Immunoreactivity In Tissues Research Articles

Loss of microfibril-associated protein 5 (MFAP5) expression in colon cancer stroma.

MFAP5, a 25-kD microfibril-associated glycoprotein that is involved in elastic microfibril assembly, has been demonstrated to be significantly downregulated in tumor stroma by previous gene expression study. The aim of this study was to confirm the reduced expression of MFAP5 in colonic tumor stroma using immunohistochemistry and to explore the utility of MFAP5 as a marker to facilitate diagnosing an invasive component versus pseudoinvasion in colon polyps. In all 19 colon cancer resection cases evaluated, while there was intact MFAP5 immunoreactivity in the uninvolved normal connective tissue, there was marked reduction of MFAP5 immunoreactivity in the desmoplastic stroma surrounding the invasive component. The difference in MFAP5 expression levels was most pronounced within the tumor, while a more heterogeneous expression pattern was observed at the tumor invasive front. Reduction of MFAP5 staining was also observed in the stroma around mucin pools in 6 out of 9 sections from mucinous adenocarcinomas and in areas with high-grade dysplasia. For the polypectomy cases, intact expression of MFAP5 was seen in the stroma surrounding the displaced adenomatous glands in 9 out of 12 polyps with pseudoinvasion. Loss of expression of MFAP5 was observed in the stroma surrounding small foci of invasive adenocarcinoma in 8 of 10 malignant polyps. MFAP5 is a useful marker that may help distinguish normal connective tissue from stroma within invasive colonic adenocarcinoma. MFAP5 may facilitate the distinction between pseudoinvasion and true invasive cancer in colonic adenomatous polyps with a sensitivity of 80% (confidence interval 44-96%) and a specificity of 75% (confidence interval 43-93%) in this small cohort.

Race-associated expression of MHC class I polypeptide-related sequence A (MICA) in prostate cancer

Despite the lack of a complete understanding of the disparities involved, prostate cancer (PCa) has both higher incidence and death rates in African American Men (AAM) relative to those of Caucasian American Men (CAM). MHC class I polypeptide related sequence A (MICA) is an innate immunity protein involved in tumor immunoevasion. Due to a lack of reports of race-specific expression of MICA in PCa, we evaluated MICA expression in patients' tumors and in cell lines from a racially diverse origin. Immunohistochemistry was done on a tissue microarray (TMA) with antibodies against MICA. Tumor MICA mRNA was assessed by data mining using Oncomine and PROGeneV2. Surface MICA and release rate of soluble (s) MICA was evaluated in PCa cell lines originally derived from African American (MDA-PCa-2b) or Caucasian (LNCaP and DU-145) PCa patients. Prostate tumor tissue had a 1.7-fold higher MICA expression relative to normal tissue (p < .0001). MICA immunoreactivity in PCa tissue from AAM was 24% lower (p = .002) compared to CAM. Survival analysis revealed a marginal association of low MICA with poor overall survival (OS) (p = .058). By data mining analysis, a 2.9-fold higher level of MICA mRNA was evidenced in tumor compared to normal tissue (p < .0001). Tumors from AAM had 24% lower levels of MICA mRNA compared to tumors from CAM (p = .038), and poor prognosis was found for patients with lower MICA mRNA (p = .028). By flow cytometry analysis, cell fraction positive for surface MICA was of 3% in MDA-PCa-2b cells, 54% in DU-145 cells, and 67% in LNCaP cells (p < .0001). sMICA was detected in DU-145 and LNCaP cells, but was not detected in MDA-PCa-2b cells. Both LNCaP and DU-145 cells were sensitive to cytolysis mediated by Natural killer (NK) cells. MDA-PCa-2b cells, however were between 1.3-fold at 10:1 Effector:Target (E:T) ratio (p < .0001) and 2-fold at 50:1 E:T ratio (p < .0001) more resistant to NK-mediated cytolysis relative to cells from Caucasian origin.These results suggest that MICA expression may be related to the aggressive nature of PCa. Our findings also demonstrate for the first time that there are variations in MICA expression in the context of racial differences. This study establishes a rationale for further investigation of MICA as a potential race-specific prognostic marker in PCa.

