Positive Immunoreactivity Research Articles

CD30 Expression and Its Correlation with Clinicopathologic Features in Indonesian Diffuse Large B-Cell Lymphoma

Background: Diffuse large B-cell lymphoma (DLBCL) is the most common subtype (68.2%) of B cells Non Hodgkin Lymphoma in Indonesia. This tumor heterogeneity is characterized by a variety of clinical conditions, morphology, genetic profiles, therapeutic response, prognosis and survival. Recent studies have shown that CD30 immunohistochemical staining also plays an important role in determining the therapy and prognosis of DLBCL disease. CD30 can also be expressed in DLBCL in approximately 9.5-40%. However, CD30 expression and clinicopathological characteristics of Indonesian DLBCL remain unknown. This study aimed to determine the prevalence of CD30 expression and its correlation with clinicopathological characteristics of Indonesian DLBCL patients. Methods: During a study period of four years, a total of 104 FFPE of DLBCL cases were collected from Anatomical Pathology Department, Sardjito Hospital, Special Region of Yogyakarta, Indonesia. CD30 expression was studied using immunohistochemical techniques (Mouse monoclonal antibody MoAb CD30 cell marque Ber-H2). Correlations between positive CD30 immunoreactivity and clinicopathological characteristics in DLBCL patients were statistically analyzed using chi-square tests. Result: Positivity rate of CD30 expression in 104 DLBCL samples was 13.5% (14/104) using cutoff value of 0% while using a 20% cutoff, it was 1.9% (2/104). Statistical associations of positive CD30 expression and clinicopathological characteristics (age, sex, Ann Arbor stage, extranodal involvement and morphological variants) were not significant (p > 0.05).Conclusions: The prevalence of positive CD30 expression in Indonesian DLBCL patients is 13.5%. There was no statistically significant associations between positive CD30 expression and clinicopathological characteristics.

POSSIBLE TOXICOLOGICAL EFFECTS ASSOCIATED WITH CHRONIC DERMAL EXPOSURE TO PARA-PHENYLENEDIAMINE (PPD) ON PANCREAS OF ADULT MALE ALBINO RATS

Paraphenylenediamine (PPD) is a chemical compound that is commonly used as dark cosmotics and in hair coloring. The aim of this work is to evaluate toxic effect of chronic dermal exposure to PPD on pancreas. Methods: Forty adult male albino rats were used in this study divided into; control group (10 rats) painted on their dorsal sides with 2 ml distilled water and PPD- exposed group (30 rats) was divided into 3 groups; groups I, II and III which were exposed to PPD dermally by painting their dorsal sides with 1, 2, and 3 mg/kg of PPD respectively (dissolved in 2 ml distilled water) daily for 12 weeks. At the end of the study, all rats were tested for intraperitoneal glucose tolerance test (IPGTT), Hemoglobin A1c (HbA1C) and plasma insulin levels. Histopathological examination and immunohistochemical studies for PCNA and insulin were also performed. Results: The results revealed significant glucose intolerance with decreased level of plasma insulin level in PPD-exposed rats when compared with control group. On histopathological examination, PPD groups showed vacuolations in cells of islets of Langerhans, irregular distorted acini, and congested blood vessels. Immunohistochemical staining revealed weak positive immunoreaction for insulin with significant decrease in area percentage of positive insulin immunoreaction and strong positive immunoreaction for PCNA in groups II and III. Conclusion: It was concluded that chronic dermal exposure to PPD resulted in dose-dependent toxic effects on pancreas, with recommendations to avoid chronic dermal exposure to hair dyes containing PPD and adherence to wear protective gloves when chronic handling of these dyes is necessary.

