Positive Immunoreactivity Research Articles

Histological and immunohistochemical study on the effect of zinc oxide nanoparticles on cerebellar cortex of adult male albino rats

Background: Zinc oxide nanoparticles (ZnONPs) are one of metal nanoparticles that have widespread use in many fields.Objective: To investigate the effect of ZnONPs on cerebellar cortex of rats through histological and immunohistochemical study.Materials and methods: Thirty adult male albino rats were divided into three groups; group I (control), Group II (ZnONP-1 treated group) which received orally 50 mg/kg of ZnONP for two months and Group III (ZnONP-1I treated group) which received orally 200 mg/kg of ZnONP for two months. Specimens of the cerebellar cortex were processed for histological and immunohistochemical study. Morphometric and statistical analysis were carried out.Results: Group II showed Purkinje cells were crowded in many layers surrounded by perineuronal vacuoles and had pyknotic nuclei. They had cytoplasmic vacuoles and perinuclear Golgi apparatus revealed fragmented dilated cisternae. Nearby Bergmann astrocyte cells had highly vacuolated cytoplasm and the nerve fibers were also affected and showed dysmyleination (disrupted myelin sheath). Immunohistochemical study of the same group showed Purkinje cell cytoplasm had positive immunoreactions for calretinin proteins. In group III, there was a wide spread of neuronal affection to the degree of loss of many of Purkinje cells leaving empty spaces. Ultrastructurly, their cytoplasm appeared with multiple variable sized and had dilated mitochondria with disrupted cristae. The Bergmann astrocytes revealed nuclei with disrupted nuclear envelope and nearly absence their cytoplasmic organelle and there was more affection to the nerve fibers in the form of vacuolated axoplasm and demyelination (areas of myelin loss) beside dysmyleination. Immunohistochemical study of group III showed Purkinje cells cytoplasm with negative immunoreactions for calretinin proteins.Conclusion: Intake of ZnONPs induced various adverse alterations in the histological and immunohistochemical structures of cerebellar cortex indicating the occurrence of neurotoxicity. These changes were exaggerated with increasing the dose of their intake.

Allyl-isothiocyanate ameliorates the pre-neoplastic changes induced by the fern Dryopteris nigropalaceae on experimental feeding in Guinea pigs

Enzootic bovine haematuria, caused by long-term ingestion of ferns, is a chronic disease of hill cattle characterized by neoplastic lesions in the urinary bladder. Objectives of this study were to investigate the toxicity potential of long-term feeding of the fern Dryopteris nigropalaceae and effect of allyl isothiocyanate (AITC) to ameliorate fern toxicity and the associated pathological changes. The LC-MS analysis of the fern showed presence of ptaquiloside (4.5 ± 1.0 μg/g) and pterosin B (39 ± 9.1 μg/g). Groups of animals were fed dried fern powder at the dose of 20% w/w in normal feed and treated with and without AITC at graded doses. Long term feeding of fern induced inflammatory and pre-neoplastic lesions in urinary bladder. The important lesions included cystitis, squamous metaplasia and high-grade dysplasia. Urothelium showed positive immunoreactions for nuclear expression of H-ras and p53. However, no mutation suggestive of neoplastic change was observed on partial mRNA sequences analyses of exon 2 of H-ras and 5 or 7&8 of p53 genes. Strikingly, AITC showed dose-dependent amelioration of pre-neoplastic changes in fern-fed animals. In conclusion, AITC is shown to limit pre-neoplastic changes caused by D. nigropalaceae feeding in guinea pigs.

LIPOPHAGY: a novel form of steroidogenic activity within the LEYDIG cell during the reproductive cycle of turtle

