BIIB021, an orally available and small-molecule inhibitor of HSP90, activates intrinsic apoptotic pathway in human cervical adenocarcinoma cell line (HeLa).

OBJECTIVE: Heat shock protein 90 (HSP90) is a highly conserved ATP-dependent chaperone protein that plays a vital role in tumorigenesis. This study aims to investigate the apoptosis inducer role of BIIB021 (orally available HSP90 inhibitors) compound via inhibition of HSP90 activity in the human cervical cancer cell line (HeLa). PATIENTS AND METHODS: The anticancer potential of BIIB021 was determined by XTT [2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)] cell proliferation assay against the human cervical cancer cell line (HeLa). ATPase and luciferase aggregation assays were carried out to detect the HSP90 inhibitor potential of BIIB021. To determine the antiproliferative mechanism of the BIIB021, the expression level of the pro-apoptotic and antiapoptotic markers was determined by reverse transcription polymerase chain reaction (RT-PCR) and ELISA experiments. RESULTS: BIIB021 exhibited a cytotoxic effect on HeLa cell proliferation and the inhibitory concentration (IC)50 dose of BIIB021 was found to be 14.79 nM at 48 h. BIIB021 decreased the ATP hydrolysis rate of HSP90 and blocked the refolding of the desaturated luciferase in the presence of ATP. To understand the antiproliferative mechanism of the BIIB021 in HeLa cells, the mRNA and protein expression levels of the apoptotic markers [BCL-2 associated X (BAX), B-cell lymphoma 2 (BCL-2), cytochrome-c (CYT-c), and caspase-3 (CAS-3)] were determined by RT-PCR and ELISA experiments. The results obtained indicated that BIIB021 decreased BCL-2 levels and increased BAX, CYT-c, and CAS-3 levels in human cervical cancer cells. CONCLUSIONS: These results confirmed that BIIB021 inhibited the chaperone activity of HSP90, resulting in anti-proliferating effects in cervical cancer cells via the induction of the intrinsic apoptotic pathways.

Evaluation of second trimester uterine artery doppler indices in women diagnosed with COVID-19 during the first trimester of their pregnancy.

OBJECTIVE: This study aimed at evaluating the hypothesis that 6-14 months pregnant women diagnosed with COVID-19 may have abnormal placental function detectable by increased uterine artery Doppler indices in the second trimester and whether these women could benefit from treatment. PATIENTS AND METHODS: A total of 63 women were diagnosed with COVID-19 during the first trimester of their pregnancy and 68 healthy women were involved according to the exclusion criteria. In both groups, Doppler measurements were performed for the determination of high-risk pregnancy by increased uterine artery Doppler indices in the second trimester. RESULTS: It was observed that uterine artery Doppler indices (PI and RI) were significantly increased in second trimester women with COVID-19 infection compared to those without COVID-19. Furthermore, the number of women with a PI value exceeding the 95th percentile and the number of patients with early diastolic notch were higher in the COVID group compared to the control group. CONCLUSIONS: Doppler Ultrasound measurement may be a potential method for the management of high-risk pregnancies after asymptomatic/mild COVID-19 infection.

Evaluation of extremity vascular injuries and treatment approaches.

BACKGROUND: : Vascular injuries are commonly seen in both emergency services and forensic medicine practise. They are often life-threatening, with high morbidity and mortality rates. AIMS: This study aimed to retrospectively evaluate extremity vascular injuries and the associated treatment approaches. METHODS: After obtaining approval from the ethics committee of the university, those patients admitted to the emergency department of Adiyaman between 1 February 2013 and 31 August 2018 were included in this study. The patients' data were obtained through the electronic records system, and the cases were evaluated according to the age, gender and cause of injury, including blunt force trauma injuries (accidents, traffic accidents, crush injuries and occupational accidents) and penetrating injuries (stabbing, gunshot wounds, suicide attempts and assaults). Additionally, the injuries were evaluated based on the extremity, according to the anatomical location and whether the injury was life-threatening. RESULTS: This study included 76 patients with extremity vascular injuries; 65 were males (85.52%), 11 were females (14.48%) and their average age was 33.24 ± 15.85 years. Forty-five (59.2%) of the patients had upper extremity vascular injuries, and 31 (40.3%) had lower extremity vascular injuries. In addition to arterial injuries, 26 (34.21%) of the patients had venous injuries and 22 (28.94%) had nerve injuries. Nine of these patients had neurological deficits due to their nerve injuries. All of the patients were revascularized within 3-5 hours, and none of the patients required amputations. CONCLUSION: The primary goals in extremity vessel injury cases are to prevent mortality, especially after major vascular injuries, and save the extremity from amputation. With a fast, effective and multi-disciplinary approach, an accurate diagnosis and effective surgical intervention can prevent morbidity and mortality as well as reduce the rate of undesirable complications.

