Adult Organs Research Articles

Pimozide Inhibits Type II but Not Type I Hair Cells in Chicken Embryo and Adult Mouse Vestibular Organs

Background: Pimozide is a conventional antipsychotic drug of the diphenylbutylpiperidine class, widely used for treating schizophrenia and delusional disorders and for managing motor and phonic tics in Tourette’s syndrome. Pimozide is known to block dopaminergic D2 receptors and various types of voltage-gated ion channels. Among its side effects, dizziness and imbalance are the most frequently observed, which may imply an effect of the drug on the vestibular sensory receptors, the hair cells. Amniotes possess two classes of vestibular hair cells, named type I and type II hair cells, which differ in terms of signal processing and transmission. We previously reported that Pimozide [3 μM] significantly increased a delayed outward rectifying K+ current (IK,V). Methods and Results: In the present study, using the whole-cell patch-clamp technique we additionally show that Pimozide decreases the inward rectifying K+ current (IK,1) and the mixed Na+/K+ current (Ih) of chicken embryo type II hair cells, whereas it does not affect type I hair cells’ ionic currents. Since ion channels’ expression can vary depending on age and animal species, in the present study, we also tested Pimozide in adult mouse vestibular hair cells. We found that, like in the chicken embryo, Pimozide significantly increases IK,V and decreases IK,1 and Ih in type II hair cells. However, in the adult mouse, Pimozide also slightly increased the outward rectifying K+ current in type I hair cells. Conclusions: While providing a possible explanation for the vestibular side effects of Pimozide in humans, its inhibitory action on mammalian hair cells might be of interest for the local treatment of vestibular disorders characterized by altered vestibular input, like Ménière’s disease.

The Production of Fairy Shrimp as Live Food for Aquatic Animals and Transfer to Farmers

Fairy shrimps are zooplankton, a small crustacean with high nutritional value and carotenoids. Fairy shrimps are aquatic animals that feed on both living and non-living food by filtering. In Thailand, there are 3 species of fairy shrimps, namely Streptocephalus sirindhornae, Branchinella thailandensis, and Streptocephalus siamenesis. The three species of fairy shrimps exhibit distinct characteristics in terms of size, color, and egg morphology. There are 3 types of fairy shrimp culture in Thailand, namely, cement ponds, clay ponds, and hapas, which are adult fairy shrimp cultures. Both living and non-living forms of fairy shrimps are used by freezing. The usage of fairy shrimps come in both living and non-living, frozen forms. Fairy shrimps are fed as food for beautiful aquatic animals such as goldfish, cichlids, etc. They are also used as food pellets to feed aquatic animals such as giant freshwater prawns, Rad tilapia fish, etc. Therefore, it can be concluded that fairy shrimps have a significant potential as a valuable food source for both larval and adult aquatic organisms. Consequently, the widespread dissemination of knowledge about fairy shrimp aquaculture and utilization can greatly enhance their adoption in the aquaculture industry, thereby promoting sustainability.

Systematic review of mobile health applications in transplant patients.

Poor medication management can lead to adverse outcomes for transplant patients, including acute rejection and graft loss. In recent years, mobile applications have been proposed as innovative tools to improve patient treatment management. This review aimed to systematically evaluate the available research evidence on the relationship between mobile applications and treatment management in transplant patients. The following databases were systematically searched for relevant publications on April 8, 2022, using the PRISMA method: PubMed, Embase, and Google Scholar. The Cochrane risk of bias tool was used to assess the included studies. Observational or interventional studies focusing on the use of mobile applications in adult solid organ transplant patients were included for analysis. A total of 28 articles met the inclusion criteria. The overall methodological quality of the evidence was assessed as low. Most studies were monocentric (n = 23, 82%). The majority of follow-ups focused on kidney transplants (n = 12, 42.9%) with small sample sizes (54%, n < 99), including participants < 60 years old (n = 26, 93%) with follow-up ≤ 6 months (61%). Medication adherence rates showed significant improvements in seven out of 13 trials compared to standard care or placebo. Several features were reported to be most effective in improving patient treatment management, such as self-registration and monitoring, medication reminders based on alerts, and caregiver monitoring to check patients' health indicators or medication adherence. Mobile applications tended to improve medication management in transplant patients compared to standard care. However, due to the heterogeneity of the objectives of the analyzed studies, which do not allow for meta-analysis, further high-level evidence studies evaluating the effects of mobile applications in this area are needed to support effective interventions.

