The role of protein phosphorylation in regulation of carbon catabolite repression in Bacillus subtilis

CCR ist eines der bestuntersuchtesten Signaltransduktionssysteme in Bakterien. Es erlaubt Bakterien, sich an Veränderung im Vorhandensein von Kohlenstoffquellen anzupassen. Die CCR wird definiert als die Umsetzung der bevorzugten Kohlenstoffquelle, welche die Expression von Gene für die Umsetzung sekundärer Kohlenstoffquellen reprimiert. Auf der molekularen Ebene wird die CCR in B. subtilis durch den globalen Transkriptionsregulator CcpA erreicht. Wenn bevorzugte Kohlenstoffquellen wie Glukose vorhanden sind, bildet CcpA mit den phosphorylierten HPr- oder Crh- Proteinen einen Komplex. Dieser Komplex bindet an Operatorstellen der DNA, die als cre-Stellen bezeichnet werden. So werden viele Gene und Operons reprimiert, die an der Verwertung von sekundären Kohlenstoffquellen beteiligt sind. Die regulatorischen Phosphorylierungen von HPr und Crh wird durch das bifunktionale Enzym HPrK/P erreicht.In dieser Arbeit wurde die Wirkung von verschiedenen Kohlenstoffquellen auf die CCR untersucht. Für viele Kohlenstoffquellen konnte eine CcpA-abhängige Kataboliten-Repression im Reportersystem gezeigt werden. Des Weiteren konnte eine Hierarchie der Kohlenstoffquellen anhand ihres reprimierenden Potentials erstellt werden. Die verschiedenen Stärken der Repression durch die verschiedenen Kohlenstoffquellen könnten durch unterschiedliche Mengen des CcpA/Ko-Repressor-Komplexes erklärt werden. Die Mengen an CcpA und HPr ändern sich bei verschiedenen Kohlenstoffquellen nicht. Deshalb wurde das Phosphorylierungsmuster von HPr untersucht. Dazu konnte gezeigt werden, dass HPr aber nicht Crh für die Repression durch CcpA relevant ist. Deshalb wurde nur die Menge von HPr(Ser-P) in der Zelle untersucht. Stark reprimierende Kohlenstoffquellen führen zu einer hohen intrazellulären Konzentration von HPr(Ser-P). Die Särken der Katabolitenrepression lassen sich zurückführen auf die unterschiedliche Fähigkeit der Kohlenstoffquellen HPr(Ser-P) herzustellen.Bei Kohlenstoffquellen, die nur eine schwache Katabolitenrepression hervorrufen, konnten neben einer geringen Menge HPr(Ser-P) auch HPr-Proteine detektiert werden, die am Histidin phosphoryliert waren. HPr ist ebenfalls am PTS beteiligt. Das PTS kombiniert die Aufnahme von Kohlenstoffquellen mit deren Phosphorylierung. Im PTS wird das HPr-Protein durch EI am Histidinrest 15 phosphoryliert. Diese Phosphorylierung könnte die Menge an verfügbarem HPr als Substrat für die HPrK/P verringern. Deshalb wurde die Möglichkeit überprüft, ob HPr(His-P) die Phosphorylierung von HPr durch HPrK/P negativ reguliert. In einer EI-Mutante kann kein HPr(His-P) mehr gebildet werden. Trotzdem bleibt die Stärke der Katabolitenrepression in dieser Mutante bei nicht-PTS-Zuckern gleich. Daraus kann geschlossen werden, dass die Phosphorylierung von HPr am Ser46 allein durch die Aktivität von HPrK/P reguliert wird und unabhängig vom PTS ist. Die niedrigen Mengen HPr(Ser-P) bei Kohlenstoffquellen mit schwacher Katabolitenrepression resultieren aus einer geringen Kinaseaktivität der HPrK/P. Diese Hypothese wurde mit einer Mutante belegt, die eine HPrK/P-Mutation trägt, die dazu führt, dass das Enzym keine Phosphorylaseaktivität mehr besitzt. In dieser Mutante kommt es immer zur Katabolitenrepression.Es wird vermutet, dass die Aktivität der HPrK/P durch eine allosterische Regulation durch Metabolite wie FBP oder Pi erreicht wird. Deshalb wurde die FBP-Konzentration in vivo bestimmt. Mit dem meisten Zuckern ist der intrazelluläre FBP-Spiegel ausreichend hoch, um vollständige Kinaseaktivität der HPrK/P zu erreicht. Es müssen also weitere Faktoren vorhanden sein, die die Aktivität in vivo beeinflussen. Um zu überrpüfen, ob der HPr(Ser-P)-Pool durch andere Enzyme außer der HPrK/P beeinflußt wird, wurde die Rolle von PrpC untersucht. PrpC, eine Ser/Thr-Phosphatase, kann HPr(Ser-P) in M. pneumoniae dephosphorylieren. Das PrpC aus B. subtilis kann HPr(Ser-P) in vitro dephosphorylieren, aber es konnte kein Einfluß auf die Katabolitenrepression in vivo festgestellt werden.Zusammenfassend wurde in dieser Arbeit gezeigt, dass die HPrK/P eine zentrale Rolle bei der Katabolitenrepression in B. subtilis spielt. Die verschieden starken Effekte auf die Katabolitenrepression durch verschiedene Kohlenstoffquellen konnten durch einen unterschiedlichen HPr(Ser-P)-Spiegel erklärt werden. Außerdem konnte der fundamentale Unterschied im Mechanismus der Katabolitenrepression zwischen E. coli und B. subtilis weiter verdeutlicht werden. Die Aktivität des PTS ist in E. coli der ausschlaggebende Faktor für die globale sowie für die Operon-spezifische Katabolitenrepression. Ohne HPr(His-P) das EIIAglu aus E. coli unphosphoryliert und übt eine starkt Katabolitenrespression aus. In Gegensatz dazu wird die globale Katabolitenrepression in B. subtilis nicht direkt von der PTS-Aktivität beeinflußt

