Rajarshi Kar

Clove Oil Reverses Learning and Memory Deficits in Scopolamine-Treated Mice

The present study was performed to examine the effect of Eugenia caryophyllata (Myrtaceae) on learning and memory, and also evaluate whether it can modulate oxidative stress in mice. Passive avoidance step-down task and elevated plus-maze were used to assess learning and memory in scopolamine-treated mice. Oxidative stress parameters were also assessed in brain samples by estimating the malondialdehyde (MDA) and reduced glutathione (GSH) levels at the end of the study. Scopolamine (0.3 mg/kg, i. p.) produced impairment of acquisition memory as evidenced by a decrease in step-down latency and an increase in transfer latency on day 1, and also impairment of retention of memory on day 2. Pretreatment with clove oil (0.05 mL/kg and 0.1 mL/kg) for 3 weeks significantly reversed the increase in acquisition latency and all the doses (0.025, 0.05, 0.1 mL/kg, i. p.) reversed the increase in retention latency induced by scopolamine (0.3 mg/kg, i. p.) in elevated plus-maze. However, 0.05 mL/kg clove oil attenuated memory deficits in the passive avoidance step-down task. Brain samples showed a significant decrease in MDA levels in the group treated with clove oil (0.05 and 0.025 mL/kg). GSH levels were also increased in clove oil-treated mice though the results were not significant. Thus, it can be concluded that clove oil can reverse the short-term and long-term memory deficits induced by scopolamine (0.3 mg/kg, i. p.) and this effect can, to some extent, be attributed to decreased oxidative stress.

Cadmium exposure during lactation causes learning and memory-impairment in F1 generation mice: amelioration by quercetin

Abstract Cadmium (Cd) is a known pollutant present in the environment at low levels and is reported to affect reproduction in many ways. The present study was undertaken to explore the effect of Cd in F1 generation mice on cognitive parameters, and to further investigate whether quercetin could modulate these effects. In this study, female lactating mice were exposed to cadmium for seven days just after delivery. The new born pups in their adulthood were tested for learning and memory parameters by passive avoidance task and Morris water maze (MWM) test. It was observed that pups exposed to Cd showed significant impairment of memory in step down latency test, which was reversed by quercetin (100 mg/kg). In MWM test for spatial memory, animals exposed to Cd exhibited increased escape latency, which was reversed by quercetin (50 mg/kg) significantly. Quercetin alone (50 and 100 mg/kg) also demonstrated improved spatial memory, and showed improved retention memory in the passive avoidance paradigm at dose 50 mg/kg. On testing oxidative stress parameters, we observed significantly increased malondialdehyde (MDA) levels in brain tissue of Cd-treated mice. Moreover, co-treatment with quercetin (50 mg/kg) and Cd significantly reduced these MDA levels. The other doses (25 and 100 mg/kg) also showed reduction in MDA levels as compared to the group exposed to Cd alone, though the difference was not statistically significant. Hence, this study highlights the possibility of cognitive impairment in adulthood if there is Cd exposure during lactation and oxidative stress could possibly attribute to this effect.

Response of primary culture of human ovarian cancer cells to chemotherapy: In vitro individualized therapy.

OBJECTIVE This study focused on whether primary cultures of ovarian cancer (OC) cells established from ascites can be used to evaluate response to chemotherapeutic agents and if curcumin could enhance the efficacy of these agents. MATERIALS AND METHODS We established five primary cultures of ascitic cells from OC patients and treated them with curcumin, carboplatin, and paclitaxel singly and in combinations. The percentage of apoptotic cells was determined by flow cytometry. RESULTS There was a wide variation in the response of individual primary cultures to treatment with the chemotherapeutic agents. Curcumin by itself was as good as carboplatin or paclitaxel in inducing apoptosis in the primary OC cells. Curcumin was not able to affect the carboplatin mediated cell killing. However, a combination of curcumin and paclitaxel was additive and was equally effective as a combination of carboplatin and paclitaxel. A combination of curcumin carboplatin, and paclitaxel was also found to be additive and, in fact, turned out to be the best combination that gave the highest percentage of apoptosis in vitro. CONCLUSION This study highlights the fact that primary cultures of OC cells can be used to detect response to chemotherapeutic agents and help to individualize the treatment offered to OC patients.

Expression of Lipoprotein associated phospholipase A2 enzyme in medical undergraduate students with metabolic syndrome

AIMS Metabolic syndrome (MS) and atherosclerosis are chronic inflammatory conditions. Lipoprotein-associated phospholipase A2 (LpPLA2) is a circulatory marker of systemic inflammation and a risk predictor for cardiovascular diseases. This study aims to evaluate the expression of this enzyme in an effort to understand the underlying mechanism of atherosclerosis in MS. METHODS This study included twenty patients of MS and same number of healthy controls. Anthropometry and clinical examination were carried out in both groups. Real time PCR was performed for LpPLA2 mRNA and relative expression was calculated using ΔΔCT method keeping β2 microglobin and β-actin as internal controls. RESULTS LpPLA2 mRNA expression was higher in patients of MS. Fold change was 5.7 when β2 microglobin was used as normaliser and 4.97 when β-actin was used. mRNA levels of LpPLA2 correlated significantly with waist circumference (r=0.462, p=0.003) and systolic blood pressure (r=0.392, p=0.015) as well as high density lipoprotein cholesterol (r=-0.453, p=0.003). CONCLUSIONS High expression of LpPLA2 mRNA indicates that systemic inflammation has role in pathogenesis of atherosclerosis in patients of MS. This is evident from its direct correlation with blood pressure. The study also suggests that expression of LpPLA2 may be associated with obesity. Therefore, LpPLA2 mRNA expression levels may develop as an important risk predictor for vascular complications in MS.