Neural Crest-Derived Mesenchymal Cells Support Thymic Reconstitution After Lethal Irradiation.

Reconstitution of the thymus is essential for assessing thymic function following injury. However, the currently employed cytoreductive regimes unvaryingly affect the thymic microenvironment, thereby impeding the recovery of T lymphopoiesis. The thymic stroma is composed of epithelial and mesenchymal cells. Thymic mesenchymal cells originate from the Neural crest (NC) and mesoderm and contribute to thymus organogenesis, yet their role in thymic regeneration is unclear. In this study, using transgenic mice expressing NC-specific Cre and Cre-driven DT receptors, we investigated the role of NC-derived mesenchymal cells in thymic regeneration following total body irradiation. We revealed that NC-derived mesenchymal cells have reduced susceptibility to irradiation and induce the upregulation of hematopoietic factors that promote thymus regeneration after irradiation. Additionally, using adult thymic organ culture and renal capsule transplantation, depletion of NC-derived mesenchymal cells resulted in a reduction of DN1-like early T-cell progenitors (ETP) and impaired thymic regeneration. Furthermore, among the numerous factors upregulated by NC-derived mesenchymal cells, Periostin and Flt3L were markedly increased after irradiation and promoted abundance of DN1-like ETPs during thymic reconstitution. Collectively, these findings highlight the importance of NC-derived mesenchymal cells in thymic regeneration.

Unusual modes of cell and nuclear divisions characterise Drosophila development.

The growth and development of metazoan organisms is dependent upon a co-ordinated programme of cellular proliferation and differentiation, from the initial formation of the zygote through to maintenance of mature organs in adult organisms. Early studies of proliferation of ex vivo cultures and unicellular eukaryotes described a cyclic nature of cell division characterised by periods of DNA synthesis (S-phase) and segregation of newly synthesized chromosomes (M-phase) interspersed by seeming inactivity, the gap phases, G1 and G2. We now know that G1 and G2 play critical roles in regulating the cell cycle, including monitoring of favourable environmental conditions to facilitate cell division, and ensuring genomic integrity prior to DNA replication and nuclear division. M-phase is usually followed by the physical separation of nascent daughters, termed cytokinesis. These phases where G1 leads to S phase, followed by G2 prior to M phase and the subsequent cytokinesis to produce two daughters, both identical in genomic composition and cellular morphology are what might be termed an archetypal cell division. Studies of development of many different organs in different species have demonstrated that this stereotypical cell cycle is often subverted to produce specific developmental outcomes, and examples from over 100 years of analysis of the development of Drosophila melanogaster have uncovered many different modes of cell division within this one species.

Prosaposin/Saposin Expression in the Developing Rat Olfactory and Vomeronasal Epithelia.

Prosaposin is a glycoprotein widely conserved in vertebrates, and it acts as a precursor for saposins that accelerate hydrolysis in lysosomes or acts as a neurotrophic factor without being processed into saposins. Neurogenesis in the olfactory neuroepithelia, including the olfactory epithelium (OE) and the vomeronasal epithelium (VNE), is known to occur throughout an animal's life, and mature olfactory neurons (ORNs) and vomeronasal receptor neurons (VRNs) have recently been revealed to express prosaposin in the adult olfactory organ. In this study, the expression of prosaposin in the rat olfactory organ during postnatal development was examined. In the OE, prosaposin immunoreactivity was observed in mature ORNs labeled using olfactory marker protein (OMP) from postnatal day (P) 0. Immature ORNs showed no prosaposin immunoreactivity throughout the examined period. In the VNE, OMP-positive VRNs were mainly observed in the basal region of the VNE on P10 and showed an adult-like distribution from P20. On the other hand, prosaposin immunoreactivity was observed in VRNs from P0, suggesting that not only mature VRNs but also immature VRNs express prosaposin. This study raises the possibility that prosaposin is required for the normal development of the olfactory organ and has different roles in the OE and the VNE.

