Metagenomic analysis of rats with diarrhea treated with mixed probiotics: response to consecutive and alternate-hour supplementation.

Background: Diarrhea is the leading contributory factor of sickness and mortality among children under five and an economic burden for families. This study aimed to investigate the effects of mixed probiotics supplementation at different times (consecutive and alternate-hour) on intestinal microecology in Sprague-Dawley (SD) rats with acute diarrhea. Methods: A total of 40 SD rats were randomly assigned to four groups, including the control group, model group, probiotic group A, and probiotic group B. An acute diarrhea model was induced by administration of 5% dextran sulfate sodium. Rats in probiotic group A and probiotic group B were fed with Clostridium butyricum (C. butyricum), Bifidobacterium infantis (B. infantis), and Saccharomyces boulardii (S. boulardii) for a total of 7 days. Probiotic group A was fed with all probiotics simultaneously. Probiotic group B was fed with C. butyricum and B. infantis simultaneously, and then after a 2-hour interval, with S. boulardii. Metagenomic next-generation sequencing was used to analyze the fecal samples from every rat. The metagenomic sequencing used in this experiment was used to evaluate the effect of probiotics on the composition as well as function of the gut microbiota in order to gain a deeper comprehension of probiotic-host interactions on health and disease. Results: The structure of the gut microbiota in probiotic group A showed significant changes. Compared to the model group, the abundance of some beneficial bacteria had increased, including Actinobacteria (P=0.048), Lactobacillus (P=0.050), and Lactobacillus johnsonii (P=0.042), and many opportunistic pathogenic bacteria has decreased, such as Ruminococcus (P=0.001). Compared to the control group, the abundance of some beneficial bacteria had increased, including Fusobacteria (P=0.02) and Phascolarium (P=0.002), and there was a reduction in the abundance of many opportunistic pathogenic bacteria such as Roseburia (P=0.03), Lachnoclosterium (P=0.009), and Oscillibacter_sp_1-3 (P=0.002). In addition, metagenomic analysis showed that as well as an up-regulation of glycoside hydrolase expression, amino acid and inorganic ion transport, and metabolism-related pathways, there was a down-regulation of cell motility. Conclusions: Simultaneous administration of probiotics may have more positive implications in improving the gut microbiota of acute diarrhea rats.

Application of the full-width-at-half-maximum image segmentation method to analyse retinal vascular changes in patients with internal carotid artery stenosis.

Background: To investigate the correlation between retinal vascular changes and ICA stenosis by measuring retinal vessels using full-width-at-half-maximum (FWHM) and intelligent image recognition. Methods: This research selected patients who were admitted to the Vascular Surgery Department of Quzhou People's Hospital from January 2018 to December 2020 and were preparing for Carotid Artery Stenting (CAS). Participants were divided into two groups: without ICA stenosis (Group 0) and with ICA stenosis (Group 1). A total of 109 cases were included in the study, with 50 cases in Group 1 and 59 cases in Group 0. Vascular images of superior temporal zone B of the retina were obtained by spectral domain optical coherence tomography (SD-OCT). The edges of retinal vessels were identified by FWHM. Each vessel of all subjects was measured three times with the FWHM, and the average value was taken to obtain the retinal arteriolar lumen diameter (RALD), retinal arteriolar outer diameter (RAOD), retinal venular lumen diameter (RVLD), and retinal venular outer diameter (RVOD),Arterial Wall Thickness (AWT),Venular Wall Thickness (VWT)=(RVOD-RVLD)/2,Arteriovenous Ratio (AVR) = RAOD/RVOD. Results: We found that compared to Group 0, Group 1 had smaller RALD (P < 0.001) and RAOD (P < 0.001), and wider RVOD (P < 0.001), with thicker VWT (P < 0.001). When compared with the contralateral eye in Group 1, the ipsilateral eye exhibited even smaller RALD,RAOD and AVR (P < 0.001, P < 0.001, P < 0.001). After CAS, the RALD,RAOD and AVR in Group 1 increased (P < 0.001, P < 0.001, P < 0.001),while the RVLD and RVOD decreased (P < 0.05, P < 0.001). Our research reveals a significant correlation between retinal vascular changes and internal ICA stenosis. Conclusion: Utilizing SD-OCT in conjunction with the FWHM,we achieved a non-invasive, intelligent, stable, and precise acquisition of data pertaining to retinal vessels. These findings underscore a significant correlation between alterations in retinal vascular structure and the presence of ICA stenosis, as demonstrated by our research.

