Functional block of IL-17 cytokine promotes bone healing by augmenting FOXO1 and ATF4 activity in cortical bone defect model.

We determine the effect of interleukin (IL)-17 neutralizing antibody on new bone regeneration. Anti-IL-17 antibody promoted new bone regeneration in cortical bone defect model by augmenting FOXO1 and ATF4 activity thereby decreasing oxidative stress. Our study demonstrates the bone healing and regeneration potential of neutralizing IL-17antibody in osteoporotic fractures. INTRODUCTION: The immune system plays important role in the fracture healing process. However, fracture healing is prolonged in disorders associated with systemic inflammation. Fracture healing is decelerated in osteoporosis, condition linked with systemic inflammation. Bone regeneration therapies like recombinant human BMP2 are associated with serious side effects. Studies have been carried out where agents like denosumab and infliximab enhance bone regeneration in osteoporotic conditions. Our previous studies show the osteoprotective and immunoprotective effects of neutralizing IL-17 antibody. Here, we determine the effect of IL-17 neutralizing antibody on new bone regeneration and compare its efficacy with known osteoporotic therapies. METHODS: For the study, female BALB/c mice were ovariectomized or sham operated and left for a month followed by a 0.6-mm drill-hole injury in femur mid-diaphysis. The treatment was commenced next day onwards with anti-IL-17, anti-RANKL (Receptor activator of nuclear factor kappa-B ligand), parathyroid hormone (PTH), or alendronate for a period of 3, 10, or 21 days. Animals were then autopsied, and femur bones were dissected out for micro-CT scanning, confocal microscopy, and gene and protein expression studies. RESULTS: Micro-CT analysis showed that anti-IL-17 antibody promoted bone healing at days 10 and 21, and the healing effect observed was significantly better than Ovx, anti-RANKL antibody, and ALN, and equal to PTH. Anti-IL-17 also enhanced new bone regeneration as assessed by calcein-labeling studies. Additionally, anti-IL-17 therapy enhanced expression of osteogenic markers and decreased oxidative stress at the injury site. CONCLUSION: Overall, our study demonstrates bone healing and regeneration potential of neutralizing IL-17 antibody in osteoporotic fractures.

The role of the male cloacal gland in reproductive success in Japanese quail (Coturnix japonica).

The adult male Japanese quail has a well developed cloacal gland that produces meringue-like white foam. The physiological significance of the cloacal gland and its foam is still unclear. Therefore, we conducted two experiments to establish the functional role of the cloacal gland and its foam in natural mating and oviducal sperm transport. In the first experiment, artificial insemination of equal numbers of spermatozoa diluted in foam extract and normal saline once in a week were used to determine the role of foam in sperm transport in the female oviduct. After artificial insemination, eggs were collected to measure fertility, the duration of the fertile period, sperm holes and attached spermatozoa in the perivitelline membrane. Higher (P<0.05) fertility and greater duration of the fertile period were observed when semen was inseminated along with foam extract compared with normal saline. Further, the sperm holes and trapped spermatozoa in the perivitelline membrane were also higher (P<0.05) in the presence of foam extract. In the second experiment, two males with bigger and smaller cloacal gland areas were allowed to mate with a female. The mating attempts of males with larger cloacal gland were more successful (P<0.05) than males with smaller cloacal glands. Our results indicated that cloacal foam improves sperm transport in the female oviduct and that males with larger cloacal gland areas are preferred during mating.

Cytokines Expression and Nitric Oxide Production under Induced Infection to Salmonella Typhimurium in Chicken Lines Divergently Selected for Cutaneous Hypersensitivity.

In the present study, the impact of Salmonella Typhimurium on cell-mediated immunity (CMI) was investigated in 5 week-old immuno divergent broiler lines selected for the high and low response to phytohemagglutinin-P. The immune response was assessed in peripheral-blood mononuclear cells (PBMCs) induced with Salmonella Typhimurium at different time intervals (0 h, 0.5 h, 2 h, 4 h, 6 h, 12 h and 24 h). The differential mRNA expression patterns of IFN-γ, IL-2 and iNOS were evaluated by quantitative real time PCR. In-vitro production of nitric oxide (NO) was also estimated in the culture supernatant and correlated with iNOS mRNA expression. Present study showed higher production of NO in the high cell-mediated line (HCMI) as compared to the low cell-mediated line (LCMI) upon stimulation with Salmonella Typhimurium. Correspondingly, higher mRNA expression of iNOS and IFN-γ were observed in high response birds (HCMI); but IL-2 was down regulated in this line compared to the low response birds (LCMI). Significantly (p<0.05) higher expression of iNOS, IFN-γ and higher production of NO in high line indicated that the selection for PHA-P response might be employed for increasing the immune competence against Salmonella Typhimurium in chicken flocks.