The effects of leptin on F-actin remodelling in type 1 diabetes.

BACKGROUND: The aim of the current study is to investigate the effect of leptin on cytoskeleton structures in both in vivo and in vitro model of diabetes. MATERIALS AND METHODS: For in vivo studies, leptin in different doses (240, and 480 mg/kg) was injected to the diabetic rats after 1-week of streptozotocin (STZ, 55 mg/kg) treatment. Leptin levels were analysed in serum, liver, and pancreas samples. Hepatic and pancreatic F- and G-actin expressions were determined by Western blotting. For in vitro studies, hepatic and pancreatic primary cell lines were obtained from the control rats. To these cultures, STZ (15 and 30 mM), leptin (50, 60 and 100 ng/mL), and their combinations were applied for 1, 3, and 4 weeks. After the treatment period, F-actin was visualised by the Alexa-fluor fluorescent dye. RESULTS: Streptozotocin decreased the G-actin in both tissues in vivo. However, leptin caused a dose-dependent increase in G-actin levels while F-actin decreased in both tissues. Moreover, leptin caused the perimembranous condensation of actin filaments and amelioration of F-actin structures in vivo. A dose-dependent corruption of F-actin filament structures was observed in leptin-treated primary cells in vitro, while STZ also caused corruption of these filaments. Co-exposure of STZ and leptin caused the amelioration of F-actin filaments, while the peri- membranous condensation was also observed as was in vivo study. CONCLUSIONS: Leptin therapy could be a candidate for diabetes, but it should not be ruled out as being important the severity of diabetes and leptin doses.

Placement of a vena cava filter in term pregnancy: case report and review of the literature.

During pregnancy there are hemostatic changes that may result in a hypercoagulable state producing thrombotic consequences. This condition may be aggravated in women who are carriers of congenital thrombophilic factors. These factors may increase obstetric complications such as miscarriages, fetal growth restriction, placental abruption and preeclampsia. Trombophilic factors may also cause venous thromboembolism, which is the leading cause of maternal morbidity and mortality. We report a case of a 22-year-old woman with factor V Leiden mutation, whose pregnancy was complicated with deep venous thrombosis requiring placement of a vena cava filter.

Relation between Doppler findings and perinatal outcomes in fetuses with intrauterine growth restriction.

PURPOSE OF INVESTIGATION: To find the relationship between fetal Doppler findings and perinatal outcomes in intrauterine growth restriction. METHODS: Eighty-two cases with a prenatal diagnosis of intrauterine growth restriction between November 2008 and July 2009 were included in this prospective study at Ege University School of Medicine. Fetuses were grouped according to Doppler parameters: those with normal Doppler findings (n = 43), and those with impaired arterial (n = 27) and venous systems (n = 12). RESULTS: Out of 82 growth restricted cases, 43 (52.4%) had normal Doppler findings, while 27 (32.9%) displayed impaired arterial parameters and 12 (14.6%) had impaired venous parameters. The mean first minute Apgar scores were 7.57 +/- 1.53 for the group with normal Doppler flows, 6.8 +/- 2 for the group with an impaired arterial system, and 4 +/- 1.94 for the group with an impaired venous system. Two cases from the normal Doppler flow group (n = 42), four cases from the impaired arterial flow group (n = 27), and 11 cases from the impaired venous flow group (n = 11) had fifth minute Apgar scores under 6. Evaluation of the umbilical artery blood gas revealed acidosis in two cases from the normal Doppler flow group (n = 42), three cases from the impaired arterial system group (n = 27), and five cases from the impaired venous system group (n = 11). CONCLUSION: A Doppler spectrum from normal to venous system impairment correlated with poor fetal outcomes including fetal acidosis, fetal mortality and morbidity, decreased Apgar scores at 1 and 5 min, and neonatal morbidity.

