Mild Immunoreactivity Research Articles

Adropin maypromote insulin stimulated steroidogenesis and spermatogenesis in adult mice testes.

Adropin is a versatile peptide which was discovered as a novel metabolic hormone that is involved in the regulation of lipid and glucose homeostasis. However, its possible role in the testicular function is not yet understood. The aim of our study was to explore the distribution pattern of adropin and GPR19 in various cell types and its possible role in testicular functions of adult mice. Immunohistochemical study revealed the intense immunoreactivity of adropin in the Leydig cells, while GPR19 showed intense immunoreactivity in the pachytene spermatocytes and mild immunoreactivity in Leydig cells and primary as well as secondary spermatocytes in mouse testis. Enho mRNA was also found to be expressed in the mouse testis. These findings suggested that adropin-GPR19 signaling may act in autocrine/paracrine manner to modulate testicular functions. Furthermore, to find out the direct role of adropin in the testicular function, in vitro study was performed in which testicular slices were cultured with adropin alone (10 and 100 ng/mL) and in combination with insulin (5 μg/mL). Adropin alone inhibited testicular testosterone synthesis by inhibiting the expression of P450-SCC, 3β-HSD, and17β-HSD while along with insulin stimulated the testicular testosterone synthesis by increasing the expression of GPR19, IR, StAR, P450-SCC, 3β-HSD, and 17β-HSD. Adropin alone or in combination with insulin promoted germ cell survival and proliferation by upregulating the expression of PCNA, Bcl2, and pERK1/2. Thus, it can be concluded that adropin-GPR19 signaling promotes insulin stimulated steroidogenesis and germ cell survival as well as proliferation in the mice testes in an autocrine/paracrine manner.

Evaluation of tissue-engineered human acellular vessels as a Blalock–Taussig–Thomas shunt in a juvenile primate model

ObjectivesPalliative treatment of cyanotic congenital heart disease (CCHD) uses systemic-to-pulmonary conduits, often a modified Blalock–Taussig–Thomas shunt (mBTTs). Expanded polytetrafluoroethylene (ePTFE) mBTTs have associated risks for thrombosis and infection. The Human Acellular Vessel (HAV) (Humacyte, Inc) is a decellularized tissue-engineered blood vessel currently in clinical trials in adults for vascular trauma, peripheral artery disease, and end-stage renal disease requiring hemodialysis. In addition to restoring blood flow, the engineered HAV demonstrates the capacity for host cellular remodeling into native-like vasculature. Here we report preclinical evaluation of a small-diameter (3.5 mm) HAV as a mBTTs in a non-human primate model. MethodsWe implanted 3.5 mm HAVs as right subclavian artery to pulmonary artery mBTTs in non-immunosuppressed juvenile rhesus macaques (n = 5). HAV patency, structure, and blood flow were assessed by postoperative imaging from 1 week to 6 months. Histology of HAVs and surrounding tissues was performed. ResultsSurgical procedures were well tolerated, with satisfactory anastomoses, showing feasibility of using the 3.5 mm HAV as a mBTTs. All macaques had some immunological reactivity to the human extracellular matrix, as expected in this xenogeneic model. HAV mBTTs remained patent for up to 6 months in animals, exhibiting mild immunoreactivity. Two macaques displaying more severe immunoreactivity to the human HAV material developed midgraft dilatation without bleeding or rupture. HAV repopulation by host cells expressing smooth muscle and endothelial markers was observed in all animals. ConclusionsThese findings may support use of 3.5 mm HAVs as mBTTs in CCHD and potentially other pediatric vascular indications.

One-step cell biomanufacturing platform: porous gelatin microcarrier beads promote human embryonic stem cell-derived midbrain dopaminergic progenitor cell differentiation in vitro and survival after transplantation in vivo.