Efficacy of agomelatine on sleep disorders and lateral habenula neuronal activity in chronic restraint stress depression model mice.

BACKGROUND: Sleep disorders (SD) are one of the common manifestations of depression patients. This article aimed to explore the effect of Agomelatine (Ago) on SD in chronic restraint stress (CRS) depression model mice and its effect on the activity of neurons in the lateral habenula (LHb). METHODS: 30 C57BL/6 J mice were divided into normal (C57BL/6 J) group, CRS group, and Ago group. CRS experiment was used to establish the depression model, and Ago was used to treat CRS mice. Based on behavioral tests in mice and electrophysiology record, SD and LHb neuron activity were assessed. The expression levels of brain-derived neurotrophic factor (BDNF) and nuclear phosphoprotein (c-Fos) in LHb were detected by Western blot (WB). RESULTS: As against the CRS group, the Ago group had a reduction in the immobility time during forced swimming training and an increase in the preference for sucrose in the sucrose preference test; The expression levels of c-Fos and BDNF proteins in the LHb neurons of the Ago group mice were lower than those in the CRS group (P < 0.05), and the values approached the levels of the normal control group. In both dark and light environments, the rapid eye movement (REM) sleep duration of the CRS group mice was significantly longer than that of the normal control group (P < 0.05). CONCLUSION: It was concluded that Ago may intervene in the depressive-like behavior and overall sleep patterns of CRS depression model mice by regulating the activity of LHb neurons and inhibiting the neuroinflammatory process. This provides a potential drug target for the development of new treatment strategies for depression.

Modelling a self-defined CNN for effectual classification of PCOS from ultrasound images.

BACKGROUND: Polycystic Ovary Syndrome (PCOS) is a medical condition that causes hormonal disorders in women in their childbearing years. The hormonal imbalance leads to a delayed or even absent menstrual cycle. Women with PCOS mainly suffer from extreme weight gain, facial hair growth, acne, hair loss, skin darkening, and irregular periods, leading to infertility in rare cases. Doctors usually examine ultrasound images and conclude the affected ovary but are incapable of deciding whether it is a normal cyst, PCOS, or cancer cyst manually. OBJECTIVE: To have access to the high-risk crucial PCOS and to detect the condition and the treatment aimed at mitigating health hazards such as endometrial hyperplasia/cancer, infertility, pregnancy complications, and the long-term burden of chronic diseases such as cardiometabolic disorders linked with PCOS. METHODS: The proposed Self-Defined Convolution Neural Network method (SD_CNN) is used to extract the features and machine learning models such as SVM, Random Forest, and Logistic Regression are used to classify PCOS images. The parameter tuning is done with lesser parameters in order to overcome over-fitting issues. The self-defined model predicts the occurrence of the cyst based on the analyzed features and classifies the class labels effectively. RESULTS: The Random Forest Classifier was found to be the most reliable and accurate among Support Vector Machine (SVM) and Logistic Regression (LR), with accuracy being 96.43%. CONCLUSION: The proposed model establishes better trade-off compared to various other approaches and works effectually for PCOS prediction.

Complete genome of the Medicago anthracnose fungus, Colletotrichum destructivum, reveals a mini-chromosome-like region within a core chromosome.

