Cyclooxygenase-2 Immunostaining Research Articles

A Case of Proliferative Cystitis Discovered as Protruding Lesion during Inspection of Secondary Infertility

A 42-year-old man visited our hospital complaining of secondary infertility. An abdominal ultrasonography screening incidentally revealed a protruding lesion in the bladder. As the lesion extended from the prostatic urethra and bladder neck, there was a possibility of ejaculation dysfunction after resection of the lesion. Therefore, with the patient's informed consent, sperm cryopreservation was conducted for fertility preservation, and subsequently histological examination was performed by partial transurethral resection of bladder tumor. The pathological findings were proliferative cystitis including all three subtypes (glandularis, cystica, and papillary). Cyclooxygenase-2 immunostaining was positive in cytoplasm; weakly positive in cystic and papillary lesions, and strongly positive in glandular lesions. According to a literature review of massive proliferative cystitis, the patient was the 77th case in Japan. Novel postoperative immunological pharmacotherapies with cyclooxygenase-2 inhibitors have been introduced in recent years.

Association of a Skin Dressing Made With the Organic Part of Marine Sponges and Photobiomodulation on the Wound Healing in an Animal Model.

The present study aims to characterize and to evaluate the biological effects of a skin dressing manufactured with the organic part of the Chondrilla caribensis marine sponge (called spongin-like collagen (SC)) associated or not to photobiomodulation (PBM) on the skin wound healing of rats. Skin dressings were manufactured with SC and it was characterized using scanning electron microscopy (SEM) and a tensile assay. In order to evaluate its biological effects, an experimental model of cutaneous wounds was surgically performed. Eighteen rats were randomly distributed into three experimental groups: control group (CG): animals with skin wounds but without any treatment; marine collagen dressing group (DG): animals with skin wounds treated with marine collagen dressing; and the marine collagen dressing + PBM group (DPG): animals with skin wounds treated with marine collagen dressing and PBM. Histopathological, histomorphometric, and immunohistochemical evaluations (qualitative and semiquantitative) of COX2, TGFβ, FGF, and VEGF were done. SEM demonstrates that the marine collagen dressing presented pores and interconnected fibers and adequate mechanical strength. Furthermore, in the microscopic analysis, an incomplete reepithelialization and the presence of granulation tissue with inflammatory infiltrate were observed in all experimental groups. In addition, foreign body was identified in the DG and DPG. COX2, TGFβ, FGF, and VEGF immunostaining was observed predominantly in the wound area of all experimental groups, with a statistically significant difference for FGF immunostaining score of DPG in relation to CG. The marine collagen dressing presented adequate physical characteristics and its association with PBM presented favorable biological effects to the skin repair process.

Thymoquinone attenuates doxorubicin-induced lung damage via heat shock proteins, inflammation and endoplasmic reticulum stress

The aim of this study is to determine possible therapeutic effects of THQ on doxorubicin induced lung damage in rats. Rats were divided into five groups (n = 8): control, THQ, olive oil, DOX (a single dose of 15 mg/kg intraperitoneally (i.p.) on seventh day of the experiment), and DOX+THQ (10 mg/kg THQ per day and 15 mg/kg DOX i.p. on seventh day). Animals were euthanized, and lung tissues were evaluated histopathologically. Caspase3, GRP78, GADD153, HSP90, HSP70, PCNA, COX2 and TNFα immunostaining were performed to determine the expression levels of these proteins among groups. TUNEL method was used for evaluation of apoptotic index. Moreover, total antioxidant status (TAS), total oxidant status (TOS) and IL6 in lung tissue were measured by ELISA assay. The DOX group had histopathological deterioration compared to the control group. There was an increase in apoptotic index, caspase 3, GRP78, GADD153, HSP90, HSP70, PCNA, COX2 and TNFα expressions in the DOX group. While TAS level of the DOX group decreased, IL6 and TOS level increased when compared with the other groups. However, there was improvement in lung tissue in DOX + THQ group compared to the DOX group. There was a decrease in apoptotic index, caspase 3, GRP78, GADD153, HSP90, HSP70, PCNA, COX2, TNFα expressions and TOS, IL6 in DOX+THQ group compared to the DOX group. We suggest that THQ can be used as a protective agent to reduce the toxic effects of DOX.

