Histological Investigation Research Articles

Evaluating the efficacy and mechanisms of Hua-Zhuo-Ning-Fu-Decoction on psoriasis using integrated bioinformatics analysis and metabolomics

Ethnopharmacological relevanceHua Zhuo Ning Fu Decoction (HZD) is an empirical prescription from traditional Chinese medicine that shows excellent clinical results for psoriasis patients. Uncertainty lingered over HZD's potential anti-psoriasis mechanisms. Aim of the studyThe study's objective is to investigate the pharmacological processes and therapeutic effects of HZD on psoriasis. Materials and methodsIn the initial phase of the study, an investigation was conducted to assess the effects of HZD on psoriasis-afflicted mice using an imiquimod (IMQ)-induced murine model. The experimental mice were randomly allocated to different groups, including the IMQ-induced model group, the control group, the HZD therapy groups with varying dosage levels (low, medium, and high), and Dexamethasone (DEX, the positive control medicine) group. Bioinformatics analysis and molecular docking were subsequently employed to identify the primary components and molecular targets associated with the therapeutic action of HZD in the context of psoriasis. Additionally, to find the impacts on metabolite regulation, plasma metabolomics based on ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) was used. It's interesting to note that the combined mechanisms from metabolomics were examined in tandem with the targets. In vivo tests were the last step in validating the potential mechanism. Throughout the trial, the following data were recorded: body weight, psoriasis area and severity index (PASI). The molecular targets connected to HZD's anti-psoriasis activities were revealed using histological examination, western blot (WB), and ELISA investigation. ResultsIn mice induced with IMQ, HZD shown good anti-psoriasis effects in terms of PASI score and epidermal acanthosis. 95 HZD targets and 77 bioactive chemicals connected to psoriasis were found by bioinformatics research; of these, 7 key targets (EPHX2, PLA2G2A, TBXAS1, MAOA, ALDH1A3, ADH1A, and ADH1B) were linked to the mechanisms of HZD, the combination degree of which was finally expressed by the score of docking. In addition, HZD regulated nine metabolites. In line with this, HZD modified three metabolic pathways. Additionally, a combined examination of 7 key targets and 9 metabolites suggested that the metabolism of arachidonic acid might be the key metabolic route, which was identified by ELISA analysis. The in vivo investigation shown that HZD could control cytokines associated to inflammation (IL-10, TGF-β, IL-17A, and IL-23), as well as important antioxidant system markers (ROS, GSH, and MDA). Moreover, HZD controlled iron levels and the expression of ferroptosis-related proteins (ACSL4 and GPX4), suggesting that ferroptosis played a crucial role in this process. ConclusionsOur findings demonstrated the whole mechanism and anti-psoriasis effectiveness of HZD, which will promote its clinical application and aid in the investigation of new bioactive components of HZD against psoriasis.

An Unusual Case Report of Unicystic Ameloblastoma of the Mandible

ABSTRACT There are plenty of benign lesions that can result in swelling of the mandible, and these can be classified as odontogenic and non-odontogenic lesions. Among the categories of odontogenic lesion, ameloblastoma is the most occurring lesion that takes origin from the epithelial cellular elements and dental tissues in their different stages of development. Ameloblastoma is the most serious odontogenic neoplasm due to its prevalence and clinical characteristics. Ameloblastoma is a broad class which encompasses 80% of solid multicystic type of ameloblastoma with unicystic ameloblastoma (UA) variant included as vital clinicopathological form claiming the rest 20% along with peripheral ameloblastoma variant. UA refers to cystic lesions that seem like jaw cysts clinically, radiographically, or grossly but are lined by typical ameloblastomatous epithelium, with or without luminal and/or mural tumor development, on histologic investigation. Around 5–15% of all ameloblastic lesions do not have a propensity to metastasis, and this is UA. Unicystic mural form, although slow growing overall, is very invasive locally and has a high recurrence rate. As UA tumors show very close features with dentigerous cyst, a very sharp differential diagnosis protocol need to be executed to exclude the other unicystic odontogenic lesions considering the clinical, radiological, and biological characteristics along with proper follow-up and seeing any recurrence of the lesion taking place. Here, we report the case of a twenty-one year male patient with UA of the mandible and review of the literature.