Immunohistochemical expressions of p16 and p53 proteins in cervical intraepithelial neoplasia and in benign cervical tissue.

PURPOSE OF INVESTIGATION: To evaluate the immunohistochemical expressions of p16 and p53 in cervical intraepithelial neoplasia (CIN) and do a comparison with non-neoplastic cervical lesions. METHODS: Sixty cases diagnosed as CIN after histopathological examination and 25 controls diagnosed as chronic cervicitis were included in the study. Immunohistochemical expressions for p16 and p53 were evaluated and compared in all cases. The cases in the study were defined according to the Bethesda system. Of these, 31.8% (n = 27) had a low-grade squamous intraepithelial lesion (LGSIL), and 38.8% (n = 33) had a high-grade squamous intraepithelial lesion (HGSIL). RESULTS: There was a statistically significant difference between chronic cervicitis and CIN in terms of p53 and p16 expression levels (p = 0.001). On the other hand, the level of p16 expression was statistically different between LGSIL and HGSIL (p = 0.001), while there was no significant difference in terms of p53 expression. Among the HGSIL cases (n = 33), 91% had p16 expression, while 66.7% (n = 27) of the LGSIL patients had no p16 expression. In the chronic cervicitis group, 84% (n = 21) did not reveal any p16 expression, while 96% (n = 24) did not reveal any p53 expression. Various levels of p53 expressions were detected in 59.2% (n = 16) of CIN1 cases, 69.3% (n = 9) of CIN2 cases, and 90% (n = 18) of CIN3 cases. CONCLUSION: While p16 is useful in detecting high-grade cervical lesions, p53 is not a good biomarker for distinguishing high-grade lesions from low grade ones.

Effects of laparoscopic tubal ligation on ovarian blood flow and steroidogenesis.

Changes in serum ovarian hormone and gonadotrophine levels and ovarian artery blood flow rate by Doppler ultrasonography following laparoscopic tubal sterilization were evaluated. Forty-seven cases of laparoscopic tubal sterilization by electrocoagulation were included in the study. Forty-two women who used nonhormonal contraceptive methods were randomized as a control group. Serum concentrations of estradiol, FSH, LH, progesterone and bilateral ovarian artery pulsatility indexes were measured at baseline and at three, six and 12 months. In the study group, the mean value of midluteal progesterone was significantly decreased three months postoperatively 3. and bilateral ovarian artery pulsatility indexes were increased three and six months postoperatively, compared to their preoperative values. However, there was no statistical difference between bilateral ovarian PI values calculated preoperatively and at 12 months. Laparoscopic tubal ligation by electrocoagulation may temporarily cause reduced bilateral ovarian artery flow and corpus luteum dysfunction proven by low mid-luteal progesterone levels.

Effects of simvastatin treatment on oxidant/antioxidant state and ultrastructure of diabetic rat myocardium.

In the present study we investigated the effects of simvastatin treatment on lipid metabolism and peroxidation, antioxidant enzyme activities and ultrastructure of the diabetic rat myocardium. Diabetes was induced by single injection of streptozotocin (45 mg/kg i.p.). Eight weeks after induction of diabetes, a subgroup of control and of diabetic rats was treated with simvastatin for 4 weeks (10 mg/kg/day, orally). Blood glucose, plasma cholesterol and triacylglycerol, as well as levels of cardiac thiobarbituric acid reactive substances (TBARS) were significantly increased in diabetic rats. The activities of antioxidant enzymes, catalase (CAT) and glutathione peroxidase (GSHPx), were also elevated in the diabetic myocardium. Treatment with simvastatin markedly reduced serum triacylglycerol and cholesterol, and partially controlled hyperglycemia in diabetic animals. The increased activation of antioxidant enzymes and the excess of lipid peroxidation measured by TBARS were completely reversed by simvastatin treatment. Diabetic rats displayed ultrastructural ischemia-like alterations of cardiomyocytes and capillaries, which support oxidative stress-induced tissue remodelling. In the diabetic myocardium simvastatin treatment partly attenuated angiopathic and atherogenic processes, detected by electron microscopy. These results suggest that simvastatin, known as a lipid-lowering drug, may positively affect diabetes induced cardiovascular complications via reducing risks of atherosclerotic pathological processes, such as imbalance between oxidant and antioxidant state.