Colletotrichum destructivum (Cd) is a phytopathogenic fungus causing significant economic losses on forage legume crops (Medicago and Trifolium species) worldwide. To gain insights into the genetic basis of fungal virulence and host specificity, we sequenced the genome of an isolate from Medicago sativa using long-read (PacBio) technology. The resulting genome assembly has a total length of 51.7 Mb and comprises ten core chromosomes and two accessory chromosomes, all of which were sequenced from telomere to telomere. A total of 15, 631 gene models were predicted, including genes encoding potentially pathogenicity-related proteins such as candidate-secreted effectors (484), secondary metabolism key enzymes (110) and carbohydrate-active enzymes (619). Synteny analysis revealed extensive structural rearrangements in the genome of Cd relative to the closely related Brassicaceae pathogen, Colletotrichum higginsianum. In addition, a 1.2 Mb species-specific region was detected within the largest core chromosome of Cd that has all the characteristics of fungal accessory chromosomes (transposon-rich, gene-poor, distinct codon usage), providing evidence for exchange between these two genomic compartments. This region was also unique in having undergone extensive intra-chromosomal segmental duplications. Our findings provide insights into the evolution of accessory regions and possible mechanisms for generating genetic diversity in this asexual fungal pathogen.

Risk Factors for Focal Choroidal Excavation Concurrent with Chorioretinal Disease: Evaluated by Spectral-Domain OCT.

Purpose: To investigate the risk factors for patients with focal choroidal excavation (FCE) and their correlation with chorioretinal diseases. Design: Retrospective cross-sectional study. Subjects: Patients with FCE were enrolled, while healthy subjects were recruited for the control group. Methods: The study collected demographic information, clinical features, and multimodal images. Parameters of FCE identified using spectral-domain OCT (SD-OCT) were manually measured using built-in software and subsequently analyzed statistically. Main Outcome Measures: Subfoveal choroidal thickness (SFCT), subexcavation choroidal thickness (SECT), and the greatest depth and width of each excavation were manually measured using built-in calipers in OCT software. Results: Twenty-one patients (13/8, male/female) with FCE were included in this study. The average age was 45.2 years, and their best-corrected visual acuity (BCVA) was 0.4 logarithm of the minimum angle of resolution (Snellen equivalent, 20/50). Focal choroidal excavation was present in 28 eyes of 21 patients, including isolated FCE (12 eyes) and complicated FCE (16 eyes) with choroidal neovascularization (sCNV), central serous chorioretinopathy, and other conditions. Patients with complicated FCE were significantly older than those isolated FCE (P = 0.015). The SFCT of the healthy subjects was significantly less than that of the fellow eyes of the patients with FCE (P < 0.01), as was that of the eyes with isolated FCE (P < 0.001) and complicated FCE (P < 0.001). The width of excavation was wider in eyes with complicated FCE than in those with isolated FCE (P = 0.001). Hypertransmission defect (HD) was found beneath 15 excavations and was more prevalent in the complicated FCE group than the isolated FCE group (P = 0.023). Conclusions: Focal choroidal excavation appears to be closely related to chorioretinal disorders, and the width of the excavation is a significant indicator for evaluating the risk of chorioretinal diseases. Financial Disclosures: Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.

Simultaneous estimation of paclitaxel and bortezomib via LC-MS/MS: pharmaceutical and pharmacokinetic applications.

Aim & Objective: This study evaluates the potential of combining paclitaxel (PTX) and bortezomib (BTZ) for breast cancer therapy. Materials & Methods: The nanoformulation was optimized via Box-Behnken Design (BBD), with method validation adhering to US-FDA guidelines. Results: Multiple reaction monitoring transitions for PTX, BTZ and internal standard were m/z 855.80→286.60, 366.80→226.00 and 179.80→110.00, respectively. Elution done on C18 Luna column with 0.1% FA in MeOH:10 mM ammonium acetate. The size of nanoformulation was 133.9 ± 1.97 nm, PDI 0.19 ± 0.01 and zeta potential -19.20 ± 1.36 mV. Pharmacokinetics showed higher Cmax for PTX-BTZ-NE (313.75 ± 10.71 ng/ml PTX, 11.92 ± 0.53 ng/ml BTZ) versus free PTX-BTZ (104 ± 13.06 ng/ml PTX, 1.9 ± 0.08 ng/ml BTZ). Conclusion: Future findings will contribute to the treatment of breast cancer using PTX and BTZ.

