Association of pro-fibrinolytic receptor AnnexinA2 with tissue plasminogen activator/Inhibitor-1 in pre-eclampsia.

Background: The clinical manifestations of pre-eclampsia are related to placental anti-angiogenic factor alteration. These variations are mainly due to the alteration of plasminolytic components. The study aims to compare the expression of plasminolytic components in the placenta of women with and without pre-eclampsia. Material and Methods: The study included pregnant women with pre-eclampsia as PE group (n = 30) and without pre-eclampsia as a control group (n = 30). Placental bed biopsy tissues were collected. AnxA2, tPA, PAI-1 expression in the placental villous tissue was quantitatively evaluated using immunohistochemistry, western blot, and real time-PCR analysis. Results: The results of the study showed a significant decrease in the expression of ANXA2 and increased expression of tPA and PAI-1 in PE group compared to control group (p<0.005). AnxA2 expression showed positive correlation with tPA (r=+0.895, p=0.002) and negative correlation with PAI-1(r=-0.905, p=0.020) in control group whereas in the PE group AnxA2 expression was negatively correlated with tPA ((r=-0.801, p=0.016) and PAI-1 (R=-0.831, P=0.010). Conclusion: Decreased AnxA2 with increased expression of PAI-1 and tPA may be responsible for the altered fibrinolytic activity and play a significant role in pre-eclampsia pathogenesis.

Placental Expression and Relative Role of Anti-inflammatory Annexin A1 and Animal Lectin Galectin-3 in the Pathogenesis of Preeclampsia.

Preeclampsia (PE) remains the major cause for maternal and foetal mortality and morbidity all over the world. Preeclampsia is associated with maternal, placental aggravated inflammatory response and generalized endothelial damage. AnnexinA1 (AnxA1) is glucocorticoid regulated protein regulates a wide range of cellular and molecular steps of the inflammatory response and is implicated in resolution of inflammation. Galectin-3 (Gal-3), ß-galcotoside-binding lectin participates in many functions, both intra- and extracellularly. Recently it has been shown that galectin-3 modulates the inflammation. Role of AnxA1 and Galectin-3 is poorly studied in context with human reproductive disease like Preeclampsia. Therefore, the present study examined the expression of AnxA1 and Gal-3 which are involved in modulation of inflammation and their association in the placental bed of pregnancy with and without PE. The study group consisted of placental bed biopsy tissues obtained from pregnancies with PE (n = 30) and without (n = 30) PE. The expression of AnxA1 and Gal-3 in the placental bed tissues was evaluated quantitatively using Immunohisto-chemistry (IHC), western blot and mRNA expression analysis by quantitative RT-PCR. Our IHC, western blot and RT PCR analyses showed the increase in the expression of AnxA1 and Gal-3 in PE group compared with the normotensive control group (P < 0.001). The increased expression of AnxA1 and Gal-3 in placental bed may be associated with a systemic inflammatory response in PE, suggesting role of AnxA1 and Gal-3 in PE pathogenesis.

Detection of Cognitive Impairment by Choice Auditory Reaction Time (ART) and Visual Reaction Time (VRT)s during Acute Mental Stress in Young Hypertensives: A Case Control Study.

BACKGROUND: Acute stress is known to be associated with both negative and positive influences on cognitive performance. Hypertension is one of the risk factors for lowered cognitive performance. Mental stress testing is easier to administer and can be regulated by the investigator. Mental arithmetic using serial subtraction is the most widely used method to administer stress. Reaction time (RT) is widely used to assess cognitive domains like attention, execution and psychomotor speed. Researchers have shown choice reaction times are delayed in hypertension. It is not known whether acute mental stress improves or deteriorates attention, execution and psychomotor speed in hypertension. We hypothesized in the present study that acute mental stress deteriorates cognitive function in hypertensives without overt cerebrovascular disease or other vascular risk factors. METHODS: After getting medical ethical clearance from our institution, this case-control study was carried out over eight months (January 2017 to September 2017). 60 subjects between the age group of 35 to 55 years were included in the study. They were divided into 2 groups. Group 1 consisted of 30 diagnosed cases of hypertension at least two years of duration. Group 2 consisted of 30 sex and age-matched controls. MMSE was performed to assess the cognitive function in these groups. Simple (S) and choice (C) auditory reaction time (ART) and visual reaction time (VRT)s were measured at rest and acute mental stress in these groups to assess cognitive function. Predictive value of VRTC resting and VRTC during acute mental stress among hypertensives for cognitive dysfunction was calculated using the receiver operating characteristic (ROC) curve. RESULTS: There was significant difference ART and VRT, both simple and choice, in hypertensive and nonhypertensive subjects and these reaction times further increased during mental stress (P<0.001). VRTC can be a predictor of cognitive dysfunction in hypertensives and during acute mental stress. CONCLUSION: A significant difference in cognitive functions in hypertensive and nonhypertensive subjects exists and this further deteriorates with acute mental stress.

Ubiquitin Mediated Degradation of EGFR by 17 ß-estradiol in Triple Negative MDA-MB-231 (TNBC) Breast Cancer Cells Line.