Numerous studies have shown that cell replacement therapy can replenish lost cells and rebuild neural circuitry in animal models of Parkinson's disease. Transplantation of midbrain dopaminergic progenitor cells is a promising treatment for Parkinson's disease. However, transplanted cells can be injured by mechanical damage during handling and by changes in the transplantation niche. Here, we developed a one-step biomanufacturing platform that uses small-aperture gelatin microcarriers to produce beads carrying midbrain dopaminergic progenitor cells. These beads allow midbrain dopaminergic progenitor cell differentiation and cryopreservation without digestion, effectively maintaining axonal integrity in vitro. Importantly, midbrain dopaminergic progenitor cell bead grafts showed increased survival and only mild immunoreactivity in vivo compared with suspended midbrain dopaminergic progenitor cell grafts. Overall, our findings show that these midbrain dopaminergic progenitor cell beads enhance the effectiveness of neuronal cell transplantation.

In Vitro Immunoreactivity Evaluation of H-Ferritin-Based Nanodrugs.

Biological nanoparticles, such as proteins and extracellular vesicles, are rapidly growing as nanobased drug-delivery agents due to their biocompatibility, high loading efficiency, and bioavailability. However, most of the candidates emerging preclinically hardly confirm their potential when entering clinical trials. Among other reasons, this is due to the low control of synthesis processes and the limited characterization of their potential immunoreactivity profiles. Here, we propose a combined method that allow us to fully characterize H-ferritin nanoparticles' immunoreactivity during their production, purification, endotoxin removal, and drug loading. H-Ferritin is an extremely interesting nanocage that is being under evaluation for cancer therapy due to its innate cancer tropism, favorable size, and high stability. However, being a recombinant protein, its immunoreactivity should be carefully evaluated preclinically to enable further clinical translation. Surprisingly, this aspect is often underestimated by the scientific community. By measuring proinflammatory cytokine release as a function of endotoxin content, we found that even removing all pyrogenic contaminants from the nanocage, a mild immunoreactivity was still left. When we further purified H-ferritin by loading doxorubicin through a highly standardized loading method, proinflammatory cytokine release was eliminated. This confirmed the safety of H-ferritin nanocages to be used for drug delivery in cancer therapy. Our approach demonstrated that when evaluating the safety of nanodrugs, a combined analysis of acute toxicity and immunoreactivity is necessary to guarantee the safety of newly developed products and to unveil their real translational potential.

Characterization of giant neuroendocrine cells in the pregnant New Zealand white rabbit vagina demonstrated by histological, histochemical, and immunohistochemical methods.

The vagina is part of the genitalia and constitutes part of the birth canal. Sperm is deposited into vagina, and it acts in transport of sperm. Pregnancy is a complex process involving different physiological changes in the body associated with hormonal and metabolic alterations that control maternal and fetal demands. During pregnancy, neuroendocrine cells in rabbit vagina are considered part of the diffuse neuroendocrine system observed throughout the body. Giant neuroendocrine cells in rabbit vagina during pregnancy have not been observed previously. This study detected the presence of giant neuroendocrine cells in the vagina of the pregnant rabbit. The presence of these cells was demonstrated with the use of different histological techniques, including hematoxylin and eosin, PAS, combined Alcian blue-PAS, Crossmon's trichrome, and the Grimelius silver method. Giant neuroendocrine cells were observed in late-pregnancy intraepithelial sites and on the lamina propria. These cells were characterized by vacuolated basophilic cytoplasm with PAS- and PAS-AB-positive granules. Moreover, neuroendocrine cells exhibited an argyrophilic character. Immunohistochemically, neuroendocrine cells in rabbit vagina during pregnancy demonstrated positive immunoreactivity to neuron-specific enolase (NSE) with different intensities, mild immunoreactivity to the vascular endothelial growth factor (VEGF), and negative immunoreactivity to CD68. HIGHLIGHTS: Neuroendocrine cells on the vagina of the female rabbit during pregnancy are considered part of the diffuse neuroendocrine system. Giant neuroendocrine cells on the vagina of the rabbit during pregnancy have not been observed previously. Giant neuroendocrine cells were demonstrated intraepithelial and on the lamina propria. Immunohistochemically, neuroendocrine cells demonstrated positive immunoreactivity to NSE and VEGF, and negative immunoreactivity to CD68.

