Immunohistochemical Changes Research Articles

Protective effect of vitamin D on imidacloprid-induced testicular injury in rats.

The degenerative effects of imidacloprid via oxidative stress are known. Irisin is a recently discovered peptide with energy regulator and antioxidant effects. In addition, the antioxidant potential of Vitamin D has been reported in previous studies. The current study was performed to investigate the effect of Vitamin D on testis morphology and irisin immunoreactivity in imidacloprid-treated rats. Thirty-two Wistar albino male rats were divided into groups: control (n = 6), corn oil (n = 6), Vitamin D (n = 6), imidacloprid (n = 7) and imidacloprid + Vitamin D (n = 7). Testis tissues were used to evaluate the histopathological, biochemical and immunohistochemical changes. Oxidative state in testis tissue was determined with total antioxidant and oxidant status markers, total antioxidant status (TAS) and total oxidant status (TOS) respectively. In microscopic examination, degenerative changes in the seminiferous tubule epithelium, interstitial edema and increased irisin immunoreactivity were observed in animals given imidacloprid. Also increased TOS and decreased TAS levels were measured in these animals. It was observed that Vitamin D improved the testicular damage histopathologically when compared to the imidacloprid group. However, increase in TAS levels and decrease in both TOS levels and irisin immunoreactivity were found insignificant in animals given Vitamin D. In the present study it was observed that Vitamin D ameliorated testis injury caused by imidacloprid. Furthermore, imidacloprid was found to increase the immunoreactivity of irisin. In the light of our findings, we conclude that the use of Vitamin D could be beneficial against testicular damage caused by imidacloprid.

(-)-Phenserine and the prevention of pre-programmed cell death and neuroinflammation in mild traumatic brain injury and Alzheimer's disease challenged mice

Mild traumatic brain injury (mTBI) is a risk factor for neurodegenerative disorders, such as Alzheimer's disease (AD) and Parkinson's disease (PD). TBI-derived neuropathologies are promoted by inflammatory processes: chronic microgliosis and release of pro-inflammatory cytokines that further promote neuronal dysfunction and loss. Herein, we evaluated the effect on pre-programmed cell death/neuroinflammation/synaptic integrity and function of (-)-Phenserine tartrate (Phen), an agent originally developed for AD. This was studied at two clinically translatable doses (2.5 and 5.0 mg/kg, BID), in a weight drop (concussive) mTBI model in wild type (WT) and AD APP/PSEN1 transgenic mice. Phen mitigated mTBI-induced cognitive impairment, assessed by Novel Object Recognition and Y-maze behavioral paradigms, in WT mice. Phen fully abated mTBI-induced neurodegeneration, evaluated by counting Fluoro-Jade C-positive (FJC+) cells, in hippocampus and cortex of WT mice. In APP/PSEN1 mice, degenerating cell counts were consistently greater across all experimental groups vs. WT mice. mTBI elevated FJC+ cell counts vs. the APP/PSEN1 control (sham) group, and Phen similarly mitigated this. Anti-inflammatory effects on microglial activation (IBA1-immunoreactivity (IR)) and the pro-inflammatory cytokine TNF-α were evaluated. mTBI increased IBA1-IR and TNF-α/IBA1 colocalization vs. sham, both in WT and APP/PSEN1 mice. Phen decreased IBA1-IR throughout hippocampi and cortices of WT mice, and in cortices of AD mice. Phen, likewise, reduced levels of IBA1/TNF-α-IR colocalization volume across all areas in WT animals, with a similar trend in APP/PSEN1 mice. Actions on astrocyte activation by mTBI were followed by evaluating GFAP, and were similarly mitigated by Phen. Synaptic density was evaluated by quantifying PSD-95+ dendritic spines and Synaptophysin (Syn)-IR. Both were significantly reduced in mTBI vs. sham in both WT and APP/PSEN1 mice. Phen fully reversed the PSD-95+ spine loss in WT and Syn-IR decrease in both WT and APP/PSEN1 mice. To associate immunohistochemical changes in synaptic markers with function, hippocampal long term potentiation (LTP) was induced in WT mice. LTP was impaired by mTBI, and this impairment was mitigated by Phen. In synopsis, clinically translatable doses of Phen ameliorated mTBI-mediated pre-programmed cell death/neuroinflammation/synaptic dysfunction in WT mice, consistent with fully mitigating mTBI-induced cognitive impairments. Phen additionally demonstrated positive actions in the more pathologic brain microenvironment of AD mice, further supporting consideration of its repurposing as a treatment for mTBI.