BACKGROUND: Triple Negative Breast Cancer (TNBC) commonly displays Epidermal growth factor receptor (EGFR). Effective EGFR degradation results in the suppression of tumor in various models. Studies have addressed the relevance of this strategy in the treatment of TNBC. In the present study, we examined the effect of 17 ß- estradiol on EGFR expression in MDA-MB-231 (TNBC) cell line and assessed whether 17 ß-estradiol degrades EGFR by ubiquitination pathway. OBJECTIVES: The objective of this study is to treat MDA-MB-231 cell lines with Cycloheximide with or without 17ß-estrdiol to observe whether 17ß-estradiol leads to EGFR degradation and to treat with MG-132 to assess whether degradation occurs through ubiquitination pathway. METHODS: MDA-MB-231 cells were treated with 17ß-estradiol (E2) and EGFR expression was studied by western blotting at different intervals by using Cycloheximide chase. To assess ubiquitination pathway of degradation of EGFR in MDA-MB-231 cell line, MG-132 was used. RESULTS: EGFR expression was reduced with ß-estradiol treatment in MDA-MB-231 cell line with Cycloheximide chase. Upon Treatment with MG-132 and E2, EGFR expression did not reduce, suggesting that Estrogen degrades EGFR by ubiquitination pathway. CONCLUSION: Estrogen degrades EGFR in MDA-MB-231 cells and this degradation occurs by ubiquitination.

Decreased expression of annexin A2 and loss of its association with vascular endothelial growth factor leads to the deficient trophoblastic invasion in preeclampsia.

OBJECTIVES: Preeclampsia (PE) remains the major cause for maternal and foetal mortality and morbidity. Invasion of endovascular trophoblast and remodelling of spiral artery are crucial actions of normal placental development. Non-fulfilment of these processes plays a leading role in the development of preeclampsia. Vascular endothelial growth factor (VEGF) is produced by extravillous trophoblastic tissue and decidual cell population is a well-known angiogenic growth which plays a fundamental role in placental pathogenesis of PE. Annexin A2 (ANXA2) is a profibrinolytic protein receptor required for plasminolysis, which is an important step in the formation of new blood vessel along with VEGF. Role of ANXA2 is poorly studied in context with human reproductive disease like preeclampsia. The purpose of the present study is to examine the expression and association of VEGF and ANXA2 in the term placentas of pregnancies with and without PE. METHODS: The study group comprised of placental tissues procured from gestations with PE (n=30) and without (n=20) PE. The expression of VEGF and ANXA2 in the placental villous tissue was evaluated quantitatively by means of IHC, western blotting and reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: Our IHC, western blotting and RT-PCR analysis illustrated the significant decrease in the expression of VEGF and ANXA2 in PE group compared with the normotensive control group (p<0.005). We observed statistically significant positive correlation among the expression of ANXA2 and VEGF in placentas of normotensive control group (p<0.0001). CONCLUSIONS: The diminished expression of VEGF and ANXA2 in placenta may be associated with the defective angiogenesis and which may possibly play a vital role in PE pathogenesis.

Comparative expression analysis of phospholipid binding protein annexina1 in nephrogenesis and kidney cancer.

Background The expression in the glomerular mesangial cells, papillary, and collecting duct cells demonstrated annexin A1 (AnxA1)'s role in specific renal functions. With varying concentrations of calcium (Ca2+), it is considered to regulate cellular processes such as cell proliferation, apoptosis, and clearance of apoptotic cells by forming ceramides, a key lipid mediator of apoptosis. It also participates in tumorigenesis based on its location. On account of these features, we investigated the expression of this apoptosis-associated protein in fetal kidneys at different gestational periods, mature kidneys and in kidney cancer tissues in order to localize and possibly characterize its role during nephrogenesis and renal tumors. Methods AnxA1 expression was evaluated by an immunohistochemistry technique in "paraffin-embedded" renal tissue sections from autopsied fetuses at different gestational ages, in mature kidneys and renal cancer tissues. Results The current study data demonstrated that AnxA1 is expressed in the mesangial cells and podocytes of maturing glomeruli in the developing renal cortex of fetal kidneys at 14 to 19 weeks of gestation. The expression in the mesangial cells declined in later weeks of gestation and persisted into adulthood. AnxA1 expression increased with the progression of clear cell renal cell carcinoma (CCRCC) and also in other cancer types indicating a potential role of the protein in tumorigenesis. Conclusions We presume that AnxA1 in the podocytes and mesangial cells play important roles in various signaling pathways in the functioning of the glomerulus. These results and concepts provide a framework to further dissect its biological properties and thereby develop diagnostic, prognostic, and therapeutic strategies targeting the molecule in various renal pathologies.

Differential expression pattern of annexin A2 during nephrogenesis and kidney carcinoma.