Key insights into cannabis-cancer pathobiology and genotoxicity.

Whilst mitochondrial inhibition and micronuclear fragmentation are well established features of the cannabis literature mitochondrial stress and dysfunction has recently been shown to be a powerful and direct driver of micronucleus formation and chromosomal breakage by multiple mechanisms. In turn genotoxic damage can be expected to be expressed as increased rates of cancer, congenital anomalies and aging; pathologies which are increasingly observed in modern continent-wide studies. Whilst cannabinoid genotoxicity has long been essentially overlooked it may in fact be all around us through the rapid induction of aging of eggs, sperm, zygotes, foetus and adult organisms with many lines of evidence demonstrating transgenerational impacts. Indeed this multigenerational dimension of cannabinoid genotoxicity reframes the discussion of cannabis legalization within the absolute imperative to protect the genomic and epigenomic integrity of multiple generations to come.

Human Pluripotent Stem Cell Colony Migration Is Related to Culture Environment and Morphological Phenotype.

Human pluripotent stem cells (hPSCs) are an important tool in the field of regenerative medicine due to their ability to differentiate towards all tissues of the adult organism. An important task in the study of hPSCs is to understand the factors that influence the maintenance of pluripotent and clonal characteristics of colonies represented by their morphological phenotype. Such factors include the ability of colonies to migrate during growth. In this work, we measured and analyzed the migration trajectories of hPSC colonies obtained from bright-field images of three cell lines, including induced hPSC lines AD3 and HPCASRi002-A (CaSR) and human embryonic stem cell line H9. To represent the pluripotent status, the colonies were visually phenotyped into two classes having a "good" or "bad" morphological phenotype. As for the migration characteristics, we calculated the colony speed and distance traveled (mobility measures), meandering index (motion persistence measures), outreach ratio (trajectory tortuosity characteristic), as well as the velocity autocorrelation function. The analysis revealed that the discrimination of phenotypes by the migration characteristics depended on both the cell line and growth environment. In particular, when the mTESR1/Matrigel culture environment was used, "good" AD3 colonies demonstrated a higher average migration speed than the "bad" ones. The reverse relationship between average speeds of "good" and "bad" colonies was found for the H9 line. The CaSR cell line did not show significant differences in the migration speed between the "good" and "bad" phenotypes. We investigated the type of motion exhibited by the colonies by applying two diffusion models to the mean squared displacement dynamics, one model corresponding to normal and the other to anomalous diffusion. The type of diffusion and diffusion parameter values resulting from the model fitting to data demonstrated a similar cell line, environment, and phenotype dependency. Colonies mainly showed a superdiffusive behavior for the mTESR1/Matrigel culture conditions, characterized by longer migration steps compared to the normal random walk. The specific properties of migration features and the patterns of their variation demonstrated in our work can be useful for the development and/or improvement of automated solutions for quality control of hPSCs.

Dengue fever in immunocompromised patients: A systematic review and meta-analysis

ObjectivesGiven the complex role of immunity in dengue severity, we aimed to review the clinical course of dengue infection in immunocompromised patients. MethodsWe conducted a systematic review of studies reporting outcomes among immunocompromised patients with laboratory-confirmed dengue infection. Meta-analysis using the Mantel-Haenszel method (fixed effects) was performed for studies with control groups. We registered the study with PROSPERO (No. CRD42021258930). ResultsWe included 115 studies. Among these, 30 studies compared immunocompromised (cases) and nonimmunocompromised (control) patients, focusing mainly on children (n = 22 studies) with malnutrition (n=18). Immunocompromised patients had a higher likelihood of dengue complications (OR 1.87; 95% CI: 1.04-3.35]) but a lower likelihood of severe dengue (OR 0.83; 95% CI: 0.69-1.00]. No significant difference in mortality was observed. In the 85 studies focused solely on immunocompromised patients, severe dengue and mortality rates were 9% and 4%, respectively, mostly among adult solid organ transplant recipients and those with inflammatory diseases. Immunosuppressive treatment alterations and temporary graft dysfunction were reported. ConclusionImmunocompromised patients have an increased risk of dengue-related complications. However, definitive conclusions about the comparative severity of dengue across different immunocompromised patient groups are limited by a lack of robust data, highlighting the need for well-designed future studies.