Application of Intracerebral Aquaporin-4 Expression in Differential Diagnosis of Drowning: an Immunohistochemical Study Related to Timing of Autopsy.

The role of forensic pathologists is pivotal in definitively diagnosing drowning cases. Further differentiation becomes essential for distinguishing between freshwater drowning (FWD) and saltwater drowning (SWD). Aquaporins are small integral membrane proteins that serve as major water transport pathways in various cell types. AQP4 appears to be involved in mechanisms related to cerebral volume regulation. Our study aims to examine the expression of AQP4 in the brain as a potential marker for differentiating between FWD and SWD relating to autopsy-performing timing. A total of 23 cases were classified into three groups: FWD, SWD, and controls. All samples were classified upon autopsy-performing timing into two subgroups: within and after 72 hours of death. The samples were then processed for histological and immunohistochemical investigations. For autopsies performed within 72 hours of death, we found a significantly higher value of AQP4-positive astrocytes in cases of FWD compared to SWD and control groups. We also found a significantly lower AQP4 expression in SWD cases compared to the control group. For autopsies conducted after 72 hours, the immunohistochemical staining does not reveal the peripheral terminations of astrocytes, which appear blurred and only recognizable as halos. In conclusion, the data aligns with existing literature about autopsies performed within 72 hours. However, in autopsies conducted after 72 hours, uncertain and even opposed results are observed. The difference can be ascribed to the post-mortem transformative processes that take place upon the cessation of vital functions.

Особенности выявления костных включений в мясных изделиях микроструктурными методами.

Mechanically separated poultry meat (MSPM), being a valuable food raw material, is widely used in the meat industry. Therefore, its detection in finished products is an urgent task in controlling their quality and composition. The article presents the results of the application of histochemical staining methods to identify bone fragments as the main indicator of the presence of MSPM in meat products. To ensure accuracy and first of all, comparability of results of histological investigations, it is important that all laboratories adhere to the principles of the microscopic laboratory practice and use the uniform methodology for preparation of histological specimens and interpretation of obtained results.

In vivo anticancer study of sodium 2‐[(4‐oxidobenzylidene)amino]‐6H‐1,3,4‐thiadiazine‐5‐olate against Ehrlich ascites carcinoma via targeting PI3K/mTOR pathway

Abstract1,3,4‐Thiadiazine‐based components have gained the attention as they afford significant intervention for cancer therapy. We aimed at assessing the in vivo antitumor activity of sodium 2‐[(4‐oxidobenzylidene) amino]‐6H‐1,3,4‐thiadiazine‐5‐c (SOTA) versus Ehrlich ascites carcinoma (EAC) cells in female mice's peritoneal cavity. An advantageous interaction of the test compound with the receptors p53 (2J1X), Caspase‐3 (3KJF), mTOR (3QAR), and PI3K (4JPS) was revealed by the docking study. The in vivo study of the test compound against EAC, equated with the EAC control mice, EAC bearing mice that received the test compound by i.p. injection proven a substantial decrease in the viability of tumor cell by 70%. Treatment with the tested compound increased total antioxidants capacity and decreasing malondialdehyde levels. Also, the apoptotic activity of the test compound was confirmed by up regulation of caspase‐3 and p53 and downregulation of PI3K and mTOR. Through the liver, kidney, and heart function assessments, the examined component has no adverse effects on either of the two organs, which were supported by a histological study. Significant positive and negative associations among variables were discovered, according to our research. Through the induction of apoptosis and antioxidant effects, the test compound show potential anticancer activity against EAC cells. The treatment with SOTA eliminated most of the pathological abnormalities brought on by EAC cells in mice, according to the results of tests on the liver, kidney, and heart as well as the histological investigation. Taking together, the tested compound SOTA is promising chemotherapeutic agent as supported by targeting PI3k/mTOR pathway.

Hyaluronic Acid Treatment of Post-Extraction Tooth Socket Healing in Subjects with Diabetes Mellitus Type 2: A Randomized Split-Mouth Controlled Study.