Estimation of instant solar radiation by using of instant temperature

An attempt has been made to correlate the recorded instant temperature and corresponding instant solar radiation of differentdays in hot season of Lucknow, India. The obtained graph for these data and also the obtained equation relating temperatureand corresponding instant radiation have good correlation and are useful in predicting of solar radiation from instant temperature

Role of vibroacoustic stimulation test in prediction of fetal outcome in terms of Apgar score

Background: Role of vibroacoustic stimulation test in prediction of fetal outcome in terms of Apgar score. Aim: To know the efficacy of vibroacoustic stimulation test in prediction of fetal outcome.Methods: The study was conducted in department of obstetrics and gynecology at Guwahati Medical College, in duration of 2007-2009. Total 200 high risk patients underwent VAST.Results: VAST done among all these patients and according to fetal response divided into VAST reactive and non-reactive. In 162 (81%) patients VAST was reactive and 38 (19%) non-reactive. Among VAST (162) reactive patients, 126 (77.77%) went for spontaneous vaginal delivery and for 36 (22.22%)   induction planned. Induction failed in 9 patients and cesarean section done. Total babies shifted to NICU 13 (6.5%), 4 babies expired (2%) and 9 (4.5%) improved. Thus VAST has good correlation in prediction of fetal outcome.Conclusions: It is simple, rapid and non-invasive tool for detection of fetal well-being. It is efficacious in detection of early fetal jeopardy as correlated with Apgar score. There are currently no randomized controlled trials that address the safety and efficacy of VAST used to assess fetal well-being.

Correlation of Pap smear and visual inspection with acetic acid for screening of premalignant and malignant lesion of cervix

Background: Cervical cancer still remains a leading cause of morbidity and mortality among women worldwide. Despite attempts to increase screening over the past several decades, incidence and mortality rates of cancer cervix have not shown substantial reduction. The conventional method of screening by cervical cytology/pap smear has failed to reduce the disease burden due to lack of patient compliance for repeated testing and a relatively sophisticated infrastructure. Therefore, alternative methods such as visual inspection after application of acetic acid (VIA), visual inspection with Lugol's Iodine (VILI) and human papillomavirus (HPV) DNA testing have been developed. The aim of present study was to compare Pap smear and VIA and evaluate their usefulness as tools for screening of premalignant and malignant lesions of cervix.Methods: This was a crossectional study over 5 months from 1 Jan 2015 to 31 may 2015 in which 212 patients attending the obstetrics and gynecology department, BHU were enrolled. Pap smear was taken followed by VIA. Pap smears were sent to pathology department, BHU where they were reported as per Bethesda System, 2001. Cervical biopsies were done in positive cases. Data obtained and statistically analyzed.Results: The present study was conducted over a period of 6 months among 212 patients age of 18-60 years screened. Positive results obtained from cytology were 26, VIA was positive in 28 women. Cervical biopsy was done in 34 women who had positive results by either test. Histology in 31 cases was suggestive of cervical intraepithelial carcinoma (CIN).Conclusions: VIA, though less specific has comparable sensitivity to Pap smear and may be used as a primary screening tool for cervical cancer. In combination both the tests have a higher predictive accuracy.

