A Descriptive Study of Histopathological of Chronic Brain Ischemia of Rat Tissue

Worldwide, cerebrovascular accidents (stroke) are the second leading cause of death and the third leading cause of disability. However, not many the histopathological study of progression in chronic stroke has been published so far. This study gives the detail explanation of mechanism of recovery and might give the idea of new timeline when to set up the treatment to regenerate restoration of damaged cells. Fourteen male Wistar rats (15–20 weeks, weighing 250-400 g) were used in this study. Prior to 7 days of adaptation to the laboratory environment, rats were divided into four groups. Sham group (n=2), rats that sacrificied 4th week (n=2), 8th week (n=5), 12th week(n=5). 90 minutes temporary MCAO procedures were performed using the Indonesian modified technique. CD31 and Doublecortin markers were used to evaluate angiogenesis and neurogenesis. The results showed that ventricle size of ipsilateral brain was not so affected as in week 12th compared to 8th week. Gliosis as a response to damage to the central nervous system was more dense in week 12th as oppose to week 4th. Regarding angiogenesis and neurogenesis, there is significant improvement of angiogenesis and neurogenesis within weeks, however 4th week post MCAO shows prominent recovery. We summarized that rat’s brain shows spontanenous improvement in chronic phase of stroke ischemia and angiogenesis and neurogenesis still happends until week 12th.

Preparation of Affinity Purified Antibodies against ε-Glutaryl-Lysine Residues in Proteins for Investigation of Glutarylated Proteins in Animal Tissues

The glutarylation of lysine residues in proteins attracts attention as a possible mechanism of metabolic regulation, perturbed in pathologies. The visualization of protein glutarylation by antibodies specific to ε-glutaryl-lysine residues may be particularly useful to reveal pathogenic mutations in the relevant enzymes. We purified such antibodies from the rabbit antiserum, obtained after sequential immunization with two artificially glutarylated proteins, using affinity chromatography on ε-glutaryl-lysine-containing sorbents. Employing these anti(ε-glutaryl-lysine)-antibodies for the immunoblotting analysis of rat tissues and mitochondria has demonstrated the sample-specific patterns of protein glutarylation. The study of the protein glutarylation in rat tissue homogenates revealed a time-dependent fragmentation of glutarylated proteins in these preparations. The process may complicate the investigation of potential changes in the acylation level of specific protein bands when studying time-dependent effects of the acylation regulators. In the rat brain, the protein glutarylation, succinylation and acetylation patterns obtained upon the immunoblotting of the same sample with the corresponding antibodies are shown to differ. Specific combinations of molecular masses of major protein bands in the different acylation patterns confirm the selectivity of the anti(ε-glutaryl-lysine)-antibodies obtained in this work. Hence, our affinity-purified anti(ε-glutaryllysine)-antibodies provide an effective tool to characterize protein glutarylation, revealing its specific pattern, compared to acetylation and succinylation, in complex protein mixtures.

Desoxyrhapontigenin Modulates Immunity and Inflammation in an Ovalbumin-Induced Rat Model of Asthma

Asthma is a state of hyperresponsiveness of airways with associated inflammation and obstruction to air track. Present report evaluated the protective effect of desoxyrhapontigenin against ovalbumin-induced rat model of asthma. In asthmatic rats, there was an increase in serum IL-4, IL-5, bronchoalveolar lavage fluid IgE, IL-4, IL-13, IL-17, transforming growth factor β1, and leucocyte count. On the contrary, there was a decrease in bronchoalveolar lavage fluid IFN-γ that was reversed by desoxyrhapontigenin but not dexomethasone. Also, the mRNA expression for NF-kBp65 and vascular endothelial growth factor increased without any change in IκBα in the asthmatic rat tissue. There was a pronounced increase in the histopathological score in asthmatic rat tissue that were diminished by desoxyrhapontigenin or dexomethasone. In conclusion, study reveals that treatment with desoxyrhapontigen may protect the bronchial asthma by modulating the inflammation and immune response in the lung tissue.

HistoNet: A Deep Learning-Based Model of Normal Histology

We introduce HistoNet, a deep neural network trained on normal tissue. On 1690 slides with rat tissue samples from 6 preclinical toxicology studies, tissue regions were outlined and annotated by pathologists into 46 different tissue classes. From these annotated regions, we sampled small 224 × 224 pixels images (patches) at 6 different levels of magnification. Using 4 studies as training set and 2 studies as test set, we trained VGG-16, ResNet-50, and Inception-v3 networks separately at each magnification level. Among these model architectures, Inception-v3 and ResNet-50 outperformed VGG-16. Inception-v3 identified the tissue from query images, with an accuracy up to 83.4%. Most misclassifications occurred between histologically similar tissues. Investigation of the features learned by the model (embedding layer) using Uniform Manifold Approximation and Projection revealed not only coherent clusters associated with the individual tissues but also subclusters corresponding to histologically meaningful structures that had not been annotated or trained for. This suggests that the histological representation learned by HistoNet could be useful as the basis of other machine learning algorithms and data mining. Finally, we found that models trained on rat tissues can be used on non-human primate and minipig tissues with minimal retraining.

