Positive Immunoreactivity Research Articles

NF-κB Decoy ODN-Loaded Poly(Lactic-co-glycolic Acid) Nanospheres Inhibit Alveolar Ridge Resorption.

Residual ridge resorption combined with dimensional loss resulting from tooth extraction has a prolonged correlation with early excessive inflammation. Nuclear factor-kappa B (NF-κB) decoy oligodeoxynucleotides (ODNs) are double-stranded DNA sequences capable of downregulating the expression of downstream genes of the NF-κB pathway, which is recognized for regulating prototypical proinflammatory signals, physiological bone metabolism, pathologic bone destruction, and bone regeneration. The aim of this study was to investigate the therapeutic effect of NF-κB decoy ODNs on the extraction sockets of Wistar/ST rats when delivered by poly(lactic-co-glycolic acid) (PLGA) nanospheres. Microcomputed tomography and trabecular bone analysis following treatment with NF-κB decoy ODN-loaded PLGA nanospheres (PLGA-NfDs) demonstrated inhibition of vertical alveolar bone loss with increased bone volume, smoother trabecular bone surface, thicker trabecular bone, larger trabecular number and separation, and fewer bone porosities. Histomorphometric and reverse transcription-quantitative polymerase chain reaction analysis revealed reduced tartrate-resistant acid phosphatase-expressing osteoclasts, interleukin-1β, tumor necrosis factor-α, receptor activator of NF-κB ligand, turnover rate, and increased transforming growth factor-β1 immunopositive reactions and relative gene expression. These data demonstrate that local NF-κB decoy ODN transfection via PLGA-NfD can be used to effectively suppress inflammation in a tooth-extraction socket during the healing process, with the potential to accelerate new bone formation.

Chondroblastoma: clinicopathological analyses of 307 cases from a single institution in China and the diagnostic value of the H3F3 K36M mutant antibody

AimsTo elucidate the clinicopathological features and the diagnostic value of mutation specific antibody H3F3 K36M of chondroblastoma (CB) in China.MethodsClinicopathological profiles were retrieved, and immunohistochemistry was performed on 185 CB...

Comparative pathology and immunohistochemistry of Newcastle disease in domestic chicken (Gallus-gallus domesticus) and Alabio duck (Anas platyrhynchos Borneo).

Newcastle disease is very pathogenic in chickens, whereas in ducks, the disease does not show any clinical symptoms. To compare the clinical symptoms features, pathological lesions, viral distribution, and apoptosis response caused by the Newcastle disease virus (NDV) in domestic chicken and Alabio duck. Forty domestic chickens and 40 Alabio ducks were separated into four groups: domestic chicken and Alabio duck treatment groups, where each was infected by NDV velogenic ducks/Aceh Besar_IND/2013/eoAC080721 in 106ELD50 dosage. The domestic chicken and Alabio duck control groups were each inoculated with Phosphate Buffer Saline. The infection route was intraorbital, 0.1 ml in volume. Symptoms were observed from day 1 until day 7 post-infection (PI). Necropsy was performed on days 1, 2, 3, 5, and 7 PI for organ collection. Symptoms observed were disorders in the respiratory, gastrointestinal, and nervous systems, followed by 100% mortality in domestic chickens. In Alabio ducks, there were only depression and slight lethargy. The lesion in domestic chicken day 1 appeared by the lungs, thymus, Fabricius bursa, spleen, and kidney. On day 3 PI, lesions were also found in the heart, proventriculus, duodenum, and cecal tonsil. On days 5 and 7 PI, the trachea and brain lesions were found. In Alabio ducks, lesions were found in the lungs, thymus, spleen, and proventriculus on day 1. Afterward, on day 3, light lesions were found in the heart. On day 5, lesions were found in the trachea and brain; finally, on day 7, light lesions were only found in the thymus, spleen, and brain. Immunopositive reaction NDV in domestic chicken was highest in the proventriculus, duodenum, cecal tonsil, and lymphoreticular organs. In the Alabio duck, it was highest in the duodenum and cecal tonsil. The caspase-3 percentage in domestic chicken increased on day 3 PI; in Alabio ducks, on day 2 PI. Clinical symptoms and pathological lesions were faster and more severe in domestic chickens. The immunopositive reaction NDV in domestic chicken continued to increase, while in Alabio ducks, it decreased until the last observation day. Apoptosis percentage increased earlier in the Alabio duck than in domestic chicken.

