cells carrying NUBPL mutations were subjected to cell viability assays, DNA damage and mitochondrial stressors. Our qRT-PCR and Western blot analyses confirmed decreased mRNA and protein levels of NUBPL and PINK1, emphasising an age-related reduction in these proteins. PINK1 mutants exhibited significant mitochondrial dysfunction, whereas the PINK1B9, NUBPL double mutant partially rescued mitochondrial function, indicating the activation of compensatory mechanisms. RNA sequencing discovered the enrichment of several pathways, including innate immune defence pathways and mitochondrial bioenergetics pathways, highlighting genes such as Defensin, Metchnikowin, Attacin-A, mitochondrial aconitase and blw. Human fibroblast cells with NUBPL mutations showed no changes in the treatment with mitochondrial stressors and DNA damage toxins, indicating cell adaptation and resilience against toxins. Overall, the research outlines that the NUBPL and PINK1 mutations cause previously unpredicted effects on mitochondria and compensatory responses in order to ensure cell survival

Colitis and Colon Cancer in WASP-Deficient Mice Require Helicobacter Species

Background: Wiskott–Aldrich syndrome protein–deficient patients and mice are immunodeficient and can develop inflammatory bowel disease. The intestinal microbiome is critical to the development of colitis in most animal models, in which Helicobacter spp. have been implicated in disease pathogenesis. We sought to determine the role of Helicobacter spp. in colitis development in Wiskott–Aldrich syndrome protein–deficient (WKO) mice. Methods: Feces from WKO mice raised under specific pathogen-free conditions were evaluated for the presence of Helicobacter spp., after which a subset of mice were rederived in Helicobacter spp.–free conditions. Helicobacter spp.–free WKO animals were subsequently infected with Helicobacter bilis. Results: Helicobacter spp. were detected in feces from WKO mice. After rederivation in Helicobacter spp.–free conditions, WKO mice did not develop spontaneous colitis but were susceptible to radiation-induced colitis. Moreover, a T-cell transfer model of colitis dependent on Wiskott–Aldrich syndrome protein–deficient innate immune cells also required Helicobacter spp. colonization. Helicobacter bilis infection of rederived WKO mice led to typhlitis and colitis. Most notably, several H. bilis–infected animals developed dysplasia with 10% demonstrating colon carcinoma, which was not observed in uninfected controls. Conclusions: Spontaneous and T-cell transfer, but not radiation-induced, colitis in WKO mice is dependent on the presence of Helicobacter spp. Furthermore, H. bilis infection is sufficient to induce typhlocolitis and colon cancer in Helicobacter spp.–free WKO mice. This animal model of a human immunodeficiency with chronic colitis and increased risk of colon cancer parallels what is seen in human colitis and implicates specific microbial constituents in promoting immune dysregulation in the intestinal mucosa.National Institutes of Health (U.S.) (R01OD011141)National Institutes of Health (U.S.) (R01CA067529)National Institutes of Health (U.S.) (P01CA026731)National Institutes of Health (U.S.) (P30ES02109

Integrating Data Analytics and Decision Support Systems in Public Health Management

For better data-driven decision-making and better health results, it is important for public health management to include data analytics and decision support systems (DSS). This abstract talks about why combining these tools is important and how they might affect public health management.Data analytics is an important part of public health because it uses big sets of data to find useful information for making decisions. By looking at patterns, trends, and connections in health data, public health managers can find new health problems, make good use of resources, and keep an eye on how well measures are working.As an addition to data analytics, decision support systems offer tools and models that make the decision-making process easier. Algorithms and models are used by these systems to look at data, make suggestions, and weigh possible results. This helps public health managers make smart choices in settings that are complicated and changeable.There are several perks to using both data analytics and DSS together in public health management. It makes decisions more accurate and reliable by giving real-time data and suggestions based on proof. It also helps plan and allocate resources better by finding groups at high risk and directing actions more effectively.Putting these tools together also helps public health managers handle public health situations better, like disease attacks or natural disasters. Using data analytics and DSS, public health agencies can quickly figure out what\u27s going on, put resources where they\u27re needed most, and keep real-time track of how measures are working. DOI: https://doi.org/10.52710/seejph.49