[Box: see text].

Age-Related Retinal Layer Thickness Changes Measured by OCT in APPNL-F/NL-F Mice: Implications for Alzheimer's Disease.

In Alzheimer's disease (AD), transgenic mouse models have established links between abnormalities in the retina and those in the brain. APPNL-F/NL-F is a murine, humanized AD model that replicates several pathological features observed in patients with AD. Research has focused on obtaining quantitative parameters from optical coherence tomography (OCT) in AD. The aim of this study was to analyze, in a transversal case-control study using manual retinal segmentation via SD-OCT, the changes occurring in the retinal layers of the APPNL/F-NF/L AD model in comparison to C57BL/6J mice (WT) at 6, 9, 12, 15, 17, and 20 months of age. The analysis focused on retinal thickness in RNFL-GCL, IPL, INL, OPL, and ONL based on the Early Treatment Diabetic Retinopathy Study (ETDRS) sectors. Both APPNL-F/NL-F-model and WT animals exhibited thickness changes at the time points studied. While WT showed significant changes in INL, OPL, and ONL, the AD model showed changes in all retinal layers analyzed. The APPNL-F/NL-F displayed significant thickness variations in the analyzed layers except for the IPL compared to related WT. These thickness changes closely resembled those found in humans during preclinical stages, as well as during mild and moderate AD stages, making this AD model behave more similarly to the disease in humans.

Changes in Macular Pigment Optical Density after Intravitreal Faricimab in Neovascular Age-Related Macular Degeneration: A Pilot Study.

Background: This study aimed to evaluate the effects of faricimab intravitreal injections in patients with exudative age macular degeneration (nAMD) after the loading dose using spectral domain optical coherence tomography (SD-OCT) and macular pigment optical density (MPOD). Methods: In this observational prospective study, we enlisted a total of 12 consecutive eyes of 12 patients (six females, six males; mean age 70.47 ± 2.46 years) affected by nAMD who consecutively presented to the Eye Clinic of the University of Naples "Federico II" and Monaldi Hospital of Naples, from June 2023 to December 2023. All patients received four once-monthly intravitreal injections of faricimab (6 mg/0.05 mL) (loading phase). At baseline and 1 month after the fourth faricimab monthly injection, all patients underwent assessment of best correct visual acuity (BCVA) and ophthalmic examination, including slit-lamp biomicroscopy, intraocular pressure (IOP), fundus biomicroscopy, SD-OCT, and MPOD. Results: A total of 12 eyes of 12 patients (six women, six men; mean age 70.47 ± 2.46 years) were included in this study. A statistically significant raise in BCVA and MOPD parameters was shown between baseline and after the loading phase (p < 0.001). Conclusions: Intravitreal injections of faricimab led in the short term to a significant functional and MPOD improvement along with a decrease in central macular thickness (CMT) and thus appears to be an effective treatment option without relevant adverse effects. MOPD may be considered as a prognostic factor associated with a good visual prognosis after intravitreal injections treatment.

Prenatal exposure to bisphenol AF causes toxicities in liver, spleen, and kidney tissues of SD rats.