BackgroundSteroidogenesis is an indispensable process that is indirectly associated with spermatogenesis in the Leydig cell (LC) to utilize the lipid droplets (LDs) that are critical to maintaining normal testosterone synthesis. The regulation of LD mobilization, known as lipophagy, in the LC is still largely unknown.MethodIn the present study, the LC of the Chinese soft-shelled turtle was investigated to identify the steroidogenic activity and lipophagy during the annual reproductive cycle by light microscopy, immunohistochemistry (IHC), immunofluorescence (IF), and transmission electron microscopy (TEM).ResultsThe LC showed a dynamic steroidogenic function with strong activity of 3β-HSD, vimentin and tubular ER during hibernation by IHC and TEM. The tubulo-vesicular ER had a weak immunopositive reaction for 3β-HSD in the LC during reproductive phase, suggesting persistent steroidogenic activity. ORO staining and TEM demonstrated that a larger number of LDs had accumulated in the LC during hibernation than in the reproductive phase. These LDs existed in close association with mitochondria and lysosomes by being dynamically surrounded by intermediate filaments to facilitate LD utilization. Lysosomes were found directly attached to large LDs, forming an autophagic tube and engulfing LDs, suggesting that micro-lipophagy occurs during hibernation. Furthermore, the IHC of ATG7 (Autophagy Related Gene 7) and the IF of the LC3 (Microtubule-associated protein light chain 3), p62 (Sequestosome-1 (SQSTM1) and LAMP1(Lysosomal-associated membrane protein 1) results demonstrated strong expression, and further confirmation by TEM showed the existence of an autophagosome and an autolysosome and their fusion during the hibernation season.ConclusionIn conclusion, the present study provides clear evidence of LD consumption in the LC by lipophagy, lysosome and mitochondria during the hibernation period, which is a key aspect of steroidogenesis in the Chinese soft-shelled turtle.

The expression and localization of Toll-like receptors 2, 4, 5 and 9 in the epididymis and vas deferens of a adult tom cats

Toll-like receptors (TLRs) are important molecules, which provide protection against infections of the reproductive tract. This study demonstrates for the first time the expression and localization patterns of TLRs in the caput, corpus and cauda segments of the epididymal duct (ED) and the vas deferens (VD) of adult domestic cats using immunohistochemistry and western blotting. While immunoblot analyses revealed relatively similar protein levels for TLRs 2, 4, 5, and 9 in three segments of the ED, the protein levels of TLR2 and TLR4 in the VD were found to be significantly higher than those measured in the ED segments (P < 0.05). On the other hand, immunostaining showed that TLRs exhibited regional- and cell-specific localization patterns. TLR2 and TLR5 were immunolocalized to the nucleus and cytoplasm of the principal cells in all ducts. TLR4 was restricted to the stereocilia, and TLR9 was located in the cytoplasm of the principal cells. Narrow cells displayed positive immunoreactions for TLR4 and TLR5. The basal cells of the different ED segments were positive for all four TLRs. TLR2, TLR5 and TLR9 were detected in the cytoplasmic droplets of the spermatozoa. TLR4 and TLR9 were detected along the entire length of the sperm tail, whilst TLR2 and TLR5 were absent in the midpiece. TLR2 and TLR5 were also detected in the equatorial segment of the sperm head. These results suggest that TLR2, TLR4, TLR5 and TLR9 are important not only for the protection of the ED, VD and spermatozoa but also for the maturation and storage of spermatozoa in the ED and VD, respectively.

The possible protective effect of Gardenia Jasminoides extracts on the dentate gyrus changes in an Alzheimer-induced model in adult male albino rats: Histological and Immunohistochemical study

Introduction: Alzheimer's disease (AD) is the most common diagnosed form of senile dementias, a neurodegenerative disease resulting in cognitive impairment in the elderly. Geniposide, extracted of Gardenia Jasminoides, is widely used herb for brain diseases and reduce amyloid plaques deposition which is an intricate process in AD.Aim of the work: demonstrate the histological changes in the dentate gyrus of hippocampus in an Alzheimer-induced model and the possible neuroprotective effect of Gardenia Jasminoides extract.Materials and methods: Thirty six adult male albino rats(4-6 months) , weighing180-200g divided into 3 groups. Group I the control group, group II given aluminum chloride dissolved in distilled water, (300mg/kg/day) orally for one month. Group (III) given Geniposide in distilled water,at a dose of (100mg/kg/day) orally concomitant with aluminum chloride at the same dose and period of the previous group. Cognitive functions were assessed by the passive avoidance test. Temporal lobes processed for H&E , congo red stains. Immunohistochemical study for detection of (GFAP) and (INOS). Tau protein can be phosphorylated by glycogen synthase Kinase-3β(GSK-3 β). Western blot assay to investigate the total protein expression glycogen synthase Kinase-3β(GSK-3 β) . Comet assay to detect DNA damage.Results: Alzheimer-induced model group showed significantly impaired cognitive performance which was improved after treatment with Geniposide extract. Changes in the dentate gyrus apparent increase in immature neurons, disturbed granular cell layer, distortion of pyramidal cells that appeared with pyknotic nuclei. Positive immunoreaction for GFAP in astrocytes were also observed. The ultra structural study exhibited neurons with apoptotic features, heterochromatic nuclei and their cytoplasm contained dilated cisternae of rough endoplasmic reticulum. Gardenia extracts ameliorated histological alterations.Conclusion: Gardenia Jasminoides extracts significantly ameliorated the neuro- degenerative alterations induced in the Alzheimer model. Daily supplements of Gardenia Jasminoides extracts might be of benefit for the elderly people at risk of developing AD.