A Review of the Cytokine IL-17 in Ocular Surface and Corneal Disease.

Aim: To investigate the role of interleukin-17 in ocular surface and corneal disease. Ocular surface and corneal disease is a leading cause of blindness and is an ongoing challenge for the public health sector to implement effective therapies. The majority of cells in corneal lesions are derived primarily from neutrophils that induce inflammatory events that lead to tissue damage. One of the key pro-inflammatory cytokines is IL-17, and it has been investigated in order to facilitate the understanding of the pathogenesis of ocular surface lesion development. Method: A review of the literature was performed through a systematic approach. Results: IL-17 has been shown to exacerbate dry eye disease, viral and bacterial keratitis lesion severity, although it was found to be protective for Acanthamoeba. Antibodies developed to neutralize IL-17 have shown some promise in reducing the severity of some diseases. Conclusion: IL-17 plays a role in the pathogenesis of ocular surface and corneal disease and targeting this cytokine may provide a useful treatment option in the future.

Differential expression of inducible nitric oxide synthase and cytokine mRNA in chicken lines divergent for cutaneous hypersensitivity response.

Phytohemagglutinin (PHA)-induced delayed-type hypersensitivity is an immunocompetent trait considered an indicator of cell-mediated immune or T-cell responses. Divergent selection was performed to generate high and low lines for response to PHA-P. Extreme-responder birds of the F2 generation in each line were used to study possible differences in macrophage activity and the associated functional genes. To evaluate macrophage activity, nitric oxide (NO) was estimated both systemically in serum and in in vitro monocyte culture. Semi-quantitative RT-PCR was used to detect the differential mRNA expression patterns of iNOS and MIP-1beta in monocyte culture, whereas T(H)1 cytokines (IL-2 and IFN-gamma) were studied in peripheral blood mononuclear cells (PBMC) at different time intervals after lipopolysaccharide (LPS) induction. The high line showed strong systemic, as well as in vitro NO production, compared to the low line, upon stimulation with NDV and LPS, similar to early and high iNOS mRNA expression. Following the pattern of iNOS gene expression, an early strong expression of cytokines with powerful iNOS-inducing action, such as IFN-gamma and the chemokine MIP-1beta, was observed in the high line. In contrast, for response to PHA-P, low expression of IL-2 was observed in the high compared to the low line. In conclusion, the study revealed that divergent selection for response to PHA-P resulted in a divergent effect on T(H)1 cell activity, resulting in altered macrophage function in chickens. Selection, based on response to PHA-P, could lead to more resistant birds or birds with an enhanced immune response.

Evidence for the hypothalamic origin of the polycystic ovary syndrome.

The effect of suppression of gonadotropin secretion was evaluated in 21 patients with the polycystic ovary syndrome. Medroxyprogesterone acetate (MPA) was administered intramuscularly in a dose of 400 mg every 15 days for 9 months. A significant decrease in luteinizing hormone (LH) and testosterone levels (70 and 40%, respectively) was apparent after 3 months. At the end of the treatment period, the ovaries had become impalpable and hirsutism was markedly improved in 13 of 19 women. Side effects of treatment included local pain, vaginal spotting, galactorrhea, and hyperprolactinemia. Discontinuation of therapy was followed by a rapid return of follicle-stimulating hormone levels to baseline values, whereas LH and testosterone levels recovered only partially after more than 1 year. The improvement of hirsutism and ovarian shrinkage persisted for up to 2 years, and endometrial biopsy uniformly showed a pseudodecidual reaction in the stroma. After 1 year, prolactin levels declined to 52% of the baseline value and galactorrhea disappeared. The suppression of all the peripheral abnormalities of the reproductive system in polycystic ovary syndrome with MPA treatment suggests a primary hypothalamic disorder as the cause for the syndrome.

Coexistence of polycystic ovary syndrome and pelvic endometriosis.