Pathological studies on spontaneous urinary bladder lesions in Indian buffaloes

The present investigation describes the spontaneous pathological lesions in urinary bladders (n = 177) of slaughtered buffaloes collected from Northern India. Gross examination revealed congestion, haemorrhages, oedematous changes and thickening of mucosa. Microscopically, epithelial abnormalities, inflammatory and neoplastic conditions were noticed. Epithelial abnormalities included epithelial hyperplasia, von Brunn’s nests, cystitis cystica and cystitis glandularis. Inflammatory conditions such as congestion, haemorrhages and cystitis were noticed. Cystitis was classified as lymphocytic, plasmo-lymphocytic, follicular and interstitial types. However, only single case of haemangiosarcoma was observed. Polymerase chain reaction revealed the presence of DNA of only BPV-1 in one sample out of 21, while one sample showed the presence of DNA of both BPV-1 and BPV-2. On immunohistochemistry, Ki-67 expression was observed only in haemangiosarcoma and one case of plasmo-lymphocytic cystitis with immunostaining restricted to nuclei. COX-2 immunostaining was observed in one case each of flat hyperplasia, von Brunn’s nest, haemangiosarcoma and two cases of plasmo-lymphocytic cystitis. Among epithelial abnormalities, COX-2 immunostaining was restricted to epithelial component of the lesions, and it was diffused cytoplasmic type with occasional perinuclear staining particularly in the cytoplasmic membrane. However, in haemangiosarcoma the COX-2 immunoreactivity was noticed in epithelial as well as connective tissue stroma. In conclusion, it may be stated that urinary bladders of slaughtered buffaloes showed a number of epithelial abnormalities, inflammatory conditions and occasionally neoplasms. Further studies to explore the infectious as well as non-infectious etiologies should be undertaken.

High COX-2 immunostaining in papillary thyroid carcinoma is associated with adverse survival outcomes.

Thyroid carcinoma is one of the most common malignancies worldwide. More than 70%-80% are papillary thyroid carcinoma (PTC). Many factors influence the PTC pathway of development such as genetic mutations, growth factors, and radiation. More biological understanding of the genetic and molecular pathways is needed in PTC to determine tumor behavior, and initial clinical assessment. Investigate the relation of COX-2 immunostaining in thyroid carcinoma with clinicopathological parameters to assess whether immunostaining results have prognostic significance. Retrospective study SETTING: Pathology department, tertiary care center METHODS: Records of PTC were retrieved and tissue microarrays were constructed. Tissue sections were stained using anti-human COX-2 monoclonal antibody. Immunostaining results were recorded and analysed. Relationship of COX-2 immunostaining in thyroid carcinoma with clinicopathological parameters. 139 tissue samples from 139 patients RESULTS: High versus low COX-2 immunostaining showed no significant differences for most clinicopathological parameters. However, high COX-2 immunostaining showed borderline association with tumor multifocality (P=.05), lower overall (log-rank=8.739 and P=.003), and disease-free survival (log-rank=7.033, P=.008). The study showed a positive association of high COX-2 immunostaining with lower survival outcomes in PTC. COX-2 immunostaining could be a potential prognostic factor for survival in PTC. Additional molecular and clinical investigations are needed for further understanding the molecular pathways of COX-2 in PTC and the feasibility of using inhibitors of COX-2 as adjuvant therapy along with current chemotherapy. Relatively low number of PTC variants, and no testing of other thyroid carcinomas. None.

Successful Pre-Clinical Management of Irinotecan-Debilitated Animals: A Protein- Based Accessory Phytomedicine.