The present study aimed to investigate the effect of HA in improving post-extraction socket healing in subjects with diabetes mellitus (DM) type 2. DM patients requiring bilateral extraction of the homologous teeth were visited at the C.I.R. Dental School, University of Turin. After the extractions, one site was randomly assigned to the test (T) group (postoperative application of HA), while the other site was assigned to the control group (C, no treatment). Patients were then followed after 3, 7, 14, and 21 days. Primary outcomes were the healing index and socket closure. The Mann-Whitney test or the Student's t-test was used for nonparametric or parametric distributed variables. The chi-square test was used if the estimated data in any given cell were >5, otherwise the Fisher test was adopted. A p < 0.05 was considered statistically significant. In total, 36 patients (n = 36) were enrolled in this study for a total of 72 extractions (n = 72). Sockets treated with HA showed significantly (p < 0.05) better healing index values at day 7 (p = 0.01) and at day 14 (p = 0.02) and significantly (p < 0.05) better socket closure values at day 3 (p = 0.04), day 7 (p = 0.001) and day 14 (p = 0.001) compared to the C group. Based on the clinical results, HA seems to be promising in improving the timing and the quality of post-extractive wound healing in DM patients. Further clinical research, as well as histological investigations, are required to confirm the results.

Exploring Corneal Neovascularization: An Integrated Approach Using Transcriptomics and Proteomics in an Alkali Burn Mouse Model.

Corneal neovascularization (CNV) impairs corneal transparency and visual acuity. The study aims to deepen our understanding of the molecules involved in CNV induced by alkali burns, facilitate a better grasp of CNV mechanisms, and uncover potential therapeutic targets. Eighty-four mice were selected for establishing CNV models via alkali burns. On days 3, 7, and 14 after the burns, corneal observations and histological investigations were conducted. An integrated analysis of RNA sequencing (RNA-seq)-based transcriptomics and label-free quantitative proteomics was performed in both normal and burned corneas. Bioinformatics approaches, encompassing Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, were applied to discern differentially expressed genes (DEGs) and crucial signaling pathways. Four potentially CNV-related genes were validated using quantitative real-time PCR (qRT-PCR) and Western blot. Significant CNV was observed on the seventh day. Forty-one genes were differentially expressed in neovascularized corneas, with 15 upregulated and 26 downregulated at both mRNA and protein levels. Bioinformatics analysis revealed that these DEGs participated in diverse biological processes, encompassing retinol and retinoic acid metabolism, neutrophil chemotaxis, and actin filament assembly, along with significant enrichment pathways like cytochrome P450, tyrosine, and phenylalanine metabolism. The upregulation of lymphocyte cytosolic protein 1 (LCP1) and cysteine and glycine-rich protein 2 (CSRP2) genes and the downregulation of transglutaminase 2 (TGM2) and transforming growth factor-beta-induced (TGFBI) genes were confirmed. We analyzed gene expression differences in mouse corneas 7 days after alkali burns, finding 41 genes with altered expression. The exact role of these genes in CNV is not fully understood, but exploring angiogenesis-related molecules offers potential for CNV treatment or prevention.

Prenatal double-hit with aluminium and cadmium mediate testicular atrophy and hypothalamic hypoplasia: the role of oxido-nitrergic stress and endocrine perturbations.