Pelvic endometriosis was observed in 15 of 91 women (16.5%) with laparoscopically confirmed polycystic ovary syndrome. There were no significant clinical differences among those with and those without endometriosis. The groups were of similar age, parity, and ponderal indices and had similar incidences of oligomenorrhea, hirsutism, and infertility; the serum concentrations of LH, FSH, LH/FSH, prolactin, testosterone, and dehydroepiandrosterone sulfate were also similar in each group. However, women with polycystic ovaries and endometriosis presented more frequently with regular menses (40 versus 14.5%; P = .05) and less frequently with secondary amenorrhea (0 versus 38.2%; P = .05) and galactorrhea (0 versus 9.2%; P = .05) than the women with polycystic ovaries alone. Endometriosis appears to be a coincidental finding in polycystic ovary syndrome, and its development does not modify significantly the clinical picture or biochemical profiles of these patients. However, menstrual patterns seem to be affected.

Cesarean scar endometriosis: a review.

Endometriosis in the abdominal scar following cesarean section is a rare event, but may be more frequent than estimated from the literature. Five cases collected over 5 years are presented suggesting an incidence over 1 percent. Diagnosis and management are discussed and the literature regarding this entity is reviewed. Newer methods of treatment are available, but surgical excision remains the method of choice.

Clinical sequelae of Japanese encephalitis in children.

Over a five and a half year period, virological investigations for Japanese encephalitis (JE) were conducted in children admitted with acute encephalitis like illness to a large city hospital. The diagnosis of Japanese encephalitis was made by viral isolation from cerebrospinal fluid and/or a four-fold or higher rise in haemagglutination inhibiting antibodies in paired sera followed by demonstration of specific IgM antibodies by HI test after treatment with 2-mercapto ethanol. All children surviving the illness were contacted by post and followed up for sequelae. A total of 55 children could be followed up after 12-18 months and 22 of these even after 2 yr. A high rate of major sequelae (45.5%) in the form of frank motor deficits (32.7%), mental retardation (21.8%) and/or convulsions (18.2%) was observed. Neurological deficits were of diverse types and improved even after 2 yr of the illness. Fourteen patients (25.4%) had only minor deficits in the form of scholastic backwardness, behavioural problems and/or subtle neurological signs. Only 16 (29.2%) patients were completely normal on follow up. JE may therefore be an important cause of neurological handicap in this area. Sequelae of the disease were more severe if the initial illness was prolonged (P < 0.001, CI 2.45, 12.64), or associated with focal neurological deficits (P < 0.001, CI 1.97, 7.02).

Ultrastructural basis for continued steroidogenesis in the rat polycystic ovary.

Because constant light causes persistent estrus in the rat, that animal provides a suitable experimental model for studying the pathogenesis of polycystic ovary disease. We studied the scanning and transmission electron microscopic features of the rat polycystic ovary along with changes in serum estradiol and progesterone concentrations to further elucidate the mechanism of chronic anovulation during persistent estrus. The surface epithelium of the polycystic ovary showed no ovulatory stigmas and contained focal areas of degenerating and proliferating cells. The tunica albuginea was conspicuous, with many fibrils, collagenous materials and fibroblastlike cells adjoining the theca externa layer. The cytoplasm of theca interna cells contained numerous lipid vacuoles and images of extracted cholesterol crystals. The granulosa cells were atrophic and lacked the prominent Golgi apparatus typically found in granulosa cells of proestrous ovaries in cycling rats. With an increasing duration of constant light and chronic anovulation, the mean serum estradiol concentration in persistent-estrus rats was significantly higher and the mean serum progesterone concentration significantly lower than in age-matched controls. Alterations in estradiol-progesterone metabolism in the ovaries may be important etiologic factors in the pathogenesis of chronic anovulation found in polycystic ovary disease in rats.

Effect of obesity on the clinical and hormonal characteristics of the polycystic ovary syndrome.

The polycystic ovary (PCO) syndrome is frequently associated with obesity. That subset of women reportedly shows a much higher incidence of hirsutism and menstrual irregularities than do nonobese women with PCO syndrome. We evaluated the clinical features and hormonal profiles of 56 women with PCO syndrome and correlated them with the presence or absence of obesity. Thirty-eight (67.8%) of these women were obese (body mass index > or = 25 kg/m2). While presenting with the classic manifestations of PCO, they did not differ significantly from the manifestations of nonobese women with PCO syndrome. Although obese women with PCO had a lower incidence of oligomenorrhea as compared to nonobese women with PCO (57.9% vs. 83.3%, respectively) and amenorrhea was more frequent in the former group (42.1% vs. 16.6%, respectively), these findings are not statistically significant. The incidences of hirsutism and anovulatory infertility in the obese group as compared to the nonobese group were 81.6% vs. 77.8% and 28.9% vs. 27.8%, respectively (not statistically significantly different). The mean (+/- SE) serum levels of luteinizing hormone (LH), follicle stimulating hormone (FSH), LH/FSH ratios, prolactin and testosterone were not statistically significantly different among the two groups. The present study found that obesity is common in PCO syndrome but that there are no significant differences in the clinical and hormonal characteristics of obese and nonobese women with it. Further studies are warranted to clarify the impact of obesity on clinical, metabolic and hormonal changes in PCO syndrome.