Role of nitric oxide in hypoxia-induced changes in newborn rats.

In order to investigate the role of nitric oxide (NO) in hypoxic tissue damage in newborns, we studied the effects of systemic administration of an inhibitor of NO synthase, N(G)-nitro-L-arginine (L-NNA), and the precursor for the synthesis of NO, L-arginine (L-ARG), on the biochemical and histological changes in brain, heart, lung, liver, kidney, intestine, and skeletal muscle tissues. Four groups of 1-day-old Wistar rat pups were used: control, hypoxic, L-ARG, and L-NNA groups. L-ARG 100 mg/kg or L-NNA 2 mg/kg was administered as a bolus intraperitoneally 1.5 h before hypoxia. Hypoxia increased lipid peroxidation in all tissues except muscle; this increase was prevented by L-NNA and L-ARG in brain, heart, lung, kidney, and liver tissues. L-NNA in intestine and L-ARG in muscle tissue increased lipid peroxidation. The tissue-associated myeloperoxidase activity was decreased in the liver by L-NNA and L-ARG. Histopathological changes in intestines were villous epithelial separation and hyperemia in hypoxic and L-NNA groups which were not observed in control and L-ARG groups. In lungs, pulmonary hemorrhage was observed only in the hypoxic group. These data suggest that NO acts both as a destructive and a protective agent in the pathogenesis of hypoxia-reoxygenation injuries.

Enzymatic antioxidant defence mechanism in rat intestinal tissue is changed after ischemia-reperfusion. Effects of an allopurinol plus antioxidant combination.

OBJECTIVES: To establish the antioxidant status of rat intestinal tissues after ischemia-reperfusion and to determine if pretreatment with an allopurinol and antioxidant vitamin combination gives any protection against mucosal injury. EXPERIMENTAL ANIMALS: Twenty rats were divided into 4 groups of 5 animals each. METHODS: Group 1 (control) rats were not subjected to ischemia-reperfusion and received no allopurinol plus vitamin combination; group 2 rats received vitamins C (200 mg/kg) and E (100 mg/kg) and allopurinol (50 mg/kg) combination daily for 3 days preoperatively; group 3 rats were subjected to ischemia-reperfusion only; and group 4 rats were subjected to ischemia-reperfusion and received the vitamin and allopurinol combination. MAIN OUTCOME MEASURES: Activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) enzymes, the level of thiobarbituric acid-reagent substances (TBARS) and histologic grading of tissue samples. RESULTS: SOD and GSH-Px activities were decreased, but the CAT activity and TBARS level increased. Pretreatment of the rats with the allopurinol-vitamin C-vitamin E combination did not have any significant effect on the enzyme activities. However, it resulted in important reductions in the TBARS tissue levels. Histologic investigation revealed significant mucosal injury in group 3 rats compared with group 4 rats (mean [and standard deviation] for grading, 4.6 [0.5] versus 1.8 [0.4]). CONCLUSIONS: The enzymatic antioxidant defence system was significantly changed after ischemia-reperfusion and intestinal tissue was exposed to increased oxidant stress, the results of which were peroxidation of some cellular structures and increased concentrations of oxidative products. Although antioxidant treatment did not drastically affect the enzyme activities or afford complete protection of cellular structures against deformation, it apparently could eliminate oxygen radicals and prevent peroxidative reactions.

Isoflurane impairs antioxidant defence system in guinea pig kidney.