Role of insulin-like growth factor-1 in skin tags: a clinical, genetic and immunohistochemical study in a sample of Egyptian patients.

Background: Skin tags (STs) are benign connective tissue neoplasms, in which insulin-like growth factor −1 (IGF-1) has a mitogenic and antiapoptotic activity.Purpose: We aimed to study for the first time, the possible role of IGF-1 (CA) 19 and rs6214 gene polymorphisms, and its tissue immunoreactivity in the pathogenesis of STs.Patients and methods: This case–control study included 40 ST patients and 20 controls. We searched for (CA) 19 single-nucleotide polymorphism (SNP) using conversional PCR and for rs6214 gene polymorphism using real-time PCR. IGF-1 tissue immunoreactivity was investigated using polyclonal IGF-1 antibody.Results: IGF-1 immunoreactivity showed significantly strong upregulation in epidermis (p=0.002) and dermal components (endothelial cells [p=0.038] and fibroblasts [p=0.004]) of excised STs than control skin. TT and CT rs6214 genotypes and its T allele were significantly associated with STs (p=0.006 and P=0.002, respectively). Also (<192 bp) and 192–194 bp (CA) 19 genotypes were significantly predominant in ST patients than controls (p=0.013). These 4 genotypes were significantly associated with development of multiple STs and epidermal IGF-1 tissue immunoreactivity in studied patients.Conclusions: IGF-1 (CA) 19 and rs6214 gene polymorphisms may contribute to a predisposition of STs in Egyptian patients, the role of which could be mediated through local upregulation of IGF-1 in cutaneous tissues.

Immunostaining characteristics of irisin in benign and malignant renal cancers

ABSTRACTWe investigated the expression of irisin in renal cancers using immunocytochemistry. Irisin has been reported to exhibit anticancer properties. The study groups consisted of 22 cases each of control renal tissue, oncocytoma, chromophobe renal cell carcinoma (RCC), clear cell RCC (Fuhrman nuclear grades 1, 2, 3 and 4) and papillary RCC. We evaluated 10 slides for each of 176 cases. Slides were immunostained for irisin and histoscores were calculated for the prevalence and strength of immunostaining. Fuhrman nuclear grade 1, 2, 3 clear cell RCC and papillary RCC exhibited no irisin immunoreactivity. Irisin immunoreactivity was observed in some Fuhrman nuclear grade 4 RCCs. We found a significant decrease in irisin staining in chromophobe RCC compared to the control. Immunoreactivity in the oncocytoma tissue was comparable to the control group. Irisin immunoreactivity in chromophobe RCC decreased and no immunoreactivity was observed in Fuhrman nuclear grade 1, 2, 3 clear cell RCC and papillary RCC. Immunistochemical screening of irisin in renal oncocytomas and renal cancers may be useful for differential diagnosis.

The Effects of N-Acetylcysteine on MMP-2 and MMP-9 Immune Activities in Testicular Tissue of Streptezotocin Induced Diabetic Rats

Objective: This study was performed to investigate the effects of N-Acetylcysteine (NAC) on matrix metalloproteinases (Mmp-2 and Mmp-9) immunoreactivity in testicular tissue of diabetic rats. Materials and Methods: 28 male rats were allocated into four groups n (7); No treatment was applied to Control group. Diabetes was induced upon injection of a single dose streptozocin 50 mg/kg intraperitoneally on diabetes group (DM). N-Acetylcysteine group (NAC) and following diabetes development, Diabetic + NAC groups were treated with i.p.100 mg/kg NAC daily. Oxidative damage was evaluated with Total Antioxidant Status (TAS) and Total Oxidant Status (TOS) activities while the testicular damage was determined by histopathological evaluation and immunohistochemical assessment of MMP-2 and MMP-9 at the testicular tissues. Results: TAS levels were found to be increased in diabetic NAC-treated group animals whereas TOS, MMP-2 and MMP-9 levels were decreased in the same group. For the histological findings, there were no testicular changes In the NAC alone control group whereas the alterations such as marked degeneration, vacuole formation and basement membrane thickening of tubules seminiferus contortus were observed in the testicular tissues of the DM group. But, in the treatment group DM+NAC, these alterations were found to be comparatively decreased. Conclusion: Our findings suggest that administration of NAC minimize testicular damage in diabetic rats and might be a potential candidate to reduce/eliminate the negative effects of diabetes on the testicular tissue.