As a replacement for bisphenol A (BPA), bisphenol AF (BPAF) showed stronger maternal transfer and higher fetal accumulation than BPA. Therefore, concerns should be raised about the health risks of maternal exposure to BPAF during gestation on the offspring. In this study, SD rats were exposed to BPAF (0, 50, and 100 mg/kg/day) during gestation to investigate the bioaccumulation and adverse effects in liver, spleen, and kidney tissues of the offspring at weaning period. Bioaccumulation of BPAF in these tissues with concentrations ranging from 1.56 ng/mg (in spleen of males) to 55.44 ng/mg (in liver of females) led to adverse effects at different biological levels, including increased relative weights of spleen and kidneys, histopathological damage in liver, spleen, and kidney, organ functional damage in liver, spleen, and kidney, upregulated expression of genes related to lipid metabolism (in liver), oxidative stress response (in kidney), immunity and inflammatory (in spleen). Furthermore, dysregulated metabolomics was identified in spleen, with 217 differential metabolites screened and 9 KEGG pathways significantly enriched. This study provides a comprehensive insight into the systemic toxicities of prenatal exposure to BPAF in SD rats. Given the broad applications and widespread occurrence of BPAF, its safety should be re-considered.

Association Between Baseline Macular Morphologic Features on Optical Coherence Tomography and Visual Outcomes in Patients with Vogt-Koyanagi-Harada Disease.

PURPOSE: The choroidal thickening and serous retinal detachments that characterize Vogt-Koyanagi-Harada (VKH) disease can be imaged in detail using spectral domain optical coherence tomography (SD-OCT). Whether specific qualitative and quantitative SD-OCT features at presentation were associated with visual outcomes in a randomized controlled trial comparing methotrexate to mycophenolate for steroid-sparing control of uveitis were evaluated. METHODS: An exploratory subanalysis of data from the FAST trial in which SD-OCT images from VKH participants were analyzed for presence/absence of bacillary detachments, retinal pigment epithelium (RPE) folds, and internal limiting membrane (ILM) fluctuations was performed. A modified RPE undulation index was calculated to provide a quantifiable surrogate marker for choroidal folds. RESULTS: SD-OCT images were available from 158 eyes with VKH. At baseline, bacillary detachments were present in 23.5% of eyes, RPE folds in 22.8% of eyes, and ILM fluctuations in 35.2% of eyes. For each 0.1 unit increase in modified RPE undulation index, there was an associated 0.13 increase in mean logMAR BSCVA at baseline. None of the SD-OCT features were associated with BSCVA at the 6-month primary endpoint. Indeed, mean final BSCVA was similar in those with and without the SD-OCT features of interest at baseline, and was between 0.1 and 0.2 logMAR (Snellen visual acuity 20/25 to 20/30). CONCLUSIONS: While eyes with VKH may present with a variety of SD-OCT imaging pathology prior to starting immunosuppression with methotrexate or mycophenolate mofetil, final visual outcome in our study was excellent. With appropriate immunosuppression, good visual outcomes are possible in VKH.ClinicalTrials.gov Identifier NCT01829295Date of Registration: April 11, 2013.

Cross-sectional Analysis of Outer Retinal Tubulation in Inherited Retinal Diseases: A Multicenter Study.