A combined retrospective, prospective and experimental study of Ornithobacterium rhinotracheale infection in chickens

ABSTRACTThe aim of this study was to examine the histopathological and immunohistochemical changes caused by natural and experimentally-induced Ornithobacterium rhinotracheale infection in the respiratory system of chickens. To this end, three different studies were carried out. The first was a retrospective study of 82 field cases with respiratory disorders compatible with O. rhinotracheale infection. The bacterium was immunohistochemically detected in the lungs in 48 of 82 field cases, and 50 β-haemolytic (BH) and non-haemolytic (NH) strains were isolated. In the second study, an experimental model of the disease was created using 3-week-old broiler chickens, to identify possible differences of pathogenicity between the BH and NH isolates by the intravenous (IV) and intratracheal (IT) inoculation routes (IR). The group challenged with the NH isolate showed more severe lung lesions than the group challenged with the BH isolate at 7-days postinoculation (p.i.). The 14-day p.i. groups challenged with either the BH or NH isolates by the IT or IV IR had a higher histologic grade of pulmonary and hepatic lesions and a higher total histologic grade of lesions suggesting more severe pathology with longer time of exposure. A direct association between the inoculation routes and the organs affected was shown. Finally, a slaughterhouse study was carried out from October 2014 to May 2015, in which the histologic grade of lesions was significantly higher in immunohistochemically positive for O. rhinotracheale lungs of dead-on-arrival chickens.

Comparative Clinical and Histological Study of Narrow Band UVB Phototherapy Versus Combined Narrow Band UVB Phototherapy and Microneedling in Treatment of Vitiligo

AbstractBackground: Vitiligo is an acquired pigmentary disorder of unknown etiology that affects up to 1% of the general population of all skin types.It is a multifactorial disorder. There are numerous medical and surgical treatments aimed at repigmentation.Microneedling is a minimally invasive procedure that uses fine needles to puncture the epidermis. The indications for microneedling therapy have grown significantly and it is becoming a widely used treatment in Dermatology.Aim of Study: To study the clinical and histological efficacy of combined NB-UVB phototherapy and microneedling versus NB-UVB phototherapy in the treatment of vitiligo.Patients and Methods: Twenty patients were divided into two Groups (A & B), each of 10 patients. Both groups were matched regarding age, sex, skin type and have a depigmented patch at the same corresponding site. Group A was subjected to microneedling with dermapen at one depigmented patch followed by NB-UVB phototherapy sessions twice weekly for three months. Group B was subjected to NB-UVB photo-therapy sessions twice weekly for three months. Two Skin biopsies one before and one after the therapy were taken for immunohistochemical examination by HMB 45.Results: The mean percentage of improvement was sig-nificantly high in Group A (48.50%±26.46) than Group B (20.0%±19.58). In Group A 20% showed mild improvement (G1), 20% showed moderate improvement (G2), (40%) showed good improvement (G3) and 20% showed excellent improve-ment (G4). In Group B 30% showed no improvement, 50% showed mild improvement (G1), 10% showed moderate improvement (G2) and 10% showed good improvement (G3). There was statistically significant difference between both groups (p=0.027*).Immunohistochemical changes revealed marked expression of HMB45 in Group A more than Group B, the mean of color intensity in Group A was 0.05±0.04 and a median of 0.03 while in Group B was 0.01±0.01. There was statistically significant difference between both groups (p=0.002*).Conclusion: Microneedling prior to NB-UVB photother-apy sessions was clearly superior to NB-UVB phototherapy alone with accelerating response. Microneedling is a tolerable procedure, safe with minimal side effects.

Inhibition of NF-κB and the oxidative stress -dependent caspase-3 apoptotic pathway by betaine supplementation attenuates hepatic injury mediated by cisplatin in rats.

Cisplatin is a major anti-cancer drug commonly used in the treatment of various cancers; nevertheless, the associated hepatotoxicity has limited its clinical application. The aim of this investigation is to test the impact of betaine supplementation on cisplatin-induced hepatotoxicity. Animals were allocated into four groups; normal control group (control betaine group (250 mg/kg/day, po for twenty six days), cisplatin group (single injection of 7 mg/kg, ip) and betaine + cisplatin group (received betaine for twenty one days before cisplatin injection and daily after cisplatin for five days). Cisplatin-induced liver injury was confirmed by increased alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels. Cisplatin elevated lipid peroxides, and reduced the concentrations of reduced glutathione (GSH), glutathione peroxidase (GSH-Px), catalase and superoxide dismutase (SOD) in hepatic tissues. Cisplatin increased the inflammatory mediators; nitrite and tumor necrosis factor-α (TNF- α) in hepatic tissues. Increased gene expressions of the apoptotic marker, caspase-3 and nuclear factor-kappa B (NF-κB) were observed in hepatic tissues of cisplatin-treated rats. All these changes were further confirmed by histopathological findings in cisplatin group. Pre-treatment with betaine reduced serum aminotransferases (ALT and AST), and lowered hepatic concentrations of lipid peroxides, nitrite and TNF-α while increased SOD, GSH, catalase, and GSH-Px concentrations. Moreover, the histological and immunohistochemical changes were improved. The suppression of NF-κβ-mediated inflammation, oxidative stress, and caspase-3 induced apoptosis are possible mechanisms to the observed hepatoprotective effect of betaine.