Calotropis procera is a laticiferous plant (Apocynaceae) found in tropical regions all over the world. The ultrastructural characteristics of laticifers, their restricted distribution among different taxonomic groups, and in some species in each clade, as peptidases from latex, make them very attractive for biological analysis. The study aims to investigate the effects of LP-PII-IAA (laticifer protein (LP) sub-fraction II (PII) of C. procera presenting an iodoacetamide-inhibited cysteine proteinase activity) on irinotecan-induced intestinal mucositis, a serious adverse effect of this medicine for the treatment of cancer. LP-PII-IAA is composed of closely related isoforms (90%) of peptidases derived from catalysis and an osmotin protein (5%). Animals receiving co-administration of LP-PII-IAA presented a significant decrease in mortality, absence of diarrhea, histological preservation, and normalization of intestinal functions. Clinical homeostasis was accompanied by a reduction in MPO activity and declined levels of IL-1β, IL-6 and KC, while the IL-10 level increased in LP-PII-IAA-treated animals. COX-2 and NF-kB immunostaining was reduced and the levels of oxidative markers (GSH, MDA) were normalized in animals that received LP-PII-IAA. We suggest that peptidases from the latex of Calotropis procera were instrumental in the suppression of the adverse clinical and physiological effects of irinotecan.

Direct modulation of microglial function by electrical field.

Non-invasive electric stimulation (ES) employing a low-intensity electric current presents a potential therapeutic modality that can be applied for treating retinal and brain neurodegenerative disorders. As neurons are known to respond directly to ES, the effects of ES on glia cells are poorly studied. A key question is if ES directly mediates microglial function or modulates their activity merely via neuron-glial signaling. Here, we demonstrated the direct effects of ES on microglia in the BV-2 cells—an immortalized murine microglial cell line. The low current ES in a biphasic ramp waveform, but not that of rectangular or sine waveforms, significantly suppressed the motility and migration of BV-2 microglia in culture without causing cytotoxicity. This was associated with diminished cytoskeleton reorganization and microvilli formation in BV-2 cultures, as demonstrated by immunostaining of cytoskeletal proteins, F-actin and β-tubulin, and scanning electron microscopy. Moreover, ES of a ramp waveform reduced microglial phagocytosis of fluorescent zymosan particles and suppressed lipopolysaccharide (LPS)-induced pro-inflammatory cytokine expression in BV-2 cells as shown by Proteome Profiler Mouse Cytokine Array. The results of quantitative PCR and immunostaining for cyclooxygenase-2, Interleukin 6, and Tumor Necrosis Factor-α corroborated the direct suppression of LPS-induced microglial responses by a ramp ES. Transcriptome profiling further demonstrated that ramp ES effectively suppressed nearly half of the LPS-induced genes, primarily relating to cellular motility, energy metabolism, and calcium signaling. Our results reveal a direct modulatory effect of ES on previously thought electrically “non-responsive” microglia and suggest a new avenue of employing ES for anti-inflammatory therapy.

COX/iNOS dependence for angiotensin-II-induced endothelial dysfunction

Vascular dysfunction induced by angiotensin-II can result from direct effects on vascular and inflammatory cells and indirect hemodynamic effects. Using isolated and functional cultured aortas, we aimed to identify the effects of angiotensin-II on cyclooxygenase (COX) and inducible nitric oxide synthase (iNOS) and evaluate their impact on vascular reactivity.Aortic rings from mice were incubated overnight in culture medium containing angiotensin-II (100 nmol/L) or vehicle to induce vascular disfunction. Vascular reactivity of cultured arteries was evaluated in a bath chamber. Immunofluorescence staining for COX-1 and COX-2 was performed. Nitric oxide (NO) formation was approached by the levels of nitrite, a NO end product, and using a fluorescent probe (DAF). Oxidative and nitrosative stress were determined by DHE fluorescence and nitrotyrosine staining, respectively.Arteries cultured with angiotensin-II showed impairment of endothelium-dependent relaxation, which was reversed by the AT1 receptor antagonist. Inhibition of COX and iNOS restored vascular relaxation, suggesting a common pathway in which angiotensin-II triggers COX and iNOS, leading to vasoconstrictor receptors activation. Moreover, using selective antagonists, TP and EP were identified as the receptors involved in this response. Endothelium-dependent contractions of angiotensin-II-cultured aortas were blunted by ibuprofen, and increased COX-2 immunostaining was found in the arteries, indicating endothelium release of vasoconstrictor prostanoids. Angiotensin-II induced increased reactive oxygen species and NO production. An iNOS inhibitor prevented NO enhancement and nitrotyrosine accumulation in arteries stimulated with angiotensin-II.These results confirm that angiotensin-II causes vascular inflammation that culminates in endothelial dysfunction in an iNOS and COX codependent manner.