PURPOSE: This study aims to explore genetic variants that potentially lead to outer retinal tubulation (ORT), estimate the prevalence of ORT in these candidate genes, and investigate the clinical etiology of ORT in patients with inherited retinal diseases (IRDs), with respect to each gene. DESIGN: Retrospective cohort study. METHODS: A retrospective cross-sectional review was conducted on 565 patients with molecular diagnoses of IRD, confirming the presence of ORT as noted in each patient's respective spectral-domain optical coherence tomography (SD-OCT) imaging. Using SD-OCT imaging, the presence of ORT was analyzed in relation to specific genetic variants and phenotypic characteristics. Outcomes included the observed ORT frequencies across two gene-specific cohorts: non- retinal pigment epithelium (RPE)-specific genes, and RPE-specific genes; and to investigate the analogous characteristics caused by variants in these genes. RESULTS: Among the 565 patients included in this study, 104 exhibited ORT on SD-OCT. We observed ORT frequencies among the following genes from our patient cohort: 100% (23/23) forCHM, 100%(2/2) forPNPLA6, 100% (4/4) forRCBTB1, 100% formtDNA[100% (4/4) forMT-TL1and 100% (1/1) formtDNAdeletion], 100% (1/1) forOAT, 95.2% (20/21) forCYP4V2, 72.7% (8/11) forCHMfemale carriers, 66.7% (2/3) forC1QTNF5, 57.1% (8/14) forPROM1, 53.8% (7/13) forPRPH2, 42.9% (3/7) forCERKL, 28.6% (2/7) forCDHR1, 20% (1/5) forRPE65, 4% (18/445) forABCA4.In contrast, ORT was not observed in any patients with photoreceptor-specific gene variants, such asRHO(n=13),USH2A(n=118),EYS(n=70),PDE6B(n=10),PDE6A(n=4),and others. CONCLUSION: These results illustrate a compelling association between the presence of ORT and IRDs caused by variants in RPE-specific genes, as well as non-RPE-specific genes. In contrast, IRDs caused by photoreceptor-specific genes are typically not associated with ORT occurrence. Further analysis revealed that ORT tends to manifest in IRDs with milder intraretinal pigment migration (IPM), a finding that is typically associated with RPE-specific genes. These findings regarding ORT, genetic factors, atrophic patterns in the fundus, and IPM provide valuable insight into the complex etiology of IRDs. Future prospective studies are needed to further explore the association and underlying mechanisms of ORT in these contexts.

Modeling of drinking and driving behaviors among adolescents and young adults in the United States: Complexities and Intervention outcomes.

Alcohol-impaired driving is a formidable public health problem in the United States, claiming the lives of 37 individuals daily in alcohol-related crashes. Alcohol-impaired driving is affected by a multitude of interconnected factors, coupled with long delays between stakeholders' actions and their impacts, which not only complicate policy-making but also increase the likelihood of unintended consequences. We developed a system dynamics simulation model of drinking and driving behaviors among adolescents and young adults. This was achieved through group model building sessions with a team of multidisciplinary subject matter experts, and a focused literature review. The model was calibrated with data series from multiple sources and replicated the historical trends for male and female individuals aged 15 to 24 from 1982 to 2020. We simulated the model under different scenarios to examine the impact of a wide range of interventions on alcohol-related crash fatalities. We found that interventions vary in terms of their effectiveness in reducing alcohol-related crash fatalities. In addition, although some interventions reduce alcohol-related crash fatalities, some may increase the number of drinkers who drive after drinking. Based on insights from simulation experiments, we combined three interventions and found that the combined strategy may reduce alcohol-related crash fatalities significantly without increasing the number of alcohol-impaired drivers on US roads. Nevertheless, related fatalities plateau over time despite the combined interventions, underscoring the need for new interventions for a sustained decline in alcohol-related crash deaths beyond a few decades. Finally, through model calibration we estimated time delays between actions and their consequences in the system which provide insights for policymakers and activists when designing strategies to reduce alcohol-related crash fatalities.

Regulatory role of VvsB protein on serine protease activity of VvsA in Vibrio vulnificus.

BACKGROUND: Vibrio vulnificus NCIMB2137, a Gram-negative, metalloprotease negative estuarine strain was isolated from a diseased eel. A 45 kDa chymotrypsin-like alkaline serine protease known as VvsA has been recently reported as one of the major virulence factor responsible for the pathogenesis of this strain. The vvsA gene along with a downstream gene vvsB, whose function is still unknown constitute an operon designated as vvsAB. OBJECTIVE: This study examines the contribution of VvsB to the functionality of VvsA. METHOD: In this study, VvsB was individually expressed using Rapid Translation System (RTS system), followed by an analysis of its role in regulating the serine protease activity of VvsA. RESULT: The proteolytic activity of VvsA increased upon the addition of purified VvsB to the culture supernatant of V. vulnificus. However, the attempts of protein expression using an E. coli system revealed a noteworthy observation that protein expression from the vvsA gene exhibited higher protease activity compared to that from the vvsAB gene within the cytoplasmic fraction. These findings suggest an intricate interplay between VvsB and VvsA, where VvsB potentially interacts with VvsA inside the bacterium and suppress the proteolytic activity. While outside the bacterial milieu, VvsB appears to stimulate the activation of inactive VvsA. CONCLUSION: The findings suggest that Vibrio vulnificus regulates VvsA activity through the action of VvsB, both intracellularly and extracellularly, to ensure its survival.