A Study on the Effect of Vitamin E on Histomorphological and Immunohistochemical Changes Induced By Electromagnetic Field

Background: Electromagnetic fields has increased gradually due to technology expansion and thus continues exposure to it may affect the biological system, so that the present work aimed to study the biological effect of non-ionizing radiation magnetic field from power line of 50 Hz which caused by devices become widely used in human activity on the structural properties of cerebral cortex and the possible protective role of vitamin E on it through the histopathological and immunohistochemical study . Result: Thirty mice were divided into three groups, group I (control) , group II animals of this group exposed to extremely low frequency magnetic field (ELF – MF) generated by electrical device (2mT- 20G) for a total period 45 day(2h/day) and group III exposed to the same field and period after adding vitamin E (200 mg /kg) B.W. along with the drinking water. The results showed many changes such as neuronal degeneration and apoptosis in number of Purkinje cells and showing a decreased reactivity in carbohydrates , protein ,DNA and Glial Fibrillary Acidic Protein (GFAP) content in (MF) treated group as compared to control , such changes showed improvement of the histolomorpholigical and immunohistochemical changes in magnetic field (MF) and vitamin E treated group. Conclusion: Electromagnetic fields exposure led to degenerative changes on cerebellar cortex, and the data collected from histopathological and immunohistochemical study revealed a protective effect of vitamin E on cellular cortex layers of the cerebellar cortex

Differential Expression of CD3, TNF-α, and VEGF Induced by Olanzapine on the Spleen of Adult Male Albino Rats and the Possible Protective Role of Vitamin C.

Olanzapine is an antipsychotic drug effective in the treatment of stress-associated psychiatric illnesses, but its effect on the spleen remains unclear. Vitamin C is essential for the optimum function of the immune system. We aim to investigate the effect of Olanzapine on spleen structures and to assess the protective effect of vitamin C. Forty adult male albino rats were divided into four groups: group (I), a control; group (II), rats were given vitamin C at 40 mg/kg body weight; group (III), rats were given Olanzapine at 2 mg/kg body weight; and group (IV), rats were given vitamin C and Olanzapine at the same dose of group (II) and group (III) for one month. The hematoxylin and eosin (H&E) of the olanzapine treated group showed focal areas of cellular depletion and a decrease in the size of the white pulp. The red pulp was expanded and showed marked congestion and dilatation of blood sinusoids. Cluster of differentiation 3 (CD3) was significantly reduced, however both tumor necrosis factor alpha (TNF-α), and vascular endothelial growth factor (VEGF) were significantly higher. The administration of vitamin C repaired structural and immunohistochemical changes via increased CD3 and decreased TNF-α and VEGF. Therefore, the oxidative and the inflammatory pathways may be the possible mechanisms underlying olanzapine immunotoxicity. Vitamin C exerted immune modulator and antioxidant effects against olanzapine.

Enhanced Antiarthritic Efficacy by Nanoparticles of (-)-Epigallocatechin Gallate-Glucosamine-Casein.

This work aims to improve the antiarthritic activity of (-)-epigallocatechin gallate (EGCG) and glucosamine (GA) through fabrication and optimization of casein protein nanoparticles (EGC-NPs). Optimized EGC-NPs were obtained with a EGCG/GA/casein ratio of 1:2:8 (w/w/w). The EGC-NPs gave a mean size of 186 ± 3.5 nm and an entrapment efficiency of 86.8 ± 2.7%, and they exhibited a greater inhibitory activity against human fibroblast-like synoviocytes-osteoarthritis cells and human fibroblast-like synoviocytes-rheumatoid arthritis cells compared with that of the EGCG-GA mixture by 33.5% and 20.8%, respectively. Freeze-dried EGC-NPs stored at 25 °C during 12 months showed high dispersion stability. Moreover, the redispersion of the freeze-dried EGC-NPs produced almost no significant changes in their physicochemical properties and bioactivity. Rat experiments demonstrated that the antiarthritis effect of the EGC-NPs was significantly higher than that of EGCG-GA mixture, as assessed through an analysis of anti-inflammatory efficacy, radiographic images and histopathological assessments of paw joints, and immunohistochemical changes in serum cytokines. The enchanced antiarthritic activity in vivo was consistent with that in vitro. The EGC-NPs demonstrate potential as a food supplement for the treatment of arthritis.