Evaluation of Placental VEGFA mRNA Expression in Preeclampsia: A Case Control Study

ObjectiveThe aim of our case–control study was to determine expression of VEGFA mRNA in placentae of preeclamsia (PE) versus uncomplicated pregnancy to further clarify its differential expression in pregnancy hypertensive disorders.Study DesignThe PE group was subdivided into severe and non-severe; those with or without HELLP syndrome and placental VEGFA characteristics were compared for these cohorts. Additionally, the neonatal and maternal outcomes were recorded. The quantification of placental VEGFA was done using quantitative real-time PCR and results were expressed as fold change.ResultsOut of 42 PE cases, 23 (55%) were non-severe and 19 cases (45%) were severe PE. Out of 19 severe PE patients, 8 (42%) were HELLP syndrome (complete HELLP) and remaining 11 (58%) were non-HELLP severe PE. Compared to controls, the true fold change in PE, HELLP, non-HELLP, severe PE, non-severe PE was − 2.186, − 13.333, − 6.698, − 8.950 and 1.466, respectively.ConclusionsOur results showed a lowered VEGFA expression in PE placentae compared to uncomplicated controls. The finding of initial increase of VEGFA in non-severe PE and subsequent marked lowering in HELLP strengthens the existing hypothesis of decompensated VEGF being a major role player in PE.

Interrelationship between nuclear factor-erythroid-2-related factor 2, NADPH quinone oxidoreductase and lipoprotein-associated phospholipase A2 expression in young patients of metabolic syndrome

Metabolic syndrome (MS) is associated with inflammation and oxidative stress (OS). Keap1/Nrf2/ARE is a cytoprotective pathway induced by OS and inflammation. This study aims to evaluate the expression of nuclear factor-erythroid-2-related factor 2 (Nrf2) and its downstream target gene NADPH quinone oxidoreductase-1 (NQO-1) in MS. Since lipoprotein-associated phospholipase A2(LpPLA2) is an important inflammatory marker believed to have a role in complications of MS, the association of its expression with that of Nrf2 and NQO-1 was also studied. Medical students (n = 26) were categorised in two groups according to NCEP ATP III criteria with WHO criteria for obesity for South Asian region: patients of MS (n = 13) and controls (n = 13). mRNA expression of Nrf2, NQO-1 and LpPLA2 genes was evaluated by qPCR in blood using specific primers. Fold change was calculated by 2–ΔΔcT method keeping β-actin as internal control. Expression of NQO-1 and LpPLA2 was found to be higher in MS. However, Nrf2 expression was low in patients who had hypertriglyceridemia when compared with patients with normal triglyceride levels. A significant correlation was observed in expression of LpPLA2, with Nrf2 and NQO-1. Our data suggests that there may be compensatory activation of antioxidant defence mechanism in young patients of MS. Further evidence is provided by higher expression of LpPLA2 and its correlation with Nrf2 and NQO-1 in MS which suggests that inflammatory stress may induce expression of genes of cytoprotective pathways. Additionally, this study, for the first time, indicates that Nrf2 may have some role in regulating triglyceride (TG) concentration.

Prediction of preeclampsia in primigravida in late first trimester using serum placental growth factor alone and by combination model

Abstract We investigated a placental growth factor alone and combined clinical (mean arterial pressure, MAP), biophysical (uterine artery pulsability index, PI) and biochemical (placental growth factor, PLGF) model for predicting preeclampsia in late first trimester. The inclusion criteria was primigravida (<40 years) attending their first hospital visit with singleton pregnancy at 11–14 weeks of gestation. Of the enrolled and followed 291 subjects, 35 (12%) later developed PE (5.8%)/GH (6.2%). An equal number of randomised women with normotensive non-proteinuric course were considered as reference group. For preeclampsia, PLGF alone had detection rate of 40% and 51% with 5% and 10% FPR, respectively. On addition of MAP, the AUC improved to 0.937 for PE. Further, addition of mean PI slightly improved AUC to 0.965. This signifies that a model with all three markers had better prediction of preeclampsia rather than PLGF alone. Impact statement In view of high morbidity and mortality due to hypertensive disorders in pregnancy, there has been extensive research for developing markers to detect/screen the condition in early pregnancy. Several such markers have been tested in their individual capacities and in combination during early pregnancy. Most of these studies have originated from high income countries and focussed mainly on the second trimester of pregnancy. We investigated a placental growth factor alone and combined clinical (mean arterial pressure, MAP), biophysical (uterine artery pulsability index, PI) and biochemical (placental growth factor, PLGF) model for predicting preeclampsia in the first trimester in primigravida (<40 years). A nested case control model was used for our study. For preeclampsia, PLGF alone had detection rate of 40% and 51% with 5% and 10% FPR, respectively. On addition of MAP, the AUC improved to 0.937 for PE. Further, addition of mean PI slightly improved AUC to 0.965. The present study has been done in an Indian subcontinent setting (where maternal mortality related to preeclampsia are even higher) where very limited studies are available for the role of either PLGF or in combinations for prediction of preeclampsia. Our research pointed shows better predictability for PE when a combination of markers is used especially in low-risk nulligravida. These are easy, cheap and non-invasive measurements that can be taken in all women at their first routine antenatal visit.