The myocardium of human dilated nonischemic cardiomyopathy harbors unique tissue-resident lymphocyte subsets

Abstract Purpose The etiology of dilated nonischemic cardiomyopathy (NICM) is often unclear, but thought to be multifactorial, involving an interplay between genetic factors and direct myocardial damage caused by infection, autoimmunity, or toxins. Tissue-resident lymphocytes are non-circulating immune cells which are retained within various tissues and play integral roles in maintaining homeostasis, responding to infection, regulating inflammation, and mediating tissue repair. Despite their likely involvement in various cardiac pathologies, the presence of tissue-resident lymphocytes in the human heart has not yet been well-characterized, neither in health nor in disease. Methods Human left ventricular tissue and paired peripheral blood samples were obtained from six adult organ donors with normal heart function who died of noncardiac causes and from the explants of six adult patients with end-stage nonischemic cardiomyopathy. Heart tissue was processed using mechanical and enzymatic digestion. Mononuclear cells were isolated from both heart and paired blood by density centrifugation. Isolated mononuclear cells were surface stained with an antibody cocktail and interrogated using multidimensional flow cytometry. Results The adult human heart contains a diverse immune cell population (Fig 1A). Both healthy and NICM cardiac lymphocytes express distinct phenotypic profiles which resemble those of resident lymphocytes found across various tissues, distinct from those isolated from paired blood. Frequencies of effector memory T (Tem) cells (CD45RA- CCR7-) were higher in heart than in blood among both CD4+ and CD8+ T-cells. In heart compared to blood, significantly more Tem expressed CD69, a canonical marker of tissue-resident memory T (Trm) cells (Fig 1B). Cardiac Trm, especially CD8+ Trm, expressed classical Trm signature markers including integrins CD103 (epithelial binding) and CD49a (collagen binding). NICM Trm expressed more CD49a than healthy heart Trm (Fig 1C). NK-cells also demonstrated higher CD69 expression (trNK) in heart than in blood, with associated expression of CD103, CD49a, and CXCR6 (Fig 1D). Compared to healthy hearts, NICM trNK were predominantly CD56bright/CD16- (Fig 1E and 1F) and expressed less CD57 (Fig 1D), overall indicating a less mature state with lower cytotoxic potential. Confocal imaging of cardiac tissue revealed CD69-expressing tissue-resident lymphocytes within the cardiac interstitium. Conclusions The adult human heart contains diverse populations of lymphocytes which express canonical markers of tissue residency, similar to lymphocytes resident in other tissues. Compared to healthy hearts, NICM hearts have more CD49a+ Trm, perhaps related to greater fibrosis, and also harbor a distinct subset of immature tissue-resident NK cells which may shed light on pathophysiology. Future work is focused on better exploring the functional phenotypes of these resident cells and better localizing them within the myocardium.FA - Figure 1

