Morphological and immunohistochemical differentiation of neuronal and glial cells of the vascular and avascular regions of the donkey's paurangiotic retina.

INTRODUCTION: Ocular diseases pose a significant health concern for donkeys. However, studies examining the microanatomy and cell populations of the donkey retina are scarce. The current study aims to describe the vascular pattern of the donkey retina and document its cellular components. METHODS: The donkey retina specimens were obtained from different retinal regions and prepared for semithin sectioning and immunohistochemistry. RESULTS: The donkey has a paurangiotic retina in which retinal vessels are confined to a narrow area around the optic disc. Glial cells coexist with the blood vessels being very numerous in the vascular region and become scanty in the avascular ones. S-100 positive astrocytes could be observed in these avascular areas. Ganglion cells are organized in a single layer with the least population existing in the peripheral retina. Acidic fibroblast growth factor (AFGF) is immunoreactive in amacrine and ganglion cells. A subpopulation of amacrine cells reacted strongly to tyrosine hydroxylase (TH), and others reacted positively to S-100 protein. Ganglion cell nuclei exhibited a strong immunoreactivity to S-100 protein as well. Furthermore, glial fibrillary acidic protein (GFAP) is used to identify Müller cells which extend their processes across the retina from the inner to the outer limiting membrane. CONCLUSIONS: In conclusion, our findings provide novel insights into the normal retinal organization. The donkey retina shows the characteristic expression of immunohistochemical markers for the major cell types. In addition, the distribution of glial cells is comparable between the vascular and avascular regions.

Co-fractionation-mass spectrometry to characterize native mitochondrial protein assemblies in mammalian neurons and brain.

Human mitochondrial (mt) protein assemblies are vital for neuronal and brain function, and their alteration contributes to many human disorders, e.g., neurodegenerative diseases resulting from abnormal protein-protein interactions (PPIs). Knowledge of the composition of mt protein complexes is, however, still limited. Affinity purification mass spectrometry (MS) and proximity-dependent biotinylation MS have defined protein partners of some mt proteins, but are too technically challenging and laborious to be practical for analyzing large numbers of samples at the proteome level, e.g., for the study of neuronal or brain-specific mt assemblies, as well as altered mtPPIs on a proteome-wide scale for a disease of interest in brain regions, disease tissues or neurons derived from patients. To address this challenge, we adapted a co-fractionation-MS platform to survey native mt assemblies in adult mouse brain and in human NTERA-2 embryonal carcinoma stem cells or differentiated neuronal-like cells. The workflow consists of orthogonal separations of mt extracts isolated from chemically cross-linked samples to stabilize PPIs, data-dependent acquisition MS to identify co-eluted mt protein profiles from collected fractions and a computational scoring pipeline to predict mtPPIs, followed by network partitioning to define complexes linked to mt functions as well as those essential for neuronal and brain physiological homeostasis. We developed an R/CRAN software package, Macromolecular Assemblies from Co-elution Profiles for automated scoring of co-fractionation-MS data to define complexes from mtPPI networks. Presently, the co-fractionation-MS procedure takes 1.5-3.5 d of proteomic sample preparation, 31 d of MS data acquisition and 8.5 d of data analyses to produce meaningful biological insights.

Post-Exercise Hypotension: An Alternative Management Strategy for Hypertension and Cardiovascular Disease?

Cardiovascular disease (CVD), including hypertension, is a leading cause of death worldwide and imposes an enormous burden on our societies [...].

Satisfaction and anxiety level during clinical training among nursing students.