Visual Outcomes in Malignant Hypertensive Retinopathy Cases: A Clinical and Spectral Domain Optical Coherence Tomography Study.

Objective The objective is to correlate visual outcomes in malignant hypertensive retinopathy with changes in systemic causative factors and spectral domain optical coherence tomography (SD OCT) morphologic parameters. Materials and methods This is a prospective observational study including patients presenting within two weeks of acute rise of systolic blood pressure (SBP) ≥ 180 mm Hg or diastolic blood pressure (DBP) ≥ 120 mm Hg and with posterior segment involvement in both eyes. Baseline SBP, DBP, mean arterial pressure (MAP), best corrected visual acuity (BCVA), and SD OCT parameters such as central macular thickness (CMT), subfoveal choroidal thickness (SCT), and sub-retinal fluid (SRF) height were measured at presentation and followed monthly up to three months. These variables at baseline and three months were compared and correlated. Results Thirty-three patients (66 eyes) having malignant hypertension were included in the study. Diverse clinical presentations noted among patients were optic disc edema, hard exudates in the macula, peripapillary splinter hemorrhage, cotton wool spots, Elschnig spots, exudative retinal detachment, optic neuropathy, and severe exudative retinopathy. SD OCT shows hyperreflective dots and intraretinal fluid with or without SRF. At three months, the mean SBP, DBP, MAP, CMT, SRF, and SCT all decreased significantly from baseline (p<0.001). Changes in SBP, DBP, MAP, and SCT correlated significantly with changes in BCVA (p<0.001). Conclusion In malignant hypertensive retinopathy, macular edema with SRF is the major cause of mild-to-moderate decrease BCVA at presentation, but macular ischemia, exudative RD, and optic neuropathy can cause a significant decrease in vision. A decrease in SBP, DBP, MAP, and SCT correlate significantly with visual outcomes.

Tetrameric, active PKM2 inhibits IP3 receptors, potentially requiring GRP75 as an additional interaction partner.

Pyruvate kinase M2 (PKM2) is a key glycolytic enzyme interacting with the inositol 1,4,5-trisphosphate receptor (IP3R). This interaction suppresses IP3R-mediated cytosolic [Ca2+] rises. As PKM2 exists in monomeric, dimeric and tetrameric forms displaying different properties including catalytic activity, we investigated the molecular determinants of PKM2 enabling its interaction with IP3Rs. Treatment of HeLa cells with TEPP-46, a compound stabilizing the tetrameric form of PKM2, increased both its catalytic activity and the suppression of IP3R-mediated Ca2+ signals. Consistently, in PKM2 knock-out HeLa cells, PKM2C424L, a tetrameric, highly active PKM2 mutant, but not inactive PKM2K270M or the less active PKM2K305Q, suppressed IP3R-mediated Ca2+ release. Surprisingly, however, in vitro assays did not reveal a direct interaction between purified PKM2 and either the purified Fragment 5 of IP3R1 (a.a. 1932-2216) or the therein located D5SD peptide (a.a. 2078-2098 of IP3R1), the presumed interaction sites of PKM2 on the IP3R. Moreover, on-nucleus patch clamp of heterologously expressed IP3R1 in DT40 cells devoid of endogenous IP3Rs did not reveal any functional effect of purified wild-type PKM2, mutant PKM2 or PKM1 proteins. These results indicate that an additional factor mediates the regulation of the IP3R by PKM2 in cellulo. Immunoprecipitation of GRP75 using HeLa cell lysates co-precipitated IP3R1, IP3R3 and PKM2. Moreover, the D5SD peptide not only disrupted PKM2:IP3R, but also PKM2:GRP75 and GRP75:IP3R interactions. Our data therefore support a model in which catalytically active, tetrameric PKM2 suppresses Ca2+ signaling via the IP3R through a multiprotein complex involving GRP75.