There is limited experimental evidence on the biochemical consequences of aluminium (Al) and cadmium (Cd) co-exposures during pregnancy and postnatal life.This study investigated the impacts of perinatal Al chloride (AlCl3) and Cd chloride (CdCl2) co-exposures on neuroendocrine functions in mice offspring during postnatal life. The study comprised of four pregnant experimental groups. Group 1 received AlCl3 (10mg/kg), group 2 were administered CdCl2 (1.5mg/kg), while group 3 received both AlCl3 (10mg/kg) and CdCl2 (1.5mg/kg) (AlCl3+CdCl2), and group 4 received saline (10mL/kg) only and served as control group. All experimental animals were chemically exposed once daily from gestation days 7-20. Upon delivery, male pups were regrouped based on maternal chemical exposure on postnatal day 21 (PND 21) and allowed to grow to adulthood until PND 78, after which they were sacrificed for assessment of neuroendocrine markers and histological investigations. There was no statistical significance (p > 0.05) on follicle stimulating hormone, testosterone, estrogen and progesterone, thyroid stimulating hormone, thyroxine (T4) in all treatment groups relative to controls|. However, AlCl3 and AlCl3-CdCl2 significantly (p < 0.05) reduced triiodothyronine (T3) levels, with a profound increase in T3:T4 ratio by AlCl3, and AlCl3+CdCl2 compared to control. Furthermore, pups from pregnant mice treated with CdCl2 and AlCl3+CdCl2 demonstrated increased testicular malondialdehyde concentration with increased catalase activity relative to controls, suggesting oxidative imbalance. In addition, AlCl3, CdCl2, and AlCl3+CdCl2 exposures induced testicular and hypothalamic architectural disruption compared to controls, with marked architectural derangement in the AlCl3+CdCl2 group. Our findings suggest that prenatal co-exposures to Alcl3 and CdCl2 induce testicular and hypothalamic alterations in offspring via a testicular oxidative stress and thyrotoxicosis-dependent mechanisms.

The impact of chronic Trimethylamine N-oxide administration on liver oxidative stress, inflammation, and fibrosis

TMAO, a gut microbiota derived byproduct, has been associated with various cardiometabolic diseases by promoting oxidative stress and inflammation. The liver is the main organ for TMAO production and chronic exposure to high doses of TMAO could alter its function. In this study, we evaluated the effect of chronic exposure of high TMAO doses on liver oxidative stress, inflammation, and fibrosis.TMAO was administered daily via gastric gavage to laboratory rats for 3 months. Blood was drawn for the quantification of TMAO, and liver tissues were harvested for the assessment of oxidative stress (MDA, GSH, GSSG, GPx, CAT, and 8-oxo-dG) and inflammation by quantification of IL-1α, TNF-α, IL-10, TGF-β, NOS and COX-2 expression. The evaluation of fibrosis was made by Western blot analysis of α-SMA and Collagen-3 protein expression. Histological investigation and immunohistochemical staining of iNOS were performed in order to assess the liver damage. After 3 months of TMAO exposure, TMAO serum levels enhanced in parallel with increases in MDA and GSSG levels in liver tissue and lower values of GSH and GSH/GSSG ratio as well as a decrease in GPx and CAT activities. Inflammation was also highlighted, with enhanced iNOS, COX-2, and IL-10 expression, without structural changes and without induction of liver fibrosis.

Anti-Alzheimer potential of Solanum lycopersicum seeds: in vitro, in vivo, metabolomic, and computational investigations

Abstract Background Solanum lycopersicum Linn. (Tomato, Family Solanaceae) is one of the fruits that are most consumed worldwide. The current research intends to emphasise the possibility of positive and therapeutic effects of S. lycopersicum seed extract (SLSE) on Alzheimer's disease's neurodegeneration effects being reversed in a study utilising rats exposed to aluminium chloride. Investigations were done on the cholinesterase and antioxidant in vitro activity of SLSE. Rats with Alzheimer's disease were given SLSE, and donepezil (500, and 10 mg/kg.b.wt., daily for six weeks, respectively) to test SLSE biological activity. Beam-balance and T-maze tests, as well as serum levels of AChE, norepinephrine, dopamine, serotonin, IL-6, glycated end product, BDNF, MDA, TAC, and GSH were assessed, accompanied with histological investigation. To impact the effectiveness of this extract, bioinformatics study was validated. Results Crude SLSE showed in vitro DPPH scavenging and AChE inhibition activities, indicating the extract might have anti-Alzheimer potential, which was validated using an aluminium-intoxicated rat model, in vivo. In Alzheimer's rats, in vivo studies showed considerable improvements, as seen by improved beam balance, and T-maze tests and decreased serum levels of AChE, norepinephrine, dopamine, serotonin, IL-6, glycated end product, BDNF, and MDA, with increasing in TAC, and GSH levels. Brain tissue histological tests revealed a largely typical pattern of collagen fibre distribution. LC–HRESIMS metabolomic profiling of crude SLSE identified 33 compounds. Furthermore, the bioinformatics study discovered 378 targets related to the major identified compounds, of which only 133 were related to Alzheimer's and memory disorders, with APP, AChE, and PSEN2 targets which were marked as the top genes. Gene enrichment analysis identified the arachidonic acid metabolism and PPAR signalling pathway as the biological pathways enriched by all the gene sets under investigation. Conclusion As a result, the study findings are expected to pave the way for the creation of dietary supplements for Alzheimer's disease management. Graphic abstract