Pathomorphology of fetal and mature liver under the lead intoxication and after the correction: the review of experimental data

Lead is one of the most widespread pollutant which alters the mature liver and is especially harmful for the fetal liver. The typical alterations in the mature liver after the lead exposure are the hypertrophy and vacuolization of hepatocytes, circulatory disorders, mononuclear cellular infiltration. The morphological changes in the liver during prenatal development under the maternal lead treatment are inhibition of hematopoiesis, dystrophy of hepatocytes, disturbances in the liver architecture, its vessels and stroma with a gradient of pathological changes toward the peripheral parts of the organ. The manifestations of liver alteration after birth become deeper with age and develop to the necrosis, edema and inflammation. The biochemical disturbances in the liver are the decrease in the activity of enzyms of energy metabolism, inhibition of protein and nucleic acids synthesis, imbalance of the lipid peroxide oxidation system, and the increasing of oxidative stress with the further alteration of the membranes of the endothelium, red blood cells, hepatocytes, as well as mitochondrial membrane. The changes in expression of the immunohistochemical markers can differentiate the processes in the liver, which are relatively stable or sensitive to lead, especially in prenatal development, whereas the biochemical parameters are valuable for estimation of the liver damage in postnatal life. The immunohistochemical changes under the lead treatment reflect the inhibition in protein synthesis such as albumin and cytokeratins, as well as growth factors, nitric oxide synthases and matrix metalloproteinases expression; whereas the expression of apoptotic markers increases. The search for the natural products, dietary supplements and drugs with the protective properties is ongoing and covers the wide spectrum of agents, including vitamins, micro- and macroelements, antioxidants, chelating agents, natural extracts, proteins, sorbents and complex-producing drugs. The immunohistochemical markers as well as biochemical parameters can be used to prove the efficacy of protectants for chronic lead intoxication.

Metastatic bone lesions of men in combination with an increase in serum PSA from the point of view of diagnosing carcinomas of unknown primary localization

In cases of “malignant neoplasm of unknown origin” of men with bone metastases, it is the serum PSA level that can be crucial for further treatment. The literature describes carcinomas with bone metastases and an increase in the prostate-specific antigen, which were twice refuted by a prostate biopsy but had a positive dynamic during anti-androgen therapy (bicalutamide) in combination with chemotherapy (zoledronic acid). The aim of the work is to improve diagnostic algorithms, to investigate a complex of morphological, morphometric and immunohistochemical characteristics of metastatic foci in the bones, which are represented by adenocarcinomas of unknown primary localization in men with increased serological marker PSA, compared with primary carcinomas of the prostate gland. Materials and methods. A retrospective analysis of clinical data, histological and immunohistochemical characteristics of 21 observations of biopsy material from patients with primary carcinoma of the prostate gland (group 1, men aged 56 to 82 years, average 66.00 ± 7.76; median 65) and 10 patients with bone carcinoma metastases without primary localization and increased serum marker PSA (group 2, men aged 43 to 57 years, mean 50.00 ± 9.89; median 46.5) on the basis of the morphological department of diagnostic center “Pharmacy of Medical Academy” for the period 2015–2017. Results. Morphological, morphometric and immunohistochemical changes of carcinomas of unknown primary site in men with metastatic bone lesions and increased serum marker PSA associated with the acquisition of more aggressive properties were identified in comparison with the primary carcinomas of the prostate gland, which tend to local distribution. There were prevalence of poorly differentiated histological forms of metastatic adenocarcinomas, an objective reduction in the area and perimeter of nuclei (P = 0.00002, P < 0.0001, respectively), possible loss of immunohistochemical markers AMACR (p504s) and AR (androgen receptors) specific for acinar prostate carcinomas (P = 0.008). Conclusion. The use of minimal primary immunohistochemical panel from cytokeratin, Pan AE1/AE3 (+) / Vimentin(-) / CD45(-) / S100(-) and secondary – cytokeratin 8(+) / PSA(+) / AMACR (p504s)(+) panels / AR(+/-) will allow to prove the conformity of the carcinoma phenotype with unknown primary site in men with metastatic bone lesions and an increase in the PSA serological marker to the form of disseminated prostate carcinoma with justification for the use of appropriate therapy in cases of non-informative prostate biopsy.