Cu/Zn superoxide dismutase enzyme immunoreactivity in the stomach tissue of rats fed with mussels (Mytilus galloprovincialis)

Objectives: Mussels accumulate heavy metals in their tissues. Although there are few data about the toxicity of seafood that is exposed to environmental pollution, there are no animal studies about the gastric toxicity of mussels grown in the Dardanelles. The antioxidant Cu/Zn superoxide dismutase (Cu/Zn SOD) enzyme catalyzes the hydrogen peroxide dismutation of superoxide radicals and removes the effects of free radicals which cause oxidative stress. The purpose of the study was to demonstrate the Cu/Zn SOD in the stomach tissues of rats which are fed with mussels that are collected from the Çamburnu region of the Dardanelles. Methods: A total of 24 male Wistar albino rats were randomly divided into four groups: Group 1 (n = 6), control group fed with standard rat food; Group 2 (n = 6), 75% mussels and 25% standard rat food daily; Group 3 (n = 6), 75% mussels and 25% standard rat food every two days; and Group 4 (n = 6), 75% mussels and 25% standard rat food every three days. To detect Cu/Zn SOD localization in the tissues, the LAB-SA Detection System was used. Results: Cu/Zn SOD enzyme immunoreactivity was not detected in Group 1 and in samples without Cu/Zn SOD primer antibody. Cu/Zn SOD enzyme immunoreactivity was detected 82% in Group 2, 79% in Group 3, and 61% in Group 4. There was statistically significant difference between the Cu/Zn SOD immunoreactivity of epithelial cells in the gastric mucosa of the rats in the experimental and control groups (p &amp;lt; 0.05). Conclusions: Determination of the increase of Cu/Zn SOD enzyme in the gastric mucosa of mussel-fed rats that we used in our study suggests that it triggers the antioxidant defense mechanism against mussel toxicity.

Comparison of irisin hormone expression between thyroid cancer tissues and oncocytic variant cells.

Objective: The incidence of thyroid cancer has been continuously increasing. The main objective of this study was to investigate irisin expression in various thyroid pathologies and to compare these expression patterns with irisin expression in healthy thyroid tissues.Methods: The study groups consisted of 20 cases each of control thyroid tissue, Hashimoto’s thyroiditis, thyroid papillary carcinoma, oncocytic papillary carcinoma, follicular thyroid carcinoma, oncocytic follicular thyroid carcinoma, medullary thyroid carcinoma, anaplastic thyroid carcinoma. Irisin expression was evaluated using immunohistochemistry. Irisin levels in thyroid tissue supernatants were measured using ELISA.Results: Patients with HT showed increased irisin expression compared with controls (p<0.05). In addition, mild immunoreactivity was observed in the thyroid tissues of patients with papillary carcinoma while significantly increased irisin immunoreactivity was observed tissues of patients with oncocytic papillary carcinoma (p<0.05). There was no difference in irisin immunoreactivity in thyroid tissues between patients with follicular carcinoma and controls. However, irisin immunoreactivity was higher in tissues of patients with oncocytic follicular carcinoma than in tissues of patients with follicular carcinoma (p<0.05). No irisin immunoreactivity was observed in tissues of patients with medullary carcinoma, a malignant tumor the thyroid; however, irisin expression was significantly increased in tissues of patients with anaplastic carcinoma compared with that in tissues of controls (p<0.05). Furthermore, in all thyroid tissues with irisin expression, irisin immunoreactivity was observed in follicular cells, indicating that irisin is produced by these cells.Conclusion: Irisin is a novel potential immunohistochemical marker for differentiating oncocytic variants of papillary and FTCs from papillary and follicular thyroid cancers.