Distribution of estrogen receptor in the rabbit cervix during pregnancy with special reference to stromal elements: an immunohistochemical study

The estrogen plays a critical role during pregnancy through their receptors. Although the rabbit is one of the most important lab animal estrogen receptor alpha (ERA) localization on basic cells, newly discovered cells including telocyte and neuroendocrine cells, vascular compartments and interstitium during pregnancy not been described. At 0 day pregnancy, the most prominent immunoreactivity was moderate to ERA and observed on the ciliated cells, secretory cells, blood plasma, and interstitium. The smooth muscles and the endothelial cells showed mild immunoreactivity to ERA. Lymphocytes only exhibited strong immunoreactivity to ERA. At 7 days pregnancy moderate immunoreactivity to ERA observed on ciliated cells, secretory cells, smooth muscles, interstitium, and lymphocytes. Strong immunoreactivity to ERA detected on endothelial cells and blood plasma. At 14 days of pregnancy, the most prominent immunoreactivity was strong and detected on ciliated cells, smooth muscles, lymphocytes, and interstitium. Moderate immunoreactivity detected on endothelial cells and blood plasma. Secretory cells only exhibited mild immunoreactivity to ERA. At 21 days of pregnancy, the immunoreactivity to ERA ranged between mild on ciliated cells, smooth muscles, blood plasma and interstitium and negative on secretory cells, endothelial cells and lymphocytes. Our results indicated that the frequency and intensity of ERA immunostaining in the rabbit cervix varied on different structural compartments of the cervix during different pregnancy stages.

Prognostic Role of Aryl Hydrocarbon Receptor Interacting Protein (AIP) Immunohistochemical Expression in Patients with Resected Gastric Carcinomas.

Aryl hydrocarbon receptor (AHR) interacting protein (AIP) is a chaperone which binds to inactive AHR in the cell cytoplasm. AHR is best known for mediating the toxicity of halogenated aromatics, but it has also been linked to carcinogenesis and tumor progression in several tumor types. Our aims are to assess the features of AIP immunohistochemical (IHC) staining and to evaluate its possible role as a prognostic marker in gastric cancer (GC). Retrospective study of 147 cases of resected GC. Clinicopathological features were collected, tissue microarrays were constructed for AIP IHC and statistical analysis were performed. AIP staining was observed in 50.3% of tumors. All AIP-positive cases exhibited cytoplasmic or membranous staining, variably associated with nuclear co-staining. 93.2% of AIP-positive tumors showed AIP immunoreactivity in 100% of cells. Staining intensity was mild, moderate and intense in 33.8%, 13.5% and 52.7% of cases. Tumors were stratified according to AIP staining intensity into low expression (no or mild AIP immunoreactivity) and high expression (moderate or intense AIP immunoreactivity). 36.6% of our cases showed high AIP expression. High AIP expression was significantly and independently correlated to tumor progression and cancer death. Tumors with high AIP expression showed lower survival and higher progression rates. AIP expression might be useful for determining GC prognosis. More studies are needed to clarify the role of AHR pathway in GC, AIP expression and its potential use as a surrogate marker for selecting patients for AHR modulation therapy.

Renoprotective effect of celecoxib against gentamicin-induced nephrotoxicity through suppressing NFκB and caspase-3 signaling pathways in rats