A Novel Murine Model of a High Dose Brachytherapy-Induced Actinic Proctitis.

BackgroundRadiation proctitis affects 1-20% of cancer patients undergoing radiation exposure due to pelvic malignancies, including prostate, gynecological and rectum cancers. The patients manifest rectal discomfort, pain, discharge, and bleeding. Notably, the efficacy of prophylactic measures remains controversial due to the lack of adequate animal models that mimic this condition.ObjectiveThe present study then aimed to develop a murine model of high-dose-rate (HDR) brachytherapy-induced proctitis.Material/MethodsC57BL/6 male mice were subjected to HDR (radiation source: iridium-192 [Ir-192]) through a cylindrical propylene tube inserted 2 cm far from the anal verge into the rectum. The animals received radiation doses once a day for three consecutive days (fractions of 9.5 Grays [Gy]), 3.0 mm far from the applicator surface. The sham group received only the applicator with no radiation source. The survival rate was recorded, and a colonoscopy was performed to confirm the tissue lesion development. Following euthanasia, samples of the rectum were collected for histopathology, cytokines dosage (IL-6 and KC), and immunohistochemical analysis (TNF-α and COX-2).ResultsHDR significantly reduced animals’ survival ten days post first radiation exposure (14% survival vs. 100% in the non-irradiated group). Day seven was then used for further investigation. Mice exposed to radiation presented with rectum injury confirmed by colonoscopy and histopathology (P < 0.05 vs. the control group). The tissue damage was accompanied by an inflammatory response, marked by increased KC and IL-6 tissue levels, and immunostaining for TNF-α and COX-2 (P < 0.05 vs. control group).ConclusionsWe established a novel animal model of actinic proctitis induced by HDR brachytherapy, marked by inflammatory damage and low animal mortality.

Elucidation of the Metabolite Profile of Yucca gigantea and Assessment of Its Cytotoxic, Antimicrobial, and Anti-Inflammatory Activities.

The acute inflammation process is explained by numerous hypotheses, including oxidative stress, enzyme stimulation, and the generation of pro-inflammatory cytokines. The anti-inflammatory activity of Yucca gigantea methanol extract (YGME) against carrageenan-induced acute inflammation and possible underlying mechanisms was investigated. The phytochemical profile, cytotoxic, and antimicrobial activities were also explored. LC-MS/MS was utilized to investigate the chemical composition of YGME, and 29 compounds were tentatively identified. In addition, the isolation of luteolin-7-O-β-d-glucoside, apigenin-7-O-β-d-glucoside, and kaempferol-3-O-α-l-rhamnoside was performed for the first time from the studied plant. Inflammation was induced by subcutaneous injection of 100 μL of 1% carrageenan sodium. Rats were treated orally with YGME 100, 200 mg/kg, celecoxib (50 mg/kg), and saline, respectively, one hour before carrageenan injection. The average volume of paws edema and weight were measured at several time intervals. Levels of NO, GSH, TNF-α, PGE-2, serum IL-1β, IL-6 were measured. In additionally, COX-2 immunostaining and histopathological examination of paw tissue were performed. YGME displayed a potent anti-inflammatory influence by reducing paws edema, PGE-2, TNF-α, NO production, serum IL-6, IL-1β, and COX-2 immunostaining. Furthermore, it replenished the diminished paw GSH contents and improved the histopathological findings. The best cytotoxic effect of YGME was against human melanoma cell line (A365) and osteosarcoma cell line (MG-63). Moreover, the antimicrobial potential of the extract was evaluated against bacterial and fungal isolates. It showed potent activity against Gram-negative, Gram-positive, and fungal Candida albicans isolates. The promoting multiple effects of YGME could be beneficial in the treatment of different ailments based on its anti-inflammatory, antimicrobial, and cytotoxic effects.