Association of PD-L1 and p16 expression with clinicopathological parameters in oral cavity and oropharyngeal squamous cell carcinoma.

High expression of PD-L1 in tumour cells is associated with a higher response rate to immune checkpoint inhibitors, which target T regulatory pathways. p16 is a surrogate marker for human papilloma virus associated cancers. The present study was conducted to evaluate PD-L1 and p16 expression in head and neck squamous cell carcinomas and its correlation with clinicopathological parameters. Histologically confirmed cases of head and neck squamous cell carcinomas were evaluated for PD-L1 and p16 expression along with its correlation with grade and site of the tumour. Out of 40 cases, 21 (52.5 %) showed positive immunoreactivity for PD-L1 while 13 (32.5 %) were positive for p16. A significant association was observed between PD-L1 and p16 expression in the oropharynx (p value= 0.035). No significant association could be established between clinicopathological parameters and PD-L1/ p16 expression or between PD-L1 and p16 in the oral cavity. PD-L1 expression shows a positive association with p16 in oropharyngeal squamous cell carcinomas. Immune checkpoint inhibitors against PD-L1 can be used in carcinomas with positive expression for PD-L1.

Expression of lysophosphatidic acid receptors in the rat uterus: cellular distribution of protein and gestation-associated changes in gene expression.

Though lysophosphatidic acid (LPA) shows a variety of regulatory roles in reproduction, its action mechanisms in the gestational organs are still largely unknown. We here characterized cellular distribution of its six kinds of specific receptors (LPA1-6) in rat uteri by immunohistochemistry and quantitatively analyzed changes in Lpar1-6 mRNAs expression throughout pregnancy. Among LPA1-6, evident expression of LPA3, LPA4, and LPA6 was immunologically detected and less expression of immunoreactive LPA1 and LPA2 was also found. Luminal and glandular epithelial cells, stromal cells, and myometrial cells are sites of positive immunoreactions, and they are all likely to express three or more subtypes. All of Lpar1-6 mRNAs were expressed, and their alterations were variable depending on subtypes and gestational age. The present information suggests that diverse actions of LPA in the uterus involve varied expression of LPA receptors dependent on tissue/cell types, receptor subtype(s), and organ reproductive states and helps to understand uterine biology of LPA.

Solitary extramedullary plasmacytoma of the gingiva—A case report with literature review

Abstract Extramedullary solitary plasmacytoma (EMP) is a plasma cell neoplasm involving the soft tissues without the involvement of the bone marrow. In this case report, we present an isolated soft tissue growth on the lingual gingiva of the left mandibular canine-premolar region diagnosed as EMP based on clinical, histological, and laboratory results. Orthopantomogram and occlusal radiographs showed no osteolytic lesions. Hematoxylin and Eosin staining showed sheets of inflammatory cells, predominantly atypical plasma cells with varying degrees of differentiation showing features of pleomorphism, and multinucleated giant cells with intervening collagen fibers. Positive immunoreactivity for CD56, CD138, and kappa light chain restriction were observed. Serum protein electrophoresis demonstrated an increase in IgA levels and immunofixation electrophoresis confirmed the presence of the M band in the IgG and kappa lanes, indicating monoclonal gammopathies. Based on the above findings, the case was diagnosed as EMP; the consultant pathologist, however, recommended excluding multiple myeloma.

Cytoarchitecture of greater cane rat (Thryonomys swinderianus) habenulae and mammillary body