Gentamicin-induced nephrotoxicity has been well documented, although the causing mechanisms and preventative measures need further investigation. The current study aimed to explore the potential protective impacts of celecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, on gentamicin-induced nephrotoxicity and the potential mechanisms in rats. Rats were randomly divided into four groups as follows: group1: normal control, group 2: received gentamicin only (100 mg/kg intraperitoneally), group 3: concurrently received gentamicin and celecoxib (30 mg/kg, orally) and group 4: received celecoxib.Celecoxib administration decreased gentamicin-induced rise in kidney weight, renal somatic index (RSI), blood urea nitrogen (BUN), serum creatinine (Cr), protein in urine, lactate dehydrogenase (LDH), nitric oxide (NOx), meanwhile, it increased serum albumin, urine Cr level and creatinine clearance (CCr), increased the renal endogenous antioxidant status, revealed by decreased malondialdehyde (MDA) and increased superoxide dismutase (SOD) and reduced glutathione (GSH). Gentamicin-induced elevated nuclear factor kappa B-P65 subunit (NFκB-p65), tumor necrosis factor-alpha (TNF-α) and apoptotic markers (tumor suppressor protein (p53) and caspase-3) protein levels were significantly decreased upon celecoxib treatment. Moreover, celecoxib suppressed renal myeloperoxidase (MPO) activity and posed improvement of histological features. In immunohistochemistry, celecoxib-treated rats showed decreased immunoreactivity against COX-2 in tubular cells and a mild positive immunoreactivity against heat shock protein 70 (HSP70) in renal interstitial cells. These findings propose that celecoxib treatment mitigates renal dysfunction via decreasing renal inflammation, oxidative/nitrosative stress, and apoptosis.

Histology and Brain Derived Neurotrophic Factor immunoreaction of the neurons in the corpora quadrigemina of the African grasscutter (Thryonomys swinderianus – Temminck, 1827)

Abstract The African grasscutter is the second largest rodent in Africa, thus, a key component of the minilivestock industry. The study described the histological features and probed the distribution of Brain Derived Neurotrophic Factor (BDNF) in the corpora quadrigemina of the African grasscutter at foetal and postnatal developmental periods. Brain samples from foetuses explanted on foetal days 60 (F60) and 90 (F90) and extracted from 3 and 6-days-old pups (P3 and P6, respectively), 72-days-old juveniles (P72) and 450-days-old adults (P450) were used. They were prepared for histology and immunohistochemistry. Three laminae were distinct in the rostral colliculi at the foetal and postnatal periods; an outermost stratum zonale, middle stratum griseum superficiale and an inner stratum griseum profundum. Stratum griseum intermediale and stratum medullarium were not distinct. On F60 and F90, the stratum zonale was made of immature neurons, devoid of neurites; the central nucleus of the caudal colliculus was also made of immature migrating neurons. On P3, the neurons were already mature. The stratum zonale was made of medium-sized neuronal cells and thick processes. The thickness of this layer decreased with age. On P3, the caudal colliculus was made of all the components typical of a developed caudal colliculus. There was no BDNF immunoreactive cell in the stratum zonale at any postnatal period; a moderate BDNF-immunoreactivity in the stratum griseum superficiale on P3, a mild immunoreactivity on P6, none reactivity on P72 and mild on P450. The dorsal and lateral cortices of the caudal colliculus were none reactive to BDNF at any postnatal period. The results suggest a better auditory than visual capacity in the rodent.

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.

Immunoexperssion of cancer stem cell marker (CD44) in ameloblastoma.

Background:Ameloblastoma is the most frequently encountered benign, locally invasive tumor. Attempts to surgically resect the tumor often leave small islands of tumor, which later result in recurrence in 50%–90% of cases. This has raised questions regarding the tumor cell populations that are responsible for tumor growth and recurrence. In ameloblastoma, whether or not cancer stem-like cells are present remains undetermined. However, if cancer stem-like cells are present in ameloblastoma, it is important to identify which type of cell possesses the stem-like characteristics and is responsible for ameloblastoma progression and recurrence.Aim:Our study aims at analyzing immunohistochemical staining to detect the expression of cancer stem cell (CSC) marker CD44 in relation to proliferative activity of tumor cells in histopathologically diagnosed cases of ameloblastoma variants and to derive a correlation between the CD44 expression and biologic behavior of the lesion.Materials and Methods:A retrospective study, was conducted on total 25 cases ameloblastoma and were immunostained for CD44 expression. Results obtained were statistically analyzed.Results:A positive correlation was observed between staining intensity of CD44 marker and the known biological behavior of the lesion. Intense staining reaction was found to be only in 8% cases, whereas 76% cases demonstrated moderate intensity and remaining 16% displayed mild immunoreactivity to CD44 marker. Staining location was more to be in stellate reticulum-like (SR-like) cells when compared to ameloblast-like (AB-like) cells. Intense immunostaining was localized in the small tumor follicles, especially in SR-like cells situated in close vicinity of peripheral AB-like cells whereas mild intensity of staining was observed in keratinizing areas.Conclusion:CSCs marker positive expression in benign tumor like ameloblastoma may be responsible for its aggressiveness and recurrence. CD44 marker may be of great value in predicting the biological behavior and growth potential of ameloblastoma.