PURPOSE: To investigate whether free radical metabolism is changed due to isoflurane treatment and, if so, to elucidate the role of changed free radical metabolism in the nephrotoxicity. MATERIALS AND METHODS: Fifteen guinea pigs were used in the study. Five were treated with isoflurane in oxygen, five with oxygen and five were controls. Animals were exposed to isoflurane and oxygen three times. Each treatment was performed for 30 min once a day for three consecutive days. Activities of free radical enzymes, superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px); values of antioxidant parameters, antioxidant potential (AOP), non-enzymatic superoxide radical scavenger activity (NSSA) and oxidation resistance (OR) and, level of an oxidant parameter namely, malondialdehyde (MDA) were determined in the renal tissues of the groups. Blood was also obtained for serum creatinine and urea analyses. RESULTS: AOP, NSSA, SOD and CAT activities were decreased; (0.0188 +/- 0.0026 vs 0.0156 +/- 0.0015, P < 0.025; 8.72 +/- 1.80 vs 6.40 +/- 1.22, P < 0.05; 76.71 +/- 18.54 vs 52.79 +/- 11.68, P < 0.025; 71.26 +/- 15.58 vs 55.39 +/- 8.83; P < 0.05, respectively) but, MDA level, OR value and GSH-Px activities increased (10.89 +/- 1.57 vs 15.87 +/- 2.97, P < 0.01; 0.84 +/- 0.34 vs 2.28 +/- 1.39, P < 0.05; 1.45 +/- 0.83 vs 3.45 +/- 1.20, P < 0.01, respectively) in kidney tissues from isoflurane-treated group compared with controls. No differences were observed between control and oxygen groups with regard to all analysis parameters except GSH-Px. CONCLUSION: Isoflurane impairs the antioxidant defence system and this oxidant stress may play a part in the isoflurane-induced renal toxicity.

Fertility of ejaculated and testicular megalohead spermatozoa with intracytoplasmic sperm injection.

In this study the fertility and outcome of intracytoplasmic sperm injection (ICSI) using megalohead spermatozoa from the ejaculates and testicles was evaluated. Seventeen males with megalohead and pinhead sperm forms in their ejaculate were studied in 22 cycles. A high number of sperm heads without tails and abundant round spermatid forms were commonly observed. Round-headed spermatozoa were seldom accompanied by these severely abnormal spermatozoa. The majority of megalohead spermatozoa were observed to have multiple tails, were predominant in the sample, and were used for ICSI. Ejaculated megalohead spermatozoa were used for ICSI in 15 cycles, while testicular spermatozoa were used in seven cycles where there were no vital spermatozoa or spermatozoa of low vitality in the ejaculate. The same abnormal morphology was observed in the testicles as in the ejaculated spermatozoa in the same males. Mean (+/- SD) low motility 4.7 +/- 5.6% and sperm count (3.8 +/- 4.19 x 10(6)) were common findings in these severely teratozoospermic patients. A low fertilization rate (43.2%) was achieved by using megalohead sperm forms (group I, n = 17) in comparison with the control group (60.2%) which had zero normal sperm morphology according to strict criteria (group II, n = 30) (P <0.01). Furthermore, a low pregnancy rate (9.1%) was obtained in the megalohead sperm group in comparison with the control group (40%) (P <0.05). Low fertilization and pregnancy rates may be due to a high incidence of chromosomal abnormalities from severely defective spermatozoa in the ejaculate. Couples should be counselled and warned about possible low fertilization and pregnancy rates with ICSI when only pinhead and megalohead forms with a high number of sperm heads without tails are present in the ejaculate.

Effects of intra-articular vitamin E and corticosteroid injection in experimental hemarthrosis in rabbits.

The effects of intra-articular injections of vitamin E and corticosteroid were investigated in skeletally mature New Zealand white rabbits in which experimental hemarthrosis was induced for 14 days. The rabbits were divided into three groups composed of eight rabbits each: the first group comprised of animals with hemarthrosis, the second group animals with hemarthrosis and intra-articular injections of 20 mg vitamin E, and the third group animals with hemarthrosis and intra-articular injections of 10 mg of triamcinolone acetonide (TCA). Proteoglycan levels in the joint cartilage of the hemarthrosis group were found to be increased significantly compared with the controls (P < .01), whereas in the vitamin E-injected group they were significantly decreased (P < .05). In the TCA-injected group, proteoglycan levels were not found to be significantly different from those in the hemarthrosis group (P > .05). Histopathological evaluation showed that the cartilage structure in the joint of the control limbs was identical to that in the vitamin E- and TCA-injected limbs. In the hemarthrosis group, in comparison with the controls, the joint surface was roughened and fibrillated. In the superficial areas of the cartilage tissue, chondrocytes were decreased in number. These findings suggest that in this model, vitamin E and TCA may be helpful in preventing the joint cartilage changes seen in hemarthrosis.