Toxicity of crude oil: a systematic review of the acute impacts on Cnidaria

Cnidarians are ecosystem engineers that play a fundamental role in food webs. However, they are increasingly threatened by anthropogenic activities, such as oil spills, which can cause acute sublethal effects and in some cases, mortality. The literature on this theme is limited and fragmented, with diverse objectives and methodologies, making it challenging to identify consistent patterns and knowledge gaps. This article aims to provide a systematic review of the acute impacts of crude oil and petroleum hydrocarbons on members of the Phylum Cnidaria, to synthesize and integrate the available data. The research followed the PRISMA 2020 guidelines, and employed the search terms: “crude oil” AND “acute” AND (Cnidaria OR coral), along with the names of each Cnidarian class. The search was restricted to articles published in English from 2013 to 2023, excluding studies that did not directly address the core topic. The number of records collected was relatively low, with only 30 articles identified from 20 studies. The highest number of publications occurred in 2022, and the USA was the most represented country. The Class Anthozoa, particularly corals (Order Scleractinia) was the most studied. Research focused on the effects of crude oil on gametes, embryos, larvae and adults, with most studies examining adult organisms. The majority (56.6%), utilized mixtures of crude oil hydrocarbons, while a smaller proportion (36.6%) focused on pure aromatic hydrocarbons. Most experiments employed Water Accommodated Fractions (36.6%) and continuous flow system (20%). The most common exposure durations (30%) were 96 and 48h, with exposure times ranging from 18h to several days. There was significant variation in methodological approaches, underscoring the need for standardized protocols. Research on adult cnidarians primarily assessed health and morphology, lethality, genetic alterations, and symbiont efficiency. In contrast, studies on early life stages focused on lethality, larval metamorphosis and settlement. Overall, the results indicate that responses vary considerably, particularly concerning exposure time and oil concentration, with sublethal effects at different levels. Further studies incorporating a broader range of species, novel approaches, and the combined effects of elevated temperature and UV radiation are crucial to advancing our understanding of oil impacts on cnidarians.

Identification of the proliferative activity of germline progenitor cells in the adult ovary of the bat Artibeus jamaicensis.

Until a few years ago, it was assumed that oocyte renewal did not take place in the ovary of adult organisms; however, the existence of germline progenitor cells (GPCs), which renew the ovarian follicular reserve, has now been documented in mammals. Specifically, in the adult ovary of bats, the presence of cells located in the cortical region with characteristics similar to GPCs, called adult cortical germ cells (ACGC), has been observed. One of the requirements that a GPC must fulfil is to be able to proliferate mitotically, so the evaluation of cell proliferation in ACGC is of utmost importance in order to be able to relate them to a parental lineage. Currently, there are several methods to determine cell proliferation, including BrdU labelling or the use of endogenous proliferation markers. Thus, the aim of this work was to evaluate the proliferative activity of ACGC in the adult ovary of the bat Artibeus jamaicensis, using different proliferation markers and correlating these with the protein expression of the transcription factor Oct4 and the germ line marker Ddx4. We found that the expression pattern of the proliferation markers BrdU, PCNA, Ki-67 and pH3 occurs at different times of the cell cycle, so co-localization of two or more of these markers allows us to identify proliferating cells. This allowed us to identify ACGC with proliferative capacity in the adult ovary of A. jamaicensis, suggesting that GPCs renew the follicle reserve during the adult life of the organism.

Five-factor theory of aging and death due to aging

This new theory of aging explains that aging and death due to aging are due to five factors, and also explains how these factors are interconnected and jointly lead to aging and death of the organism, pointing to many facts that strongly support it. The first factor is the harmful changes that occur in cellular structures. The second factor is the cessation of cell division in adult organisms, which leads to the inability to restore cellular structures. The third factor is the feature that cells do not die due to the accumulation of harmful changes that occur in the cells during the life of the organism. The fourth factor is the inability of stem cells to regenerate tissue by replacing such cells with new ones, because somatic cells do not die and there are no signals that stimulate the proliferation of stem cells and their differentiation into new ones that would replace dead cells. The fifth factor is that all cells die suddenly, due to the cessation of one of the vital functions of the organism, and not gradually during life, due to a decrease in the functionality of cells caused by the introduction of harmful changes in cellular structures, which would allow stem cells to regenerate tissues and keep the body young. Also, to show that this aging theory is valid, the theory gives its view of the evolution of five factors, which according to this theory lead to aging, which gives strong support to this theory.

Insights into human norovirus cultivation in human intestinal enteroids.