Pathology and immunohistochemical evaluation of Vibrio alginolyticus infection in Macrobrachium rosenbergii

Vibriosis is one of the common bacterial diseases plaguing the prawn and shrimp industry. However, the sequential histopathological changes associated with Vibrio alginolyticus-induced vibriosis is scanty in literature. This study investigated the histological alterations in groups of Macrobrachium rosenbergii experimentally injected intramuscularly between the second and third abdominal segment with 50 μL of 107 CFU of V. alginolyticus. Twenty-four hours post inoculation, the organism was isolated and molecularly characterized while infected giant freshwater prawns were observed for histological alterations and immunoreactivity in different tissues due to the experimental challenge. The lesions observed in the organs include incipient haemocytic infiltration into the interstitial space of the tubules of the hepatopancreas and loss of epithelial layer, muscular necrosis and disruption of muscular layer with haemocytic infiltration, hyperplasia of epithelial cells and degeneration of the epithelium, deformed and necrotizing lamellae, and enlargement of the lamellar sinus of the gill, and the heart is characterized by localized nodular haemocytic reaction and melanization. Organs such as the heart, gill, hepatopancreas and muscle following injection of the putative bacteria showed immunohistochemical activity. Hence, histology and immunohistochemistry still remain important tools for vibriosis disease diagnoses in giant freshwater prawns.

Tau immunoreactivity in peripheral tissues of human aging and select tauopathies

Many studies have been directed at understanding mechanisms of tau aggregation and therapeutics, nearly all focusing on the brain. It is critical to understand the presence of tau in peripheral tissues since this may provide new insights into disease progression and selective vulnerability. The current study sought to determine the presence of select tau species in peripheral tissues in elderly individuals and across an array of tauopathies. Using formalin fixed paraffin embedded sections, we examined abdominal skin, submandibular gland, and sigmoid colon among 69 clinicopathologically defined cases: 19 lacking a clinical neuropathological diagnosis (normal controls), 26 progressive supranuclear palsy (PSP), 21 Alzheimer’s disease (AD), and 3 with corticobasal degeneration (CBD). Immunohistochemistry was performed using antibodies for “total” tau (HT7) and two phosphorylated tau species (AT8 and pT231). HT7 staining of abdominal skin revealed immunoreactivity of potential nerve elements in 5% of cases (1 AD, 1 AD/PSP, and 1 CBD out of 55 cases examined); skin sections lacked AT8 and pT231 immunoreactive nerve elements. Submandibular glands from all cases had HT7 immunoreactive nerve elements; while pT231 was present in 92% of cases, and AT8 in only 3 cases (2 AD and one AD/PSP case). In sigmoid colon, HT7 immunoreactivity was present in all but 2 cases (97%), pT231 in 54%, and AT8 was present in only 5/62 cases (8%). These data suggest select tau species in CNS tauopathies do not have a high propensity to spread to the periphery and this may hold clues for the understanding of CNS tau pathogenicity and vulnerability.

HDAC2 dysregulation in the nucleus basalis of Meynert during the progression of Alzheimer's disease.