Sodium Butyrate Inhibits the Inflammation of Lipopolysaccharide-Induced Acute Lung Injury in Mice by Regulating the Toll-Like Receptor 4/Nuclear Factor κB Signaling Pathway.

Bacterial pneumonia is a common disease in dairy herds worldwide, which brings great economic losses to farmers. Sodium butyrate (SB), an inhibitor of histone deacetylase, plays an important role in limiting inflammation. The purpose of this study was to investigate the protective effects of SB on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice and explore the potential mechanism of SB protection. A total of 30 ICR mice were randomly divided into three groups ( n = 10): a phosphate-buffered saline (PBS) intratracheal instillation group, a LPS intratracheal instillation group, and a SB gavage group (SB was given 1 h before the LPS stimulation). After 12 h, samples of the blood and lung tissue were collected from the mice for experimental analysis. The results showed that the concentration of inflammatory cytokines [interleukin 1β (IL1β) and tumor necrosis factor α (TNF-α)], myeloperoxidase (MPO) activity in the lung tissue and blood, protein abundance of toll-like receptor 4 (TLR4), nuclear factor κB (NF-κB, p65), phosphorylated p65 (p-p65), inhibitor κBα (IκBα), and phosphorylated IκBα (p-IκBα), and relative mRNA expression of genes associated with inflammation, such as TLR4, NF-κB, IL1β, interleukin 6 (IL6), and TNF-α, were significantly upregulated in the LPS group compared to the PBS group. However, the SB addition markedly downregulated the levels of these parameters in the LSB group compared to those in the LPS group. Furthermore, the structure of the lung tissue from the LPS group was severely disrupted in comparison to that of the PBS group. However, with SB administration, the severe structural disruption was relieved. In addition, an immunohistochemical analysis showed that positive immunoreactions to TLR4, p65, and TNF-α were significant in the LPS group; however, SB addition markedly attenuated this phenomenon. In conclusion, the ALI mouse model was successfully established with an intratracheal instillation of LPS. Furthermore, gavage with SB inhibited inflammation in LPS-induced ALI.

Comparison of Surgically Excised Premacular Membranes in Eyes with Macular Pucker and Proliferative Vitreoretinopathy

ABSTRACTPurpose: To describe and compare characteristics of premacular membranes in idiopathic macular pucker (iMP) and proliferative vitreoretinopathy (PVR) using immunofluorescence and transmission electron microscopy.Materials and methods: For immunocytochemical and ultrastructural analyses, premacular membranes were harvested during vitrectomy from 16 eyes with iMP and 12 eyes with PVR. All specimens were processed as flat mounts for phase-contrast and fluorescence microscopy. We used 19 different primary antibodies such as anti-α-smooth muscle actin (α-SMA), anti-integrin-αv, anti-galectin, anti-IBA-1, anti-EMMPRIN (CD147), anti-ricinus (RCS) and anti-collagen-type I. Eight of 28 eyes were also prepared for transmission electron microscopy.Results: In all eyes with iMP and PVR, positive immunoreactivity of integrins, especially αvβ3 was found. There was also a strong staining of anti-α-SMA, anti-galectin, anti-EMMPRIN, anti-RCS, anti-IBA1 and anti-collagen-type I. Transmission electron microscopy showed that premacular membrane of iMP composed of myofibroblasts, glial cells and fibroblasts. In eyes with PVR, retinal pigment epithelial cells and myofibroblasts were seen as predominant cell types.Conclusion: Premacular membranes of iMP and PVR presented with similarities in cell distribution and immunoreactivity, but showed differences in cell composition. Herein, we demonstrate immunocytochemical characteristics involved in fibrotic processes. Cell transdifferentiation into myofibroblasts represents an important process in pathogenesis of both entities. In order to address future anti-fibrotic treatment strategies, we emphasize that both fibrotic diseases share distinct immunocytochemical and ultrastructural features.