Human noroviruses (HuNoVs) are a significant cause of epidemic and sporadic acute gastroenteritis worldwide. The lack of a reproducible culture system hindered the study of HuNoV replication and pathogenesis for almost a half-century. This barrier was overcome with our successful cultivation of multiple HuNoV strains in human intestinal enteroids (HIEs), which has significantly advanced HuNoV research. We optimized culture media conditions and generated genetically modified HIE cultures to enhance HuNoV replication in HIEs. Building upon these achievements, we now present new insights into this culture system, which involve testing different media, unique HIE lines, and additional virus strains. HuNoV infectivity was evaluated and compared in new HIE models, including HIEs generated from different intestinal segments of individual adult organ donors, HIEs from human intestinal organoids produced from directed differentiation of human embryonic stem cells that were then transplanted and matured in mice before making enteroids (H9tHIEs), genetically engineered (J4FUT2 knock-in [KI], J2STAT1 knockout [KO]) HIEs, as well as HIEs derived from a patient with common variable immunodeficiency (CVID) and from infants. Our findings reveal that small intestinal HIEs, but not colonoids, from adults, H9tHIEs, HIEs from a CVID patient, and HIEs from infants support HuNoV replication with segment and strain-specific differences in viral infection. J4FUT2-KI HIEs exhibit the highest susceptibility to HuNoV infection, allowing the cultivation of a broader range of genogroup I and II HuNoV strains than previously reported. Overall, these results contribute to a deeper understanding of HuNoVs and highlight the transformative potential of HIE cultures in HuNoV research.IMPORTANCEHuman noroviruses (HuNoVs) cause global diarrheal illness and chronic infections in immunocompromised patients. This paper reports approaches for cultivating HuNoVs in secretor positive human intestinal enteroids (HIEs). HuNoV infectivity was compared in new HIE models, including ones from (i) different intestinal segments of single donors, (ii) human embryonic stem cell-derived organoids transplanted into mice, (iii) genetically modified lines, and (iv) a patient with common variable immunodeficiency disease. HIEs from small intestine, but not colon, support HuNoV replication with donor, segment, and strain-specific variations. Unexpectedly, HIEs from one donor are resistant to GII.3 infection. The genetically modified J4FUT2 knock-in (KI) HIEs enable cultivation of a broad range of GI and GII genotypes. New insights into strain-specific differences in HuNoV replication in HIEs support this platform for advancing understanding of HuNoV biology and developing potential therapeutics.

Evidence of Peripheral Vestibular Impairment Among Adults With Chronic Moderate-Severe Traumatic Brain Injury.

Traumatic brain injury (TBI) is a leading cause of death and disability among adults in the United States. There is evidence to suggest the peripheral vestibular system is vulnerable to damage in individuals with TBI. However, there are limited prospective studies that describe the type and frequency of vestibular impairment in individuals with chronic moderate-severe TBI (> 6 months postinjury). Cervical and ocular vestibular evoked myogenic potentials (VEMPs) and video head impulse test (vHIT) were used to assess the function of otolith organ and horizontal semicircular canal (hSCC) pathways in adults with chronic moderate-severe TBI and in noninjured comparison (NC) participants. Self-report questionnaires were administered to participants with TBI to determine prevalence of vestibular symptoms and quality of life associated with those symptoms. Chronic moderate-severe TBI was associated with a greater degree of impairment in otolith organ, rather than hSCC, pathways. About 63% of participants with TBI had abnormal VEMP responses, compared to only ~10% with abnormal vHIT responses. The NC group had significantly less abnormal VEMP responses (~7%), while none of the NC participants had abnormal vHIT responses. As many as 80% of participants with TBI reported vestibular symptoms, and up to 36% reported that these symptoms negatively affected their quality of life. Adults with TBI reported vestibular symptoms and decreased quality of life related to those symptoms and had objective evidence of peripheral vestibular impairment. Vestibular testing for adults with chronic TBI who report persistent dizziness and imbalance may serve as a guide for treatment and rehabilitation in these individuals.

YAP Overcomes Mechanical Barriers to Induce Mitotic Rounding and Adult Cardiomyocyte Division.