Alzheimer's disease (AD) is characterized by degeneration of cholinergic basal forebrain (CBF) neurons in the nucleus basalis of Meynert (nbM), which provides the major cholinergic input to the cortical mantle and is related to cognitive decline in patients with AD. Cortical histone deacetylase (HDAC) dysregulation has been associated with neuronal degeneration during AD progression. However, whether HDAC alterations play a role in CBF degeneration during AD onset is unknown. We investigated global HDAC protein levels and nuclear HDAC2 immunoreactivity in tissue containing the nbM, changes and their association with neurofibrillary tangles (NFTs) during the progression of AD. We used semi-quantitative western blotting and immunohistochemistry to evaluate HDAC and sirtuin (SIRT) levels in individuals that died with a premortem clinical diagnosis of no cognitive impairment (NCI), mild cognitive impairment (MCI), mild/moderate AD (mAD) or severe AD (sAD). Quantitative immunohistochemistry was used to identify HDAC2 protein levels in individual cholinergic nbM nuclei and their colocalization with the early phosphorylated tau marker AT8, the late-stage apoptotic tau marker TauC3 and Thioflavin-S, amarker of β-pleated sheet structures in NFTs. In AD patients, HDAC2 protein levels were dysregulated in the basal forebrain region containing cholinergic neurons of the nbM. HDAC2 nuclear immunoreactivity was reduced in individual cholinergic nbM neurons across disease stages. HDAC2 nuclear reactivity correlated with multiple cognitive domains and with NFT formation. These findings suggest that HDAC2 dysregulation contributes to cholinergic nbM neuronal dysfunction, NFT pathology, and cognitive decline during clinical progression of AD.

HNK-1 in Morphological Study of Development of the Cardiac Conduction System in Selected Groups of Sauropsida.

Human natural killer (HNK)-1 antibody is an established marker of developing cardiac conduction system (CCS) in birds and mammals. In our search for the evolutionary origin of the CCS, we tested this antibody in a variety of sauropsid species (Crocodylus niloticus, Varanus indicus, Pogona vitticeps, Pantherophis guttatus, Eublepharis macularius, Gallus gallus, and Coturnix japonica). Hearts of different species were collected at various stages of embryonic development and studied to map immunoreactivity in cardiac tissues. We performed detection on alternating serial paraffin sections using immunohistochemistry for smooth muscle actin or sarcomeric actin as myocardial markers, and HNK-1 to visualize overall staining pattern and then positivity in specific myocyte populations. We observed HNK-1 expression of various intensity distributed in the extracellular matrix and mesenchymal cell surface of cardiac cushions in most of the examined hearts. Strong staining was found in the cardiac nerve fibers and ganglia in all species. The myocardium of the sinus venosus and the atrioventricular canal exhibited transitory patterns of expression. In the Pogona and Crocodylus hearts, as well as in the Gallus and Coturnix ones, additional expression was detected in a subset of myocytes of the (inter)ventricular septum. These results support the use of HNK-1 as a conserved marker of the CCS and suggest that there is a rudimentary CCS present in developing reptilian hearts. Anat Rec, 302:69-82, 2019. © 2018 Wiley Periodicals, Inc.

Alteration of surfactant protein A expression in renal cell carcinoma

Surfactant protein-A (SP-A) belongs to a family of collagen-containing C-type lectins called collectins. SP-A is expressed by renal tubule epithelial cells. We investigated the distribution of SP-A in renal cell carcinomas (RCC) using immunohistochemical techniques and western blotting. We used 35 formalin fixed, paraffin embedded (FFPE) RCC tissue samples. We compared results with clinico-pathological parameters of RCC including age, sex, Fuhrman grade, tumor volume, tumor node metastasis (TNM) and clinical stage. SP-A was localized in the glomerulus and renal tubule epithelium in nontumor tissue and strong SP-A immunoreactivity was observed in tumor tissue. SP-A was expressed in the RCC tumor cells (64%) and nontumor cells (34%) in males and RCC tumor cells (90%) and nontumor cells (30%) in females. There was a significant correlation between SP-A immunoreactivity in tumor cells and gender, age, tumor diameter, Fuhrman grade and tumor diameter. Western blot analysis supported the immunohistochemical findings. We present evidence for involvement of SP-A in RCC and suggest that increased SP-A expression in RCC is associated with favorable prognosis.

Cognitive Deficits Are Attenuated in Neuroglobin Overexpressing Mice Exposed to a Model of Obstructive Sleep Apnea.