This study was conducted to investigate expression of neurons, glia and connective tissue markers in the habenulae and mammillary body of the Greater Cane Rat (GCR) using histological and immunohistochemical techniques. Four apparently healthy males GCR obtained from Lagos Nigeria were used for this experiment. The brains were harvested after perfusion, carefully dissected and post-fixed in 10% neutral-buffered-formalin (NBF) at 40C for 48 hours. Subsequently, tissue sections were prepared and stained using Haematoxylin and Eosin (H&E), Cresyl-violet, ionized calcium-binding-adapter molecule 1 (Iba1; microglia), glial fibrillary acidic protein (GFAP; astrocytes), neuronal nuclei (NeuN; neurons) and collagen type 1 (Collagen1) antibodies. H&E and Cresyl-violet staining showed the neuronal nuclei and their fibres in the habenulae, mammillary body, and pial capsule that surround the mammillary body ventrally. Astrocytes in the habenulae showed positive immunoreaction to anti-GFAP while the mammillary body revealed expression of perivascular astrocytes and capsular astrocytes. Also, in the mammillary body, the perivascular microglia and meningeal microglia in the pial capsule showed immunopositive reaction to anti-Iba1 while the fibres within blood vessels and capsule were immunopositive to collagen type 1 antibody in the same structure. This work has provided the first cytoarchitectural features and clear description of expression of these specific markers in habenulae and mammillary body in the GCR, which are similar to other rodents but with slight variations.

In vitroSodium Fluoride Treatment Significantly Affects Apoptosis and Proliferation in the Liver of Embryonic Chickens

Sodium fluoride (NaF), although helpful in preventing dental decay, may negatively affect the body. The aim of this study was to examine the effects of a 6-hin vitrotreatment of livers isolated from 14-day-old chicken embryos with NaF at doses of 1.7 (D1), 3.5 (D2), 7.1 (D3) and 14.2 mM (D4), with regard to apoptosis, cell proliferation and tissue structure. The mRNA expression of the apoptosis regulatorsCYCS,APAF1,BCL2,CASP3,CASP9andTMBIM1was analysed by the qPCR method. Apoptotic cells were detected by a TUNEL assay. The tissue and DNA structure were also analysed by histological staining (H&E, Feulgen). The number of proliferating cells was determined and the apoptosis regulatory proteins were localised by the immunohistochemical staining of PCNA, CASP3 and APAF1. The results showed that the mRNA expression ofCYCS,BCL2,CASP3,CASP9andAPAF1increased significantly in the D1 group, as did that ofCASP9in the D3 group and ofBCL2andAPAF1in the D4 group. The number of apoptotic cells increased significantly in the D4 group, where they increased from 18% to 49%. On the other hand, the number of proliferating cells decreased gradually, in a dose-dependent manner, from 84% in the control group to 5.5% in the D4 group. The expression of apoptosis-regulating factors also increased: in the D3 and D4 groups, the CASP3 immunopositive reaction was more intensive in single cells in the embryonic livers, whereas that of APAF1 increased in the hepatocytes as well as in the hepatic blood vessel walls. The mechanism of the effect of NaF on apoptosis in the embryonic liver is very complex. In the groups exposed to higher doses of NaF, apoptosis was significantly stimulated, while proliferation was inhibited and the tissue structure was damaged. The expression of apoptosis regulators at the mRNA and protein levels increased, but the mRNA expression did not depend on the NaF dose. These results reveal that NaF, by changing the balance between apoptosis and the proliferation of hepatocytes, may disturb the development and function of the liver in embryonic chickens. Therefore, the risk of exposure to NaF should be considered when determining the standards for human and animal exposure to this compound.

Expression of vasopressin and its receptors in migraine-related regions in CNS and the trigeminal system: influence of sex

BackgroundHypothalamus is a key region in migraine attacks. In addition, women are disproportionately affected by migraine. The calcitonin gene-related peptide (CGRP) system is an important key player in migraine pathophysiology. CGRP signaling could be a target of hormones that influence migraine. Our aim is to identify the expression of vasopressin and its receptors in the brain and in the trigeminovascular system with focus on the migraine-related regions and, furthermore, to examine the role of sex on the expression of neurohormones in the trigeminal ganglion.MethodsRat brain and trigeminal ganglia were carefully harvested, and protein and mRNA levels were analyzed by immunohistochemistry and real-time PCR, respectively.ResultsVasopressin and its receptors immunoreactivity were found in migraine-related areas within the brain and, in the trigeminal ganglion, predominantly in neuronal cytoplasm. There were no differences in the number of positive immunoreactivity cells expression of CGRP and vasopressin in the trigeminal ganglion between male and female rats. In contrast, the number of RAMP1 (CGRP receptor), oxytocin (molecular relative to vasopressin), oxytocin receptor and vasopressin receptors (V1aR and V1bR) immunoreactive cells were higher in female compared to male rats. Vasopressin and its receptors mRNA were expressed in both hypothalamus and trigeminal ganglion; however, the vasopressin mRNA level was significantly higher in the hypothalamus.ConclusionsA better understanding of potential hormonal influences on migraine mechanisms is needed to improve treatment of female migraineurs. It is intriguing that vasopressin is an output of hypothalamic neurons that influences areas associated with migraine. Therefore, vasopressin and the closely related oxytocin might be important hypothalamic components that contribute to migraine pathophysiology.