Many specialized cells in adult organs acquire a state of cell cycle arrest and quiescence through unknown mechanisms. Our limited understanding of mammalian cell cycle arrest is derived primarily from cell culture models. Adult mammalian cardiomyocytes, a classic example of cell cycle arrested cells, exit the cell cycle postnatally and remain in an arrested state for the life of the organism. Cardiomyocytes can be induced to re-enter the cell cycle by YAP5SA, an active form of the Hippo signaling pathway effector YAP. We performed clonal analyses to determine the cell kinetics of YAP5SA cardiomyocytes. We also performed single-cell RNA sequencing, marker gene analysis, and functional studies to examine how YAP5SA cardiomyocytes progress through the cell cycle. We discovered that YAP5SA-expressing cardiomyocytes divided efficiently, with >20% of YAP5SA cardiomyocyte clones containing ≥2 cardiomyocytes. YAP5SA cardiomyocytes re-entered cell cycle at the G1/S transition and had an S phase lasting ≈48 hours. Sarcomere disassembly is required for cardiomyocyte progression from S to G2 phase and the induction of mitotic rounding. Although oscillatory Cdk expression was induced in YAP5SA cardiomyocytes, these cells inefficiently progressed through G2 phase. This is improved by inhibiting P21 function, implicating checkpoint activity as an additional barrier to YAP5SA-induced cardiomyocyte division. Our data reveal that YAP5SA overcomes the mechanically constrained myocardial microenvironment to induce mitotic rounding with cardiomyocyte division, thus providing new insights into the in vivo mechanisms that maintain cell cycle quiescence in adult mammals.

EUROPEAN PAPER BEES, POLISTES DOMINULA AND POLISTES NIMPHA (CHRIST, 1791) (HYMENOPTERA: VESPIDAE) PATHOGENS PRESENCE AND THEIR POTENTIAL INSECTICIDAL EFFECTS ON HONEYBEES ADULTS OF APIS MELLIFERA CAUCASIA (POLLMANN, 1889)

Honeybee (Apis mellifera) is an important element of biodiversity and terrestrial ecosystems. Any pathogenic infection in this beneficial insect can lead to major undesirable disasters. This study investigated the pathogenic bacteria and fungi from Polistes dominula and Polistes nimpha wasps and their potential insecticidal effects on Apis mellifera caucasia. For this purpose, bacteria and fungi were isolated from dead and diseased bees collected from Terme district of Samsun province in Türkiye in May and June 2020. In the study, Granulicatella adiacens, Staphylococcus xylosus, Sphingomonas paucimobilis bacteria and Cryptococcus laurentii and Candida famata fungi were obtained from the internal tissues and organs of Polistes dominula paper wasp adults. Staphylococcus xylosus and Sphingomonas paucimobilis were found to be common bacteria in both bee species. Serratia marcescens and Enterococcus faecalis bacterial species were found to have a very lethal effect on honeybees. Bioassay experiments were performed on the detected fungi, and it was observed that Cryptococcus laurentii and Candida famata fungi species also had lethal effects on honeybees. It has been revealed that entomopathogenic bacteria, which are known to be very effective in biological control against harmful insects, can cause unwanted infections in honeybees.

Reprogramming with Atoh1, Gfi1, and Pou4f3 promotes hair cell regeneration in the adult organ of Corti

Abstract Cochlear hair cells can be killed by loud noises, ototoxic drugs, and natural aging. Once lost, mammalian hair cells do not naturally regenerate, leading to permanent hearing loss. Since the mammalian cochlea lacks any intrinsic ability to regenerate, genetic reprogramming of cochlear supporting cells that lie adjacent to hair cells is a potential option for hearing restoration therapies. We targeted cochlear supporting cells with three hair cell transcription factors: Atoh1, or Atoh1 + Gfi1, or Atoh1 + Gfi1 + Pou4f3 and found that 1- and 2-factor reprogramming is not sufficient to reprogram adult supporting cells into hair cells. However, activation of all three hair cell transcription factors reprogrammed some adult supporting cells into hair cell-like cells. We found that killing endogenous hair cells significantly improved the ability of supporting cells to be reprogrammed and regenerated numerous hair cell-like cells throughout the length of the cochlea. These regenerated hair cell-like cells expressed myosin VIIa and parvalbumin, as well as the mature outer hair cell protein prestin, were innervated, expressed proteins associated with ribbon synapses, and formed rudimentary stereociliary bundles. Finally, we demonstrate that supporting cells remained responsive to transcription factor reprogramming for at least 6 weeks after hair cell damage, suggesting that hair cell reprogramming may be effective in the chronically deafened cochlea.