Polyphenol-Standardized Aphanamixis polystachya Leaf Extract Ameliorates Diabetes, Oxidative Stress, Inflammation, and Fibrosis in Streptozotocin-Induced Diabetic Rats

Background. Diabetes is a rising disorder that affects millions of people annually. It also creates more complications, such as neuropathy, oxidative stress, and hepatic and kidney impairment. Aphanamixis polystachya plant, which possesses multiple medicinal values, is used in this study to explore its potential in treating diabetes. Methods. A single dose (65 mg/kg) of intraperitoneal streptozotocin injection was utilized to mediate diabetes in Sprague-Dawley rats. Diabetic animals were treated orally with 250 or 500 mg/kg of standardized leaves’ extract of A. polystachya (AP) for 28 days to evaluate the antidiabetic and organ-protective effects of the plant. Different biochemical and histological markers are measured according to the established protocol. Results. Our results demonstrated a significant decrease in blood glucose (p &lt;0.001) and HbA1c (p &lt;0.05) levels in the diabetic animal after administering AP (250 and 500 mg/kg doses) compared to the control groups. AP can also regularize lipids, glycogen, alanine aminotransferase, and aspartate aminotransferase. Furthermore, serum urea nitrogen and creatinine decreased after treatment with AP in diabetic rats. AP also reduced oxidative stress markers and showed a substantial elevation in antioxidant enzymes in diabetic animals. Overall, AP at 500 mg/kg revealed comparable results against the standard antidiabetic drug glyburide. Besides, the histological investigation showed the AP’s efficacy in attenuating kidney tissue inflammation and damage. HPLC data elucidated that the AP leaf extract contains polyphenols with potential antioxidant, antidiabetic, and organ protective agents: gallic acid, rutin hydrate, and quercetin hydrate. Conclusion. Taken together, AP may be one of the potential sources of antidiabetic agents.

A Large Mandibular Anterior Swelling—A Diagnostic Dilemma: A Case Report

AbstractHybrid lesions are lesions that combine characteristics of many pathologies. The literature has shown a rare relationship between aneurysmal bone cysts, fibro-osseous lesions, and a central giant cell lesion. In this case, we describe an unusual hybrid lesion in the mandibular anterior area. As a result, the goal of this article is to highlight the significance of comprehensive clinical, radiographic, and histological investigation in addition to presenting a rare instance of hybrid lesion. This will lessen the likelihood of misdiagnosis in these situations and aid in prompt treatment planning of the lesion.

Oral cancer and salivary biomarkers in its early diagnosis: A review of literature

Oral cancer (OC), the sixth most common cancer, is one of the leading public health problems worldwide, which is mainly represented by oral squamous cell carcinoma. Smoking and alcohol consumption are considered risk factors for OC. The major hindrance regarding OC is delayed diagnosis at the advanced clinical stages (i.e., III or IV), which affects its morbidity and mortality rates with a minimum 5-year survival rate of 20%–25%. Recurrences and metastasis often occur in more than 80% of patients within the first 2 years, even after primary treatment. Although a histological investigation is available as the gold standard of OC diagnosis, the application of noninvasive methods, including liquid biopsy, has been the focus of recent studies. In cancer diagnosis and health surveillance, saliva as a noninvasive and cost-effective bio-fluid containing several biomarkers, including metabolites, circulating tumor and tissue-derived cells, cytokines, DNA, RNA, and exosomes, has promising potential. Therefore, the improvement of salivary biomarkers is an utmost need for early cancer detection, particularly in the oral cavity and oropharynx, which gained the interest of most researchers and clinicians. The current review aimed to discuss the emerging issues of the most recent data on salivary biomarkers with particular attention to the potential early detection markers in OC. In addition, we report our previous findings on salivary biomarkers and their clinical significance as potentials in diagnosis, prognosis, and monitoring of OC.