Comparative Evaluation of Laminin 1 Expression in Odontogenic Keratocyst, Dentigerous Cyst, and Ameloblastoma: An Immunohistochemical Study

Introduction: Odontogenic keratocysts, dentigerous cysts, and ameloblastoma are the most common forms of cystic pathology that affect the jaw bones. The purpose of this study was to assess the immunohistochemical expression of laminin 1 in these lesions and to study the probable role of laminin 1 in their biologic behavior. Materials and Methods: Serial sections of 4 µm were obtained from the archival blocks using standard microtome. One set of sections was used for immunohistochemical study. Presence of brown colored end product at the site of target antigen was considered as positive immunoreactivity. Results: Of the 10 ameloblastomas, 9 were positive. Among the 10 odontogenic keratocysts, 9 showed a positive immunoreactivity for laminin 1 and 1 case was negative. Among the 10 dentigerous cysts, all of them showed a positive immunoreactivity for laminin 1. A statistical analysis was carried out using chi-square test and the statistical significance was evaluated. Discussion: All the lesions that showed positive immunoreactions for laminin 1 had basement membrane immunolocalization. The laminin 1 in odontogenic keratocysts showed that it is not involved in the formation of daughter cysts and the separation of epithelium and surrounding connective tissue. Conclusion: Expression of laminin 1 in the stellate reticulum can be a sign of the aggressiveness of ameloblastoma. Laminin 1 expression in the lining epithelium of dentigerous cyst can be used to explain its potential to transform to ameloblastoma. It can be concluded that the nature and aggressiveness of odontogenic cysts and tumors can be explained using laminin 1.

Evaluation of MUC-1, MUC-4, CDX-2 and OCT-1 expression profiles in gastric carcinomas and precancerous lesions

Aim: We investigated the diagnostic and prognostic utility of MUC-1, MUC-4, CDX2 and OCT-1 expression profiles in high-mortality gastric carcinomas and their precancerous lesions.Material and Methods: Areas in proximity of dysplasias and gastric carcinomas that were suspected of having intestinal metaplasia were stained with PAS / Alcian Blue at pH 2.5 and pH 0.5; Giemsa staining was applied for evaluation of presence of H. Pylori.Results: Presence of MUC-1 expression may exclude the diagnosis of low-grade dysplasia. MUC-1 and MUC-4 overexpression may be used for determination of groups at high risk of development of carcinomas, primarily in cases of high-grade dysplasia, while overexpression of MUC-1 in gastric carcinomas may be used as a favorable prognostic factor. Presence of CDX2 expression in various histological types of carcinomas gives rise to a suggestion that intestinal differentiation occurs in all histological types at least to some degree. OCT-1 plays a role in CDX2-mediated carcinogenesis and its regulation. Conclusion: In cases of dysplasia occurring with intestinal metaplasia, CDX2 was found to be expressed at a significantly higher degree (95.5%, p=0.011). Expression of CDX2 and OCT-1 was observed in all high-grade dysplasia cases, and was significantly higher in comparison to low-grade dysplasia cases (p=0.042; p=0.006, respectively). MUC-1 and MUC-4 expression significantly increased with each progressing stage of precancerous lesions in all lesions following low-grade dysplasia (p=0.0001; p=0.027, respectively). MUC-1 expression was the lowest (53.6%) in weak cohesive type carcinomas. Positive immunoreactivity with MUC-1 and CDX2 was higher in cases with a smaller number of metastatic lymph nodes compared to the cases with a larger number thereof.