Application of hsp60 amplicon sequencing to characterize microbial communities associated with juvenile and adult Euprymna scolopes squid.

The symbiotic relationship between Vibrio (Aliivibrio) fischeri and the Hawaiian bobtail squid, Euprymna scolopes, serves as a key model for understanding host-microbe interactions. Traditional culture-based methods have primarily isolated V. fischeri from the light organs of wild-caught squid, yet culture-independent analyses of this symbiotic microbiome remain limited. This study aims to enhance species-level resolution of bacterial communities associated with E. scolopes using hsp60 amplicon sequencing. We validated our hsp60 sequencing approach using pure cultures and mixed bacterial populations, demonstrating its ability to distinguish V. fischeri from other closely-related vibrios and the possibility of using this approach for strain-level diversity with further optimization. This approach was applied to whole-animal juvenile squid exposed to either seawater or a clonal V. fischeri inoculum, as well as ventate samples and light organ cores from wild-caught adults. V. fischeri accounted for the majority of the identifiable taxa for whole-animal juvenile samples and comprised 94%-99% of amplicon sequence variants (ASVs) for adult light organ core samples, confirming that V. fischeri is the dominant, if not sole, symbiont typically associated with E. scolopes light organs. In one ventate sample, V. fischeri comprised 82% of reads, indicating the potential for non-invasive community assessments using this approach. Analysis of non-V. fischeri ASVs revealed that Bradyrhizobium spp. and other members of the Rhodobacterales order are conserved across juvenile and adult samples. These findings provide insight into the presence of additional microbial associations with the squid host tissue outside of the light organ that have not been previously detected through traditional culture methods.

Heterogeneity of synaptic NMDA receptor responses within individual lamina I pain-processing neurons across sex in rats and humans.

Excitatory glutamatergic NMDA receptors (NMDARs) are key regulators of spinal pain processing, and yet the biophysical properties of NMDARs in dorsal horn nociceptive neurons remain poorly understood. Despite the clinical implications, it is unknown whether the molecular and functional properties of synaptic NMDAR responses are conserved between males and females or translate from rodents to humans. To address these translational gaps, we systematically compared individual and averaged excitatory synaptic responses from lamina I pain-processing neurons of adult Sprague-Dawley rats and human organ donors, including both sexes. By combining patch-clamp recordings of outward miniature excitatory postsynaptic currents with non-biased data analyses, we uncovered a wide range of decay constants of excitatory synaptic events within individual lamina I neurons. Decay constants of synaptic responses were distributed in a continuum from 1-20ms to greater than 1000ms, suggesting that individual lamina I neurons contain AMPA receptor (AMPAR)-only as well as GluN2A-, GluN2B- and GluN2D-NMDAR-dominated synaptic events. This intraneuronal heterogeneity in AMPAR- and NMDAR-mediated decay kinetics was observed across sex and species. However, we discovered an increased relative contribution of GluN2A-dominated NMDAR responses at human lamina I synapses compared with rodent synapses, suggesting a species difference relevant to NMDAR subunit-targeting therapeutic approaches. The conserved heterogeneity in decay rates of excitatory synaptic events within individual lamina I pain-processing neurons may enable synapse-specific forms of plasticity and sensory integration within dorsal horn nociceptive networks. KEY POINTS: Synaptic NMDA receptors (NMDARs) in spinal dorsal horn nociceptive neurons are key regulators of pain processing, but it is unknown whether their functional properties are conserved between males and females or translate from rodents to humans. In this study, we compared individual excitatory synaptic responses from lamina I pain-processing neurons of male and female adult Sprague-Dawley rats and human organ donors. Individual lamina I neurons from male and female rats and humans contain AMPA receptor-only as well as GluN2A, GluN2B- and GluN2D-NMDAR-dominated synaptic events. This may enable synapse-specific forms of plasticity and sensory integration within dorsal horn nociceptive networks. Human lamina I synapses have an increased relative contribution of GluN2A-dominated NMDAR responses compared with rodent synapses. These results uncover a species difference relevant to NMDAR subunit-targeting therapeutic approaches.