BACKGROUND: Quality is a primary concern of health care agencies worldwide. A conducive clinical training environment is essential for nursing students to be capable of enhancing their learning experiences and achieving the desired training outcomes. AIM: This study aimed to examine the satisfaction and anxiety levels during clinical training among nursing students. TYPE OF STUDY: A descriptive -analytical cross-sectional study design was utilized. The research was conducted at the Faculty of Nursing, Assiut University and Colleges of Applied Medical Sciences in Alnamas and Bisha, University of Bisha. Sampling method: A convenience sampling technique was used. SAMPLE SIZE: a sample of 1052 undergraduate nursing students. The data was gathered via a structured questionnaire including the socio-demographic characteristics and nursing students' satisfaction with the hospital and laboratory training. Additionally, Self-Rating Anxiety Scale (SAS) was adopted to measure the anxiety level. RESULTS: The mean age of the studied sample was 21.9 ± 1.83 years, and 56.9% are females. Moreover, 90.1% & 76.4% of the nursing students were satisfied with their hospital and laboratory training. Furthermore, 61.1% & 54.8% of the students had mild levels of anxiety regarding their hospital training and laboratory training, respectively. CONCLUSION: The undergraduate nursing students had a high level of satisfaction with their clinical training at the hospitals and laboratories. Moreover, they had mild anxiety related to hospital and laboratory clinical training. RECOMMENDATIONS: Developing clinical orientation and training programs and improvement strategies to enhance the effectiveness of the clinical training environment. The establishment of a modern, tastefully designed, and fully stocked skill lab for the college's student training should receive more attention. CLINICAL RELEVANCE: Through the provision of ongoing education about different method of practice, nursing was intended to shape future professional nurses who master core competencies of the profession. Organizations may benefit from developing a comprehensive strategy to achieve an effective teaching program.

Sanitation enzymes: Exquisite surveillance of the noncanonical nucleotide pool to safeguard the genetic blueprint.

Reactive oxygen species (ROS) are common products of normal cellular metabolism, but their elevated levels can result in nucleotide modifications. These modified or noncanonical nucleotides often integrate into nascent DNA during replication, causing lesions that trigger DNA repair mechanisms such as the mismatch repair machinery and base excision repair. Four superfamilies of sanitization enzymes can effectively hydrolyze noncanonical nucleotides from the precursor pool and eliminate their unintended incorporation into DNA. Notably, we focus on the representative MTH1 NUDIX hydrolase, whose enzymatic activity is ostensibly nonessential under normal physiological conditions. Yet, the sanitization attributes of MTH1 are more prevalent when ROS levels are abnormally high in cancer cells, rendering MTH1 an interesting target for developing anticancer treatments. We discuss multiple MTH1 inhibitory strategies that have emerged in recent years, and the potential of NUDIX hydrolases as plausible targets for the development of anticancer therapeutics.

Triggers of Atrial Fibrillation in the Geriatric Medical Intensive Care Unit: An Observational Study.

Background: Atrial fibrillation (AF) is a common arrhythmia in the non-cardiac intensive care unit (ICU). However, data concerning AF incidence and predictors in such populations are scarce and controversial. The study aimed to investigate the contributing factors of new-onset AF in elderly patients within the medical intensive care setting. Methods: Patients admitted to ICU during a 6-month period were prospectively studied. Patients admitted for short period postoperative monitoring and patients with chronic or paroxysmal AF were excluded. The conditions involved as AF risk factors or "triggers" from demographic data, history, and echocardiography were recorded. Acute Physiology and Chronic Health Evaluation II score was calculated. Electrolytes including some trace elements (zinc, copper, and magnesium) were analyzed. Results: The study included 142 patients (49% females). Mean age was 69.5 ± 7.3 years. AF was observed in 12%. Diagnosis of pneumonia (P < 0.001), low copper (P < 0.0001) and low zinc levels (P < 0.0001) was significantly associated with the occurrence of AF. By multivariate analysis, they remained statistically significant (odds ratio, 7.0; 95% confidence interval, 2.0 - 24.6; P < 0.01). Conclusions: A significant fraction of ICU elderly patients manifests AF. The relevant factors contributing to AF incidence in the elderly are pneumonia and low zinc and low copper.

The Complexity of Peripheral Arterial Disease and Coronary Artery Disease in Diabetic Patients: An Observational Study.