Background: Obstructive sleep apnea (OSA) is a highly prevalent disease manifesting as intermittent hypoxia during sleep (IH) and is increasingly recognized as being independently associated with neurobehavioral deficits. These deficits may be due to increased apoptosis in the hippocampus and cerebral cortex, as well as increased oxidative stress and inflammation. It has been reported that neuroglobin (Ngb) is upregulated in response to hypoxia-ischemia insults and exhibits a protective role in ischemia-reperfusion brain injury. We hypothesized that transgenic overexpression of Ngb would attenuate spatial learning deficits in a murine model of OSA.Methods:Wild-type mice and Ngb overexpressing male mice (Ngb-TG) were randomly assigned to either IH or room air (RA) exposures. The effects of IH during the light period on performance in a water maze spatial task were assessed, as well as anxiety and depressive-like behaviors using elevated plus maze (EPM) and forced swim tests. Cortical tissues from all the mice were extracted for biochemical studies for lipid peroxidation.Results:Ngb TG mice exhibited increased Ngb immunoreactivity in brain tissues and IH did not elicit significant changes in Ngb expression in either Ngb-TG mice or WT mice. On a standard place training task in the water maze, Ngb-TG mice displayed preserved spatial learning, and were protected from the reduced spatial learning performances observed in WT mice exposed to IH. Furthermore, anxiety and depression levels were enhanced in WT mice exposed to IH as compared to RA controls, while alterations emerged in Ngb-TG mice exposed to IH. Furthermore, WT mice, but not Ngb-TG mice had significantly elevated levels of malondialdehyde in cortical lysates following IH exposures.Conclusions:In a murine model of OSA, oxidative stress responses and neurocognitive and behavioral impairments induced by IH during sleep are attenuated by the neuroprotective effects of Ngb.

HOXB4 Immunoreactivity in Endometrial Tissues From Women With or Without Endometriosis.

Endometriosis is a common disease characterized by the presence of ectopic endometrial tissue. Although the pathogenesis of endometriosis remains unclear, several factors have been implicated, including the dysregulation of homeobox ( HOX) genes. Our objective was to investigate the localization and immunoreactivity of HOXB4 in endometrial tissues from women with or without endometriosis. We studied samples of eutopic endometrium (EE), endometriomas (Eoma), superficial endometriosis (SE), and deep infiltrating endometriosis (DIE) from 34 women with endometriosis, as well as eutopic endometrium from 38 women without endometriosis (EC). HOXB4 localization and immunoreactivity was assessed using immunohistochemistry and histoscore analysis. Data were analyzed with and without stratification by menstrual cycle phase. HOXB4 protein was present in the nuclei of endometrial glandular epithelial cells but not in stromal cells. HOXB4 immunoreactivity was reduced in DIE samples compared to all other groups. A smaller reduction in HOXB4 immunoreactivity was observed in SE samples compared to EC samples. HOXB4 immunoreactivity was significantly greater in proliferative compared to secretory phase samples in the EC group but not in EE, Eoma, or DIE groups. Among only proliferative phase samples, HOXB4 immunoreactivity was reduced in EE, Eoma, and DIE groups compared to EC. Based on these data, we suggest that an impaired capacity of eutopic and ectopic endometrial tissue to upregulate levels of HOXB4 during the proliferative phase may play a role in the pathogenesis of endometriosis and that further downregulation of HOXB4 may enhance ectopic implant invasiveness.

INOS IMMUNOREACTIVITY IN THE OVARIES OF RATS FEEDING WITH MUSSEL (Mytilus galloprovincialis)