Clinical Routine Application of the Second-generation Neuroendocrine Markers ISL1, INSM1, and Secretagogin in Neuroendocrine Neoplasia: Staining Outcomes and Potential Clues for Determining Tumor Origin

Neuroendocrine neoplasms (NENs) have traditionally been identified via expression of proteins associated to the regulation of secretory vesicles and granules. We report the clinical usage of the “second-generation” proteins ISL LIM homeobox 1 (ISL1), INSM transcriptional repressor 1 (INSM1), and secretagogin (SECG) as immunohistochemical markers of neuroendocrine differentiation since their introduction in clinical routine and compare the results with the established proteins chromogranin A (CGA) and synaptophysin (SYP). In total, 161 tumors, including 139 NENs and 22 “non-NENs” (unrelated tumors with an initial suspicion of NEN), were informatively stained for ISL1, and subsets were also interrogated for INSM1 and/or SECG. Diffuse or focal positive immunoreactivity was noted for ISL1 in 91/139 NENs (65%) and in 6/22 (27%) non-NENs, for INSM1 in 76/85 NENs (89%) and in 2/5 (40%) non-NENs, and for SECG in 49 out of 64 NENs (77%) and in 0/5 non-NENs (0%). Generally, ISL1, INSM1, and SECG exhibited sensitivities in line with or slightly below that of CGA and SYP—largely attributable to tissue-specific patterns regarding tumoral origin. Moreover, for pancreatic and small intestinal NENs, the two largest subgroups, ISL1 staining results were consistent irrespectively of tumor source and WHO grade. We verify previously suggested immunohistochemical schemes of neuroendocrine markers of first- and second-generations to facilitate the diagnostic process for NENs and confirm that the second-generation neuroendocrine markers display tissue-specific patterns. We therefore recommend their implementation in tertiary endocrine pathology centers, not least to aid in the identification of primary tumors when analyzing metastases.

Biodistribution Analysis of Oncolytic Adenoviruses in Canine Patient Necropsy Samples Treated with Cellular Virotherapy

Oncolytic immunotherapy with competent viruses is an emerging approach in cancer treatment. The clinical safety of many types of oncolytic viruses (OVs) has been demonstrated. However, there is a lack of information about viral biodistribution in patients. The available data about oncolytic adenovirus biodistribution in human subjects treated intravenously consists of virus detection in body fluids, a few tumor biopsies, and a single report of patient necropsy samples. There is no information about adenoviral biodistribution in patients treated intravenously with cellular vehicles carrying an oncolytic adenovirus. We previously published reports regarding the efficacy and clinical safety of infusing mesenchymal stem cells (MSCs) infected with an OV in human and canine patients. In this study, we performed necropsies on 12 canine patients treated with dCelyvir, canine MSCs infected with ICOCAV17, a canine oncolytic adenovirus. The prevalence of microscopic lesions, especially chronic inflammatory responses in different organs, was higher than expected. Concomitantly, we found a positive immunoreaction to ICOCAV17 in analyzed samples. These findings support a possible role of the virus in development of histopathological alterations and ongoing systemic viral replication of ICOCAV17 in the period after therapy administration.

Abstract 5041: Inhibition of RUNX-CBFβ binding reduces RSPO3 expression and EMT features in breast cancer cells

Abstract We have recently identified R-spondin3 (RSPO3) as a novel key modulator of breast cancer development that may become a potential target for treatment of the basal subtype, which lacks efficient therapeutic options to date. RSPO3 is expressed in the basal stem cell-enriched compartment of mouse mammary glands whereas it is absent in the mature luminal cells. RSPO3 knockdown in mouse mammary tumor cells reduces canonical Wnt signaling pathway, epithelial-to-mesenchymal transition (EMT)-like features, migration capacity, and tumor formation in vivo. To evaluate the relevance of RSPO3 in human breast cancer, we analyzed samples from 75 patients with ductal infiltrating breast carcinomas, where we found a high prevalence of positive immunoreactivity for RSPO3 (70%). Expression data of The Cancer Genome Atlas Network from 1985 human breast carcinoma samples showed that RSPO3 is primarily expressed in Claudin-low breast cancer subtype, which is characterized for having stem cells features. In culture assays revealed that knockdown of RSPO3 expression reduced SOX2 expression and impaired mammary tumor cell ability to form mammospheres. Previous reports suggested the potential involvement of RUNX1-CBFβ on RSPO3 transcription in mammary tumor cells. Therefore, we evaluated if pharmacologic inhibition of RUNX1 activity would reduce RSPO3 expression and activity in the cancer stem cell compartment. Treatment of breast cancer cells with AI-10-104, a small molecule that inhibits CBFβ-RUNX interaction, reduced RSPO3 mRNA and protein levels, and reduced the migration ability of MDA-MB231 breast cancer cells, ability that was recovered upon treatment with recombinant RSPO3. In addition, AI-10-104 inhibited the expression of canonical Wnt pathway targets, EMT and stem-cell markers. Therefore, our results demonstrate that inhibition of RUNX-CBFβ transcriptional activity leads to a down-regulation of RSPO3 expression, constituting a new molecular target for therapeutic modulation in breast cancer treatment. Citation Format: Carla M. Felcher, Johanna M. Tocci, Martin E. Garcia Sola, John H. Bushweller, Lucio H. Castilla, Edith C. Kordon. Inhibition of RUNX-CBFβ binding reduces RSPO3 expression and EMT features in breast cancer cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5041.

