Molecular insights into codon usage analysis of mitochondrial fission and fusion gene: relevance to neurodegenerative diseases.

Mitochondrial dysfunction is the leading cause of neurodegenerative disorders like Alzheimer's disease and Parkinson's disease. Mitochondria is a highly dynamic organelle continuously undergoing the process of fission and fusion for even distribution of components and maintaining proper shape, number, and bioenergetic functionality. A set of genes governs the process of fission and fusion. OPA1, Mfn1, and Mfn2 govern fusion, while Drp1, Fis1, MIEF1, and MIEF2 genes control fission. Determination of specific molecular patterns of transcripts of these genes revealed the impact of compositional constraints on selecting optimal codons. AGA and CCA codons were over-represented, and CCC, GTC, TTC, GGG, ACG were under-represented in the fusion gene set. In contrast, CTG was over-represented, and GCG, CCG, and TCG were under-represented in the fission gene set. Hydropathicity analysis revealed non-polar protein products of both fission and fusion gene set transcripts. AGA codon repeats are an integral part of translational regulation machinery and present a distinct pattern of over-representation and under-representation in different transcripts within the gene sets, suggestive of selective translational force precisely controlling the occurrence of the codon. Out of six synonymous codons, five synonymous codons encoding for leucine were used differently in both gene sets. Hence, forces regulating the occurrence of AGA and five synonymous leucine-encoding codons suggest translational selection. A correlation of mutational bias with gene expression and codon bias and GRAVY and AROMA signifies the selection pressure in both gene sets, while the correlation of compositional bias with gene expression, codon bias, protein properties, and minimum free energy signifies the presence of compositional constraints. More than 25% of codons of both gene sets showed a significant difference in codon usage. The overall analysis shed light on molecular features of gene sets involved in fission and fusion.

Synthetic biology approach revealed enhancement in haeme oxygenase-1 gene expression by codon pair optimization while reduction by codon deoptimization.

Haem oxygenase-1 (HO-1) is a ubiquitously expressed gene involved in cellular homoeostasis, and its imbalance in expression results in various disorders. To alleviate such disorders, HO-1 gene expression needs to be modulated. Codon usage bias results from evolutionary forces acting on any nucleotide sequence and determines the gene expression. Like codon usage bias, codon pair bias also exists, playing a role in gene expression. In the present study, HO-1 gene was recoded by manipulating codon and codon pair bias, and four such constructs were made through codon/codon pair deoptimization and codon/codon pair optimization to reduce and enhance the HO-1 gene expression. Codon usage analysis was done for these constructs for four tissues brain, heart, pancreas and liver. Based on codon usage in different tissues, gene expression of these tissues was determined in terms of the codon adaptation index. Based on the codon adaptation index, minimum free energy, and translation efficiency, constructs were evaluated for enhanced or decreased HO-1 expression. The analysis revealed that for enhancing gene expression, codon pair optimization, while for reducing gene expression, codon deoptimization is efficacious. The recoded constructs developed in the study could be used in gene therapy regimens to cure HO-1 over or underexpression-associated disorders.

Implications of endoplasmic reticulum stress and beta-cell loss in immunodeficient diabetic NRG-Akita mice for understanding monogenic diabetes.