Benign Spindel Cell Neoplasam: A Central Myofibroma of Maxilla in Male Child of Seven Years of Age: A Case Report

Myofibroma is a rare benign spindle cell neoplasm which occurs predominantly in infants and young children. It can occur as a solitary mass or as a multicentric lesion, consisting of myofibroblasts. In oral cavity, the sites most commonly involved are the mandible, tongue, lips and buccal mucosa however, it rarely involves the maxilla. It has aggressive clinical presentation mimicking malignancies. We report a case of a male child seven years of age presented with intraoral swelling on the left palatal aspect of the maxilla that extended from deciduous upper left second molar to the distal of upper permanent left first molar causing the mobility of these teeth. Radiologically, the lesion was osteolytic causing marked resorption of alveolar bone and displacement of permanent maxillary left second molar. A differential diagnosis of peripheral giant cell granuloma, central giant cell granuloma, fibroma, aggressive fibromatosis, nodular fasciitis, peripheral ossifying fibroma and spindle cell neoplasm was made. After the excluding malignant and vascular involvement, the surgical excision of the lesion was planned and was carried out under general anesthesia. Histologically, benign proliferation of spindle cells was seen giving the diagnosis of benign spindle cell neoplasm. Reports also showed positive immunoreactivity with vimentin and αSMA while non-responsive to desmin, validates the diagnosis of myofibroma. KEYWORDS: spindle cell neoplasm; myofibroma; maxilla

INNV-19. THE DETECTION OF DISSEMINATED PINEAL PARENCHYMAL TUMOR OF INTERMEDIATE DIFFERENTIATION (PPTID) IN CEREBROSPINAL FLUID (CSF) USING CELL CAPTURE AND IMMUNOCYTOCHEMISTRY

Abstract BACKGROUND Pineal parenchymal tumors of intermediate differentiation (PPTID) account for 21% to 54% of pineal parenchymal tumors and may be complicated by cerebrospinal dissemination (most commonly at time of reoccurrence; up to 4-10 years post resection). The integral membrane protein synaptophysin is expressed in neuroendocrine cells and virtually all neuraxial neurons that contribute to synaptic transmission. PPTID frequently demonstrates synaptophysin independently of other neural differentiation markers. Cerebrospinal fluid (CSF) cell capture assays have demonstrated utility for assessing disseminated intracranial neoplasms, thus we explore this technology for monitoring a patient with PPTID. METHODS A 30-year-old female patient with PPTID diagnosed two years prior was suspected for intracranial dissemination and underwent five CSF collections over the course of 64 days. Per collection, approximately 7 mL of CSF was submitted for each cytology and cell capture. Cytology was performed at Providence Saint John’s Health Center. The CNSide™ platform (Biocept, Inc.) was used to capture and stain cells for DAPI, synaptophysin, and CD45. Multiple capture antibody cocktails (termed 1986, 1822, and gTP1—originally developed for use in carcinoma, melanoma, and glioma) were tested. Cells of interest (COI) were defined by positive immunoreactivity for both DAPI and synaptophysin and no immunoreactivity for CD45. Results were quantified and compared longitudinally. RESULTS CSF analysis on days 0, 9, 21, 36, and 64 demonstrated negative results by cytology and 51, 96, 170, 130, and 43 COI/mL by the CNSide platform, respectively. Cumulative mean capture rates for cocktails 1986, 1822, and gTP1 were 47, 14, and 36 COI/mL/sample, respectively. DISCUSSION A significant amount of PPTID patients will develop cerebrospinal dissemination. The importance of craniospinal control in this setting has been previously demonstrated. Our work suggests that cell capture assays paired with immunocytochemistry can be used as sensitive means to monitor disseminated PPTID and may impact treatment decisions.