EXPRESSION OF VASCULAR ENDOTHELIAL GROWTH FACTOR (VEGF) IN HEMANGIOMAS AND INTERMEDIATE VASCULAR LESIONS

INTRODUCTION: Vascular tumors of intermediate malignancy encompass a group of tumors whose biologic potential is intermediate between hemangiomas and angiosarcomas. Angiogenesis is characterized by development of new blood vessels through the division or migration of pre-existing vasculature. VEGF deserves attention because it is associated with the stimulation of endothelial cell proliferation and vascular permeability and promotion of angiogenesis.OBJECTIVES: to evaluate the immunohistochemical expression of vascular endothelial growth factor (VEGF) in pyogenic granulomas,hemangiomas and intermediate vascular tumors (hemangioendotheliomas, hemangiopericytomas) and to highlight the possible role of VEGFin diagnosis of each of these tumorsMATERIALS AND METHODS: Immunohistochemical analysis of 20 cases of vascular and intermediate vascular tumors, with one section from each specimen (20 sections for VEGF antibody) and 5 control cases. Immunohistochemical staining was performed using a Labeled Strept-Avidin Biotin method (LSAB).RESULTS: Normal gingival tissue showed mild immunoreactivity for VEGF. All the examined cases showed strong positive expression for VEGF antibody, with different intensities.CONCLUSIONS: The marker VEGF was overexpressed in vascular and intermediate vascular tumors than normal gingival tissueKEYWORDS: Vascular tumors, Intermediate vascular tumors, Immunohistochemistry, VEGF.

Comparative analysis of p53 and p21 proteins in normal cervix and HPV associated precancerous and cancerous lesions of cervix

IntroductionHPV-E6 oncoprotein of High-Risk HPV plays critical role in the degradation of p53 protein for acquisition of malignant phenotype in cervix cells. The present study, was sought to analyze the expression of HPV-E6, p53 and p21 in cervical cancer to explore possible impact of HPV-E6 in the modulation of these proteins. MethodsThe expression of HPV16/18-E6, p53 and p21 proteins in cervix tissues [normal (n = 100), cervical intraepithelial neoplasia (CIN; n = 67), squamous cell carcinoma (SCCs; n = 153)] was analyzed by immunohistochemistry and revalidated by Western Blotting. The p53 protein functionality was determined by using phosphospecific p-p53-ser46 antibody. ResultsThe negative expression of HPV-E6 and p53 but, mild immunoreactivity of p21 was noticed in normal stratified squamous epithelium. In CIN and SCCs, the HPV-E6 was found to be predominantly over-expressed in 80.0% and 84.9% of cases respectively. Significant nuclear accumulation of p53 was observed in 77.6% of precancerous and 83.0% of cancerous tissues whereas, 74.6% of CIN and 73.8% of SCCs represented loss of nuclear p21 expression as compared to normal (p = 0.0001, p = 0.0001). Pearson's correlation test revealed significant inverse association between p53 and p21 proteins both in CIN (p = 0.0001) and SCCs (p = 0.0001). Interestingly, significant positive association of HPV-E6 with p53 and negative association with p21 were also detected in CIN (p = 0.0001) and SCCs (p = 0.0001). No positive expression of p-p53-ser46 was detected in any of the cases. DiscussionThe present study provides the evidences of inhibition of p53 and p21 transactivity by HPV-E6 resulting alteration of cell growth inhibitory signals in cervical cancer.