Hypoglycemic Assessment of Aqueous Leaf Extract of Moringa oleifera on Diabetic Wistar Rats.

Background: Moringa oleifera leaf is used for diabetes due to its pharmacologic effects. Patients with hyperglycemia experience beta cell destruction. However, no research on risk awareness has been done to ascertain its safety. The present study describes the antidiabetic effect of Moringa oleifera leaf, such as the protection of pancreatic beta cells and the induction of glycogen synthesis, before addressing the secondary effects of diabetes, such as hepatic and renal toxicity. Methods: Forty-five Wistar rats weighed 160 ± 10 g were divided into nine groups. All animal operations complied with the National Institute of Health (NIH) guidelines for the care and use of laboratory animals as approved by the Animal Ethical Committee, University of Jos. Group I was normal control and Group II was diabetic animals induced with alloxan. Insulin and extract doses of 200, 400, and 800 mg/kg were given to diabetic Groups III-VI. Normal animals in Groups VII-IX were given extract at doses of 200, 400, and 800 mg/kg for 28 days. Tissues were retrieved for biochemical and histological investigations using standard techniques. Results: There was decrease relative body weight of diabetic animals (95.50 ± 5.50) when compared to normal control (142.75 ± 20.08) with increased levels of urea (control 6.13 ± 0.523 and diabetes 29.23 ± 1.267) and creatinine (control 0.70 ± 0.057 and diabetes 2.13 ± 0.185). Histology of the liver and pancreas also points to organ damage due to hyperglycemia. However, oral administration of extract showed antidiabetic effect with protection of pancreatic beta cells and the induction of glycogen synthesis, no glycogen was deposited in the liver, addressing the secondary effects of diabetes, such as hepatic and renal toxicity. Further discovery revealed that extract elevated antioxidant enzyme expression. Conclusion: Leaf extract from Moringa oleifera reduces blood sugar and lessens the damage caused by hyperglycemia in the pancreas and liver.

Histological and Immunohistochemical Investigation of Smoking-Induced Changes in Human Gingival Tissue: A Focus on p16 and CD34 Expression

Histological and Immunohistochemical Investigation of Smoking-Induced Changes in Human Gingival Tissue: A Focus on p16 and CD34 Expression

In vitro and in vivo evaluation of genetically corrected RPE cells for autologous cell therapy of hereditary retinal dystrophies

Purpose: The generation of genetically corrected RPE cells from patient‐derived human induced pluripotent stem cells (hiPSCs) may provide a source for autologous therapeutically‐relevant therapy for this rare retinal dystrophy as well to create the human cellular disease models. Recently we generated a faithful RPE model from patient bearing mutation in MERTK gene and genetically corrected these cells using CRISPR/Cas9 system. For further application in humans is necessary in vivo validation for survival.Methods: The RPE cells from the genetically corrected patient‐derived hiPSC were characterized using immunocytological methods for the differentiation and polarization markers, electronic microscopy, RT‐PCR, TEER measurement, ELISA test, Western Blot and phagocytic assay. The implantations of RPE‐scaffolds were carried out into the minipig eyes. Due to the xenogeneic transplantation of human cells into minipig eyes, tacrolimus immunosuppression therapy was used. At the 2‐week follow‐up from transplantation, non‐invasive OCT and fundus camera examinations of the implanted RPE‐scaffolds were performed. Consequently, euthanasia, histological and immunohistochemical investigations of the implanted retina were performed.Results: All generated RPE cells exhibit typical features of RPE cells. We demonstrated the reestablishment of the expression of full‐length MERTK protein as well as the reversion of lost phagocyte function of hiPSC‐RPE in vitro, which represents the first example of its kind in this field. Histological and immunohistochemical investigation of the RPE implants and adjacent retina showed structurally healthy RPE‐scaffold implant and neuroretinal cells above the implant.Conclusions: The generated hiPSC‐derived RPE in clinical grade conditions display the typical characteristics of mature RPE cells re‐establishing the function of phagocytosis in genetically corrected patient‐derived RPE cells in vitro. Functional in vivo studies in minipigs confirmed the RPE survival as well as survival of adjacent photoreceptors and thus can be employed as a potential treatment for patients.