BACKGROUND: Immunodeficient mice models have become increasingly important as in vivo models engrafted with human cells or tissues for research. The NOD-Rag1null Ins2Akita Il2rnull (NRG Akita) mice is a model combined with immunodeficient NRG and monogenic diabetes Akita mice that develop spontaneous hyperglycemia with progressive loss of pancreatic insulin-producing beta-cells with age. This model is one of the monogenic diabetic models, which has been providing a powerful platform for transplantation experiments of stem cells-generated human ß-cells. This research aimed to provide insights into the mechanisms underlying this monogenic diabetes, which remains incompletely understood. METHODS: Histological and immunofluorescence analyses were conducted on endocrine pancreatic islets to compare NRG wild-type (Wt) controls with NRG-Akita mice. Our investigation focused on assessing the expression of endocrine hormones, transcription factors, proliferation, ER stress, and apoptosis. RESULTS: Histological analyses on NRG-Akita mice revealed smaller islets at 6-weeks-old, due to fewer ß-cells in the islets, compared to NRG-Wt controls, which further progressed with age. The proliferation rate decreased, and apoptosis was abundant in ß-cells in NRG Akita mice. Interestingly, our mechanistic analyses revealed that ß-cells in NRG-Akita mice progressively accumulated the endoplasmic reticulum (ER) stresses, leading to a decreased expression of pivotal ß-cell transcriptional factor PDX1. CONCLUSIONS: Altogether, our mechanistic insight into ß-cell loss in this model could shed light on essential links between ER stress, proliferation, and cell identity, which might open the door to new therapeutic strategies for various diseases since ER stress is one of the most common features not only in diabetes but also in other degenerative diseases.

Advancements and mechanisms of stem cell-based therapies for spinal cord injury in animals.

Spinal cord injury (SCI) is a neurodegenerative disorder of the central nervous system (CNS) that can lead to permanent loss of sensation and voluntary movement beyond the affected area. Extensive preclinical and clinical trials have been conducted to evaluate the safety and effectiveness of stem cells for the treatment of various CNS diseases or disorders, including SCI. However, several challenges hinder nerve cell regeneration in the injured spinal cord, such as extensive cell loss, limited neural cell regeneration capacity, axonal disruption, and the presence of growth-inhibiting molecules, particularly astroglial scarring or glial scars at the injury site in chronic cases. These obstacles pose significant challenges for physicians in restoring normal motor and sensory nerve function in both humans and animals following SCI. This review focuses on SCI pathogenesis, the mechanisms underlying the therapeutic potential of Mesenchymal Stem Cells (MSCs) in SCI, and the potential of stem cell-based therapies as promising avenues for treatment. This review article also included relevant preclinical and clinical data from animal studies.

Posthatch developmental changes in the ovary with emphasis on follicular development and atresia in the native chicken (Uttara fowl) of Uttarakhand, India.

This experiment was designed to investigate the postnatal development of the ovary in the Uttara fowl chicken and was conducted on 54 apparently healthy female birds divided into different age groups, namely Day 1 and Weeks 1, 4, 8, 12, 16, 20, 24, 28 with six birds each. During postnatal development, the left ovary gradually increased in size and complexity. The segmentation of the ovary started by 4 weeks, follicular eruption by 8 weeks, small liquor follicles (1-5 mm) appeared by 16 weeks, pre-hierarchical follicles by 20 weeks and hierarchical follicles by 24 weeks of age. The cortex was distinctly differentiated from the medulla in the early stage of ovarian development. However, the division between cortex and medulla was gradually obscured with age (transitional stage) and distinction was completely lost in the mature ovary. The different stages of follicular development in the chicken ovary were classified as primordial, primary, growing at Stage I, II and III stromal follicles besides pre-hierarchical and hierarchical surface follicles. The primordial and primary follicles showed cytoplasmic sudanophilic substances, especially in the Balbiani's yolk body, indicating the presence of lipids (Sudan Black B) with no activity for neutral polysaccharides (periodic acid Schiff method). It was observed that apoptotic changes may affect any stage of developing follicle resulting in arrested growth and atrophy. An early form of follicular atresia was the fate of the growth-arrested primordial and primary follicles, whereas the glandular form of atresia was commonly observed in growing follicles arrested at Stages I and II. The scanning electron micrographs unveiled the follicles as hollow oval structures with a follicular lumen lined by the perivitelline membrane (glycoprotein membrane) having lacunae giving a honeycomb-like appearance.

Pecten oculi of kestrel (Falco tinnunculus rupicolaeformes) and little owl (Athene noctua glaux): Scanning electron microscopy and histology with unique insights into SEM-EDX elemental analysis.