Testicular effects of vasectomy in rats: an ultrastructural and immunohistochemical study.

OBJECTIVES: The correlation between infertility and morphofunctional alterations following vasal occlusion is not clearly understood. Although a correlation has been found between the expression of a high titer of antisperm antibodies and the status of infertility, the relationship between the immunoglobulin (Ig) depositions in the testis and ultrastructural alterations of the peritubular structures has not been clearly established. The objective of this study is to explain the etiopathogenesis of diffuse and focal testicular lesions caused by vasal obstruction. METHODS: Unilateral vasectomy was performed on adult male rats. Ultrastructural changes of testicular tissues and immunostaining affinity of peritubular structures with anti-actin, anti-vimentin, anti-laminin, and anti-immunoglobulin G (anti-IgG) antibodies were analyzed. RESULTS: In an ultrastructural study, vasectomized animals presented abundant collagen fibril accumulation in the peritubular area. In testis with intense IgG antibodies, staining revealed weak immunostaining of peritubular myoid cells with anti-actin antibodies, but intense immunostaining with anti-vimentin antibodies. The tubules of rats with no IgG deposition on peritubular structures revealed intense immunostaining with anti-actin antibodies but scant immunostaining with anti-vimentin antibodies. Anti-laminin deposits decreased severely in most of the tubules demonstrating intense IgG deposition. CONCLUSIONS: Our findings suggest that spermatogenesis deteriorates more severely in testes with dense IgG deposition. The sclerosis of the lamina propria in cases with vasal occlusion is secondary to alterations in the peritubular myoid cells. With the progressive changes that occur in the peritubular myoid cells or differentiation of the peritubular myoid cells that acquire active cell characteristics, collagen accumulation adjacent to these cells increases markedly. The alterations of the peritubular myoid cells reported here may be caused by alterations in basement membrane structures.

Histopathological and ultrastructural examination of the rat conjunctiva after exposure to tobacco smoke.

Exposure to tobacco smoke can cause irritation of the conjunctiva. We conducted this study to identify the effect of tobacco on rat conjunctiva. Animals were divided into experimental and control groups and we exposed the experimental group to tobacco smoke. Control group rats inhaled only room air. Spectrophotometric analysis of the smoke-air mixture revealed that many toxic substances were present in this compound. We found very high levels of plasma thiocyanate after exposure to smoke in experimental group animals but no increase in the control group. So, this data indicates that these animals inhaled smoke effectively in our method. After 3 months conjunctivas were examined by light and electron microscopy. In the experimental group, conjunctivas were thinned, atrophied and microvillous projections and desmosomal connections were absent in comparison with the control conjunctivas. This pathologic change is very similar to conjunctival response to chronic irritants of any type.

Protection by Iloprost (stable analogue of prostacyclin) of endothelial damage due to chronic vasospasm in dogs: an electron microscope study.

The resolution of cerebral vasospasm and protection of endothelial damage by Iloprost was evaluated with multicisternal injections. Sixteen adult mongrel dogs (18-20 kg) were assigned to one of three experimental groups. All animals received a total amount of 15 ml fresh unheparinized arterial blood via three injections into the cisterna magna. Selective vertebral angiography was performed on day 0 and subsequently blood injections were performed on the 2nd and 3rd days after the first injection. On the 7th day angiography was reperformed to determine the chronic vasospasm. The first group (5 dogs) was the control group and received intrathecal saline which was equal to the amount of saline in which Iloprost was diluted. The second group (5 dogs) did not receive any treatment until the 7th day. The third group (6 dogs) received Iloprost intrathecally (total 10 micrograms kg-1). In the first two groups angiographic vasospasm was prominent. For the second group intraarterial Iloprost was given on the 7th day in order to evaluate its acute effect. However there was no evidence of resolution of vasospasm. In the third group, resolution of vasospasm was verified on angiograms. Electron microscope studies of basilar arteries of the first two groups revealed degenerative changes of the endothelial cells which were separated from each other and the elastic lamina was irregularly arranged. In the intrathecal Iloprost-treated group there was little thickening in the elastic lamina and the endothelial cells were almost normal in structure. These results can be considered as the evidence of the prophylactic effect of Iloprost given by the intrathecal route in the prevention of chronic cerebral vasospasm.