Pathological studies and detection of co-infection of bovine papilloma viruses in cattle cutaneous and teat warts

Bovine papillomas are benign tumors of the cutaneous and mucosal epithelia and are commonly found in cattle. The productivity loss and economic impact depends on the location and degree of infection. The present study was undertaken to investigate the pathology and association of different bovine papilloma virus (BPV) types in cattle cutaneous and teat warts. Grossly, the warts were of variable size and shape (rice grain, cauliflower and finger-like horny growths or irregular). Histopathologically, the warts were diagnosed as fibropapilloma, papilloma, fibrosarcoma and hyperplasia. Fibropapilloma was the most frequent histological type observed and was characterized by hyperkeratosis, parakeratosis and acanthosis. PCR revealed the presence of either BPV-1, -2 and -5 DNA or their co-infections. Transmission electron microscopy on negative staining showed BPV-like particles. Varied copy numbers of viral DNA of BPV-1, -2 and -5 were detected by real-time PCR. Immunohistochemistry revealed the expression of Ki-67 mainly in the proliferating cells of stratum spinosum and a few basal cells in papilloma and fibropapilloma. Cyclooxygenase-2 immunostaining was observed in the cytoplasm and cell membrane of suprabasal cells. In conclusion, cutaneous and teat warts in cattle in India are more frequently associated with BPV-1/ -2 and their mixed infections, with the rare presence of BPV-5. The DNA of BPV-5 was detected for the first time in warts in India. Co-infection with two or three different viral types demonstrated the diversity of BPV types involved in warts. The frequent expression of Ki-67 in suprabasal layers may be indicative of its association with viral replication and that they are as proliferation sites.

Effect of high fat diet on structure of liver and gallbladder of adult male mice – an experimental study

Introduction. High fat diet (HFD) intake induces obesity and adversely affects different body organs including liver and gallbladder. Aim. It was to clarify the effects of HFD on the liver and gallbladder structure using light microscopic (LM) examination. Material and methods. 16 healthy adult male mice were equally divided into 2 groups. Control group mice were fed normal diet. HFD group was fed using HFD. At the end of the 8-week experiment, mice were anesthetized. Liver and gallbladder were removed and prepared to histological processing. Sections were stained with hematoxylin and eosin (H&amp;E) and immunostaining for cyclooxygenase-2 (COX-2) cellular localization. Oil Red O (ORO)-stained frozen liver sections were prepared. Results. H&amp;E-stained sections of HFD group revealed rounded swollen hepatic cells with pale cytoplasm suggesting cellular ballooning. Dilated congested sinusoids and portal vein, cellular degeneration and collection of inflammatory cells were observed between hepatic cells and in portal region. Gallbladder sections showed epithelial stratification and cellular vacuolation. Strong immunoexpression of COX-2 was observed in Kupffer and hepatic cells of the liver and gallbladder mucosal epithelial cells. Conclusion. HFD is suggested to alter the normal histological features of liver and gallbladder represented by fatty liver and gallbladder epithelial hyperplasia and inflammatory reaction.

Developmental Expression of Cyclooxygenase isoforms in the hepatocytes of Albino Rat

AbstractBackground: The liver represents the major organ that participates in the elimination and degradation of arachidonic acid of systemic origin. This study is one of few types of research that studied the detailed expression of cyclooxygenase in the hepatocytes and other cells in liver tissue at different stages of maturation. Material and methods: A total number of 8 embryos and 40 rats at different ages postnatal were used. The prenatal rats were 18 days old of gestation while the postnatal rats were at 1, 7, 14, 21, and 28 days old. Aim of the work: The present study is concerned with exploring and compare the developmental expression of cyclooxygenase (COX-1, COX-2, and COX-3) in the liver tissue using immunohistochemical techniques. This will open the future for a new rationale of the optimal therapeutic use of COX in liver diseases.Results: The results showed that, in liver cells, the expression of COX isoforms was scientifically changed with the development of the liver tissue. COX-1 and COX-2 immunostaining were noticed to be the same in some ages however, the COX-1 expression resembled the COX-3 expression in other ages. Conclusion: It can conclude the expression of three types of COX isoforms in liver tissue played an important role in the development and maturation of different cells population either through prostaglandin production or a novel gene production. Nuclear localization of COX in the hepatocytes arises a lot of questions about the role of COX in the enhancement of liver division and maturation.

Ameliorative effect of nicorandil in ovarian ischemia-reperfusion-induced injury in rats: role of potassium channel.