Date palm fruit extract ameliorated pancreatic apoptosis, endocrine dysfunction and regulatory inflammatory cytokines in Streptozotocin-induced diabetes in rats.

The current work studied the mechanism(s) and ability by which date palm (Phoenix dactylifera L.) fruit extract (DPE) inspired a glucose-lowering impact in rats suffering from diabetes. Forty-eight albino rats were divided into six various experimental treatments after induction of diabetes by intraperitoneal infusion of streptozotocin (45mg/kg bwt) as follows: normal control, DPE, diabetic control, diabetic glibenclamide (GLI), diabetic DPE, and diabetic GLI plus DPE-treated groups. In animals euthanized after 8weeks, blood and pancreatic tissue samples were assembled to assess different biochemical and histopathological changes. The expressions of insulin, B cell lymphoma-2 (Bcl-2), and cysteine aspartate-specific protease-3 (caspase-3) in islet β cells were also evaluated using immunohistochemical assessment. Diabetic rats exhibited hyperglycemia; increment of pancreatic malondialdehyde (lipid peroxidation biomarker), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β); and decrement of plasma insulin and pancreatic antioxidants: glutathione, superoxide dismutase, and catalase values. Also, the pancreatic islets exhibited histopathological and morphometric alternations associated with weak positive insulin and Bcl-2 immunoreactivity and strong positive caspase-3 immunoreactivity. DPE and/or GLI, an anti-diabetic drug, improved the pancreatic histoarchitecture and improved β cell function and structure, which increased insulin levels and improved the insulin, Bcl-2, and caspase-3 immunoreactivity in diabetic rats. Nevertheless, the combined DPE and GLI therapy revealed a significant recovery and restoration of β cells' structure and function. The date palm fruit has anti-apoptotic, anti-inflammatory, and antioxidant activities and hypoglycemic effects, which in turn play a pivotal role in avoiding the progression of diabetes mellitus. Moreover, it could potentiate the glucose-lowering activity of anti-diabetic drugs.

Mapping of spinal interneurons involved in regulation of the lower urinary tract in juvenile male rats

Coordination between the urinary bladder (BL) and external urethral sphincter (EUS) is necessary for storage and elimination of urine. In rats interneuronal circuits at two levels of the spinal cord (i.e., L6-S1 and L3-L4) play an important role in this coordination. In the present experiments retrograde trans-synaptic transport of pseudorabies virus (PRV) encoding fluorescent markers (GFP and RFP) was used to trace these circuits. To examine the relative localization of EUS-related and BL-related interneuronal populations we injected PRV-GFP into the EUS and PRV-RFP into the BL wall. The PRV infected populations of spinal interneurons were localized primarily in the dorsal commissure (DCM) of L6/S1 and in a hypothesized lumbar spinal coordinating center (LSCC) in L3/L4 above and lateral to central canal (CC). At both sites colocalization of markers occurred in a substantial number of labeled interneurons indicating concomitant involvement of these double-labelled neurons in the EUS- and BL-circuits and suggesting their role in EUS-BL coordination. Intense GFP or RFP fluorescent was detected in a subpopulation of cells at both sites suggesting that they were infected earlier and therefore likely to represent first order, primary interneurons that directly synapse with output neurons. Larger numbers of weakly fluorescent neurons that likely represent second order interneurons were also identified. Within the population of EUS-related first order interneurons only 3–8 % exhibited positive immunoreaction for an early transcription factor Pax2 specific to GABAergic and glycinergic inhibitory neurons suggesting that the majority of interneurons in DCM and LSCC projecting directly to the EUS motoneurons are excitatory.