There is scanty data about the comparative morphological features between the pecten oculi of two carnivorous birds with different visual active clock hours: the diurnal common kestrel and the nocturnal little owl. This study illustrated the comparative gross, scanning electron microscopy, and histological characteristics between pecten oculi of kestrel and little owl. This study first attempts to describe the scanning electron microscopy-energy dispersive x-ray (SEM-EDX) elemental analysis at the parts (apex, middle, and base) of the pecten oculi of these two birds. The present study results observed the same position, origin, directions, parts, convoluted outer pleat surfaces, and SEM-EDX elemental analysis, but there were some minor variations due to the different visual active clock hours. These minor variations were summarized in the following points: pleat number (21-23 in the kestrel and 10-11 in the owl), shape (fan rhomboid in the kestrel and accordion in the owl), inter-pleat spaces (wider in the kestrel than in the owl), pigmentations (highly black pigmented in the owl than in the kestrel), hyalocyte cell aggregations (highest in the middle and dwindling at the apex and base in the kestrel, while highest in the middle and base and dwindling at the apex in the owl), and SEM-EDX elemental analysis percentage. SEM/EDX elemental analysis confirmed the presence of oxygen (the highest one), carbon, nitrogen (the second one), nitrogen (the third one), and aluminum (the lowest one) in varying percentages within the pecten oculi; these findings contribute to our understanding of its structural, adaptations with different visual active clock hours, and functional characteristics. RESEARCH HIGHLIGHTS: This study compared the pecten oculi of two carnivorous birds with different visual active clock hours: the diurnal common kestrel and the nocturnal little owl. Anatomically, the characteristic features were similar in both the birds, but some minor variations were observed adapted to their visual active clock hours. The pecten oculi of both birds were analyzed using SEM-EDX for elemental analysis, and it revealed that oxygen was the highest elemental concentration, followed by carbon and nitrogen. Aluminum concentrations were small as per SEM-EDX analysis. The study suggested that the pecten oculi of these birds are related to their active visual clocks and adaptive nutritional mechanisms.

Tongue of the Egyptian Endemic Bridled Skink (Heremites vittatus; Olivier, 1804): Gross, Electron Microscopy, Histochemistry, and Immunohistochemical Analysis.

The present study used light and scanning electron microscopy to describe the integrative morphological description of the tongue and laryngeal mound of Heremites vittatus, an endemic lizard of Saharan Africa. Additionally, ultrastructure, histology, histochemistry, and immunohistochemical approaches were used to characterize the lingual apparatus adaptations. In the present study, Heremites vittatus consisted of a complex lingual papillary system in which the ventral apical surface of the foretongue comprised conical papillae. The dorsal surface consisted of different filiform papillary (papillae filiformes) types: the anterior section had two types (bifid and pointed), and the posterior section had four types (triangular, trifid, quadrifid, and pentafid) papillae. The dorsal midtongue surface exhibits scale-like, serrated filiform papillae with anterior gland openings. The hindtongue consisted of two overlapping filiform papillae: scale-like, board-serrated papillae on the median portion and finger-like papillae on the wings. The dorsal surface of the laryngeal mound had 18 longitudinal folds with glandular openings. Histologically, the foretongue was covered by a slightly keratinized layer that was absent in the mid- and hindtongue. The lingual glands were absent from the foretongue but present in the interpapillary space in the mid- and hindtongues. We observed a few rounded taste buds in the conical papilla epithelium. Histochemical analysis revealed strong glandular Alcian Blue (AB)-positive and Periodic Acid-Schiff (PAS)-positive reactions. Immunohistochemistry showed strong cytokeratin immunopositivity in all parts of the tongue. In conclusion, the obtained data about the lingual characterizations have been consistent with the active foraging behavior of the species and its environmental conditions.

Osteometric and topographic measurement of the skull and mandible of Siirt colored Mohair goat with three-dimensional (3D) modeling technique.