Background: Atherosclerosis is a systemic disease that causes luminal narrowing. Patients with peripheral arterial disease (PAD) also exhibit an increased risk of death from cardiovascular complications. This risk is the same for symptomatic or asymptomatic patients. Over a 5-year period, patients with PAD have a 20% chance of suffering from a stroke or myocardial infarction. Additionally, their mortality rate is 30%. This study aimed to assess the relationship between coronary artery disease (CAD) complexity using SYNTAX score and PAD complexity using Trans-Atlantic Inter-Society Consensus II (TASC II) score. Methods: The study was designed as single-center cross-sectional observational and included 50 diabetic patients referred for elective coronary angiography and peripheral angiography was done. Results: Most of the patients were males (80%) and smokers (80%) with mean age of 62 years. The mean SYNTAX score was 19.88. There was a significant negative correlation between SYNTAX score and ankle brachial index (ABI) (r = -0.48, P = 0.001) and a significant positive correlation with glycated hemoglobin (HbA1c) level (R2 = 26, P = 0.004). Complex PAD was found in nearly half of the patients with 48% having TASC II C or D classes. Those with TASC II classes C and D had higher SYNTAX scores (P = 0.046). Conclusions: Diabetic patients with more complex CAD had more complex PAD. In diabetic patients with CAD, those with worse glycemic control had higher SYNTAX scores and the higher the SYNTAX score, the lower the ABI.

Impact of COVID-19 on youth's mental health in Egypt.

Background: Youth represent 21% of the Egyptian population; such proportion can create a leading demographic power for economic development and transition. However, with the current COVID-19 pandemic, everyone is exposed to more than usual stressors, adding a burden to their mental health and well-being. Aim: This study aims to understand the pandemic's effect on youth's mental health in Egypt to strengthen the intervention areas needed to tackle such issues. Methods: This observational, analytical, cross-sectional study employed internet platforms of Facebook & WhatsApp groups for a web-based survey that included 412 respondents between 15 and 30 years old. RESULTS The median age of the respondents was 22 years. At least 30% reported increased violence in the street and/or household, and 27.4% of the respondents have considered visiting a psychiatrist during the last period. Conclusion: It is evident that the current situation is unprecedented and challenging for everyone; however, some populations are more vulnerable than others. Thus, it's important to support young people to ensure that the whole community can withstand the pandemic. The governments should support and mitigate some of the stresses that can be directly amended, like the education and job security concerns.

Insights into SACS pathological attributes in autosomal recessive spastic ataxia of Charlevoix-Saguenay (ARSACS)☆.

Autosomal recessive spastic ataxia of Charlevoix-Saguenay (ARSACS) is a rare early-onset neurodegenerative disease caused by mutations in the SACS gene, encoding Sacsin. Initial functional annotation of Sacsin was based on sequence homology, with subsequent experiments revealing the Sacsin requirement for regulating mitochondrial dynamics, along with its domains involved in promoting neurofilament assembly or resolving their bundling accumulations. ARSACS phenotypes associated with SACS loss-of-function are discussed, and how advancements in ARSACS disease models and quantitative omics approaches can improve our understanding of ARSACS pathological attributes. Lastly in the perspectives section, we address gene correction strategies for monogenic disorders such as ARSACS, along with their common delivery methods, representing a hopeful area for ARSACS therapeutics development.

A computational approach to rapidly design peptides that detect SARS-CoV-2 surface protein S.

The coronavirus disease 19 (COVID-19) caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) prompted the development of diagnostic and therapeutic frameworks for timely containment of this pandemic. Here, we utilized our non-conventional computational algorithm, InSiPS, to rapidly design and experimentally validate peptides that bind to SARS-CoV-2 spike (S) surface protein. We previously showed that this method can be used to develop peptides against yeast proteins, however, the applicability of this method to design peptides against other proteins has not been investigated. In the current study, we demonstrate that two sets of peptides developed using InSiPS method can detect purified SARS-CoV-2 S protein via ELISA and Surface Plasmon Resonance (SPR) approaches, suggesting the utility of our strategy in real time COVID-19 diagnostics. Mass spectrometry-based salivary peptidomics shortlist top SARS-CoV-2 peptides detected in COVID-19 patients' saliva, rendering them attractive SARS-CoV-2 diagnostic targets that, when subjected to our computational platform, can streamline the development of potent peptide diagnostics of SARS-CoV-2 variants of concern. Our approach can be rapidly implicated in diagnosing other communicable diseases of immediate threat.

The evaluation of the effect of performing guided lid surgery with enucleation of a cystic lesion; a case report.