Toxoplasmosis-associated abortion in an alpaca (Vicugna pacos) fetus.

A near-full-term alpaca (Vicugna pacos) was stillborn 2 days before the expected date of delivery; necropsy examination was conducted within 6 hr of delivery. Gross lesions were enlarged liver and hydrocephalus. On histologic examination, mild inflammatory lesions were identified in the placenta, liver, and lungs, although no etiology was recognized. Within the brain, there was a mild nonsuppurative meningoencephalitis, hydrocephalus, and hydromyelia. Both kidneys had inflammatory foci in cortex and medulla associated with protozoal tissue cysts. Protozoa in brain and kidneys were identified as Toxoplasma gondii based on immunoreactivity to T. gondii polyclonal antibodies that stain both tachyzoites and tissue cysts and BAG1 T. gondii antibodies that are bradyzoite specific. The tissue cysts exhibited intense positivity to T. gondii and mild immunoreactivity to Neospora caninum antibodies. The dam had a high antibody titer (1 : 12,800) to T. gondii and low titer (1 : 100) to N. caninum using their respective agglutination tests. This is the first report of toxoplasmosis-associated abortion in alpaca.

Immunolocalization of estrogen and progesterone receptors in ewe mammary glands

Immunopresence of estrogen receptor α (ERα), β (ERβ) and progesterone receptor (PR) were examined in the ewe mammary gland from prepubertal stage to involution. Immunolocalization of ERα revealed a strong positive staining in nuclei of cells composing terminal ductular units (TDUs) in prepubertal ewes. A mild immunoreactivity was identified in early lactating gland. During late lactation immunoreactive product to ERα was observed in the cytoplasm of glandular epithelial cells in all alveoli. In mammary glands at involution ERα positivity was clearly nuclear, with weak to moderate cytoplasmic staining. Cytoplasmic strong immunostaining for ERβ was detected in cells of TDUs, whereas some stromal cells exhibited nuclear staining. A nuclear ERβ immunostaining was observed at early lactation, instead during late lactation, the positivity for ERβ showed only a moderate cytoplasmic distribution. At involution, ERβ positivity was very moderate and detected just in the cytoplasm of shrunken alveoli. Scattered nuclear staining of PR was observed just in mammary glands at early lactation. These results showed that in the mammary glands of sheep both estrogen receptor isoforms were displayed during lactation cycle and that PR appeared just at early lactation, reflecting their regulatory role in alveolar cells.

Effects of acute treatment with dexamethasone on hemodynamic and histopathological changes in rats

We assessed the time-dependent effects of intraperitoneal (i.p.) and intravenous (i.v.) application of dexamethasone (Dexa) on the mean arterial blood pressure (MAP), heart rate (HR) and total blood volume (TBV). We evaluated also the relation between the effects and immunoreactivities of transforming growth factor-beta (TGF-β), epithelial nitric oxide synthase (eNOS), interleukin-1 beta (IL1-β) and vascular endothelial growth factor (VEGF) in rat brain, lung and kidney tissues. Rats were anesthetized and while still breathing spontaneously, a tracheotomy and femoral vein and artery catheterizations were performed. To determine TBV using the hemodilution method, 2 ml albumin-electrolyte solutions were applied by i.v. injection. Group 1 (control group) received a 1 ml bolus injection of physiologic saline, Group 2 received 15 mg/kg and Group 3 received 75 mg/kg Dexa i.p. The hematocrit was measured at 10, 20, 60 and 120 min. For each animal, the values of MAP, HR and TBV were measured within 2 h. For immunohistochemical evaluation, anti-TGF-β, anti-eNOS, anti-IL1-β and anti-VEGF primary antibodies were tested using the avidin-biotin-peroxidase method. TBV was decreased in Group 1 and the increase in MAP was statistically significant. HR values increased slightly. None of the values changed significantly in Group 2. Although TBV was unchanged in Group 3, the decrease in MAP was statistically significant. HR values increased, but the increase was not statistically significant. Mild IL1-β immunoreactivity and moderate TGF-β, eNOS and VEGF immunoreactivities were observed in the brain, lung and kidney samples in Group 1. Increased eNOS immunoreactivity in the kidney samples were observed in Group 2. eNOS immunoreactivity was as strong in the brain and the kidney samples in Group 3. Decreased VEGF immunoreactivity was observed in the lung and kidney tissues in Group 3. Significantly decreased TGF-β immunoreactivity was observed in all tissue samples in Group 3. The decreased MAP values in Group 3 differed from those in Groups 1 and 2. Despite increased eNOS immunoreactivity, especially in brain and kidney, the decrease in VEGF immunoreactivity in Group 3, especially lung and kidney, were consistent with a drop in blood pressure.