Lacrimal gland adenoma in a sheep

The lacrimal gland is a diamond-shaped, tubuloalveolar gland that secretes the serous component of tears. A four-year-old female crossbreed sheep suffering from left eye protrusion was referred to a Veterinary Hospital. Ophthalmic examination revealed epiphora, superficial ulcerative keratitis, corneal edema and neovascularisation. Moreover, ultrasound examination showed a large heterogeneous mass with variable reflectivity in the intraconal and extraconal spaces. Grossly, a 2.5×1.5×0.5 cm oval firm grayish mass was observed. Histopathologically, the mass was composed mainly by tubules with two cell types including cuboidal luminal epithelial cells and peripheral myoepithelial cells. The tubular structures were separated by proliferating myoepithelial cells. Mitotic figures, cellular pleomorphism and atypia were not seen. Immunohistochemically, most of the luminal epithelial cells showed an immunopositive reaction with a cytokeratin (AE1/AE3) marker. On the basis of these findings, the mass was diagnosed as a lacrimal gland adenoma.

Alzheimer’s disease-like pathological lesions in an aged bottlenose dolphin (Tursiops truncatus)

Alzheimer’s disease (AD) is the most common neurodegenerative disorder causing dementia. AD-like pathological lesions have been described in human and in several other animal species. In this study, we report...

Immunohistochemical localization of adropin in the small intestine of rats

The aim of this study was to determine the occurrence and distribution of adropin in the small intestine in control and gastrectomized rats. The study was carried out on 12 Wistar rats. Six control rats (SHO) underwent a sham operation. Six rats were subjected to gastrectomy (Gx); i.e. resection of the antrum and the glandular part of the stomach. Six weeks after the surgery, the rats were sacrificed. The duodenum and middle part of the jejunum were collected for immunohistochemical and immunofluorescence procedures. An immunopositive reaction to ADR was detected in the duodenum and jejunum, in crypts, and in enterocytes located along the entire length of the villi. Furthermore, more intense ADR immunoreactivity was observed in the crypts and villi of the duodenum than in the jejunum. The presence of adropin was also detected in the submucosa and muscularis externa of the duodenum and jejunum, in the cells of Brunner’s gland of the duodenum, and in myenteric and submucosal plexus nerve. The ADR immunoreactivity in the crypts and villi in the duodenum and jejunum was lower in the Gx rats compared to the SHO rats. The ADR distribution in the duodenum and jejunum has also been confirmed in immunofluorescence studies. In conclusion, our data demonstrated ADR expression in all histological layers of the small intestine in the rats. Gx reduced ADR immunoreactivity in the crypts and enterocytes. This may suggest a potential impact of ADR on intestine function such as absorbability and immune reactions. However, further research is needed to explain the ADR role in gastrointestinal functions

Postnatal and aging-related changes of the retina in albino rats: a histological, immunohistochemical, and quantitative study

Background Retina is the best structural model for histological study of the nervous system. Aging is a biological phenomenon that involves gradual degradation of the structure and subsequent deterioration of the retina. Aim This study aimed to clarify the postnatal changes in the albino rat retina as well as the effect of aging on its structure. Materials and methods Twenty albino rats were divided into four groups (five rats each) according to their ages into 1, 3, 18, and 24 months. The retinae of all groups were prepared for light microscopic examination (histological and immunohistochemical), electron microscopic examination, and quantitative study including statistical analysis of the number of retinal ganglion cells. Results All layers of the retina were highly organized at 1 and 3 months with strong positive immunoreactivity of the ganglion cells to tubulin β III stain at 3 months. Ultrastructurally, oval light cones, irregular dark rods, and bipolar cells were noticed and the nuclei of ganglion cells layer exhibited finely dispersed chromatin at 1 month and at 3 months these cells were arranged in several layers of variable sizes and shapes. Aging caused degenerative changes to all layers of the retina at the age of 18 months which became marked with complete loss of the outer layers at 24 months. Ganglion cells showed weak immunoreactivity to tubulin β III stain and statistically, their number showed a highly significant increase from 1 to 3 months and then showed a highly significant reduction with aging. Conclusion Retina became highly organized postnatally and aging caused its deterioration with significant reduction in the number of ganglion cells.