Distinct Molecular Landscape of Epstein-Barr Virus Associated Pulmonary Lymphoepithelioma-Like Carcinoma Revealed by Genomic Sequencing.

Pulmonary lymphoepithelioma-like carcinoma (LELC) is a subtype of non-small cell lung cancer (NSCLC) characterized by marked lymphocytic infiltration and association with Epstein–Barr virus (EBV). The molecular basis underlying the disease remains unclear. We sought to study the molecular landscape by multiple approaches including whole genomic sequencing, capture-based targeted sequencing, fluorescent in situ hybridization and immunohistochemistry. Tumor cells from 57 EBV-positive pulmonary LELCs were isolated by careful microdissection prior to genomic sequencing. Integrated analysis revealed a distinct genomic landscape of low TP53 mutation rate (11%), low incidence of known drivers in the RTK/RAS/RAF (11%) and PI3K/AKT/mTOR pathways (7%), but enriched for loss-of-function mutations in multiple negative regulators of the NF-κB pathway. High level programmed cell death ligand-1 (PD-L1) expression was shown with 47% and 79% of the cases showing positive PD-L1 immunoreactivity at ≥50% and ≥1% tumor proportion score, respectively. Subsets of the patients with actionable fibroblast growth factor receptor 3 (FGFR3) aberrations (4%) and mismatch repair deficiency (4%) were potentially eligible for precision medicine. Pulmonary LELC showed a distinct genomic landscape, different from major NSCLC subtypes but resembled that of EBV-associated nasopharyngeal carcinoma. Our work facilitated the understanding of molecular basis underlying pulmonary LELC to explore potential therapeutic options.

Structural Investigation of Epididymal Microvasculature and Its Relation to Telocytes and Immune Cells in Camel.

The vascular and perivascular cells, including telocytes (TCs) and immune cells, play an important role in male fertility. The current study intended to describe in detail the microvascular structures harboring special regulatory devices in addition to the interstitial cellular components of the one-humped camel epididymis. The samples were collected from 10 clinically healthy mature camels (Camelus dromedarius). The distribution and characteristics of TCs, peripheral blood vessels of the epididymis, and immune cells were investigated using the light, immunohistochemistry, immunofluorescence, and transmission electron microscopy analyses. Frequent occlusive or throttle arterioles were demonstrated in the epididymal interstitium and their tunica media consisted of glomus cells. In addition, some vein walls consisted of one or two layers of glomus cells. TCs, fibroblasts, muscle cells, and tunica media of the blood vessels, that present in the loose connective tissue surrounding the intertubular interstitium of camel epididymis, showed a positive reaction with vimentin. The endothelium of blood vessels and veins showed positive immunoreactivity for CD34 and vascular endothelial growth factor (VEGF). Furthermore, VEGF, CD34, and S100 proteins were expressed in dendritic cells (DCs) as well as TCs. The current data suggest the involvement of DCs and TCs in angiogenesis and a possible role for the interstitial components in creating an appropriate milieu for the full maturation of sperms.

Immunohistochemical detection of prolactin in clinically non-functioning pituitary adenomas.