Riboflavin attenuates tartrazine toxicity in the cerebellar cortex of adult albino rat

Tartrazine is a synthetic yellowish dye considered one of the most common food colorants. Extensive usage of tartrazine in humans led to harmful health impacts. To investigate the impact of tartrazine administration on the cerebellum and to assess the potential role of riboflavin co-administration in the adult male albino rat. Four groups of adult albino rats were included in this study. Group I was supplied with distilled water. Group II was supplied tartrazine orally at a dose of 7.5 mg/kg BW dissolved in distilled water. Group III was supplied with tartrazine at the same previously mentioned dose and riboflavin orally at a dose of 25 mg/kg BW dissolved in distilled water. Group IV was supplied with riboflavin at the same previously mentioned dose. The study was conducted for 30 days then rats were sacrificed, weighted and the cerebella extracted and handled for light, ultrastructural and immunohistochemical evaluation. It was found with tartrazine treatment focal areas of Purkinje cell loss leaving empty spaces, a broad spread of neuronal affection to the degree of the disappearance of some of the granular cells, reduced the thickness of the molecular and granular layers, and strong positive GFAP immunoreactions. With riboflavin coadministration restored continuous Purkinje layer with normal appeared Purkinje cells, but some cells were still shrunken and vacuolated as well as the molecular and granular cell layers appeared normal. Tartrazine had deleterious effects on the cerebellar cytoarchitecture, and riboflavin co-administration alleviated these neurotoxic effects.

A Scalp Metastatic Follicular Thyroid Carcinoma with a Silent Thyroid History and Normal Thyroid Uptake by PET-CT

Abstract Introduction/Objective Follicular thyroid carcinoma (FTC) accounts for 10%-15% of all primary thyroid cancers. Follicular thyroid cancer is known to metastasize hematogenously, with distant metastasis occurring in 6-20% of cases. Scalp metastasis with an underlying thyroid carcinoma is an exceedingly rare clinical entity; there are less than 15 case reports in the literature. 1-2. Here, we present a case of widely invasive FTC with skull metastases located on the parietal calvarium. The scalp mass was confirmed as follicular thyroid carcinoma via microscopic morphology and positive immunoreactivity for Vimentin, TTF1, thyroglobulin, and PAX8. We present this case not only because of the diagnostic challenge and rare scalp metastasis, but also because of the patient’s unusual “silent” thyroid medical history and no abnormal thyroid uptake by PET-CT. Methods/Case Report 81-year-old female presenting 2-year history of right parietal scalp mass, which was previously suspected for lipoma with attempted resection in Mexico but interrupted due to severe hemorrhage. No palpable thyroid nodule was noted. PET-CT showed avid lesions of the right breast with nipple retraction, as well as avid suspicious lesions of bilateral lungs, calvarium, xiphoid process, right acetabulum, T spine most c/f widely metastatic cancer, most likely a breast primary; evaluation of the neck demonstrates physiologic FDG-activity in the thyroid. Thyroid studies showed hyperthyroidism: TSH (L) <0.010 uIU/mL; Free T3 (H) 4.7 pg/mL and Free T4 1.00 ng/d. The patient was performed Ultrasound-guided FNA and core biopsy of the large parietal scalp lesion, however, FNA of the cystic component was nondiagnostic (no malignant cells seen). Results (if a Case Study enter NA) NA. Conclusion Core biopsy of the large parietal mass the tumor cells tend to form follicular or microfollicular architecture with colloid present, but no papillary nuclear features. Rare mitotic figures are seen. IHC stain of tumor cells are positive for AE1/AE3, Cam5.2, CK7, TTF1, thyroglobulin, PAX8, and Vimentin, which show strong evidence for metastatic follicular thyroid carcinoma (FTC) origin. Tumor cells are negative for Napsin-A, P63, Synaptophysin, Ck5/6, Ck20, EMA, WT1, CDX2, GATA3, Mammaglobin, ER, PR, and BRST2, which helps rule out the primary malignancy from lung, breast, neuroendocrine or other adenocarcinomas.