INTRODUCTION AND IMPORTANCE: Dentigerous cysts are benign odontogenic cysts of developmental origin. Enucleation and marsupialization are still considered the blueprint of cystic lesion treatment. CASE PRESENTATION: A 23-year-old male patient presented complaining of a minimal swelling in his upper jaw with slight tenderness in his upper anterior teeth. Cone Beam Computed Tomography (CBCT) on the maxilla was requested. The cystic lesion was found to be minimally expansile with intact cortical plates of the maxilla in the affected area. The CBCT was used to fabricate a cutting guide to determine the exact location of the bony window to fully access the lesion. Root canal treatment was done for the affected non-vital teeth. The cuts were done using a piezo-electric device. Complete enucleation was done for the lesion followed by fixation of the cortical bone lid using micro-plates and screws. The case was followed up after 6 months for new bone deposition using CBCT and 1 week, 1 month, and 6 months postoperatively for postoperative pain using the Visual Analogue Scale (VAS). CLINICAL DISCUSSION: Piezo-electric surgery was used due to the selective cutting merit to cut through bone while preserving the cystic lining intact. Lid surgery aims to maximize the volume of bone deposited in place of the defect by converting the cavity of the cystic lesion into a contained defect. CONCLUSION: Guided lid surgery using a piezo-electric device could be a useful technique for cystic enucleation regarding the new bone formation and pain level.

Auxotrophic and prototrophic conditional genetic networks reveal the rewiring of transcription factors in Escherichia coli.

Bacterial transcription factors (TFs) are widely studied in Escherichia coli. Yet it remains unclear how individual genes in the underlying pathways of TF machinery operate together during environmental challenge. Here, we address this by applying an unbiased, quantitative synthetic genetic interaction (GI) approach to measure pairwise GIs among all TF genes in E. coli under auxotrophic (rich medium) and prototrophic (minimal medium) static growth conditions. The resulting static and differential GI networks reveal condition-dependent GIs, widespread changes among TF genes in metabolism, and new roles for uncharacterized TFs (yjdC, yneJ, ydiP) as regulators of cell division, putrescine utilization pathway, and cold shock adaptation. Pan-bacterial conservation suggests TF genes with GIs are co-conserved in evolution. Together, our results illuminate the global organization of E. coli TFs, and remodeling of genetic backup systems for TFs under environmental change, which is essential for controlling the bacterial transcriptional regulatory circuits.

Morphological, Immunohistochemical, and Ultrastructural Studies of the Donkey's Eye with Special Reference to the AFGF and ACE Expression.

The donkey is mainly used as a working animal for riding and pack transport, as well as for dairy and meat production. Eye afflictions are common in donkeys, thus requiring a detailed study. A few studies had focused on the donkey's eye, and most of them had considered it, merely, a horse's eye. This study aimed to investigate the anatomy, histology, ultrastructure, and immunohistochemical features of the donkey's eye. The results were recorded and compared to those of horses in certain dimensions. Unlike horses, the donkey's eye is more circular in the contour of the cornea, has smaller lenticular thickness, and has longer anterior and vitreous chambers. Positive immunoreactivity to acidic fibroblast growth factor in the basal cell layers of the cornea was observed, indicating their role in cell differentiation and the renewal of the epithelium. Moreover, the corneal keratocytes expressed angiotensin-converting enzyme, which plays a role in corneal homeostasis and wound healing. Additionally, telocytes, hyalocytes, and other immune cells were observed within the iris and ciliary processes. Hence, this work is an updated detailed study of the morphology and ultrastructure of the donkey's eye and reveals some similarities and dissimilarities to the horse's eyes, which should be considered in clinical practice.

The conserved Tpk1 regulates non-homologous end joining double-strand break repair by phosphorylation of Nej1, a homolog of the human XLF.