Approximately one third of pituitary adenomas are manifested neither by specific symptoms of hormone overproduction nor by elevated blood levels of pituitary hormones. However, these tumours, diagnosed before surgical intervention as clinically non-functioning pituitary adenomas (CNFPAs) express in majority different pituitary hormones, as can be revealed by means of immunohistochemical examination. One of the pituitary hormones which may be expressed in CNFPAs is prolactin (PRL) but the clinical and pathological data on this condition are very scarce. Sixty two pituitary adenomas, diagnosed before surgery as CNFPAs, were immunoassayed with antibodies against PRL, growth hormone (GH), luteinizing hormone (LH), follicle stimulating hormone (FSH), thyrotropin (TSH), alpha subunit (alpha-SU), corticotropin (ACTH) and dopamine receptor type 2. In a proportion of the patients the presurgical concentrations of insulin-like growth factor 1 (IGF-1) were estimated by means of enzyme-amplified chemiluminescence assay. Twenty-three (37.1%) of the examined CNFPAs presented the positive immunoreaction with anti-PRL antibody. Most cases concerned women. Only in two cases (one woman and one man), PRL was the unique hormone expressed in the tumour. In the remaining adenomas PRL immunopositivity was accompanied by GH expression - 17, LH or free bLH - 13, FSH - 2, free a subunit - 4 or by ACTH - 5 tumours. Seven (30.43%) of them were recurrent in comparison with 12.8% PRL-immunonegative recurrent CNFPAs. Dopamine receptors were positively immunostained in all the investigated PRL-immunopositive and all PRL-immunonegative adenomas. Our data confirm the observations that monohormonal silent prolactinomas are very rare but frequently silent PRL often co-expressed with GH or LH. Although in the whole population of patients with CNFPAs both sexes are equally represented, in the case of silent prolactinomas the female sex is prevalent. The observation of the higher rate of recurrent tumours within PRL-immunopositive adenomas versus PRL-immunonegative CNFPAs has to be confirmed on the larger material.

Role of Heat Shock Protein 70 Expression in Identification of Death Due to Fatal Thermal Exposure

Heat shock proteins (HSPs), are a group of proteins that function as intra-cellular chaperones for other proteins. They are expressed by cells in response to exposure to stress conditions. The aim of the present study was to evaluate the role of heat shock protein 70 (HSP70) expression as vitality marker in identification death due to fatal thermal exposure (fire). This study was carried out on 21 cases of fire related fatalities (group I), 21 cases of acute traumatic death without any evidence of antemortem thermal exposure (group II). All autopsy cases of group I were examined regarding the degree of burn, carbon soot aspiration and survival time. Carboxy hemoglobin (COHb) blood level was assessed. Immunohistochemical examination of lungs, kidneys, heart and liver in all cases of both groups was done. Cases of group I showed significant strong positive HSP70 immunoreactivity (grade 3) in the lung and kidney sections either with short or long survival time exposure when compared with cases of group II. Sections from heart and liver in both groups showed weak HSP70 immunoreactivity. There was no association between the degree of HSP70 expression and degree of burn, presence or absence of carbon soot and COHb concentration in blood in group I. It was concluded that the immunohistochemical investigation of HSP 70 expression in lungs and kidneys can support the proof of vitality in death due to fatal thermal exposure.

Syndecan-1, CD117 and CD56 Immunohistochemical Evaluation of Plasmacytoma

Background and Objective: World Health Organization (WHO) describes two types of plasmacytomas which are solitary bone plasmacytoma (SBP) and extramedullary plasmacytoma (EMP). 17% to 33% of plasmacytomas, will progress into multiple myeloma (MM). The early diagnosis is very important as the plasmacytoma may be primary or metastatic lesion of MM. Prognosis of solitary plasmacytoma could be worse if it recurred as in case of its evolution toward systemic MM. Methodology: 10 patients with plasmacytoma in the National Cancer Institute and Histopathological laboratory of Minia University were analyzed immunohistochemically for detection of syndecan-1 as a diagnostic marker for plasmacytoma and detection of plasmacytoma behavior through interpretation the expression of both CD117 and CD56. Results: All cases showed strong positive immunoreaction for Syndecan-1(grade 3) according to scoring system for immunohistochemical expression. The expression of CD117 in plasmacytoma was similar to the expression of CD56, (60%) showed grade 1, while (40%) were grade 0. Conclusion: We concluded that the importance of histopathological biopsy for early diagnosis and prognostic evaluations are important in that the plasmacytoma may be a primary or metastatic lesion of MM. and Syndecan-1can be used as diagnostic marker for plasma cells, while CD56 and CD117 expressed only in neoplastic plasma cells and their expression indicate good prognosis.