Presence and localization of apelin and its cognate receptor in canine testes using immunohistochemical and RT-PCR techniques

Apelin, a member of the adipokine family, is a novel endogenous peptide which regulates the male reproductive system of mammals by interacting with a specific receptor. Recent studies have highlighted that apelin may play a role in the regulation of reproduction by reducing testosterone production and inhibiting LH secretion. To the best of our knowledge, there is no available data on the presence of the apelin and its receptor in canine testes. Therefore, the aim of this study was to reveal the presence of apelin and evaluate its distribution in the canine testes using immunohistochemical and RT-PCR techniques. For this purpose, five fertile and healthy male dogs were subjected to elective orchiectomy. The immunohistochemical reaction revealed the presence of apelin and its receptor in the canine testes. Apelin was localized in spermatids and spermatozoa with a positive signal in the “acrosomal bodies”. As regards the apelin receptor, a positive immunoreaction was detected in the cytoplasm of the cells localized near to the basal membrane of the seminiferous tubules and in the cytoplasm of Leydig cells. The RT-PCR analysis showed the presence of transcripts for apelin and apelin receptor in all of the samples under study. A 35kDa band confirmed apelin receptor protein expression in all of the samples analysed. In conclusion, the paracrine and endocrine role of apelin and its cognate receptor on male reproduction reported in humans and laboratory animals could also be hypothesized in dogs.

Diagnostic utility of Bcl-2 immunohistochemical expression in pediatric functional bowel obstruction cases with ganglionated specimens

BackgroundFunctional disturbances of the gastrointestinal tract are caused by a number of neurodysplastic conditions, including diseases that are rarer than Hirschsprung’s disease (HSCR), such as ganglion cell immaturity and intestinal neuronal dysplasia (IND). Bcl-2 shows positive immunoreactivity in degenerative and immature ganglion cells (IGCs). This work evaluates the implication of the immunohistochemical expression of Bcl-2 in cases of pediatric intestinal pseudo-obstruction (IPO) with ganglionated specimens.This 2-year prospective observational study was conducted from January 2017 to December 2019 on all intestinal specimens for cases referred from a pediatric surgery department with clinical data suggesting IPO (abdominal distension > 3 months, intermittent bilious vomiting, no radiological evidence of the transitional zone of HSCR or evidence of mechanical obstruction). The exclusion criteria were patients with IPO related to myopathic disorders, aganglionic biopsies, and specimens with inadequate tissue blocks. The same number of intestinal specimens of cases without IPO was used as a control group. All specimens underwent a histologic examination of ganglion cells and nerve bundles and were also evaluated for Bcl-2 immunohistochemical expression.ResultsTwenty-one specimens were analyzed, including six colonic resection specimens and 15 intestinal biopsies taken by full-thickness transanal biopsy (n = 12) or incisional biopsy from inadequately functioning stomas (n = 3). The mean ± standard deviation age of the patient cohort was 22 ± 7.4 months (range, 19–153 months), and there were 13 (61.9%) male patients. Bcl-2 protein was strongly positive in 57.1% of the cases, weakly positive in 19%, and negative in 23.8% as well as in all 21 control specimens. A highly significant correlation was observed between Bcl-2 expression and ganglion cell number (P < 0.001), where all hyperganglionic specimens showed strong positivity compared with the hypoganglionic and adequately ganglionic specimens. A positive association was also found between Bcl-2 expression and IND compared with other disorders of dysganglionosis (P = 0.04) and post-HSCR cases (P = 0.002).ConclusionBcl-2 immunohistochemistry is a valuable tool to diagnose allied disorders of HSCR through its expression in IGCs, which are difficult to identify by conventional hematoxylin and eosin staining.

Morphological, Immunohistochemical, and Ultrastructural Studies of the Donkey's Eye with Special Reference to the AFGF and ACE Expression.