Establishment of Primary Cell Culture From Ascitic Fluid and Solid Tumor Obtained From Epithelial Ovarian Carcinoma Patients

Objective Ovarian cancer is the seventh leading cause of cancer death worldwide. This is mainly due to late diagnosis and high rate of relapse and resistance following chemotherapy. In the present study, we describe simple and cost-effective method to establish primary culture from ascitic fluid and solid tumor obtained from epithelial ovarian carcinoma patient, which may provide a better tool for in vitro testing of drug sensitivity and designing individualized treatment protocol. Methods Complete Dulbecco modified Eagle medium (DMEM) was prepared by supplementing DMEM with 10% fetal bovine serum and antibiotics (ciprofloxacin and amphotericin B). Establishment of primary culture of ovarian cancer cells from ascites fluid and solid tumor was done by using complete DMEM media. Results Primary cultures of ovarian cancer cells were established from ascitic fluid and solid tumor tissue. Of the 7 ascitic fluid samples, we were able to establish 5 primary cultures of ovarian cancer cells. All the 7 samples were diagnosed as serous papillary adenocarcinoma. Some fibroblasts were also attached to culture flask on day 4; they were removed by exposing them to trypsin for a brief period. On day 7, grape-like clusters were visualized under inverted microscope. The cells became confluent on the 10th and 11th day and showed cobblestone appearance, which is a hallmark of ovarian cancer cells. Senescent irregularly shaped cells that have ceased dividing were seen after 8 to 10 passages. Conclusion This study highlights the fact that establishing primary cultures from ascitic fluid or solid tumor tissue may help us to understand the molecular profile of the cancer cells, which allow us to select the best chemotherapeutic agent for ovarian cancer patients and thus take a step toward patient-tailored therapy so that patients are not exposed to drugs to which they are not likely to respond.

Cadmium Enhances Locomotor Behaviour in F1 Generation Mice Following Maternal Exposure During Lactation: Modulation by Quercetin

The present study was designed to investigate the effects of Cd exposure on locomotor behaviour in F1 mice whose dams were exposed to low dose cadmium for a short duration during the lactation period and whether quercetin could modulate the effects. The locomotor behaviour and musculoskeletal activity were tested using photoactomter and rota rod test respectively. We observed that at the given dose of Cd (1.2mg/kg) for a short duration, the muscle coordination on rota rod showed significant improvement as compared to the control group. Cotreatment of Cd with quercetin (100mg/kg) further enhanced the rota rod activity. Similarly the spontaneous motor activity as tested on the photoactometer also exhibited improvement in Cd treated group though this change was not statistically significant. However, Cd when cotreated with quercetin (50 mg/kg) showed significant enhancement in photoactometer activity compared to control. Quercetin administered alone (dose 25 mg/kg) also demonstrated significantly improved rota rod and photoactometer activity as compared to control. Thus cadmium exposure to dams at dose (1.2mg/kg) during lactation may

Pitfalls of currently practiced approach for teaching medical students: opinions & options

Introduction: During a didactic lecture, information is given to the students by the teacher, where students are passive listeners and the teacher is the narrator. Interaction between the students and the teacher, though intended, is not always possible because of several reasons. Other formats for teaching (small group discussions, seminars by students etc.) have been utilized but in a limited way, particularly in India. In many medical institutes, students are expected to attend several lectures and there is no scheduled respite between lectures. This study has been done to understand the shortcomings of the current format and duration of lecture from the point of view of students. Method: Feedback was obtained from first year MBBS students under anonymity and was analyzed. There were 94 responses on the format of lectures and 98 for duration of lecture. Their responses were analyzed and the percentage of response for each choice was noted. Results: 57.14% students were of the opinion that didactic lectures must be supplemented with small group discussions and 46.81 % of students feel that the optimum duration of a lecture should be between 30 to 45 minutes. Discussion: Our study shows that majority of the students are of the opinion that duration of lecture should not be more than 45 minutes and more focus should be on supplementation of didactic lectures with small group discussion. Incorporating the students view point while forming educational policies may improve their performance to great extent. Key words: Didactic lectures, Educational policy, Educational program planning, Medical students, Performance improvement