Ovarian torsion is a gynecological emergency that leads to serious outcomes. Nicorandil (NIC) is an ATP-sensitive potassium (KATP) channel activator that protects the heart from ischemia. The current study aimed to investigate the role and mechanism of action of NIC in ovarian ischemia-reperfusion (OIR) and possible KATP participation. Twenty-four female albino rats were classified into 4 groups: sham control, OIR, OIR + NIC, OIR + NIC+ glibenclamide (GLB) groups. Serum anti-Müllerian hormone (AMH), ovarian malondialdehyde (MDA), total nitrites (NOx) contents, and superoxide dismutase (SOD) activity were evaluated. Bax and Bcl2 mRNA were also assessed. Histological and immunohistochemical (anti-COX-2 and anti CD68) studies were done. The OIR non-treated group showed histopathological ovarian injury with decreased AMH level. Ovarian MDA, NOx, and Bax mRNA and the expression of COX-2 and CD68 were increased; however, SOD activity and Bcl2 mRNA level were decreased by OIR. NIC significantly ameliorated the histopathological ovarian injury with the restoration of AMH level. NIC significantly corrected oxidative stress and apoptotic biomarkers with decreased COX-2 and CD68 immunostaining. GLB co-administration significantly decreased the protection afforded by NIC. These results imply that NIC has a protective role against OIR via antioxidant, anti-inflammatory, and anti-apoptotic effects and such protection relies, at least partially, on the KATP channel.

Cyclooxygenase-2 expression as a prognostic factor in pediatric classical Hodgkin lymphoma.

Cyclooxygenase-2 (COX-2) is an inflammation-related enzyme that has been shown to have a role in tumor initiation, angiogenesis, and proliferation. It has been demonstrated that COX-2 expression is increased in many tumors and is a negative prognostic parameter. Our objective is to investigate the prognostic value of COX-2 expression in pediatric patients with classical Hodgkin lymphoma (CHL). This was a retrospective analysis in pediatric patients (n = 127) diagnosed with CHL and treated at the pediatric oncology department, National Cancer Institute, Cairo University, January 2005-June 2013. We correlated COX-2 immunostaining in Reed-Sternberg (RS) cells with clinical variables and outcome. COX-2 was expressed on 38.6% of RS cells. The median follow-up time was 48.4months (range 4-114months). The 5-year OS and PFS, in COX-2(+ve) versus COX-2(-ve) was 85.3% versus 96.0% (p = 0.248) and 78.6% versus 84.3% (p = 0.354), respectively. A multivariate analysis showed that COX-2(+ve) was not significantly associated with the 5-year OS (HR = 2.9; 95% CI 0.7-12.4, p = 0.149) or with the 5-year PFS (HR = 1.4; 95% CI 0.6-3.2, p = 0.490). High-risk patients in the COX-2(+ve) group had a significantly lower 5-year OS (p = 0.021). The 5-year PFS was significantly lower in the COX-2(+ve) group with B symptoms (p = 0.023) and bulky disease (p = 0.028). Radiotherapy was given only to high-risk patients; survival was much better in radiation-treated children in both the Cox-2(+ve) and Cox-2(-ve) groups. The magnitude of the radiotherapy effect was also greater in the Cox-2(+ve) group, but this difference was not statistically significant. COX-2 expression showed a tendency to be a poor prognostic factor, but it failed to provide meaningful independent information. Further larger studies are needed to investigate COX-2 as a prognostic factor and potential therapeutic target.

Endometritis in the bitch: Immunohistochemical localization of cyclooxygenase 2.

Background:In several mammals, subfertility or infertility associated with endometritis was reported. Although there have been studies about endometritis in bitches, the pathophysiological mechanisms are not completely known.Aim:This study aimed to evaluate the immunohistochemical expression of Cyclooxygenase 2 (COX2) in clinically healthy bitches with normal uterine tissue and bitches with endometritis. Methods:Forty-eight mixed breed bitches in diestrus were used. Uterine biopsies were collected for diagnosis [normal endometrium (n = 15; NE), cystic endometrial hyperplasia (n = 1), atrophy (n= 2), acute endometritis (n = 9; AE), subacute endometritis (n = 7; SE), and chronic endometritis (n = 14; CE)]. Immunostaining and quantification of positively stained cells was performed on full-thickness uterine biopsies. Data were analyzed by the GLIMMIX procedure of SAS. Results:COX2 immunostaining was scattered and restricted to cells in the stroma in bitches with NE. However, in bitches with endometritis, strong staining was observed in the luminal epithelium, glandular epithelium, and stromal cells. Staining was also observed in inflammatory cells localized in the stroma as well as inside of the glands. The percentage of COX2 positive stromal cells in bitches with AE, SE, and CE was significantly higher compared with NE (p < 0.005). In addition, the percentage of COX2 positive stromal cells in bitches with SE, and CE was significantly lower compared with AE (p < 0.003). Conclusion:COX2 could be involved in the pathophysiological mechanisms producing endometritis without the presence of cystic endometrial hyperplasia in bitches. However, further researches on this topic are required.