Utility of the WHO dengue guidelines in pediatric immunological studies.

Dengue is a significant health problem due to the high burden of critical infections during outbreaks. In 1997, the World Health Organization (WHO) classified dengue as dengue fever (DF), dengue hemorrhagic fever (DHF), and dengue shock syndrome (DSS). It was revised in 2009 (updated in 2015), and the new guidelines recommended classifying patients as dengue without warning signs (DNS), dengue with warning signs (DWS), and severe dengue (SD). Although the utility of the revised 2009 classification for clinical studies is accepted, for immunological studies it needs to be clarified. We determined the usefulness of the 2009 classification for pediatric studies that analyze the circulating interleukin (IL)-6 and IL-8, two inflammatory cytokines. Plasma levels of IL-6 and IL-8 were evaluated in the acute and convalescent phases by flow cytometry in children with dengue classified using the 1997 and 2009 WHO guidelines. The plasma levels of IL-6 and IL-8 were elevated during the acute and decreased during convalescence, and both cytokines served as a good marker of acute dengue illness compared to convalescence. There were no differences in the plasma level of the evaluated cytokines among children with different clinical severity with any classification, except for the IL-8, which was higher in DWS than DNS. Based on the levels of IL-8, the 2009 classification identified DWS plus SD (hospital-treated children) compared to the DNS group [area under the curve (AUC): 0.7, p = 0.028]. These results support the utility of the revised 2009 (updated in 2015) classification in studies of immune markers in pediatric dengue.

Corrigendum: Current phenotypic and genetic spectrum of syndromic deafness in Tunisia: paving the way for precision auditory health.

[This corrects the article DOI: 10.3389/fgene.2024.1384094.].

Pathological Changes in the Gastrocnemius Muscle throughout Hind Limb Ischemia Modeling in Rats.

We present a two-stage model for the study of chronic hind limb ischemia in rats. In the area of ischemia, sclerotic changes with atrophic rhabdomyocytes and reduced vascularization were revealed. CD31 expression in the endothelium increased proportionally to the number of vessels in the ischemic zone, and at the same time, focal expression of ßIII-tubulin was detected in the newly formed nerve fibers. These histological features are equivalent to the development of peripheral arterial disease in humans, which allows using our model in the search for new therapeutic strategies.

Physiotypic variations lead to variations within the normal range for red blood cells and haemoglobin levels in a healthy human population: An evaluation using generalised additive modelling and hierarchical structure analysis.

OBJECTIVES: Haematological parameters have been used for a long time for clinical evaluations, however the dynamics of these parameters has not been studied at length, in healthy populations. We aim to understand the dependence of haematological parameters on human physiotypes. DESIGN AND METHODS: Using an age and gender restricted healthy human (male) population (n = 100), we attempt to analyse the dynamics of haemoglobin and red blood cells, with reference to age, height and weight of individuals. Using advanced generalised additive modelling and classical hierarchical structural analysis we aim to establish relationships between these parameters and human physiotypes. RESULTS: We demonstrate that definitive relationships can be established for number of red blood cells, haemoglobin levels, RDW-CV, RDW-SD and weight, height and age of individuals. CONCLUSION: This study provides a proof of principle, that haematological parameters are dependent on physiotypic variation, within the normal ranges in a healthy population. It may also be noted that there is a definitive influence of height, weight and age on normal ranges and stratification by these factors might therefore make reference intervals narrower, in turn, possibly allowing more precise clinical decisions based on the complete blood count (CBC).