Milk-Derived Exosomes and Metabolic Regulation

Exosomes are natural nanoparticles that play an important role in cell-to-cell communication. Communication is achieved through the transfer of cargos, such as microRNAs, from donor to recipient cells and binding of exosomes to cell surface receptors. Exosomes and their cargos are also obtained from dietary sources, such as milk. Exosome and cell glycoproteins are crucial for intestinal uptake. A large fraction of milk exosomes accumulates in the brain, whereas the tissue distribution of microRNA cargos varies among distinct species of microRNA. The fraction of milk exosomes that escapes absorption elicits changes in microbial communities in the gut. Dietary depletion of exosomes and their cargos causes a loss of circulating microRNAs and elicits phenotypes such as loss of cognitive performance, increase in purine metabolites, loss of fecundity, and changes in the immune response. Milk exosomes meet the definition of bioactive food compounds

Interface-guided phenotyping of coding variants in the transcription factor RUNX1.

Single-gene missense mutations remain challenging to interpret. Here, we deploy scalable functional screening by sequencing (SEUSS), a Perturb-seq method, to generate mutations at protein interfaces of RUNX1 and quantify their effect on activities of downstream cellular programs. We evaluate single-cell RNA profiles of 115 mutations in myelogenous leukemia cells and categorize them into three functionally distinct groups, wild-type (WT)-like, loss-of-function (LoF)-like, and hypomorphic, that we validate in orthogonal assays. LoF-like variants dominate the DNA-binding site and are recurrent in cancer; however, recurrence alone does not predict functional impact. Hypomorphic variants share characteristics with LoF-like but favor protein interactions, promoting gene expression indicative of nerve growth factor (NGF) response and cytokine recruitment of neutrophils. Accessible DNA near differentially expressed genes frequently contains RUNX1-binding motifs. Finally, we reclassify 16 variants of uncertain significance and train a classifier to predict 103 more. Our work demonstrates the potential of targeting protein interactions to better define the landscape of phenotypes reachable by missense mutations

Exploring Therapeutic Strategies for Infantile Neuronal Axonal Dystrophy (INAD/PARK14)

Infantile neuroaxonal dystrophy (INAD) is caused by recessive variants in PLA2G6 and is a lethal pediatric neurodegenerative disorder. Loss of the Drosophila homolog of PLA2G6, leads to ceramide accumulation, lysosome expansion, and mitochondrial defects. Here, we report that retromer function, ceramide metabolism, the endolysosomal pathway, and mitochondrial morphology are affected in INAD patient-derived neurons. We show that in INAD mouse models, the same features are affected in Purkinje cells, arguing that the neuropathological mechanisms are evolutionary conserved and that these features can be used as biomarkers. We tested 20 drugs that target these pathways and found that Ambroxol, Desipramine, Azoramide, and Genistein alleviate neurodegenerative phenotypes in INAD flies and INAD patient-derived neural progenitor cells. We also develop an AAV-based gene therapy approach that delays neurodegeneration and prolongs lifespan in an INAD mouse model

The ‘microflora hypothesis’ of allergic diseases

Increasingly, epidemiologic and clinical data support the hypothesis that perturbations in the gastrointestinal (GI) microbiota because of antibiotic use and dietary differences in ‘industrialized’ countries have disrupted the normal microbiota-mediated mechanisms of immunological tolerance in the mucosa, leading to an increase in the incidence of allergic airway disease. The data supporting this ‘microflora hypothesis’ includes correlations between allergic airway disease and (1) antibiotic use early in life, (2) altered fecal microbiota and (3) dietary changes over the past two decades. Our laboratory has recently demonstrated that mice can develop allergic airway responses to allergens if their endogenous microbiota is altered at the time of first allergen exposure. These experimental and clinical observations are consistent with other studies demonstrating that the endogenous microbiota plays a significant role in shaping the development of the immune system. Data are beginning to accumulate that a ‘balanced’ microbiota plays a positive role in maintaining mucosal immunologic tolerance long after post-natal development. Other studies have demonstrated that even small volumes delivered to the nasopharynx largely end up in the GI tract, suggesting that airway tolerance and oral tolerance may operate simultaneously. The mechanism of microbiota modulation of host immunity is not known; however, host and microbial oxylipins are one potential set of immunomodulatory molecules that may control mucosal tolerance. The cumulative data are beginning to support the notion that probiotic and prebiotic strategies be considered for patients coming off of antibiotic therapy.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/73451/1/j.1365-2222.2005.02379.x.pd