The yeast cyclic AMP-dependent protein kinase A (PKA) is a ubiquitous serine-threonine kinase, encompassing three catalytic (Tpk1-3) and one regulatory (Bcy1) subunits. Evidence suggests PKA involvement in DNA damage checkpoint response, but how DNA repair pathways are regulated by PKA subunits remains inconclusive. Here, we report that deleting the tpk1 catalytic subunit reduces non-homologous end joining (NHEJ) efficiency, whereas tpk2-3 and bcy1 deletion does not. Epistatic analyses revealed that tpk1, as well as the DNA damage checkpoint kinase (dun1) and NHEJ factor (nej1), co-function in the same pathway, and parallel to the NHEJ factor yku80. Chromatin immunoprecipitation and resection data suggest that tpk1 deletion influences repair protein recruitments and DNA resection. Further, we show that Tpk1 phosphorylation of Nej1 at S298 (a Dun1 phosphosite) is indispensable for NHEJ repair and nuclear targeting of Nej1 and its binding partner Lif1. In mammalian cells, loss of PRKACB (human homolog of Tpk1) also reduced NHEJ efficiency, and similarly, PRKACB was found to phosphorylate XLF (a Nej1 human homolog) at S263, a corresponding residue of the yeast Nej1 S298. Together, our results uncover a new and conserved mechanism for Tpk1 and PRKACB in phosphorylating Nej1 (or XLF), which is critically required for NHEJ repair.

From fuzziness to precision medicine: on the rapidly evolving proteomics with implications in mitochondrial connectivity to rare human disease.

Mitochondrial (mt) dysfunction is linked to rare diseases (RDs) such as respiratory chain complex (RCC) deficiency, MELAS, and ARSACS. Yet, how altered mt protein networks contribute to these ailments remains understudied. In this perspective article, we identified 21 mt proteins from public repositories that associate with RCC deficiency, MELAS, or ARSACS, engaging in a relatively small number of protein-protein interactions (PPIs), underscoring the need for advanced proteomic and interactomic platforms to uncover the complete scope of mt connectivity to RDs. Accordingly, we discuss innovative untargeted label-free proteomics in identifying RD-specific mt or other macromolecular assemblies and mapping of protein networks in complex tissue, organoid, and stem cell-differentiated neurons. Furthermore, tag- and label-based proteomics, genealogical proteomics, and combinatorial affinity purification-mass spectrometry, along with advancements in detecting and integrating transient PPIs with single-cell proteomics and transcriptomics, collectively offer seminal follow-ups to enrich for RD-relevant networks, with implications in RD precision medicine.

Mitochondria under the spotlight: On the implications of mitochondrial dysfunction and its connectivity to neuropsychiatric disorders.

Neuropsychiatric disorders (NPDs) such as bipolar disorder (BD), schizophrenia (SZ) and mood disorder (MD) are hard to manage due to overlapping symptoms and lack of biomarkers. Risk alleles of BD/SZ/MD are emerging, with evidence suggesting mitochondrial (mt) dysfunction as a critical factor for disease onset and progression. Mood stabilizing treatments for these disorders are scarce, revealing the need for biomarker discovery and artificial intelligence approaches to design synthetically accessible novel therapeutics. Here, we show mt involvement in NPDs by associating 245 mt proteins to BD/SZ/MD, with 7 common players in these disease categories. Analysis of over 650 publications suggests that 245 NPD-linked mt proteins are associated with 800 other mt proteins, with mt impairment likely to rewire these interactions. High dosage of mood stabilizers is known to alleviate manic episodes, but which compounds target mt pathways is another gap in the field that we address through mood stabilizer-gene interaction analysis of 37 prescriptions and over-the-counter psychotropic treatments, which we have refined to 15 mood-stabilizing agents. We show 26 of the 245 NPD-linked mt proteins are uniquely or commonly targeted by one or more of these mood stabilizers. Further, induced pluripotent stem cell-derived patient neurons and three-dimensional human brain organoids as reliable BD/SZ/MD models are outlined, along with multiomics methods and machine learning-based decision making tools for biomarker discovery, which remains a bottleneck for precision psychiatry medicine.

HERG channel and cancer: A mechanistic review of carcinogenic processes and therapeutic potential.