Platelet-derived growth factor receptor-β and epidermal growth factor receptor in pulmonary vasculature of systemic sclerosis-associated pulmonary arterial hypertension versus idiopathic pulmonary arterial hypertension and pulmonary veno-occlusive disease: a case-control study

IntroductionSystemic sclerosis (SSc) complicated by pulmonary arterial hypertension (PAH) carries a poor prognosis, despite pulmonary vascular dilating therapy. Platelet-derived growth factor receptor-β (PDGFR-β) and epidermal growth factor receptor (EGFR) are potential therapeutic targets for PAH because of their proliferative effects on vessel remodelling. To explore their role in SScPAH, we compared PDGFR- and EGFR-mmunoreactivity in lung tissue specimens from SScPAH. We compared staining patterns with idiopathic PAH (IPAH) and pulmonary veno-occlusive disease (PVOD), as SScPAH vasculopathy differs from IPAH and sometimes displays features of PVOD. Immunoreactivity patterns of phosphorylated PDGFR-β (pPDGFR-β) and the ligand PDGF-B were evaluated to provide more insight into the patterns of PDGFR-b activation.MethodsLung tissue specimens from five SScPAH, nine IPAH, six PVOD patients and five controls were examined. Immunoreactivity was scored for presence, distribution and intensity.ResultsAll SScPAH and three of nine IPAH cases (P = 0.03) showed PDGFR-β-immunoreactivity in small vessels (arterioles/venules); of five SScPAH vs. two of nine IPAH cases (P = 0.02) showed venous immunoreactivity. In small vessels, intensity was stronger in SScPAH vs. IPAH. No differences were found between SScPAH and PVOD. One of five normal controls demonstrated focally mild immunoreactivity. There were no differences in PDGF-ligand and pPDGFR-b-immunoreactivity between patient groups; however, pPDGFR-b-immunoreactivity tended to be more prevalent in SScPAH small vasculature compared to IPAH. Vascular EGFR-immunoreactivity was limited to arterial and arteriolar walls, without differences between groups. No immunoreactivity was observed in vasculature of normals.ConclusionsPDGFR-β-immunoreactivity in SScPAH is more common and intense in small- and post-capillary vessels than in IPAH and does not differ from PVOD, fitting in with histomorphological distribution of vasculopathy. PDGFR-β immunoreactivity pattern is not paralleled by pPDGFR-β or PDGF-B patterns. PDGFR-β- and EGFR-immunoreactivity of pulmonary vessels distinguishes PAH patients from controls.