Increased COX-2 Immunostaining in Urothelial Carcinoma of the Urinary Bladder Is Associated with Invasiveness and Poor Prognosis.

Background Urothelial carcinoma of the urinary bladder (UCB) is the commonest bladder tumor. Cyclooxygenase-2 (COX-2) mediates angiogenesis, cell survival/proliferation, and apoptosis. This study investigates the relation of COX-2 immunostaining in UCB to clinicopathological parameters in Saudi Arabia. Methods The study population includes 123 UCB and 25 urothelial mucosae adjacent to UCB. UCB samples were collected before any local or systemic therapy. Tissue microarrays were designed and constructed, and TMA blocks were sliced for further immunohistochemical staining. Immunohistochemical staining was done using a mouse anti-human COX-2 monoclonal antibody. A cutoff point of 10% was chosen as the threshold to determine low and high COX-2 immunostaining. Results COX-2 immunostaining is higher in UCB than in the adjacent urothelium (p = 0.033). High COX-2 immunostaining is associated with high-grade UCB (p = 0.013), distant metastasis (p = 0.031), lymphovascular invasion (p = 0.008), positive muscle invasion (p = 0.017), pT2 and above (p = 0.003), and high anatomical stages (stage II and above). High COX-2 immunostaining is an independent predictor of higher tumor grade (p < 0.001), muscle invasion (p = 0.015), advanced pathological T (p = 0.014), lymphovascular invasion (p = 0.011), and distant metastasis (p = 0.039). High COX-2 immunostaining is associated with lower overall survival rate (p = 0.019). Conclusion COX-2 immunostaining is associated with the invasiveness of UCB which may be used as an independent prognostic marker. COX-2 may be a significant molecule in the initiation and progression of UCB. Molecular and clinical investigations are required to explore the molecular downstream of COX-2 in UCB and effectiveness of COX-2 inhibitors as adjuvant therapy along with traditional chemotherapy.

Chronic treatment with zoledronic acid alters the expression levels of inflammatory, bone, and apoptotic markers and Toll-like receptors 2 and 4 in rat dental pulp

The aim of this study was to evaluate the immunostaining of inflammatory, apoptotic, and bone markers, as well as Toll-like-receptors (TLRs) 2 and 4 in the dental pulp in rats treated with zoledronic acid (ZA). We administered 4 intravascular infusions of saline (control group) or 0.20 mg.kg-1 ZA in Wistar rats (n = 6/group). After 70 days, the 3 rights molars (n = 18/group) were microscopically evaluated (presence of ectasic/dilated blood vessels and inflammatory cells). Immunohistochemistry was performed for tartrate resistant acid phosphatase 5 (TRAP; cell counting), cyclooxygenase-2 (COX-2), interleukin-6 (IL-6), TLR2, TLR4, receptor activator of nuclear kappa B ligand (RANKL), osteoprotegerin (OPG), and caspase-3 (scored 0-3 in odontoblast and nonodontoblast dental pulp cells). Mann-Whitney and Fisher's exact tests and Spearman's correlation were used (GraphPad Prism 5.0). There was no alteration in ectasic/dilated blood vessels (P = .101) or inflammatory cells (P = .500), but the number of TRAP-positive cells was reduced in the ZA-group (P = .027). In ZA-group odontoblasts, immunostaining for COX-2 (P = .044), TLR4 (P = .003), OPG (P = .035) and caspase-3 (P = .039) increased, and that for RANKL (P = 0.045) decreased. In nonodontoblast dental pulp cells, RANKL immunostaining decreased (P = .009). In the ZA group, the RANKL/OPG ratio decreased in odontoblast (P = .022) and nonodontoblast dental pulp cells (P = .007). IL-6 did not differ between the groups. ZA increases the expression levels of inflammatory, apoptotic markers, and TLR4 and alters bone makers in the dental pulp of rats.