The human ether-à-go-go related gene (HERG) encodes the alpha subunit of Kv11.1, which is a voltage-gated K+ channel protein mainly expressed in heart and brain tissue. HERG plays critical role in cardiac repolarization, and mutations in HERG can cause long QT syndrome. More recently, evidence has emerged that HERG channels are aberrantly expressed in many kinds of cancer cells and play important roles in cancer progression. HERG could therefore be a potential biomarker for cancer and a possible molecular target for anticancer drug design. HERG affects a number of cellular processes, including cell proliferation, apoptosis, angiogenesis and migration, any of which could be affected by dysregulation of HERG. This review provides an overview of available information on HERG channel as it relates to cancer, with focus on the mechanism by which HERG influences cancer progression. Molecular docking attempts suggest two possible protein-protein interactions of HERG with the ß1-integrin receptor and the transcription factor STAT-1 as novel HERG-directed therapeutic targeting which avoids possible cardiotoxicity. The role of epigenetics in regulating HERG channel expression and activity in cancer will also be discussed. Finally, given its inherent extracellular accessibility as an ion channel, we discuss regulatory roles of this molecule in cancer physiology and therapeutic potential. Future research should be directed to explore the possibilities of therapeutic interventions targeting HERG channels while minding possible complications.

Misconnecting the dots: altered mitochondrial protein-protein interactions and their role in neurodegenerative disorders.

Introduction: Mitochondria (mt) are protein-protein interaction (PPI) hubs in the cell where mt-localized and associated proteins interact in a fashion critical for cell fitness. Altered mtPPIs are linked to neurodegenerative disorders (NDs) and drivers of pathological associations to mediate ND progression. Mapping altered mtPPIs will reveal how mt dysfunction is linked to NDs.Areas covered: This review discusses how database sources reflect on the number of mt protein or interaction predictions, and serves as an update on mtPPIs in mt dynamics and homeostasis. Emphasis is given to mRNA expression profiles for mt proteins in human tissues, cellular models relevant to NDs, and altered mtPPIs in NDs such as Parkinson's disease (PD), Amyotrophic lateral sclerosis (ALS) and Alzheimer's disease (AD).Expert opinion: We highlight the scarcity of biomarkers to improve diagnostic accuracy and tracking of ND progression, obstacles in recapitulating NDs using human cellular models to underpin the pathophysiological mechanisms of disease, and the shortage of mt protein interactome reference database(s) of neuronal cells. These bottlenecks are addressed by improvements in induced pluripotent stem cell creation and culturing, patient-derived 3D brain organoids to recapitulate structural arrangements of the brain, and cell sorting to elucidate mt proteome disparities between cell types.

A Gaussian process-based definition reveals new and bona fide genetic interactions compared to a multiplicative model in the Gram-negative Escherichia coli.

MOTIVATION: A digenic genetic interaction (GI) is observed when mutations in two genes within the same organism yield a phenotype that is different from the expected, given each mutation's individual effects. While multiplicative scoring is widely applied to define GIs, revealing underlying gene functions, it remains unclear if it is the most suitable choice for scoring GIs in Escherichia coli. Here, we assess many different definitions, including the multiplicative model, for mapping functional links between genes and pathways in E.coli. RESULTS: Using our published E.coli GI datasets, we show computationally that a machine learning Gaussian process (GP)-based definition better identifies functional associations among genes than a multiplicative model, which we have experimentally confirmed on a set of gene pairs. Overall, the GP definition improves the detection of GIs, biological reasoning of epistatic connectivity, as well as the quality of GI maps in E.coli, and, potentially, other microbes. AVAILABILITY AND IMPLEMENTATION: The source code and parameters used to generate the machine learning models in WEKA software were provided in the Supplementary information. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Histomorphometric analysis of the choroid of donkeys, buffalos, camels and dogs.

AIM: The present study was carried out to investigate the morphological and histomorphometric characters of choroid in donkeys, buffalos, camels and dogs. RESULTS: The findings of the study revealed that, macroscopically, the choroid was consisted of two areas in all studied animals, except in camel which consists of one area. Histologically, the choroid consists of five layers. Interestingly, the anterior borders of all investigated animals were free of pigments except in camel. Morphometric analysis revealed significant species differences in the mean total thickness of the choroid and its different layers. In addition, significant differences were also found between the ratios of the means of different layers to the total thickness of the choroid. CONCLUSION: In conclusion, these variations might be related to the different lifestyles and visual behavior of the investigated animals.