The ovary is an endocrine and exocrine gland that plays primary role in female development and reproductive activities. The seamen are collecting a lot of heavy metals by serving as filters in the seas. Since the last decades of the nineteenth century and until today, the study of the consequences of human exposure to heavy metals has risen as a central research area in the toxicological field. Among the group of metals with proven human toxicity aluminium (Al) and lead (Pb) are known to be highly neurotoxic. The Mediterranean mussel (Mytilus galloprovincialis) is a species of bivalve, a marine mollusc in the family Mytilidae. In our previous researches, we found Al, zinc (Zn) and iron (Fe) values were higher in mussels taken from Çamburnu region of the Dardanelles. In many tumors, inducible nitric oxide synthase (iNOS) expression is high, however, the role of iNOS during tumor development is very complex and quite perplexing, with both promoting and inhibiting actions having been described. The purpose of the study is to demonstrate the iNOS immunoreactivity in the ovarian tissues of rats which are fed with mussels that are collected from the Çamburnu region of the Dardanelles. The mussels given as food to the rats were removed from the Çamburnu region of the Dardanelles. Average 100±10 g weight were selected. After the beaks were overcooked, the meat broke off and the meat at 100 degrees was dried. Two groups of rats are included in the study, group 1 (n=6), control group fed with standard rat food, group 2 (n=6), 90% mussels and 10% standard rat food daily. To detect iNOS localization in the tissues, the LAB-SA Detection System was used. iNOS immunoreactivity was detected in the interstitial cell cytoplasm of the ovaries of rats fed with mussel. iNOS immunoreactivity was also observed in germ cell cytoplasm in some primordial follicles. There was statistically significant difference between the iNOS immunoreactivity of the interstitial cells in the ovarian parenchyma of the rats in the experimental and control groups (p&gt; 0.05).

INOS IMMUNOREACTIVITY IN THE OVARIES OF RATS FEEDING WITH MUSSEL (Mytilus galloprovincialis)

The ovary is an endocrine and exocrine gland that plays primary role in female development and reproductive activities. The seamen are collecting a lot of heavy metals by serving as filters in the seas. Since the last decades of the nineteenth century and until today, the study of the consequences of human exposure to heavy metals has risen as a central research area in the toxicological field. Among the group of metals with proven human toxicity aluminium (Al) and lead (Pb) are known to be highly neurotoxic. The Mediterranean mussel (Mytilus galloprovincialis) is a species of bivalve, a marine mollusc in the family Mytilidae. In our previous researches, we found Al, zinc (Zn) and iron (Fe) values were higher in mussels taken from Camburnu region of the Dardanelles. In many tumors, inducible nitric oxide synthase (iNOS) expression is high, however, the role of iNOS during tumor development is very complex and quite perplexing, with both promoting and inhibiting actions having been described. The purpose of the study is to demonstrate the iNOS immunoreactivity in the ovarian tissues of rats which are fed with mussels that are collected from the Camburnu region of the Dardanelles. The mussels given as food to the rats were removed from the Camburnu region of the Dardanelles. Average 100±10 g weight were selected. After the beaks were overcooked, the meat broke off and the meat at 100 degrees was dried. Two groups of rats are included in the study, group 1 (n=6), control group fed with standard rat food, group 2 (n=6), 90% mussels and 10% standard rat food daily. To detect iNOS localization in the tissues, the LAB-SA Detection System was used. iNOS immunoreactivity was detected in the interstitial cell cytoplasm of the ovaries of rats fed with mussel. iNOS immunoreactivity was also observed in germ cell cytoplasm in some primordial follicles. There was statistically significant difference between the iNOS immunoreactivity of the interstitial cells in the ovarian parenchyma of the rats in the experimental and control groups (p> 0.05).

Identification of immunohistochemical localization of irisin in the dwarf hamster (Phodopus roborovskii) tissues.

Irisin is mainly secreted by heart and skeletal muscle cells. It is an exercise-induced protein that converts white adipose tissue to brown. Increased irisin expression was lead to weight loss and improved glucose tolerance. We investigated irisin immunoreactivity in various tissues of the dwarf hamsters (Phodopus roborovskii). Tissues were processed, embedded in paraffin, sectioned at 5μm and stained immunohistochemically for irisin. In the retina, irisin was found almost all layers, except outer nuclear layer. Also, irisin immunoreactivity was observed in the skin, cornea, striated muscle, parotid gland, tongue, oesophagus, stomach and small intestine. The findings from this study support the notion that skeletal muscle is not the primary source of irisin.