Siirt-colored Mohair goat is one of the breeds that contributed significantly to the existence of Mohair goats reared in Turkey. Morphological and morphometric characteristics of the Siirt-colored Mohair goat remained vague owing to a lack of studies. Recent advances in high-tech imaging have replaced conventional two-dimensional anatomical structures with three-dimensional (3D) models. In our study, morphometric features were determined by 3D modeling from computed tomography images obtained from the skull and mandibular bones of Siirt-colored Mohair goats. For this purpose, the skulls and mandibular bones of 20 Siirt-colored Mohair goats (10 females and 10 males) were used. The images were reconstructed with the help of a particular software program. The craniometric data were analyzed in terms of sexual dimorphism, and statistically significant difference was found in the A5, A18, and A31 measurement parameters (P<0.05) and Skull index (P<0.01) parameters. In the mandible measurements, there was a statistically significant difference between the sexes in C5, C10 measurement points (P<0.05), C2, C8, C12, C18, C21 measurement points (P<0.001) and surface area parameter (P<0.01). The morphometric data obtained is a resource in the fields of zoo archaeology, anatomy, forensics, anesthesia, surgery, and treatment.

Nanovaccines: A game changing approach in the fight against infectious diseases.

The field of nanotechnology has revolutionised global attempts to prevent, treat, and eradicate infectious diseases in the foreseen future. Nanovaccines have proven to be a valuable pawn in this novel technology. Nanovaccines are made up of nanoparticles that are associated with or prepared with components that can stimulate the host's immune system. In addition to their delivery capabilities, the nanocarriers have been demonstrated to possess intrinsic adjuvant properties, working as immune cell stimulators. Thus, nanovaccines have the potential to promote rapid as well as long-lasting humoral and cellular immunity. The nanovaccines have several possible benefits, including site-specific antigen delivery, increased antigen bioavailability, and a diminished adverse effect profile. To avail these benefits, several nanoparticle-based vaccines are being developed, including virus-like particles, liposomes, polymeric nanoparticles, nanogels, lipid nanoparticles, emulsion vaccines, exomes, and inorganic nanoparticles. Inspired by their distinctive properties, researchers are working on the development of nanovaccines for a variety of applications, such as cancer immunotherapy and infectious diseases. Although a few challenges still need to be overcome, such as modulation of the nanoparticle pharmacokinetics to avoid rapid elimination from the bloodstream by the reticuloendothelial system, The future prospects of this technology are also assuring, with multiple options such as personalised vaccines, needle-free formulations, and combination nanovaccines with several promising candidates.

ChatGPT for Veterinary Anatomy Education: An Overview of the Prospects and Drawbacks / ChatGPT para la Educación en Anatomía Veterinaria: Una Descripción General de las Perspectivas y los Inconvenientes

SUMMARY: The integration of artificial intelligence in veterinary medical education has the potential to revolutionize the way students learn veterinary anatomy. ChatGPT, launched by OpenAI in November 2022, has been the fastest-growing artificial intelligence (AI) application as a learning tool in the past few months. ChatGPT is a virtual assistant that provides students with detailed and relevant anatomical knowledge based on internet sources. Apart from the advantages of ChatGPT in veterinary anatomy education, the challenges and limitations must also be considered, as highlighted in this letter. Further research and evaluation are necessary to ensure the optimal integration of the ChatGPT tool into veterinary anatomy education. ChatGPT can provide students with valuable insights and educational support but cannot replace hands-on experiences; dissection-based laboratory sessions remain essential for developing practical skills and spatial understanding in veterinary anatomy education.