Gestational diabetes and neonatal macrosomia in the polycystic ovary syndrome.

Since patients with polycystic ovary syndrome (PCOS) commonly have insulin resistance, albeit with normal glucose tolerance, we evaluated glucose tolerance in PCOS patients exposed to the diabetogenic effect of pregnancy. The clinical material was obtained from two centers, in Springfield, Illinois (22 patients), and New York, New York (31 patients), and the results were compared with a control population with 2,306 consecutive general pregnancies. There were no differences between PCOS patients from the two centers in regard to age or ponderal index (P greater than .1). A review of the medical records showed that the incidence of gestational diabetes in the PCOS patients was 7.5%, similar (P greater than .1) to the 6.6% frequency of gestational diabetes in the controls. The overall incidence of neonatal macrosomia (birth weight greater than 4,000 g) was 7% (4 of 57) among infants born to PCOS women. That was similar to the 12.4% incidence of neonatal macrosomia among infants born to women with normal glucose tolerance and to the 14.5% incidence among infants born to women with gestational diabetes. Preexisting PCOS does not appear to increase the risk of developing gestational diabetes or neonatal macrosomia.

Clomiphene-dexamethasone treatment of clomiphene-resistant women with and without the polycystic ovary syndrome.

We used clomiphene and dexamethasone in 40 infertile women to treat chronic anovulation resistant to the use of clomiphene alone. Eighteen (45%) of the women had the polycystic ovary (PCO) syndrome; the remaining 22 (55%) had clomiphene-resistant anovulation from idiopathic causes. Both groups of women were similar in regard to age, parity, duration of infertility and absence of other causes of infertility besides chronic anovulation. Ovulation could be induced in approximately 90% of the women in each group. Altogether, 19 of 36 women (52.8%) conceived without any side effects or complications. The cumulative probability of conception at nine months of treatment was 87.5% in PCO patients and 46% in the non-PCO group. Clomiphene plus dexamethasone was highly effective in the treatment of clomiphene-resistant anovulation associated with infertility in women with and without the PCO syndrome.

Characteristics of indigent women with ruptured and unruptured tubal pregnancies.

The diagnosis of tubal pregnancy, whether ruptured or unruptured, often requires a surgical procedure, such as laparoscopy or laparotomy, for confirmation. We compared women with ruptured and unruptured tubal pregnancies to determine whether the clinical presentations, morbidity and surgical complications in the two groups were significantly different. We compared the demographic characteristics, clinical presentations, laboratory findings, morbidity and complications from surgical management in the two groups. Women with ruptured tubal pregnancies had a higher incidence of abdominal pain lasting less than 24 hours, adnexal tenderness and positive culdocentesis from hemoperitoneum as compared to women with unruptured tubal gestations. Abnormal uterine bleeding was observed less frequently in women with ruptured tubal pregnancies as compared to women with unruptured ones despite similar gestational ages at presentation. All the patients with a tubal pregnancy were managed surgically. The morbidity and surgical complication rates in the two groups were not significantly different.

Comparative ABR profile in high risk infants.

Auditory brainstem responses (ABR) were studied in 52 children upto the age of 3 months, suffering from neonatal jaundice, prematurity, pyogenic meningitis and septicemia. Absolute latency of wave I and interpeak latency I-V were found to be significantly delayed in cases of jaundice and absolute latency of wave V and interpeak latency I-V were prolonged in cases with prematurity. In cases following pyogenic meningitis absolute latency of wave V and interpeak latency of I-III, I-V were significantly delayed compared to septicemia where absolute latency I, V and interpeak latency I-V were significantly delayed (P < 0.05). Maximum auditory insult was seen in cases with neonatal jaundice where 30.77% each had severe SN deafness and 30.77% had moderate degree of deafness compared to prematurity, meningitis and septicemia where 14.28, 7.69 and 25% were found to have severe SN deafness and 7.14, 38.46 and 25% had moderate deafness. Overall incidence of deafness of any kind in these factors was 44.23%.

Paraganglioma of the external auditory canal.

A rare case of paraganglioma of the external auditory canal is presented. It was treated by excision and no recurrence has been noted after 18 months of follow up.