Candidate protein biomarkers in pancreatic neuroendocrine neoplasms grade 3

Pancreatic neuroendocrine neoplasms (PanNENs) are rare tumours that compose 1–2% of all pancreatic tumours.Patients with metastatic grade 3 neoplasia are usually treated with chemotherapy but have a poor progression-free and overall survival. According to the WHO 2017 classification, they are divided into neuroendocrine tumours (NETs) G3 and neuroendocrine carcinomas (NECs). Despite the new classification, new diagnostic and prognostic biomarkers are needed to sub-categorise the patients and to help guide therapy decisions. Blood from 42 patients and 42 healthy controls were screened for the presence of 92 proteins with the Immuno-Oncology panel using the Proximity Extension Assay provided by Olink Biosciences. Immunohistochemical staining of FAS ligand (FASLG) was performed on 16 patient tumour specimens using a commercial antibody. Fifty-four out of 87 evaluable proteins differed significantly in concentration between blood from patients and blood from healthy controls. FASLG was the only protein for which the concentration in blood was significantly lower in patients compared to controls and the levels correlated negatively to Ki-67 index. Seven of 14 evaluable PanNEN G3 specimens showed FASLG immunoreactivity in the tumour cells while there was scattered immunoreactivity in immune cells. Positive FASLG immunoreactivity correlated to well-differentiated morphology.FASLG concentration in blood was significantly lower in patients with pancreatic NENs G3 compared to controls, and the expression in tumour tissue was variable. Furthermore, FASLG was negatively correlated to Ki-67 and was more frequently expressed in well-differentiated tumours. Taken together, these results may suggest a role of FASLG in PanNENs.

Clinical management of extrapleural septal solitary fibrous tumor: A case report.

Solitary fibrous tumor (SFT) is a rare neoplasm arising from the pleura, although it can sometimes affect extrapleural sites, including the head and neck. The sinonasal involvement is exceptional. Recent literature defines the SFT as a single spectrum of mesenchymal tumors, including hemangiopericytoma, which is currently considered a cellular phenotypic variant. The current case describes a rare case of a nasal septal SFT. The mass was embolized and then removed through endonasal endoscopic surgery. Histopathologic examination demonstrated positive immunoreactivity for CD34, and negative for keratin and S100 protein confirming the diagnosis of benign nasal SFT. After two years of follow-up, no relapse was observed. Diagnostic assessment of SFT requires nasal endoscopy, imaging and histopathological examination and a long time follow up is mandatory.

Clinical, pathological and immunohistochemical bovine papillomatosis

The present clinical study was conducted to study incidence, clinical symptoms, histopathological and immunohistochemical findings of bovine papillomatosis. Among total of 3812 cattle presented to the clinic, 52 cattle were having papillomas. No incidence of papillomas was recorded in buffaloes. Highest incidence of bovine papillomas was observed in cattle of 3 years age. Jersey crossbred cows were the most commonly affected breed followed by HF cross breed and non – descript cows. Most of the papillomas were located on teats only (40.38%) followed by teats and udder (21.15%). Histopathologically, fibro papilloma (60%) was found to be most common papilloma followed by Angio keratotic acanthomatous papilloma (40%). Immunohistochemical studies were conducted to know the rate of proliferation of bovine papillomas. Immunopositive reaction was observed against VEGF in Angio keratotic acanthomatous papilloma. EGFR and Ki67 immunopositivity was observed in fibropapillomas.