The donkey is mainly used as a working animal for riding and pack transport, as well as for dairy and meat production. Eye afflictions are common in donkeys, thus requiring a detailed study. A few studies had focused on the donkey's eye, and most of them had considered it, merely, a horse's eye. This study aimed to investigate the anatomy, histology, ultrastructure, and immunohistochemical features of the donkey's eye. The results were recorded and compared to those of horses in certain dimensions. Unlike horses, the donkey's eye is more circular in the contour of the cornea, has smaller lenticular thickness, and has longer anterior and vitreous chambers. Positive immunoreactivity to acidic fibroblast growth factor in the basal cell layers of the cornea was observed, indicating their role in cell differentiation and the renewal of the epithelium. Moreover, the corneal keratocytes expressed angiotensin-converting enzyme, which plays a role in corneal homeostasis and wound healing. Additionally, telocytes, hyalocytes, and other immune cells were observed within the iris and ciliary processes. Hence, this work is an updated detailed study of the morphology and ultrastructure of the donkey's eye and reveals some similarities and dissimilarities to the horse's eyes, which should be considered in clinical practice.

Sex-based differences in conjunctival goblet cell responses to pro-inflammatory and pro-resolving mediators

Many conjunctival inflammatory diseases differ between the sexes and altered conjunctival goblet cells (CGCs) response is often involved. Inflammation is initiated by the release of pro-inflammatory mediators and terminated by the biosynthesis of specialized pro-resolution mediators (SPMs). Herein, we determined the sex-based difference in the responses of CGCs to inflammatory stimuli or pro-resolving lipid SPMs and their interaction with sex hormones. GCs were cultured from pieces of human conjunctiva in RPMI media. CGCs were transferred 24 h before the start of experiments to phenol red-free and FBS-free media to minimize exogenous hormones. RT-PCR, immunofluorescence microscopy (IF), and Western Blot (WB) were performed to determine the presence of sex hormone receptors. Cellular response to pro-inflammatory stimuli or SPMs was studied by measuring the increase in intracellular [Ca2+] ([Ca2+]i) using fura 2/AM microscopy. Use of RT-PCR demonstrated estrogen receptor (ER) α in 4/5 males and 3/3 females; ERβ in 2/4 males and 2/3 females; and androgen receptors (AR) in 3/3 male and 3/3 female CGCs. Positive immunoreactivity by IF and protein expression by WB was detected using antibodies for the ERα and ERβ in 3/3 males and 3/3 females, while AR were only present in males. Significantly different Ca2+ responses between sexes were found with carbachol only at 10–3 M, but not with histamine or leukotriene (LT) B4 at any concentration used. Incubation with dihydrotestosterone (DHT), estrone (E1), or estradiol (E2) at 10–7 M for 30 min significantly inhibited the LTB4-stimulated [Ca2+]i increase in male and female CGCs. Incubation with DHT, E1, and E2 overnight significantly inhibited the LTB4 response in females, while DHT and E2 significantly inhibited the LTB4 response in males. The SPM lipoxin A4 (LXA4) (10–9–10−8 M), but not the resolvins D1 or D2, induced an [Ca2+]i increase that was significantly higher in males compared to females. We conclude that male and female CGCs showed differences in the expression of sex hormone receptors. Treatment with sex hormones altered pro-inflammatory mediator LTB4-induced response. Males compared to females have a higher response to the ω-6-fatty acid derived SPM LXA4, indicating males may terminate inflammation in conjunctival goblet cells faster than females.

Immunohistochemical Expression of Ki-67, cytokeratin-18 and BCL-2 in Wistar rats testes treated with Rauwolfia vomitoria, Chlorpromazine and Co-administration of Reserpine, Ascorbate and Zinc