Iron nanoparticle-labeled mesenchymal stromal cells persist and demonstrate anti-inflammatory mechanism of action in an osteoarthritis mouse model

Background & Aim Osteoarthritis (OA) is characterized by cartilage degradation and chronic joint inflammation. Mesenchymal stromal cells (MSCs) have shown promising results in OA, but their mechanism of action is not fully understood. We hypothesize that MSCs polarize macrophages (MΦ) to more homeostatic sub-types. We tracked ferumoxytol (iron-nanoparticle)-loaded MSCs in mouse osteoarthritic joints, and investigated mechanisms of action of injected MSCs via immunohistochemical staining of homeostatic MΦ populations. Methods, Results & Conclusion 10-week-old C57/BL6 male mice were induced to undergo OA by destabilization of medical meniscus (DMM) or SHAM surgery. 3 weeks post-surgery, mice were injected intra-articularly with saline or ferumoxytol-labeled murine MSCs (50 × 103 MSCs/mouse) to yield the following groups (n=5/group): i) DMM+MSC with ferumoxytol; ii) DMM+MSC without ferumoxytol iii) DMM+Saline; iv) SHAM+MSC; and v) SHAM+Saline. MRIs of mice were taken at 1, 3 days and 1, 2, or 4 weeks after injection. At 4 weeks post-injection, mice were scored for i) OARSI to determine cartilage damage and synovitis and ii) immunohistochemical changes in CD206, F/480, Prussian Blue/Sca-1, and cleaved caspase-3 to detect homeostatic MΦ, total MΦ, ferumoxytol-labeled MSCs, and apoptotic cells respectively. Ferumoxytol-loaded MSCs persisted to time of euthanization (4 weeks) at greater levels in DMM- vs. SHAM-joints. Prussian Blue/Sca-1 co-staining confirmed the iron label resides in MSCs (at 1, 2, and 4 weeks), although some iron label was also detected in F4/80+ MΦ, likely due to phagocytosis of apoptotic MSCs. MRI analyses demonstrated a significant decrease in joint swelling at day 3 and 4 week groups in MSCs vs saline treated animals. However, OARSI synovitis scores did not capture any differences between MSCs vs saline treated animals. In depth examination by immunohistochemical staining showed for the first time that MSCs vs saline treated had increased proportions of CD206+ homeostatic MΦ (p We demonstrate that ferumoxytol-MSC can be imaged in OA mice by MRI for 4 weeks, and these mice have augmented MΦ infiltration (p

Combination antiretroviral therapy (cART)-induced hippocampal disorders: Highlights on therapeutic potential of Naringenin and Quercetin

IntroductionIn spite of the multiple benefits of combination antiretroviral therapy (cART) on HIV positive patients, prolonged usage has been reported to exacerbate oxidative stress, and induce neurological and cognitive dysfunction, thus, the need to search for an adjuvant therapy to ameliorate the oxidative and improve treatment adherence with better virological outcome. This study aimed at determining the potential therapeutic effects of Quercetin and Naringenin on cART-induced cyto-architectural, neuro-behavioral and immunohistochemical changes in the hippocampus of the adult Wister rats. Materials and MethodsThe animals were grouped as follows: Control, DMSO, 24 mg/kg cART (Tenovovir 300 mg, Lamivudine 300 mg and Efavirenz 600 mg), 50 mg/kg Naringenin, 50 mg/kg Quercetin, cART + Naringenin, cART + Quercetin were administered orally for 8 weeks. At the end of administration, neurobehavioural test was conducted, animals were euthanized and hippocampus was processed for oxidative stress markers, histology, TNF-α, and Monoamine oxidase-B expression. ResultsAt the end of 8 weeks of administration, 24 mg/kg cART decreased superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH) and increased Malondialdehyde (MDA). Whereas, 50 mg/kg quercetin, and 50 mg/kg Naringenin decreased the oxidative stress (increased SOD, CAT, GSH, and reduced MDA) induced by cART (reduced SOD, CAT, GSH, and increased MDA). In addition, hematoxylin and eosin stained hippocampus showed that quercetin and naringenin prevented neurodegenerative changes (marked cytoplasmic shrinkage and several pyknotic nuclei in the dentate gyrus and cornus ammonis regions) in cART-treated rats. Furthermore, immunohistochemical studies revealed that quercetin and naringenin attenuates cART-induced upregulation of monoamine oxidase-B (MAO-B) expression. Likewise, from the Morris water maze neurobehavioral studies, naringenin and quercetin also ameliorated cART-induced memory impairments (initial spatial memory, reversal spatial memory and probe tests). ConclusionThis study shows that Naringenin and Quercetin have a good potential in reversing cART-induced hippocampal disorders in Wistar rats.