Laryngeal malakoplakia.

We report a case of malakoplakia of the epiglottis in a 45-year-old female patient. Only three cases of laryngeal malakoplakia have been reported in the world literature, one of which was associated with tracheal malakoplakia. To our knowledge this is the first reported case of isolated malakoplakia of the epiglottis. Malakoplakia, a rare granulomatous lesion, has been seen more frequently in the urinary tract. Other organs like the genito-urinary tract, testis, epididymis, lymph nodes, middle ear, nasopharynx, tonsil and retroperitoneal tissue have been also involved though less frequently.

Pelvic urethrotomies in bulls.

Pelvic urethrotomies were attempted on 32 bulls (12 cattle and 20 buffaloes). Infra-anal urethrotomy was carried out under a tranquilliser, posterior epidural and supra-ischial blocks. Prebulbomuscular urethrotomy was performed under a tranquilliser, posterior epidural and pelvic plexus blocks. Cystoscopy and litholapaxy were attempted to make comparisons. The animals which had been operated on were maintained for one month during which postoperative signs were noted, and after they were killed post mortem, radiographic, macro- and microscopic findings were recorded. From the results it is concluded that infra-anal urethrotomy presents difficulties whereas prebulbomuscular urethrotomy, especially the closed technique, is suitable for cystoscopy, pelvic lithotomy and litholapaxy.

Changes induced by morphine administration in the hypothalmic neurosecretory system of the spotted owlet, Athene brama Temminick.

The effect of chronic administration of morphine, in gradually increasing doses, on the histology of the HNS of the spotted owlet was investigated. Administration of morphine in low doses (0.25-0.75 mg/day), for 9 days, induced marked depletion of NSM from all regions of the HNS, except the zona externa of the AME. Maximum depletion of NSM was noticed in the HNS of birds treated with morphine for 9 days. Beyond the 9th day, the HNS did not exhibit further depletion of its NSM. Administration of morphine in high doses (1.5-4 mg/day), for 18 days, did not cause depletion of NSM from the HNS. On the other hand there was noticeable increase of the NSM in the HNS over that of the controls. The histological changes in the HNS induced by low doses of morphine are comparable to the changes induced by hypertonic saline administration and are presumably indicative of augmented secretion of the ADH. The accumulation of NSM in the HNS of birds receiving high doses of morphine suggests a decrease in neurohypophysial activity. The results therefore indicate that the effect of morphine on the HNS of the spotted owlet is dose-dependent. The mechanism of action of morphine on the neurohypophysis is briefly discussed.

In vitro rapid clearance of infectious bursal disease virus in peripheral blood mononuclear cells of chicken lines divergent for antibody response might be related to the enhanced expression of proinflammatory cytokines.

Infectious bursal disease (IBD) is an acute and highly contagious viral disease of young chickens caused by infectious bursal disease virus (IBDV). An effective way to control IBDV would be to breed chickens with a reduced susceptibility to IBDV infection. In the present work, we used chickens selected for high and low specific responses to sheep red blood cells (SRBC) (H and L, respectively) to assess the susceptibility of differential immune competent animals to IBDV infection. The peripheral blood mononuclear cells (PBMCs) of high SRBC line (HL) and low SRBC line (LL) were infected with IBDV and viral RNA loads were determined at different time post-IBDV infection. Chicken orthologues of the T helper 1 (Th1) cytokines, interferon-γ (IFN-γ) and interleukin-2 (IL-2); a Th2 cytokine, IL-10; a pro inflammatory cytokine, IL-6; the CCL chemokines, chCCLi2, chCCLi4 and chCCLi7; colony stimulating factor, GM-CSF; and a anti-inflammatory cytokine, transforming growth factor ß-2 (TGFß-2) were quantified. The expression of chCCLi2, chCCLi4 and chCCLi7 was significantly higher in L line as compared to H line. However, in H line the viral RNA loads were significantly lower than in L line. Therefore, the upregulated chemokines might be associated with the susceptibility to IBDV. The expression of IFN-γ, IL-2 and IL-6 was significantly higher in H line as compared to L line. We assume that the higher proinflammatory cytokines expression in H line might be related to the rapid clearance of virus from PBMCs. Significantly higher levels of IL-10 and TGFß-2 mRNAs in L line might be related to the pathogenesis of IBDV. In conclusion, selection for antibody responses appears to influence the expression profiles of chemokines and cytokines against IBDV. Further, the selection for high SRBC response might improve the immuno-competence of chickens against IBDV.