Effects of Indomethacin on The Gastric Mucosa of Adult Male Albino Rat and The Possible Gastroprotection by Alpha Lipoic Acid

Background: Indomethacin is an effective NSAID used in inflammation treatment. However, it causes gastric mucosal lesions. Alpha lipoic acid (ALA) is an antioxidant that prevents cell damage. Aim of the work: To detect the protective role of ALA on indomethacin-induced gastric mucosal alterations. Materials and Methods: A total number of thirty adult male albino rats were randomly equally divided into three groups: control group (received distilled water), indomethacin-treated group; received 50 mg/kg b.w of indomethacin., once orally and ALA+indomethacin-treated group; received 100 mg/kg b.w. of ALA, orally for 3 days followed by once administration of 50 mg/kg b.w. of indomethacin, orally 1h after the last dose of the ALA. Four hours after indomethacin administration. The rats were anesthetized, sacrificed and the stomach immediately removed and processed for histological (light and electron microscopic) and inducible nitric oxide synthase (iNOS) immunohistochemical examination. Area % of collagen fibers and iNOS of all animal groups was measured and compared. Results: Indomethacin induced gastric mucosal erosion, degeneration, vacuolization, inflammatory cell infiltration, interstitial hemorrhage, blood vascular congestion and dilatation, deposition of collagen fibers and strong positive iNOS immunoreactivity. Area % of collagen fibers and iNOS was significantly increased in indomethacin-treated group when compared to control. ALA+indomethacin-treated group exhibited nearly normal histological, immunohistochemical and morphometric findings. Conclusion: ALA has a gastroprotective role as it can prevent the deleterious effects of indomethacin on the gastric mucosa.

THE SUB CHRONIC TOXIC EFFECT OF VOODOO EXTRACT ON THE CENTRAL NERVOUS SYSTEM IN ADULT MALE ALBINO RATS

Abstract Introduction: Voodoo is a mixture of herbs and spices that are sprayed with a synthetic compound that mimics the effects of Cannabis. The widespread over-the-counter availability of these products made these compounds the 2nd most common frequently used recreational drug after marijuana in teenagers and young adults. This study aimed to investigate the sub chronic toxic effects of Voodoo (Voodoo abuse) on the central nervous system (CNS) in adult male albino rats. Methodology: thirty adult male albino rats were divided into three equal groups; negative control group, positive control group, and Voodoo extract treated group. Behavioral tests, neurotransmitter concentration in brain tissues and histopathological examination of brain tissues were performed. Results: general activity changes, significant decrease in food intake, feces formation and weight gain were reported between control and Voodoo treated group. The behavioral Y maze test revealed significant inhibition of the spontaneous alternation while, locomotor activity and rotarod test revealed significant inhibition of the neuromuscular coordinative function. The neurotransmitters (serotonin, dopamine, adrenaline and noradrenaline) assaying in the brain tissues of Voodoo treated group revealed significant decrease. These neurological changes were supported by the histopathological findings in the form of shrunken cells with dense nuclei and vacuolated cytoplasm in addition to the positive cytoplasmic immunoreaction for caspase -3 in the brain tissues. Conclusion: Voodoo extract was found to have a functional and structural deleterious CNS effect with sub chronic exposure in adult male albino rats.

Loss of Interleukin-17RA Expression is Associated with Tumour Progression in Colorectal Carcinoma.

Interleukin-17 (IL-17) is a pro-inflammatory cytokine found in various cancers. Current evidence indicates that IL-17 plays a vital role in tumour initiation and progression in colorectal carcinoma (CRC) via binding with its receptor, IL-17RA. However, the association between clinicopathological features and presence of IL-17 and IL-17RA protein in primary CRC tissues remains unclear. This study also investigates the difference between the presence of IL-17 and IL-17RA in the paired tumour tissues versus adjacent normal tissues. The presence of IL-17RA and IL-17 protein in primary CRC tissues was determined by immunohistochemistry. Associations between clinicopathological features and IL-17RA and IL-17 immunoreactivity, were analyzed by χ2 tests. We found that both IL-17RA (p = 0.001) and IL-17 (p = 0.025) in tumour cells of primary CRC tissues was significantly lower as compared to adjacent normal tissue. Positive immunoreactivity for IL-17RA and IL-17 were detected in 51.0% and 16.8% of tumour tissues, respectively. Furthermore, negative immunoreactivity of IL-17R was significantly associated with advanced stage according to TNM classifier (p = 0.027), high grade of tumour (p = 0.019), increased depth of tumour invasion (p = 0.023) and vascular invasion (p = 0.039). Positive IL-17 immunoreactivity was associated with advanced stage (p = 0.008) and lymph node metastasis (p = 0.008). Thus, this study suggests that the loss of IL-17RA expression occurs as tumour progresses and this may predict the aggressiveness of tumour whilst expression of IL-17 promotes tumour progression and lymph node metastasis. Thus, loss of IL-17RA could be a useful prognostic biomarker for tumour progression in CRC patients.