Changes in bradykinin and bradykinin B 2-receptor during estrous cycle of mouse

The aim of the study was to investigate changes in the abundance of bradykinin and bradykinin B2-receptor in the ovary of mice during its estrous cycle. Changes in the abundance of bradykinin were correlated with changes in bradykinin B 2-receptor in order to determine the functional significance of this peptide for follicular development, ovulation and luteinization. Bradykinin immunoreactivity was mainly observed in the granulosa cells of antral follicles, especially around the oocytes and lining the antral cavity during proestrus and estrus phases of the cycle. Recently formed corpora lutea showed mild immunoreactivity for both bradykinin and bradykinin B 2-receptor. During diestrus 1 and diestrus 2, bradykinin and bradykinin B 2-receptor immunostaining was mainly found in the corpora lutea and mildly in the antral follicles. Immunoblot analysis for bradykinin and bradykinin B 2-receptor attained a peak during late evening in proestrus, which may be the time of the LH surge. Thereafter bradykinin and bradykinin B 2-receptor declined sharply during the estrus phase. When the concentration of bradykinin was correlated with bradykinin B 2-receptor throughout the estrous cycle, they showed strong positive correlation. Thus, this study indicates that the levels of bradykinin and bradykinin B 2-receptor both simultaneously regulate estrous cycle and are important components for the reproductive process.

Immunohistochemical analysis of interleukin‐1α, its type I receptor and antagonist in keratocystic odontogenic tumors

Interleukin-1 alpha (IL-1 alpha) is thought to play a crucial role in the growth of keratocystic odontogenic tumors (KCOTs) in the jaw. The function of IL-1 alpha is regulated by the local levels of IL-1 alpha, its receptor and receptor antagonist (IL-1Ra) in tissues. In this study, the expression of these proteins was investigated both before and after marsupialization in KCOTs. The expression of IL-1 alpha, IL-1 receptor type I (IL-1RI) and IL-1Ra was detected immunohistochemically in 10 specimens of KCOTs. IL-1 alpha was intensively expressed throughout the epithelium in all cases, while mild expression of IL-1 alpha was detected in the subepithelial layer endothelial cells and fibroblasts. Mild or intensive immunoreactivity for IL-1RI was also observed in the epithelial cells in all cases, and in the endothelial cells and fibroblasts in five cases respectively. The expression of IL-1Ra was detected in the epithelial cells in five cases, and in the endothelial cells and fibroblasts in three cases. After marsupialization, the immunoreactivity for IL-1 alpha and IL-1RI in the epithelial cells decreased, while the immunoreactivity for IL-1Ra in the epithelial cells increased. However, the immunoreactivity for IL-1RI and IL-1Ra in endothelial cells and fibroblasts did not change significantly. The effects of IL-1 alpha on the epithelial cells might be downregulated after marsupialization by changing the expression levels of IL-1 alpha, IL-1RI and IL-1Ra in the epithelium of KCOTs.

Localization of gonadotrophin-releasing hormone I, bradykinin and their receptors in the ovaries of non-mammalian vertebrates

GnRH I and its receptors have been demonstrated in the ovaries of various vertebrates, but their physiological significance in reproductive cascade is fragmentary. Bradykinin is a potent GnRH stimulator in the hypothalamus. In the present study, the presence of GnRH I and its receptor, and bradykinin and its receptor in the ovaries of non-mammalian vertebrates were investigated to understand their physiological significance. GnRH I immunoreactivity in the ovaries of fish, frog, reptile and bird were mainly found in the oocyte of early growing follicles and granulosa cells and theca cells of previtellogenic follicles. Vitellogenic follicles showed mild GnRH immunoreactivity. GnRH I-receptor and bradykinin were localized in the same cell types of the ovaries of these vertebrates. The presence of GnRH I, GnRH I-receptor and bradykinin in the ovaries of these vertebrates was confirmed by immunoblotting. The presence of GnRH I mRNA was demonstrated in the ovary of vertebrates using RT-PCR. The ovaries of reptiles and birds showed significantly higher intensity of immunoreactivity for GnRH I-receptor as compared with the fish and amphibian. This may have a correlation with the higher yolk content in the ovary of reptile and bird. These results suggest the possibility of GnRH I and bradykinin as important regulators of follicular development and vitellogenesis in the vertebrate ovary.