Evaluation and Comparison of the Effects of Artesunate, Dexamethasone, and Tacrolimus on Sciatic Nerve Regeneration

To examine the effect of topical artesunate treatment on peripheral nerve regeneration and compare it with the effects of topical tacrolimus and dexamethasone on nerve regeneration. Thirty-two Wistar albino rats were used in this study. The rats were divided into 6 groups: sham, saline, petrolatum, artesunate, tacrolimus, and dexamethasone. A compression injury was generated in the right sciatic nerve in all groups except the sham group. In the sham group, the nerve was dissected but compression was not applied. In the groups in which compression was applied, the agents were absorbed through resorbable gelatin sponges applied to the injured region. At the end of 4weeks, walking analysis, electromyographic measurements, and histopathologic examinations were conducted. When the sciatic function index and electrophysiologic measurements were evaluated, artesunate, tacrolimus, and dexamethasone exhibited positive effects on nerve regeneration (P<.05); there were no significant differences among these 3 agents (P>.05). Histopathologic examination showed that artesunate decreased fibrosis scores and inflammation and increased the diameter of myelinated axons; tacrolimus decreased fibroblast scores; and dexamethasone only decreased fibrosis scores (P<.05). Immunohistochemical analysis showed that the artesunate and dexamethasone groups had more positive immunoreactivity to nerve growth factor than did the saline group (P<.05). Topical artesunate treatment had a positive effect on peripheral nerve regeneration. There were no relevant differences between the topical forms of dexamethasone and tacrolimus for peripheral nerve regeneration.

Retinal Toxicity of Intravitreal Injection of Ziv-Aflibercept in Albino Rabbits.

PurposeTo evaluate retinal toxicity of ziv-aflibercept, a drug that had been approved for use for patients with colon cancer.MethodsTwenty-two albino rabbits were injected intravitreally with 0.1 mL of ziv-aflibercept solution into the experimental eye and 0.1-mL saline into the control eye. Twelve were used for electroretinogram (ERG) at 4-weeks follow-up. An additional 10 rabbits were used for testing penetration of ziv-aflibercept into the retina during follow-up. The visual-evoked potential (VEP) was recorded after 4 weeks of ERG follow-up. Glial fibrillary acidic protein (GFAP) immunocytochemistry and retinal histology were performed after the termination of the follow-up period.ResultsThe ERG responses of the experimental eyes did not show signs of permanent functional damage. The VEP responses of the experimental eyes were of normal pattern and amplitude, and were similar to those recorded by stimulation of the control eyes. Histologic studies of both experimental and control eyes did not show signs of structural damage. However, GFAP expression was increased in retinal Müller cells of the experimental eyes and not of the control eyes. Retinal penetration of ziv-aflibercept, as indicated by positive antihuman immunoreactivity, was observed 1 day postinjection and was strengthened during the next 7 days. At 14 days postinjection, ziv-aflibercept was not detected.ConclusionsZiv-aflibercept was found to be nontoxic to the retina of rabbits based on electrophysiologic testing and histologic examination. However, GFAP immunocytochemistry suggests mild retinal stress caused by the drug.Translational RelevanceIf proven safe, ziv-aflibercept may be a new affordable treatment option in conditions involving neovascularization and macular edema.

Single-stranded DNA binding protein 2 expression is associated with patient survival in hepatocellular carcinoma

BackgroundSSBP2, single-stranded DNA binding protein 2, is a subunit of the ssDNA-binding complex that is involved in the maintenance of genome stability. The majority of previous studies have suggested a tumor-suppressive role of SSBP2, which is silenced by promoter hypermethylation in several human malignancies, such as hematologic malignancies, prostate cancer, esophageal squamous cell carcinoma, ovarian cancer, and gallbladder cancer. However, an oncogenic role of SSBP2 has been suggested in glioblastoma patients. We investigated the clinicopathologic significance of SSBP2 expression in hepatocellular carcinoma.MethodsWe constructed tissue microarrays consisting of 21 normal liver parenchyma and 213 hepatocellular carcinoma tissues with corresponding adjacent non-neoplastic tissues. SSBP2 expression was investigated by immunohistochemistry, and positive expression was defined as more than 10% of the tumor cells to show nuclear staining. We then analyzed the correlations between SSBP2 expression and various clinicopathologic characteristics, and further studied the role of SSBP2 in cell growth and migration.ResultsHepatocytes were negative for SSBP2 immunohistochemistry in all normal liver samples, whereas the nuclei of normal bile duct epithelium and sinusoidal endothelium were immunoreactive. Positive immunoreactivity was found in one (0.6%) out of 180 non-neoplastic liver tissue samples adjacent to the tumor and in 16 (8.5%) out of 189 hepatocellular carcinomas. Positive SSBP2 expression was significantly correlated with tumor multifocality (P = 0.027, chi-square test), high histologic grade (P = 0.003, chi-square test), and frequent vascular invasion (P = 0.001, chi-square test). Kaplan-Meier survival curves revealed that patients with SSBP2 expression had poor prognosis in both disease-free and overall survival (P = 0.004 and P = 0.026, respectively, log-rank test). SSBP2-positive tumors also had a higher Ki-67 proliferation index (P < 0.001, t-test). Furthermore, downregulation of SSBP2 in the Huh7 cell line inhibited cell migration (P = 0.022, t-test) with altered expression of epithelial-mesenchymal transition markers.ConclusionsThe minority of hepatocellular carcinomas expressed SSBP2 by immunohistochemistry, whereas normal hepatocytes were negative. SSBP2-positive hepatocellular carcinomas were significantly associated with aggressive phenotypes and poor clinical outcome.