A prospective study to evaluate the role of routine early third trimester ultrasound in the diagnosis of intrauterine growth restriction and prediction of adverse perinatal outcome using color Doppler

Background: Intrauterine growth restriction (IUGR) is a major and silent cause of perinatal morbidity and mortality. Rate of IUGR in developing countries is 6 times higher than that of developed countries. 14 to 20 million infants are affected with IUGR in developing countries annually. The highest incidence is found in south central Asia (33%) and India (21%). Aim of study was to evaluate the role of early 3rd trimester ultrasound in diagnosis and role of color Doppler in prediction of adverse perinatal outcome in IUGR.Methods: A prospective case-control study was conducted on 720 women admitted in GMH Rewa for one year. 360 antenatal cases diagnosed as IUGR on ultrasound were taken as study group, same number of non IUGR cases were taken as control. Study cases were followed with Doppler. The umbilical artery pulsatility index (UA PI), middle cerebral artery pulsatility index (MCA PI) and cerebro-placental ratio were calculated and the results were analysed.Results: Inferences drawn from the study were: rate of caesarean section was significantly higher (31%) in study group, perinatal outcome was poor in more number of cases (10.8%) in study group than the control group (3.4%), and the sensitivity and specificity of cerebroplacental ratio was more reliable indicator in comparison to that of UA PI and MCA PI in prediction of adverse perinatal outcome.Conclusions: Looking at the high burden of IUGR cases in our country and its adverse effects on perinatal outcome, if early third trimester ultrasound is made mandatory for all pregnant women, most of the IUGR cases can be detected and managed timely and perinatal outcome can be significantly improved

Outcome and management of antenatal patients with jaundice in tertiary care centre of eastern India: a retrospective study

Background: Aim of current study was to know the etiological factors of jaundice among pregnant patients, outcome and their management.Methods: A retrospective study done at IMS, BHU in obstetrics and gynecology department among pregnant patients with jaundice admitted in obstetric wards and labour room in six months duration.  Results: Among total 1960 admissions, 78 (3.97%) patients presented with jaundice. Out of all admissions 27 (1.37%) were HBSAg, 8 (0.40%) HEV, 7 (0.35%) HCV, 6 (0.30%) of HBSAg and HEV co-infection, cholestasis with pregnancy 20 (1%) and 10 (0.51%) patients with pre-eclamptic liver disease with HELLP.Conclusions: Jaundice in pregnancy may be lethal to mother and fetus. As the course of disease is also rapid and in short period it may affect the fetus in utero also, early detection and prompt management of these cases should be done.

ZIKA VIRUS SERENE PROTEASE COMPLEX (NS2B-NS3) INHIBITION BY 2-AMINO-5-{[(1Z)-AMINO({[(Z)-BENZOYL]IMINO})METHYL]AMINO}-N-(5-AMINO-7-{[CARBAMOYL(PHENYL)METHYL]AMINO}-6-OXOHEPTYL)PENTANAMIDE, IN SILICO STUDIES

Objective: The present in silico study is taken to report 2-amino-5-{[(1Z) -amino ({[(Z) -benzoyl] imino}) methyl] amino} -N-(5-amino-7-{[carbamoyl (phenyl) methyl] amino} -6-oxoheptyl) pentanamide as Zika virus (ZIKV) NS2B-NS3 protease inhibitor.Methods: In silico studies performed on online docking servers. NS2B-NS3 serine protease from ZIKV with PDB ID: 5GJ4 a hydrolase with total structure weight of 102878.54 is selected as the target. Docking server is used for carrying out docking calculations. Lamarckian genetic algorithm and the Solis and Wets local search methods are used for performing docking simulations. Free energy calculations, hydrogen bond (HB) formation, polar and hydrophobic interactions and HB plot are studied in this study.Results: Binding pocket is found on a serine protease NS2B chain A. Binding site predictions propose NKK as the suitable ligand for binding, which has structure closely related to the proposed ligand2-amino-5-{[(1Z) -amino ({[(Z) -benzoyl] imino}) methyl] amino} -N-(5-amino-7-{[carbamoyl (phenyl) methyl] amino} -6-oxoheptyl) pentanamide. Free energy of binding is - 4.08 kcal/Mol and inhibition constant (Ki) is very less 1.02 mm. The ligand binds to chain A of NS2B and chain B of NS3 serine protease. The legend is bound to serine protease complex through strong HB, formed between THR 60 (A) and N6 of ligand, GLU62 (A) and N8 of ligand, ARG 55 (A) and N3 of ligand and ASN108 (B) and N7 of ligand apart from polar and hydrophobic interactions.Conclusion: Docking studies performed establishes the proposed ligand2-amino-5-{[(1Z)-amino ({[(Z)-benzoyl] imino}) methyl] amino} -N-(5- amino-7-{[carbamoyl (phenyl) methyl] amino}-6-oxoheptyl) pentanamide as a molecule which can be used for the inhibition of ZIKV NS2B-NS3 serine protease. Â