Dentate gyrus neurogenesis across different ages in male rats: an immunohistochemical approach

Dentate gyrus is a fundamental sub-region of the hippocampus which is directly engaged in higher memory and cognitive functions. This study was performed to describe the histological and immunohistochemical changes in dentate gyrus in experimental animals during postnatal development. Forty four male albino rats were classified into four equal groups: new born group aged one day, adult group aged 3–6 months, early senile group aged 18–20 months and late senile group aged 30–31 months. Specimens of hippocampus were processed and prepared for routine hematoxylin and eosin stains and immunohistochemical expressions of calretinin, glial fibrillary acidic protein and Ki67 (Kiel 67). Morphometric data were statistically analyzed. The present results showed significant reduction in thickness of granule cell layer of late senile group. Interestingly, the mean number of immature neurons was significantly increased in early senile group, while it was significantly reduced in late senile group. The number of mature granule cells showed marked reduction in both early and late senile groups. Furthermore, the number of astrocytes and optical density of glial fibrillary acidic protein revealed significant age-dependent increase. Measurable difference in number of calretinin positive interneurons was detected between adult and senile groups. However, mean number of Ki67 immune positive nuclei expressed significant age-dependent reduction. This study concluded that interneurons play a substantial role in modulating dentate gyrus neurogenesis which occurs throughout life and steadily decreases during aging. It is recommended to focus on the different stimuli and factors that potentiate neurogenesis to prevent or treat cognitive deficiencies associated with aging.

Histologic changes in early colonic anastomotic healing using autologous platelet-rich fibrin matrix

Anastomotic leakage represents a devastating post-surgical event with surgeon- and patient-related causes. The early lag phase of healing (day 0–4) is a time of increased vulnerability. The current role of tissue sealants as anastomotic adjuncts remains unclear, with controversial results reported. Platelet-rich plasma provides a reliable source of growth factors supporting wound healing. We aimed to assess the impact of a novel autologous platelet-rich fibrin (PRF) agent (Obsidian-ASG® matrix; GergMed Medizintechnik, Baiern, Germany) on colonic anastomotic healing in a porcine model. Left-sided colonic anastomoses were constructed in 16 pigs with reinforcement using the Obsidian-ASG® matrix in 12 animals. Animals were sacrificed on days 0, 4, 10 and 30, with histologic assessment of the anastomotic line along with evaluation of fibrotic tissue maturation, foreign body reactivity and mucin production. Collagen maturity and immunohistochemical changes of angiogenesis and infiltrating macrophage profiles were determined as well. There was no observable effect in matrix-treated animals on epithelial line thickness, granulation tissue formation, foreign body reactivity or mucosal restitution. Matrix-treated animals showed higher mucin production on day 4 accompanied by a more mature collagen and a greater degree of early angiogenic response with a higher infiltration density of M2 macrophages. Additionally, anastomotic burst pressure in matrix-treated animals was higher on day 0, 4, 10 and 30 (100, 100, 210, 160 vs. 40, 30, 60, 20 mm Hg, respectively). Obsidian-ASG® matrix may have beneficial effects on anastomotic healing by shifting the inflammatory response towards a mature and more stable collagen phenotype in the early phase.

Toxicity Induced by Zirconia Oxide Nanoparticles on Various Organs After Intravenous Administration in Rats.

ZrO₂-NPs are widely applied in industry, biomedicine and dentistry, e.g., foundry sands, refractories, ceramics dental prostheses, dental implant coatings and bone defect restorative materials. To date, little information is available on the potential adverse effects and toxic mechanism in human organs associated with exposure to ZrO₂-NPs. The biodistribution of ZrO₂-NPs and the consequent oxidative stress in the spleen, kidney, heart, brain, and lung at six time points after a single injection of ZrO₂-NPs were examined. Histopathological and immunohistochemical changes were also examined. RNA-Seq analysis was conducted in organs with high ZrO₂-NPs accumulations or obvious histopathological changes (brain and spleen). Exposure to the ZrO₂-NPs led to persistent oxidative stress and cell proliferation promotion/inhibition in various organs. RNA-Seq results of the spleen and brain point to significant gene expression changes. Metabolism was identified as leading pathways in the spleen. This study proves ZrO₂-NPs likely have negative impacts on various organs, and exhibit potential disease risks.

Cellular and Immunohistochemical Changes in Anaphylactic Shock Induced in the Ovalbumin-Sensitized Wistar Rat Model.