Human iPSC‐derived RPE cells implantation into minipig eye: Two weeks follow‐up study

Aims/Purpose: Retinal pigment epithelium (RPE) replacement is a very promising cell‐based therapy for retinal degenerative diseases including age‐related macular degeneration (AMD). RPE dysfunction is the main cause of secondary damage to the photoreceptors. We hereby present RPE implantation into the minipig eye with a 2‐week follow‐up.Methods: Cultured human iPSC‐derived RPEs (iRPE) on nanofibrous membranes, with properties very similar to Bruch's membrane, were characterized for expression of differentiation and polarization markers using immunohistochemistry. The implantations of iRPE‐scaffolds were carried out into the minipig eyes using 3‐port pars plana vitrectomy. Due to the xenogeneic transplantation of human cells into minipig eyes, tacrolimus immunosuppression therapy was used. At the 2‐week follow‐up from transplantation, non‐invasive OCT and fundus camera examinations of the implanted iRPE‐scaffolds were performed. Consequently, euthanasia, histological and immunohistochemical investigations of the implanted retina were performed.Results: OCT and fundus camera examinations revealed successfully implanted iRPE‐scaffolds into the subretinal space with a healthy neuroretina above the implant, and healed retinotomy, without immune cell reaction against the implant. Histological and immunohistochemical investigation of the iRPE implants and adjacent retina showed structurally healthy iRPE‐scaffold implant and neuroretinal cells above the implant.Conclusions: RPE cell therapy for replacement of diseased RPEs by ex vivo cultured Ipsc–RPEs on highly permeable nanofibrous membranes supports the survival of adjacent photoreceptors and thus positively affects the structural changes seen in retinal neurodegeneration.

Wound Healing and Acute Dermal Toxicity Studies of Ludwigia octovalvis (Jacq.) P. H. Raven (Onagraceae) in Sprague–Dawley Rats

Introduction. The aerial part of Ludwigia octovalvis has been used traditionally in some parts of Asia for the management of wounds owing to the presence of phytochemicals such as tannins, flavonoids, and triterpenoids among others. The incidence of wounds, their associated complications, and the cost of wound care are on the increase globally, therefore, the need to develop alternative wound care agents. The aim of this study was to scientifically investigate the wound healing potential of the ethanolic extract of L. octovalvis using the excision wound healing model in rats and also carry out an acute dermal toxicity investigation of the plant extract. Method. A 70% ethanol extract of L. octovalvis was prepared for the wound healing activity using the excision wound healing model in Sprague–Dawley rats. Aqueous creams (1, 3, and 10%) were prepared and topically applied to the wounds once daily according to the groups of animals. The wounds were assessed for rates of wound closure on days 3, 5, 7, 9, and 11. Re‐epithelialization periods were also determined. Sections of wound tissues obtained on day 13 were subjected to histological investigations. An acute dermal toxicity of the plant extract was investigated. Results. L. octovalvis treatment (1, 3, and 10%) exhibited a mean percentage wound contraction range of 85.36 ± 7.22–94.14 ± 2.23 on day 11. The extract exhibited re‐epithelialization periods of 17.3 ± 1.2, 19.8 ± 2.6, and 16.0 ± 1.7 days for the 1, 3, and 10% extract creams, respectively, whereas the cream‐only and 1% silver sulfadiazine treatments resulted in a re‐epithelialization period of greater than 28 days. Histopathological investigation revealed enhanced fibroblast infiltration and collagen deposition in the treatment groups. No adverse reaction was observed in the acute dermal toxicity study. Conclusions. Extract of L. octovalvis exhibited wound healing by enhancing wound contraction, re‐epithelialization, fibroblast infiltration, and collagen deposition at the wound site. The extract did not exhibit any toxic reaction in the acute dermal toxicity study.