La integración de la inteligencia artificial en la educación médica veterinaria tiene el potencial de revolucionar la forma en que los estudiantes aprenden anatomía veterinaria. ChatGPT, lanzado por OpenAI en noviembre de 2022, ha sido la aplicación de inteligencia artificial (IA) de más rápido crecimiento como herramienta de aprendizaje en los últimos meses. ChatGPT es un asistente virtual que brinda a los estudiantes conocimientos anatómicos detallados y relevantes basados en fuentes de Internet. Además de las ventajas de ChatGPT en la educación en anatomía veterinaria, también se deben considerar los desafíos y las limitaciones, como se destaca en esta carta. Se necesitan más investigaciones y evaluaciones para garantizar la integración óptima de la herramienta ChatGPT en la educación en anatomía veterinaria. ChatGPT puede proporcionar a los estudiantes información valiosa y apoyo educativo, pero no puede reemplazar las experiencias prácticas. Las sesiones de laboratorio basadas en disecciones siguen siendo esenciales para desarrollar habilidades prácticas y comprensión espacial en la educación de la anatomía veterinaria.

Sublingual floor of Rahmani sheep (Ovis aries): A scanning electron microscopy and histomorphology analysis.

The current study investigated the sublingual floor of Rahmani sheep (Ovis aries). Samples from nine healthy adult sheep have been analysed using morphometrical, scanning electron microscopic (SEM), and histological examination. The sublingual floor of the sheep was formed of a rostral prefrenular part (spatula-shaped) and two lateral sublingual recesses. Right and left sublingual caruncles were asymmetrical in length and each one had a serrated border, divided into three papillae. At higher magnifications, the three papillae of the sublingual caruncle's edge were variable in shape, with other two papillae were demonstrated underneath the caruncle. Each sublingual recess had a sublingual fold (plica sublingualis) extended from the level of frenulum linguae till the glossopalatine arch, bordered with caudolaterally directed papillae till the level of first lower molar cheek teeth. The sublingual salivary glands were mixed (seromucoid) type. The glandular lobule consisted of mucous acini and sporadic serous acini, as well as the presence of some mucous acini capped by acidophilic cells, serous demilunes. The mucous acinus was formed of pyramidal mucous cells with pale stained cytoplasm and basally located nuclei. Two ducts; major sublingual and submandibular ducts were observed within the submucosa of the sublingual caruncle. The sublingual ductal pseudostratified columnar epithelium was interspersed by more goblet cells than that of the submandibular duct. Collagen fibres around the glandular lobules, intralobular and interlobular excretory ducts as well as the connective core of the papillae were noticed. By SEM, two epithelium-lined pits on the surface of the sublingual floor mucosa caudal to the central incisive teeth were termed orobasal organ. As the first study to give a comprehensive analysis of the sheeps' sublingual floor, this one serves as a baseline for future clinical studies.

Antioxidant supplementation during treatment of outpatient dogs with parvovirus enteritis ameliorates oxidative stress and attenuates intestinal injury: A randomized controlled trial.

A prospective randomized controlled clinical study was conducted to determine whether antioxidant supplementation as an adjunct therapy alters hemogram, oxidative stress, serum intestinal fatty acid binding protein-2 (IFABP-2) level, fecal viral load, clinical score (CS) and survivability in outpatient canine parvovirus enteritis (CPVE) dogs. The dogs with CPVE were randomized to one of the five treatment groups: supportive treatment (ST) alone, ST with N-acetylcysteine (ST+NAC), resveratrol (ST+RES), coenzyme Q10 (ST+CoQ10) or ascorbic acid (ST+AA). The primary outcome measures were reduction of CS and fecal HA titre, and enhancement of survivability. Secondary outcome measures were reduction of oxidative stress indices and IFABP-2 level from day 0 to day 7. The mean CS and HA titre were significantly (P < 0.05) decreased from day 0 to 7 in ST and all antioxidant groups. The supplementations of NAC, RES and AA along with ST markedly (P < 0.05) reduced the concentrations of malondialdehyde, nitric oxide and IFABP-2 on day 7 as compared to ST alone. Additionally, NAC and RES supplementations markedly (P < 0.05) improved the total leukocyte count and neutrophil count in CPVE-affected dogs. NAC and RES could serve as better antioxidants for the amelioration of oxidative stress in CPVE but, the antioxidants did not confer any additional benefits in reduction of CS, fecal HA tire, or survivability when compared with ST alone.