Nuclear pseudoinclusions in melanoma cells: prognostic fact or artifact? The possible role of Golgi phosphoprotein 3 overexpression in nuclear pseudoinclusions generation.

Nuclear pseudoinclusions (NPIs) are classically found in papillary thyroid carcinoma and meningioma. Although NPIs have been described in melanocytic lesions, there is no systematic analysis of potential relationship between NPIs and other clinicopathological characteristics of melanoma. We examined the presence of NPIs in H&E-stained tissue sections form 96 melanomas and analyzed statistical associations with important clinicopathological parameters and tissue immunoreactivity for selected proteins involved in epithelial-mesenchymal transition (SPARC, N-cadherin), cell adhesion and mobility (ALCAM, ADAM-10), regulation of mitosis (PLK1), cell survival (FOXP1) and functioning of Golgi apparatus (GOLPH3, GP73). NPIs were observed in 20% of melanomas and their presence correlated with high mitotic rate and ulceration of the tumor, but not with Breslow thickness, histologic type, or presence of metastases. We observed a significant correlation with shorter cancer-specific survival, but not disease-free survival. Presence of NPIs was related to high expression of GOLPH3 in melanoma cells, whereas their absence was linked to enhanced immunoreactivity of GOLPH3 in tumor-associated macrophages. NPIs are not an uncommon finding in skin melanoma and their diagnostic and prognostic utility could be helpful in the daily routine histopathological practice. The possible explanation of NPI generation is associated with enhanced activity of Golgi apparatus in melanoma cells.

High inducible nitric oxide synthase in prostate tumor epithelium is associated with lethal prostate cancer

Objective: The aim of this study was to investigate the role of inducible nitric oxide synthase (iNOS) in lethal prostate cancer (PCa) by studying the iNOS immunoreactivity in tumor tissue from men diagnosed with localized PCa.Materials and methods: This study is nested within a cohort of men diagnosed with incidental PCa undergoing transurethral resection of the prostate (the Swedish Watchful Waiting Cohort). To investigate molecular determinants of lethal PCa, men who died from PCa (n = 132) were selected as cases; controls (n = 168) comprised men with PCa who survived for at least 10 years without dying from PCa during follow-up. The immunoreactivity of iNOS in prostate tumor epithelial cells and in cells of the surrounding stroma was scored as low/negative, moderate or high. Logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals (95% CIs) for lethal PCa according to iNOS category.Results: There was no association between iNOS immunoreactivity in stroma and lethal disease. However, when comparing high versus low/negative iNOS immunoreactivity in epithelial cells, the OR for lethal PCa was 3.80 (95% CI 1.45–9.97).Conclusion: Patients with localized PCa have variable outcomes, especially those with moderately differentiated tumors. Identifying factors associated with long-term PCa outcomes can elucidate PCa tumor biology and identify new candidate prognostic markers. These findings support the hypothesis that high iNOS in tumor epithelium of the prostate is associated with lethal disease.

Elevated TERT Expression in TERT-Wildtype Adult Diffuse Gliomas: Histological Evaluation with a Novel TERT-Specific Antibody.

Telomerase reverse transcriptase (TERT) is important for the biology of diffuse gliomas. TERT promoter mutations are selectively observed among 1p/19q-codeleted oligodendrogliomas and isocitrate dehydrogenase gene- (IDH-) wildtype glioblastoma (GBM). However, TERT transcripts range widely in various cancers including gliomas, and TERT protein expression has been rarely investigated thus far. It would be thus critical to examine the expression level of TERT in tumors in addition to its mutational status, and sensitive and specific methods are urgently needed to examine TERT protein expression for the assessment of TERT biology in gliomas. Using our newly developed TERT-specific monoclonal antibody (TMab-6) applicable to human tissue, we found an unexpected increase in TERT expression in TERT-wildtype as well as TERT-mutated gliomas and in tumor vasculature. This is the first extensive analysis on the expression of TERT immunoreactivity in human glioma tissue, suggesting that TERT protein expression may be regulated by several mechanisms in addition to its promoter mutation.