Aim: The expression of Ki-67, cytokeratin-18, and BCL-2 proteins in Wistar rats testes was studied using Rauwolfia vomitoria RV extract, Chlorpromazine (CPZ), and combination of Reserpine, Ascorbate, and Zinc (RAZ).Methods: Forty-five eight weeks old male Wistar rats (170-190 g) were selected into nine groups of five rats each. Group A was the control group, animals in groups B and C received 10 and 20 mg/kg of chlorpromazine respectively, animals in groups D and E received 2.5 and 5 mg/kg of reserpine respectively, animals in groups F and G received 150 and 300 mg/kg of RV leaf extract respectively while groups H and I animals received (2.5:5:100) mg/kg and (5:10:200) mg/kg of combination of RAZ respectively. All compounds were administered orally for 56 days.Results: Chlorpromazine and reserpine treated rats showed weak immunoreactivity to ki-67 and strong positive immunoreactivity to cytokeratin and BCL-2 proteins while RVand combination of RAZ treated rats showed weak positive immunoreactivity to cytokeratin and BCL-2 and strong immunoreactivity to ki-67. Furthermore, slight significant increase in germ cell proliferation index was seen in RV and RAZ treated groups when compared with CPZ and RES treated groups while significant decrease in germ cell apoptotic index and immature sertoli cell index were seen in RV and RAZ treated groups when compared with CPZ and RES treated groups.Conclusion: This research revealed the reproductive toxicity of synthetic antipsychotic drugs (CPZ and RES) and also unveiled the fertility potential of antipsychotic herb (RV) extract alongside RAZ by reducing the reproductive toxicity that is commonly associated with antipsychotic drugs.

Epidermal growth factor receptors and their ligands are expressed in the spleen of the Japanese Quail (Coturnix coturnix japonica) during the post-hatch period

ABSTRACT 1. The epidermal growth factor (EGF) family plays an important role in the development, differentiation, migration and apoptosis of cells, as well as in wound healing, which are all essential to the viability of multicellular organisms. The avian spleen is a principal organ of systemic immunity and its importance in disease resistance is presumably accentuated by the scarcity of avian lymph nodes. 2. The aim of this study was to determine whether EGF receptors (ErbB1-4) and their ligands (EGF, AREG and NRG) are expressed in the structural components of the quail spleen during the post-hatch period. At each selected age, from 1 d to 7, 14, 21 and 60 d, 10 quails were euthanised under ether anaesthesia and their spleens were fixed in a 10% formaldehyde-alcohol solution. Following routine histological processing, the streptavidin-biotin-peroxidase method was used for immunohistochemical examination. 3. Strong cytoplasmic immunoreactions for ErbB2, ErbB4 and NRG were observed in the ellipsoid associated cells (EAC) of the quail spleen throughout the post-hatch period. This immunoreactivity in the EAC increased after the 7th d post-hatch. ErbB1 and ErbB3 immunoreactions were relatively similar and weak in all components of the spleen during the post-hatch period. Some immune cells of the peri-arterial lymphatic sheath (PALS) and peri-ellipsoidal lymphatic sheath (PELS) showed positive immunoreactivity for the ErbB receptors and their ligands. In the vascular smooth muscle cells, immunoreactivity for ErbB2 was stronger than that for the other ErbB receptors and their ligands. 4. The data showed that ErbB receptors and their ligands (EGF, AREG and NRG) are expressed by different structural components of the quail spleen during the post-hatch period.

Astrocytes profiling in acute hepatic encephalopathy: Possible enrolling of glial fibrillary acidic protein, tumor necrosis factor-alpha, inwardly rectifying potassium channel (Kir 4.1) and aquaporin-4 in rat cerebral cortex.

Hepatic encephalopathy (HE) is a neurological disarray manifested as a sequel to chronic and acute liver failure (ALF). A potentially fatal consequence of ALF is brain edema with concomitant astrocyte enlargement. This study aims to outline the role of astrocytes in acute HE and shed light on the most critical mechanisms driving this role. Rats were allocated into two groups. Group 1, the control group, received the vehicle. Group 2, the TAA group, received TAA (300 mg/kg) for 3 days. Serum AST, ALT, and ammonia were determined. Liver and cerebral cortical sections were processed for hematoxylin and eosin staining. Additionally, mRNA expression and immunohistochemical staining of cortical GFAP, TNFα, Kir4.1, and AQP4 were performed. Cortical sections from the TAA group demonstrated neuropil vacuolation and astrocytes enlargement with focal gliosis. GFAP, TNFα, and AQP4 revealed increased mRNA expression, positive immunoreactivity, and a positive correlation to brain water content. In contrast, Kir 4.1 showed decreased mRNA expression and immunoreactivity and a negative correlation to brain water content. In conclusion, our findings revealed altered levels of TNFα, Kir 4.1, GFAP, and AQP4 in HE-associated brain edema. A more significant dysregulation of Kir 4.1 and TNFα was observed compared to AQP4 and GFAP.