Ischemia–reperfusion induced injury of rat ovary and the possible protective effect of amlodipine: Histological, immunohistochemial and biochemical study

Introduction: Ovarian torsion constitutes a gynecologic surgical emergency. However, early diagnosis of torsion may help to detorsion of the affected ovary and to preserve the fertility. Conservative therapy may lead to local and systemic deteriorating effects on the viability of the detorsioned ovary.Aim: The present study was done to evaluate the possible protective effect of amlodipine on ischemia and ischemia reperfusion (I/R) induced ovarian injury in a rat model using histological, immuno-histolochemical, biochemical and morphometric studies.Material and Methods: Forty adult female albino rats were used. They were equally divided into five groups. Group 1: Sham operation (control). Group 2: (ischemia) 3 hours duration of ischemia. Group 3: (ischemia/reperfusion) same as group 2 followed by 3 hours of reperfusion. Group 4: (ischemia and amlodipine) 5 mg amlodipine given orally one hour before ischemia then 3 hours of ischemia. Group 5: (ischemia, re-perfusion and amlodipine) 2 hours of ischemia then 5 mg amlodipine given orally then 3 hours of reperfusion. Then ovarian removal was performed and subjected to histological, immuno-histolochemical , biochemical and morphometric studies.Results: In ischemia and (I/R) groups, the ovaries revealed exfoliation and desquamation of granulosa cells into the follicular cavity of degenerated follicles, with karyolysis and karyorrhexis of the oocyte nucleus, Congested dilated vessels, hyaline degeneration and vacuolation in the interstitial region, Massive cellular infiltrate, hemosiderin deposits and massive hemorrhage of the ovarian stroma. There was positive immunoreactivity for PECAM-I and CD68 and negative for estrogen receptor immunostain. Both superoxide dismutase (SOD) and glutathione (GSH) concentrations were significantly decreased while malondialdehyde MDA (LPO) concentration was significantly increased in ischemia and I/R groups as compared with the other groups. Amlodipine addition improves both histological findings and biochemical valuesConclusion: Amlodipine attenuates ischemia and (I/R) induced ovarian tissue injury

HISTOLOGICAL AND IMMUNO-HISTOCHEMICAL STUDY ON THE POSSIBLE EFFECT OF VITAMIN K2 ON THE HEPATIC OVAL CELLS IN DIABETIC AND NON-DIABETIC ALBINO RATS

Background: Diabetes is a group of metabolic diseases characterized by hyperglycemia resulting from hypoinsulinemia, defective insulin action, or both. s-cells regeneration from stem cells or pancreatic progenitor cells is an attractive method for diabetic treatment. Hepatic oval cells (HOCs), intra-hepatic cells of bi-potent progenitor capabilities, can proliferate and differentiate into both hepatocytes and bile duct epithelial cells. They had the potentiality to transdifferentiate into pancreatic endocrine cells, as both liver and pancreas have the same origin (upper primitive foregut endoderm). Vitamin K2 can promote recovery in patients with liver cirrhosis and prevent insulin resistance reducing the risk of type 2 diabetes.Aim of the work: Evaluating the histological and immunohistochemical changes in hepatocytes and oval cells in diabetic and non-diabetic rats’ liver under treatment with vitamin K2.Materials and methods: 88 adult male albino rats were divided into 3 groups: Group I (control group), Group II (diabetic group); where diabetes was induced using single intraperitoneal (IP) dose of streptozotocin (STZ) following 2 weeks of high fat diet and Group III (treatment group) where diabetic rats were given daily oral vitamin K2 for 20 days (5 days on and 5 days off). Random blood sugar was measured for each group at day 0, 16 and then every 5 days till the end of the experiment (36 days). All groups’ sections were subjected to H&E and immunohistochemical stains for CD34, CK19 and insulin. Optical density for CD34 positive immunoreaction and mean area percent for CK19 and insulin immunoreaction were measured and data were statistically analyzed. Results and Conclusion: HOCs gave a positive reaction of changing into insulin producing cells in diabetic rats thus they may be modified to change into extra pancreatic source of insulin. Additionally, administration of vitamin K2 helped increase the proliferative ability of the HOCs in case of diabetes.