Anaphylactic shock (AS) is a life-threatening, multisystem disorder arising from sudden release of mast cell- and basophil-derived mediators into the circulation. In this study, we have used a Wistar rat model to investigate AS-associated histopathologic changes in various organs. Rats were sensitized with ovalbumin (1 mg s.c), and AS was induced by intravenous injection of ovalbumin (1 mg). Experimental groups included nonallergic rats (n = 6) and allergic rats (n = 6). Heart rate and blood pressure were monitored during one hour. Organs were harvested at the end of the experiment and prepared for histologic and immunohistochemical studies. Lung, small bowel mucosa and spleen were found to undergo heavy infiltration by mast cells and eosinophils, with less prominent mast cell infiltration of cardiac tissue. The mast cells in lung, small bowel and spleen exhibited increased expression of tryptase, c-kit and induced nitric oxide synthase (iNOS). Increased expression of endothelial nitric oxide synthase (eNOS) by vascular endothelial cells was noted principally in lung, heart and small bowel wall. The Wistar rat model of AS exhibited accumulation of mast cells and eosinophils in the lung, small bowel, and spleen to a greater extent than in the heart. We conclude that lung and gut are principal inflammatory targets in AS, and likely contribute to the severe hypotension of AS. Targeting nitric oxide (NO) production may help reduce AS mortality.

Pentoxifylline ameliorates chronic stress/high-fat diet-induced vascular wall disease: the role of circulating endothelial progenitor cells.

Depression and cardiovascular disease (CVD) are two faces of one coin. A pro-inflammatory state was previously suggested in the pathology of both diseases. We investigated the effect of chronic administration (2weeks) of imipramine (20mg/kg/day) and pentoxifylline (50mg/kg/day) on behavioral, aortic histological abnormalities, and level of circulating endothelial progenitor cells (CEPCs) in peripheral blood of male Wistar rats exposed to chronic mild stress (CMS) and high-fat diet. Exposure to CMS and high-fat diet induced a depressive-like behavior alongside aortic immunohistochemical changes associated with an increase in aortic TNF-α level. Markers of CEPCs, VEGFR-2 and CD133, were significantly disturbed in aortic sections, as compared to control animals and those exposed to CMS while fed high-fat diet, although flowcytometric analysis did not show any significant changes in the level of CEPCs in peripheral blood. Chronic pentoxifylline treatment was more effective in ameliorating the histological changes and endothelial damage compared to imipramine. Pro-inflammatory cytokines-induced disturbances in CEPCs could constitute a plausible link between depression and atherosclerotic cardiovascular disease. Current antidepressants reduce symptoms of depression without tackling the underlying link between it and cardiovascular disease. Targeting pro-inflammatory cytokines might open a new therapeutic approach to alleviate depression and reduce the risk of mortality from cardiovascular disease.

Prognostic importance of immunologochemical markers of papillary thyroid cancer

In Papillary Thyroid Cancer (PTC) in recent years an immunohistochemical markers were detected. Several authors consider their use as perspective to predict tumor behavior and the risk of recurrence and for differential diagnosis between different variants of PTC. However, the data obtained during various studies are controversial, which necessitates further research. Materials and methods: Using standard methods, the pathogistological material was obtained after surgical treatment of 273 patients with PTC. Expression of immunohistochemical markers: TTF1 and Gal3 - in 48 patients, p53 - in 50 patients, Ki-67 in 53 patients, NIS in 44 patients detected immunohistochemically. The connection between the histological type of tumor, the level of expression of immunohistochemical markers and the histological signs of aggressive tumor behavior are analyzed: multifocal, capsule invasion, extratireoidal invasion, lymph node damage. Results: In the analysis of data obtained with the use of standard histological techniques, it was found that the most aggressive variant was cylindrical cells PTC with a high incidence of extrathyroid invasion - 5.4 %, a capsule invasion -7.2 %, multifocal growth - 10.8 %, metastatic lymph nodes – 17.1 %. The high levels of TTF1 and Ki-67 expression, as well as the absence of NIS expression, were reliably associated with metastatic lymph node involvement (p <0.05). Reliable relationship with other histological signs of aggressive behavior of the tumor was not detected. No association was found between Gal3 and p53 expression levels and signs of aggressive tumor behavior. The data obtained in the study suggest that immunohistochemical markers reflect the processes characterizing the biological pathomorphism of the tumor and indicate a gradual loss of differentiation signs in more aggressive tumors. Conclusions: immunohistochemical markers changes reflect biological pathomorphism of the tumor, overexpression of ki67 and TTF1, as well as the loss of NIS, suggest a greater malignant tumor potential. The obtained data may help in individualized choice of surgical procedure