Variation of Passive Biomechanical Properties of the Small Intestine along Its Length: Microstructure-Based Characterization

Multiaxial testing of the small intestinal wall is critical for understanding its biomechanical properties and defining material models, but limited data and material models are available. The aim of the present study was to develop a microstructure-based material model for the small intestine and test whether there was a significant variation in the passive biomechanical properties along the length of the organ. Rat tissue was cut into eight segments that underwent inflation/extension testing, and their nonlinearly hyper-elastic and anisotropic response was characterized by a fiber-reinforced model. Extensive parametric analysis showed a non-significant contribution to the model of the isotropic matrix and circumferential-fiber family, leading also to severe over-parameterization. Such issues were not apparent with the reduced neo-Hookean and (axial and diagonal)-fiber family model, that provided equally accurate fitting results. Absence from the model of either the axial or diagonal-fiber families led to ill representations of the force- and pressure-diameter data, respectively. The primary direction of anisotropy, designated by the estimated orientation angle of diagonal-fiber families, was about 35° to the axial direction, corroborating prior microscopic observations of submucosal collagen-fiber orientation. The estimated model parameters varied across and within the duodenum, jejunum, and ileum, corroborating histologically assessed segmental differences in layer thicknesses.

Different Dietary N-3 Polyunsaturated Fatty Acid Formulations Distinctively Modify Tissue Fatty Acid and N-Acylethanolamine Profiles

We investigated the influence of different dietary formulation of n-3 polyunsaturated fatty acids (PUFA) on rat tissue fatty acid (FA) incorporation and consequent modulation of their bioactive metabolite N-acylethanolamines (NAE). For 10 weeks, rats were fed diets with 12% of fat from milk + 4% soybean oil and 4% of oils with different n-3 PUFA species: soybean oil as control, linseed oil rich in α-linolenic (ALA), Buglossoides arvensis oil rich in ALA and stearidonic acid (SDA), fish oil rich in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), Nannochloropsis microalga oil rich in EPA or Schizochytrium microalga oil rich in DHA. FA and NAE profiles were determined in plasma, liver, brain and adipose tissues. Different dietary n-3 PUFA distinctively influenced tissue FA profiles and consequently NAE tissue concentrations. Interestingly, in visceral adipose tissue the levels of N-arachidonoylethanolamide (AEA) and N-docosahexaenoylethanolamide (DHEA), NAE derived from arachidonic acid (AA) and DHA, respectively, significantly correlated with NAE in plasma, and circulating DHEA levels were also correlated with those in liver and brain. Circulating NAE derived from stearic acid, stearoylethanolamide (SEA), palmitic acid and palmitoylethanolamide (PEA) correlated with their liver concentrations. Our data indicate that dietary n-3 PUFA are not all the same in terms of altering tissue FA and NAE concentrations. In addition, correlation analyses suggest that NAE levels in plasma may reflect their concentration in specific tissues. Given the receptor-mediated tissue specific metabolic role of each NAE, a personalized formulation of dietary n-3 PUFA might potentially produce tailored metabolic effects in different pathophysiological conditions.

Prevention of Diabetic Complications by Walnut Leaf Extract via Changing Aldose Reductase Activity: An Experiment in Diabetic Rat Tissue

Background. Increased activity of aldose reductase (AR) is one of the mechanisms involved in the development of diabetic complications. Inhibiting AR can be a target to prevent diabetes complications. This study is aimed at evaluating the effect of cyclohexane (CH) and ethanol extracts (ET) of walnut leaves on AR activity in the lens and testis of diabetic rats. Methods. Fifty-six male rats classified into seven groups as control and treatment groups and treated for 30 days. The treatment groups were treated with different concentrations of ET and CH. The diabetic control (DC) group was exposed to streptozotocin. AR activity was measured in the lens and testis. The expression of AR in the testis was evaluated by the immunohistochemistry method. Results. Both extracts significantly reduced the AR activity (ng/mg of tissue protein) in the testis (0.034±0.004, 0.038±0.010, and 0.040±0.007 in the treatment groups vs. 0.075±0.007 in the DC group) and lens (1.66±0.09, 2.70±0.47, and 1.77±0.20 in the treatment groups vs. 6.29±0.48 in the DC group) of the treatment group compared to those of the DC group (P<0.05). AR expression in the testes of the treatment groups was decreased compared with that of the DC group (P<0.0001). Conclusion. Walnut leaf extracts can reduce the activity and localization of AR in the testes and its activity in the lens of diabetic rats.

Expression, Proliferation and Apoptosis of miR‑92b in Oral Squamous Cell Carcinoma

Background: Expression of miR‑92b in oral squamous cell carcinoma (OSCC) rat tissue and its effect on the OSCC CAL‑27 cells were investigated. Methods: The study was performed in Qingdao Stomatological Hospital, Qingdao, China on December 2018. Thirty Wistar rats were used to construct models of oral squamous cell carcinoma. CAL‑27 cells trascfected by Lipofectamine 2000 were divided into miR‑92b inhibitor, miR‑NC and blank groups. RT‑qPCR was used for the detection of the expression level of miR‑92b, and MTT and flow cytometry were carried out for the detection of the effect of miR‑92b on the proliferation and apoptosis of CAL‑27 cells, respectively. Results: The expression level of miR‑92b was significantly higher in tumor tissues than that in normal tissues (P<0.001). The miR‑92b inhibitor group had significantly lower proliferation ability but higher apoptosis rate of CAL‑27 cells than the miR‑NC and blank groups. After miR‑92b was downregulated by trans-fecting cells, the expression level of miR‑92b was significantly lower in the miR‑92b inhibitor group than that in the miR‑NC and blank groups. Conclusion: miR‑92b inhibitor can inhibit the proliferation of CAL‑27 cells and promote apoptosis, which provides certain references for clinical treatment. It is expected to be a potential target for treating OSCC.