Activation of spinal nociceptin receptors induces cardiovascular depression and antinociception in an independent manner in mice

PurposeThe nociceptin receptor (NOP) was discovered in 1994 and was designated opioid-like receptor; activation of NOP leads to reduced neuronal excitability. Although suggested by the anatomical localization of NOP in brain or spinal cord, the cardiovascular or nociceptive effects of its endogenous ligand, nociceptin, are equivocal. Taking advantage from intrathecal application of nociceptin to simultaneously activate NOP on sympathetic preganglionic neurons in the intermediolateral column (IML) and superficial laminae of dorsal horn, we investigated whether the nociceptin-induced cardiovascular effects engage the participation of baroreflex, and whether the concurrently elicited changes in blood pressure and pain responses are interrelated.MethodsNOPs in the thoracic spinal cord of ICR or C57BL/6 mice were identified with immunofluorescence staining and were activated through intrathecal administration of nocicetpin. The elicited changes in cardiovascular parameters and tail-flick nociceptive responses were measured.ResultsPositive immunoreactivity against NOP colocalized with neurons in the IML and superficial dorsal horn layers of thoracic spinal cord. Intrathecal administration of nociceptin (1, 2, or 5 nmol) elicited a significant and dose-dependent decrease in blood pressure or heart rate that was paralleled by reduced baroreflex-mediated sympathetic vasomotor tone and mirrored by augmented cardiac vagal baroreflex, alongside prolonged tail-flick latency with an efficacy of hypotension <<< antinociception. Coadministration of the specific NOP antagonist, UFP101 (10 nmol), blunted all nociceptin-elicited responses. However, restoring blood pressure to baseline level failed to affect the antinociceptive actions of nociceptin.ConclusionActivation of thoracic spinal NOP in ICR and C57BL/6 mice induces blood pressure and heart rate by decreasing the sympathetic outflow of both arms of the baroreflex arc to the blood vessels and the heart, and the antinociceptive responses to nociceptin are independent of and disproportional to its cardiovascular actions.

Primary small bowel mesentery de-differentiated liposarcoma causing torsion with no recurrence for 5 years

Rationale:Liposarcoma (LPS) is a rare malignant soft-tissue tumor. Management of LPS is relatively difficult, because there are no characteristic symptoms, or biomarkers, nor any established effective treatment. Hence, the report of the accumulation of each LPS case is necessary. We experienced an extremely rare case of torsion caused by a primary small bowel mesentery LPS.Patient's concern:A 70-year-old male consulted our hospital with the complaints of abdominal pain and sudden vomiting.Diagnosis:No lump could be palpated, and tumor markers tested were within normal limits. However, computed tomography revealed an intestinal obstruction caused by torsion of the small bowel due to an LPS tumor.Interventions:After decompression of the intestinal obstruction by use of an ileus tube, surgical treatment was performed with rapidity.Outcome:The torsion was found to be caused by the tumor that originated from the small bowel mesentery. The tumor was resected along with a portion of the small bowel. The growth of adipose tissues of various sizes and containing atypical cells was detected by histopathological examination. Also, immunohistochemical examination resulted in positive immuno-reactions for MDM2, CDK4, and p16INK4, which indicated the tumor to be a de-differentiated LPS. The patient was discharged on postoperative day 14 without any complications, and no recurrence of the tumor was observed at 5 years after the operation.Lessons:LPS should be considered in differential diagnosis of bowel torsion, and careful management is required because of the high possibility of recurrence. Patients should be followed carefully for at least 5 years, and further accumulation of data will be required in order to establish the appropriate management of LPS.