Thymoquinone Nanoparticles (TQ-NPs) in Kidney Toxicity Induced by Ehrlich Ascites Carcinoma (EAC): An In Vivo Study.

Cisplatin (Cis) is potent chemotherapy used to treating already many different types of cancer; however, it is found to correlate with nephrotoxicity and other adverse health consequences. Thymoquinone (TQ) is an antioxidant and anti-inflammatory molecule that may defend against the consequences of different chemotherapies. Thymoquinone uses, although, are negatively impacted by its weak solubility and inadequate biological availability. This investigation examined the efficacy of a new nanoparticle (NP) absorbing TQ in an Ehrlich Ascites Carcinoma (EAC) mice model to address its low solubility, enhance its bioavailability, and protect against Cis-induced nephrotoxicity. Following 4 treatment groups were included in this research: (1) control, (2) EAC, (3) EAC + Cis + Thymoquinone nanoparticle (TQ-NP) treated, and (4) EAC + Cis-treated. The study revealed that TQ-NP was efficacious in avoiding Cis-induced kidney problems in EAC mice, as well as restoring kidney function and pathology. Thymoquinone nanoparticle considerably reduced Cis-induced oxidative damage in renal tissue by augmenting antioxidant levels. According to tumor weight and histological investigation results, TQ-NP did not impair Cis's anticancer efficacy. Thymoquinone nanoparticle might be used as a potential drug along with Cis anticancer therapy to reduce nephrotoxicity and other side effects while maintaining Cis anticancer properties.

Thiazolidine-2,4-dione hybrids as dual alpha-amylase and alpha-glucosidase inhibitors: design, synthesis, in vitro and in vivo anti-diabetic evaluation.

Twelve 3,5-disubstituted-thiazolidine-2,4-dione (TZD) hybrids were synthesized using solution phase chemistry. Continuing our previous work, nine O-modified ethyl vanillin (8a-i) derivatives were synthesized and reacted with the TZD core via Knoevenagel condensation under primary reaction conditions to obtain final derivatives 9a-i. Additionally, three isatin-TZD hybrids (11a-c) were synthesized. The intermediates and final derivatives were characterized using 1H and 13C NMR spectroscopy, and the observed chemical shifts agreed with the proposed structures. The in vitro alpha-amylase and alpha-glucosidase inhibitory evaluation of newly synthesized derivatives revealed compounds 9F and 9G as the best dual inhibitors, with IC50 values of 9.8 ± 0.047 μM for alpha-glucosidase (9F) and 5.15 ± 0.0017 μM for alpha-glucosidase (9G), 17.10 ± 0.015 μM for alpha-amylase (9F), and 9.2 ± 0.092 μM for alpha-amylase (9G). The docking analysis of synthesized compounds indicated that compounds have a higher binding affinity for alpha-glucosidase as compared to alpha-amylase, as seen from docking scores ranging from -1.202 to -5.467 (for alpha-amylase) and -4.373 to -7.300 (for alpha-glucosidase). Further, the molecules possess a high LD50 value, typically ranging from 1000 to 1600 mg kg-1 of body weight, and exhibit non-toxic properties. The in vitro cytotoxicity assay results on PANC-1 and INS-1 cells demonstrated that the compounds were devoid of significant toxicity against the tested cells. Compounds 9F and 9G showed high oral absorption, i.e., oral absorption >96%, and their molecular dynamics simulation yielded results closely aligned with the observed docking outcomes. Finally, compounds 9F and 9G were evaluated for in vivo antidiabetic assessment by the induction of diabetes in Wistar rats using streptozotocin. Molecule 9G has been identified as the most effective anti-diabetic molecule due to its ability to modulate several biochemical markers in blood plasma and tissue homogenates. The results were further confirmed by histology investigations conducted on isolated pancreas, liver, and kidney.