PS02.043: OVEREXPRESSION OF HSP27 AND HSP70 ARE ASSOCIATED WITH DECREASED SURVIVAL AMONG PATIENTS WITH ESOPHAGEAL ADENOCARCINOMA

Abstract Background Heat shock protein (HSP) 27 and 70 overexpression is associated with several malignancies but reports of their incidence and association with survival among patients with esophageal carcinoma (EC) are scarce. COX-2 expression is induced by inflammation, and it has been linked to a Barrett's esophagus and a wide variety of cancers. High COX-2 expression has been linked to poor survival in EC but results regarding association to clinicopathological factors are conflicting. Methods Immunohistochemical analysis for HSP27, HSP70 and COX-2 was performed on 97 cancer cases, 19 Barrett cases and 5 controls treated between 1990 - 2007. Data was gathered from patient records and survival data was acquired in December 2015. Results Higher tumor stage was significantly associated with poorer survival (P &lt; 0001), which was independent of HSP27, HSP70 and COX-2 immunostaining. HSP27 was moderately or strongly positive in 61,8% of the cancer cases, which was significantly associated to poorer survival (25,6 and 19 months versus 65,1 in weakly positive samples) (P = 0,01). Similar results were seen for HSP70, for which 30,4% of the cases were stained moderately or strong positive, and survival was 15 and 17,3 months compared to 41,7 months in weakly stained cases. COX-2 was negative in 83,3% of the cancer cases, and none of the samples had strong COX-2 immunostaining. COX-2 did not correlate with survival. Conclusion Moderate or high HSP27 and HSP70 staining intensities were associated with poor survival among patients with esophageal adenocarcinoma, which is now reported for the first time. According to previous studies opposite conclusions for esophageal squamous cell carcinoma was found. COX-2 did not impact survival in our study even though it's been found to be a negative prognostic factor in several previous studies. Disclosure All authors have declared no conflicts of interest.

Neutrophils contribute to the pathogenesis of hemorrhagic cystitis induced by ifosfamide

Ifosfamide (IFO) is an antineoplastic drug that is commonly used to treat gynecological and breast cancers. Hemorrhagic cystitis (HC) is a common side effect associated with IFO injection, which courses with neutrophil accumulation and affects 6–50% of patients depending on dose intensity. Here, we investigated the role of neutrophils in this inflammatory process. Female Swiss mice (n = 8/group) were injected with saline, IFO (400 mg/kg, i.p.), fucoidan (a P- and L-selectins inhibitor, 100 mg/kg, i.v.) or IFO + fucoidan (1–100 mg/kg) alone or combined with mesna (80 mg/kg i.p.). Another group of mice received anti-Ly6G antibody (500 μg/mouse, once daily for 2 days) for neutrophil depletion before IFO injection. In another experimental setting, animals received granulocyte colony-stimulating factor (G-CSF, 400 μg/kg), IFO (200 mg/kg), G-CSF (25–400 μg/kg, for 5 days) + IFO (200 mg/kg, i.p.) or fucoidan + G-CSF + IFO. Bladder injury was evaluated 12 h after IFO injection. IFO 400 mg/kg significantly increased visceral hyperalgesia, bladder edema, hemorrhage, vascular permeability, MPO, IL-1β and IL-6 tissue levels, and COX-2 immunostaining and expression versus the saline group (P < 0.05). Conversely, fucoidan (100 mg/kg) significantly attenuated these parameters compared to IFO-injected mice (P < 0.05). Additionally, fucoidan potentiated mesna protective effect when compared with IFO + mesna group (P < 0.05). Accordingly, neutrophil depletion with anti-Ly6G reduced inflammatory parameters and bladder injury compared to IFO (P < 0.05). In contrast, G-CSF enhanced IFO (200 mg/kg)-induced HC, which was significantly attenuated by treatment with fucoidan (P < 0.05). Therefore, neutrophils contribute to the pathogenesis of HC.