Study of BMI in pregnancy and its correlation with maternal and perinatal outcome

Background: Maternal nutrition plays an important role in maternal and fetal outcome. The low maternal BMI or Obesity is associated with adverse outcome.Methods: A total 148 primigravida included. BMI was calculated on first visit and in each trimester, all booked patient was followed throughout pregnancy and delivery for any maternal and fetal complications.Results: (66.2%) of patients were with normal BMI, (17.56%) were underweight, (10.13%) were overweight and (6.08%) patients were obese. APH was seen in (11.1%) and (6.7%) cases in BMI grades ‘IV and III’ respectively. PIH was present in total 10 cases, (33.3%) cases belonged to BMI grades IV. Anemia was present in total 51 cases (53.8%) belonged to BMI grade I. Preterm delivery was present in total 5 cases and out of them (7.7%), (2%) and (11.1%) belonged to BMI grades I, II and IV respectively. GDM was present in 11 cases and out of them (26.7%) and (44.4%) cases belonged to BMI grade III and IV respectively. PPH was present in total 5 cases and out of them (7.7%), (2%) and (11.1%) belonged to BMI grades I, II and IV respectively. In BMI grade I group, (96.2%) babies had birth weight 2.5 (22.2%) cases of fetal macrosomia belonged to BMI grade IV.Conclusions: There is importance of pr-pregnancy counseling in maintaining weight of women during pregnancy to avoid pregnancy maternal and fetal outcomes.

Study of thrombocytopenia in pregnancy: clinical presentation and outcome at tertiary care rural institute

Background: Platelet count below 1.5 lakh/cumm is called as thrombocytopenia. After anaemia it is the second most common haematological disorder in pregnancy. It affects nearly 6 to 15%; on an average 10% of all pregnancies. Gestational thrombocytopenia is a clinically benign thrombocytopenic disorder usually occurring in late pregnancy. It resolves spontaneously after delivery.Methods: It is a hospital based prospective observational study over a period of 1 year. All pregnant women who attended OPD at the department of obstetrics and gynecology, UPUMS, Saifai for antenatal checkup were included for the study and blood sample was withdrawn.Results: Out of 263 cases enrolled for study, 90 women were found to have thrombocytopenia, and 173 had normal platelet count. Thus, incidence of thrombocytopenia was 34%. Gestational thrombocytopenia accounted for majority of cases of thrombocytopenia in pregnancy (50%) followed by hypertensive disorders (22.4%). It was further followed by ITP (11.11%) and dengue (5.5%).Conclusions: Gestational thrombocytopenia is the most common cause of thrombocytopenia during pregnancy (50%), but other underlying causes must be considered as well. A thorough history and physical examination will rule out most causes

A survey of awareness of Pap smear and cervical cancer vaccine among women at tertiary care centre in Eastern Uttar Pradesh India

Background: Cervical cancer is very common in developing countries. In Uttar Pradesh the literacy rate of women is low as compared to other states of India. Poverty is very high. Most women report with advance stage of cervical cancer. It can be prevented by early screening with Pap smear. Aim of our study is to know about the awareness of Pap smear and cervical cancer vaccine among women attending a tertiary hospital in Eastern part of Uttar Pradesh India.Methods: A questionnaire based study is done on 1500 patients attending the OPD and admitted in ward over a period of one year.Results: To our surprise only 12.06% women were aware of Pap screening test. Only 6.13% women were aware of cervical cancer vaccine. The awareness about both Pap test and cervical cancer vaccine was present in only 5.2% only. The education of the women and socioeconomic status also play an important role. The awareness among rural women was significantly less as compared to urban women.Conclusions: The awareness about Pap test and cervical cancer vaccine was very low specially in women staying in rural area. So if we really wanted to reduce cervical cancer then we have to increase the awareness among women by media, camps etc