The creation of a cancer cell could be due to reactivation of repressed gene in the process of normal embryonic development. The differences in embryonic origins and functions of various components of nephron may contribute to the diversity of morphological patterns, molecular and immunohistochemical phenotypes of common renal neoplasms. Renal cell carcinomas (RCCs) are the most common amongst the genitourinary cancers. Annexin A2 (AnxA2) is a multifunctional calcium-regulated phospholipids-binding protein found in a subset of renal neoplasms. Since the tumor cells usually recapitulate embryonic cells, we studied the ontogeny of AnxA2 in developing renal tissues and compared it with those of normal adult RCCs, to better understand their role in renal development and tumorigenesis. AnxA2 immunoexpression was evaluated by immunohistochemistry from various autopsied fetuses, mature kidney and renal cancer tissue specimens. The study showed moderate membranous AnxA2 immunoexpression in the ureteric buds and collecting tubules of fetal kidneys (in all gestational ages) and in the collecting ducts of adult normal renal tissues. It is not often expressed in the proximal convoluted tubules of normal adult kidney; however, younger fetal kidneys show moderate AnxA2 immunoexpression in the proximal convoluted tubules (thought to be the origin of RCC) and the reappearance of strong membranous AnxA2 immunoexpression in the clear cell carcinoma is suggesting a deregulation of the gene during tumorigenesis. The understanding of the AnxA2 molecular immunoexpression pattern during development, its specific function and deregulated immunoexpression in different renal carcinoma types indicates the decisive role of AnxA2 in the cancer progression.

Mean platelet volume and other platelet volume indices in patients with stable coronary artery disease and acute myocardial infarction: A case control study.

BACKGROUND: Coronary artery disease is mainly caused by atherosclerosis and its complications. Platelets and their activity have an important role in initiation of atherosclerotic lesions and coronary thrombus formation. Larger platelets are enzymatically and metabolically more active and have a higher potential thrombotic ability as compared with smaller platelets. AIMS: To study the changes in platelet volume indices and platelet count in acute myocardial infarction, stable coronary artery disease and compare them with controls to assess their usefulness in predicting coronary events. MATERIALS AND METHODS: This was a comparative study of 128 subjects; 39 patients with acute myocardial infarction (AMI), 24 patients with stable coronary artery disease (SCAD) and 65 controls. Venous sample were drawn from AMI subjects on admission (within 4 hours of chest pain) and collected in standardized EDTA sample tubes. Platelet count and volume indices were assayed within 30 minutes of blood collection, using Sysmex KX21-N autoanalyzer. Venous samples were also drawn from SCAD on who were admitted for angiography and subject attending routine checkups. RESULTS: The mean platelet volume was significantly higher in patients with AMI (9.65 ± 0.96) as compared to SCAD (9.37 ± 0.88) and controls (9.21 ± 0.58). The best cut-off values for MPV when predicting AMI and SCAD in patients were 9.25 fl (sensitivity 56.4%; specificity 45.9%) and 9.15 fl (sensitivity 54.2%; specificity 42.23%), respectively. CONCLUSIONS: Measurements of MPV may be of some benefit in detecting those patients at higher risk for an AMI and CAD.

Chronoscopic reading in whole body reaction times in detecting cognitive dysfunction in metabolic syndrome: a case control study.

BACKGROUND: The metabolic syndrome, a clustering of several commonly occurring disorders that include abdominal obesity, hypertriglyceridemia, low high-density lipoprotein level, hypertension, and hyperglycemia, has been specifically investigated as a risk factor for cognitive decline in elderly individuals. The metabolic syndrome may be a risk factor for cognitive decline because it summarizes the joint effects of these risk factors. It is known that difference between simple and choice reaction time (RT) implies time required for cognition. Though delayed choice RTs indicate involvement of cognition, they cannot quantify how much time is required for cognition. In whole body simple reaction time (WBSRT), RT is split into two chronoscopic readings: C1 and C2. C1 measures time required for central processing which requires cognition and C2 measures total RT. C2-C1 measures time required for peripheral motor response. We hypothesized that whole body choice RT chronoscopic reading 1 (WBCRTC1) will be delayed in metabolic syndrome and WBCRTC1 will have predictive value in detecting cognitive dysfunction. SETTINGS AND DESIGN: Hospital-based cross-sectional case-control study. MATERIALS AND METHODS: Study was conducted on 120 subjects using visual and WBSRT having criteria of age (40-60 years) and metabolic syndrome, compared with equal number of age- and sex-matched controls. Statistical analysis was done by independent t-test and duration of metabolic syndrome was correlated with cognition times (WBCRTC1) using Pearson's correlation. Predictive value of WBCRTC1 was calculated using receiver operating characteristic curve. RESULTS: Delayed visual simple RT, visual choice RT, WBSRT, and whole body choice RT (WBCRT) observed among subjects metabolic syndrome when compared with controls. Choice RTs were more delayed compared to simple RTs. WBCRTC1 (608.8 ± 132 ms) was more delayed than WBSRTC1 (424.05 ± 89.9 ms) among metabolic syndrome indicating cognitive dysfunction. Unfortunately, there was no significant correlation between duration of metabolic syndrome with cognition. The best cut-off value for WBCRTC1, when predicting cognitive dysfunction in metabolic syndrome was 542.5 ms (sensitivity 36.7% and specificity 31.6%). CONCLUSIONS: WBCRTC1 can be used as a tool to detect cognitive dysfunction.