Morphologic appearance of peripheral blood cells from Zovawk pigs (Sus scrofa domesticus) visualized by transmission electron microscopy.

BACKGROUND: Ultrastructural information regarding the peripheral blood cells of local (Zovawk) pigs from Mizoram, India, is not available in the scientific literature. OBJECTIVES: The present study was designed to reveal the fine structural details of the blood cells from these local pigs using a transmission electron microscope (TEM). METHODS: Blood samples were collected from 12 healthy Zovawk pigs of either sex and processed according to a standard protocol. Processed blood samples were then sent to the All India Institute of Medical Sciences, New Delhi, for further processing and imaging under TEM. Different types of blood cells were viewed under TEM, and different characteristics of these cells were assessed. RESULTS: In the present study, erythrocytes are elongated, biconcave, and nucleated without cytoplasmic organelles. Neutrophils are round with 2-5 lobed nuclei surrounded by cytoplasm with an indistinct bilayered nuclear membrane. The cytoplasm is packed with membrane bound round, oval, and elongated cytoplasmic granules. Eosinophils are round to oval with 2-3 lobed nuclei with distinct nuclear membranes. Basophils are spherical and contained small, medium, and large electron-dense granules. Lymphocytes are small, medium, and large and contained all cellular components. Monocytes are irregularly spherical with slight nuclear indentations. The platelets are elongated, oval, or rounded, with a few pseudopods at the cell surface. CONCLUSIONS: From the present study, we can conclude that the ultrastructural morphology of blood cells from Zovawk pigs resembles those of other domestic animals. However, a few differences have been observed.

Preen gland of the laughing dove (Streptopelia senegalensis aegyptiaca): Light and electron microscopic analysis.

This work reviews the microscopic anatomy of the preen gland in laughing dove (Streptopelia senegalensis aegyptiaca) using light, scanning, and transmission electron microscopes. The gland possessed two large pea-shaped lobes. The glandular lobules of each lobe were huddled in elliptical, triangle, round shapes, connected with each other by strands of connective tissue septae. The lobule was composed of glandular follicles, each follicle folded and enclosed by a sheath of connective tissue connected with the neighboring ones by interfollicular septae. The gland's parenchyma was coated with a dense connective tissue capsule composed of collagen, elastic, and reticular fibers. The secretory tubules were divided into peripheral tubules and central tubules. The central ones were located close to the major cavity and lined with thin epithelium, whereas the peripheral tubules were adjacent to the capsule and characterized by taller epithelium. The central secretory tubules were lined with four cell layers: flattened basal, large-sized polyhedral intermediate, and secretory cell layers, as well as a degenerative cells layer that formed of small cells with pale cytoplasm and pyknotic nuclei. Variable sizes and shapes of Herbst corpuscles were detected alongside the papillary duct and near the glandular lobe. Transmission electron microscopy view revealed that the cytoplasm of the intermediate cells contained a dense population of mitochondria, while the secretory and degenerative cells contained fewer mitochondria. In conclusion, these structures will be beneficial for understanding the habitat differences of laughing doves. RESEARCH HIGHLIGHTS: Grossly, the preen gland (PG) of the laughing dove formed of two large pea-shaped lobes. The glandular lobule was composed of glandular follicles, each follicle was folded and enclosed by a sheath of connective tissue connected with the neighboring ones by interfollicular septae. The central secretory tubules were lined with four cell layers: basal, intermediate, secretory, and degenerative cell layers. Variable sizes and shapes of Herbst corpuscles were detected alongside the papillary duct and near the glandular lobe of the PG. In transmission electron microscopic analysis, the cytoplasm of the intermediate cells contained a dense population of mitochondria, while the secretory and degenerative cells contained fewer mitochondria.