Immunohistochemical analysis of SOX2 expression in small‑cell neuroendocrine carcinoma of the endometrium

Small-cell neuroendocrine carcinoma (NEC) of the endometrium is an extremely rare and highly aggressive carcinoma. Sex-determining region Y-box 2 (SOX2) is a master transcription factor regulating the self-renewal, maintenance of stem cell properties and pluripotency of embryonic stem cells, and recent studies revealed that SOX2 plays important roles in cancer growth and progression in several types of carcinomas, including small-cell neuroendocrine carcinoma (NEC) of the lung and oesophagus. Few studies to date have analysed the association between SOX2 and endometrioid carcinoma, whereas the expression of SOX2 in small-cell NEC of the endometrium has not been investigated. The aim of the present study was to analyse the expression status of SOX2, p16 and paired-box gene (PAX) 8, a useful Müllerian marker, in endometrial small-cell NEC. A total of 4 patients with small-cell NEC of the endometrium were enrolled (median age, 70 years). Immunohistochemical studies revealed SOX2 expression in 3 patients and p16 expression in all patients. No patients exhibited positive immunoreactivity for PAX8. SOX2 expression has been reported to be associated with the pathogenesis of small-cell NEC of the oesophagus. Therefore, the results of the present study indicated that SOX2 expression plays an important role in the development of small-cell NEC of the endometrium and the oesophagus. Moreover, expression of p16 and loss of PAX8 do not indicate the origin of small-cell NEC of the endometrium.

Vimentin and Cytokeratin Immunostaining: The Role in Basic Diagnosis and Prognosis of Sarcomas

Introduction: Specific diagnosis which provides diagnostic, prognostic, and therapeutic information to guide patient care defines the primary goal of sarcoma management and care. The role of immunohistochemistry, using vimentin (mesenchymal linage marker) and cytokeratin (epithelial linage marker) as basic markers for diagnosis and classification of sarcomas for specific management strategies and prognosis was elucidated in the present study.
 Materials and Methods: Twenty four (24) archived paraffin wax processed tissue block sarcoma samples were randomly selected from the histopathology Laboratories and museums of the Nnamdi Azikiwe University Teaching Hospital (NAUTH) Nnewi and National hospital Abuja and necessary data obtained from records. Blocks were re-embedded with fresh paraffin wax and 3µ thick sections cut with the aid of a rotary microtome. Haematoxylin and Eosin staining method was employed to confirm diagnosis before proceeding to immunohistochemistry. Antibodies for vimentin and pancytokeratin were employed for immunohistochemistry while exposed mouse and rabbit specific horseradish peroxides/diaminobenzidine detection IHC kit was employed for immunostaining.
 Results: The mean age of patients was 26 years while the ages range from 11 to 48 years with 14 (58.3%) females and 10 (41.7%) males. Vimentin had strong positive immunoreactivity for all sarcoma samples whereas cytokeratin had positive immunoreactions for synovial sarcoma only, which also showed co-expression of both genes.
 Conclusion: Vimentin and cytokeratin may play vital role as basic biomarkers not only for diagnosis and characterization of sarcomas but for specific management regime and prognostication. However, IHC must be performed at high standard using appropriate antibodies, samples and reagents.
 Keywords: Sarcoma, immunohistochemistry, vimentin, cytokeratin, biomarker.