Formyl peptide receptor 1 signalling promotes experimental colitis in mice

Inflammatory bowel disease is characterised by intricate immune cell interactions with tissue cells and such cross-talks can become deregulated. The formyl peptide receptor 1 (Fpr1) is expressed by both immune and stromal cells including epithelial cells. We evaluated the development of the physiopathology of the DNBS induced colitis in Fpr1 KO mice on the C57BL/6 genetic background compared to C57BL/6 genetic background animals. We have assessed both macroscopic and histological markers of the diseased, together with the immunohistochemical and molecular changes. DNBS-treated Fpr1 KO mice showed a i) reduction in weight loss, ii) lower extent of colon injury and iii) an increase in MPO activity. Molecular analyses indicated that in absence of Fpr1 there was reduced NF-κB translocation into the nucleus, cytokines levels, FOXP3 and GATA3, CD4, CD8 and CD45 expression as well as a dysregulation of TGF-β signalling. In addition, the colon of DNBS-injected Fpr1 KO mice displayed a lower degree of expression of Bax and higher expression of Bcl-2 compared correspondent WT mice. Finally, intravital microscopy investigation of the microcirculation post-DNBS instillation revealed a lower degree of neutrophil-endothelial cell rolling and adhesion - mediated by P-selectin and ICAM-1 – in Fpr1 KO mice. All the main outcome in the study have a P-value, statistical significance of evidence, less than 0.05. We provide evidence for an important pathogenic role of mouse Fpr1 in experimental colitis, an outcome effected through modulation of immune cell recruitment together with a modulation of local cellular activation and survival.

Morphological, immunohistochemical and molecular features of inflammatory bowel disease associated colorectal carcinoma and associated mucosal lesions – Single institution experience

BackgroundPatients with inflammatory bowel disease (IBD) – ulcerative colitis (UC) and Crohn’s disease (CD) have an elevated risk of developing colorectal carcinoma (CRC). Major risk factor in IBD patients is the continuous chronic inflammation leading to development of dysplasia and carcinoma. Nevertheless, other types of non-conventional but suspicious mucosal changes serrated change/dysplasia, NOS and villous hypermucinous change, have also been reported in IBD patients. Preneoplastic potential of these lesions is still not well elucidated. AimsThe aim of this study was identification of IBD-associated CRCs focusing on finding related precursor lesions in the surgical specimen or in archival biopsy samples followed by a detailed morphological, immunohistochemical and molecular evaluation. For the purpose of the study the mucosal lesions were divided into conventional IBD-associated dysplasia and non-conventional lesions that were merged under a provisory term of putative preneoplastic lesions (PPL). MethodsA total of 309 consecutive IBD colectomy specimens diagnosed during a 10-year period were reviewed. Detailed morphological evaluation, immunohistochemical analysis of mismatch repair (MMR) proteins, p53 and O6-methylguanine DNA methyltransferase (MGMT) expression and molecular analysis for KRAS, NRAS and BRAF gene mutation were performed in the retrieved CRC cases as well as in the detected dysplasia and PPLs of these patients. ResultsWe identified 11 cases of morphologically heterogenous IBD-associated CRCs, occurring in 5 males and 6 females, aged 26–79 years (mean 44 years). A total of 22 mucosal lesions were revealed in 8 CRC patients comprising conventional IBD-associated dysplasia (4 lesions), PPLs as serrated change/dysplasia NOS (11 lesions), villous hypermucinous change (5 lesions), and two true serrated lesions (one sessile serrated adenoma and one traditional serrated adenoma). More than one type of lesion was found in 6 patients. Seven CRC cases harbored mutation of KRAS/NRAS and one case of BRAF. Two patients with KRAS-mutated CRC showed the same mutation in PPL in the same specimen (one serrated change NOS and one TSA with high-grade dysplasia). Similarly, one BRAF-mutated carcinoma case presented the same mutation in serrated change/dysplasia, NOS in the same specimen. Of the CRCs, two showed deficient MMR system profile, six presented with loss of MGMT expression, and six showed aberrant p53 expression. PPLs showed deficient MGMT expression (14 cases) and aberrant p53 (10 cases) as well. ConclusionIBD-associated CRCs are very heterogeneous entities. Besides conventional IBD-related dysplasia, other types of mucosal lesions may be associated with long lasting IBD and CRC e.g. villous hypermucinous change and serrated change/dysplasia, NOS. Since these lesions share certain genetic or immunohistochemical changes with the related CRC, a suspicion is raised that these lesions may also have preneoplastic potential. Awareness of these changes is necessary to prevent their missing and under-reporting, and further studies of these lesions should be carried out.