Citrate Binding to the Membrane Protein Proteorhodopsin

Proteorhodopsin (pR) is an intrinsic membrane protein with an important role in solar-energy storage of the biosphere. Earlier work in our lab has shown that polyhistidine-tagged pR can be purified by means of selective precipitation with citrate under specific conditions, as can a number of mutants based on this His6-tagged pR. Purification of a heterologously-expressed trans-membrane protein by a simple salt such as citrate is novel. However, such a phenomenon leads to several questions: How does citrate cause pR precipitation? Does the polyhistidine-tag assist in such a precipitation? Is this precipitation pH-specific? Does citrate affect the function of pR? Does citrate-induced pR precipitation have any biological significance? Are there other ions that could cause pR precipitation? This dissertation focuses on understanding the nature of the interaction of pR with citrate and other anions, and in particular on trying to take advantage of this interaction in order to develop a novel membrane protein purification method. The end goal that branches out of these two aims is to utilize the compact citrate interaction site identified in pR, by incorporating it into other membrane proteins and using it to permit their purification by similar simple procedures. In Chapter 1, I briefly provide some background information on the wide variety of structurally-similar proteins as rhodopsins that include pR. I also describe the general importance of developing purification methods for 7-helix membrane proteins, including pR. Chapter 2 focuses on the investigation of the nature of citrate-binding site of pR. To address the main question of how citrate aids in pR purification, site-directed mutagenesis technique was applied to generate several single, double, triple or quadruple mutants of pR in a histidine-tag free background, which were then tested for their reactivity to citrate. Several different anions were tested to examine if precipitation of pR was specific to citrate or whether the precipitation is susceptible to other negatively charged salts. Photocycle of pR progresses through several intermediates, each with a distinct absorption maximum (described in subsection 1.3.1). M-intermediate is detected at pH ≥ 8 with λmax = 410 nm. Flash spectroscopy involves excitation of pR at a particular wavelength that leads to transient absorption, thus, signaling the formation of the corresponding intermediate. Flash-induced transient visible absorption measurements were used to assay the effect of exposing pR to citrate on its physiological function. Chapter 3 describes the development of a method of purification of pR using simple salts, citrate and phosphate. Chapter 4 begins an exploration of a future direction. The ultimate objective is to apply the above techniques for the general purpose of 7-helix membrane protein purification, especially for the important class of pharmacological receptors known as GPCRs. An attempt at heterologous expression in E. coli, and purification, of a mammalian GPCR, is described therein. Such a method would be desirable for obtaining proteins for structural, functional and pharmacological studies

Contrast of Bayesian and Classical Sample Size Determination

Sample size determination is a prerequisite for statistical surveys. A comprehensive overview of the Bayesian approach for computation of the sample size, and a comparison with classical approaches, is presented. Two surveys are taken as example to illustrate the accuracy and efficiency of each approach, and to make recommendations about which method is preferred. The Bayesian approach of sample size determination may require fewer subjects if proper prior information is available

Demographic profile and high risk factors in morbidly adherent placenta

Background: Morbidly adherent placenta with its variants is one of the most feared complications causing high morbidity and mortality in obstetrics. The main objectives of the study were to evaluate the demographic profile and risk factors in morbidly adherent placenta.Methods: A retrospective study was done in which case records of 40 patients diagnosed to have morbidly adherent placenta were reviewed. Demographic data including age, parity, gestational age and previous caesarean delivery or other uterine surgery, details of medical and obstetric history were recorded.Results: A total of 24232 deliveries were conducted in the hospital. 40 patients were confirmed to have morbidly adherent placenta. The average age of the patients was 31.8 years and the mean parity was 1.80. 95% percent of the patients were booked in the study centre and only 5% percent were unbooked.  80% of the patients were from the rural area and 20 % from the urban region. The average gestational age of the patient at diagnosis was 35.3 weeks and average gestational age at delivery was 36.2 weeks. The risk factors included previous caesarean section in 32 patients (80%) , with history of  one C.S in 11 patients(27.5%),with two C.S in 19 patients(47.5%), and with history of three C.S in 4 patients (5%). 8 patients (20%) had history of both previous C.S and curettage and none had the history of curettage alone.Conclusion: Morbidly adherent placenta is a potentially life threatening condition. Management of such patients requires early recognition of high risk women based on their clinical risk factors, with accurate preoperative diagnosis, good maternal counselling and planning of delivery

Knowledge attitude and practice of contraceptives among married women of reproductive age group

Background: This study was conducted to assess the knowledge, attitude and practices of contraceptives among married women of reproductive age group.Methods: It was a cross sectional study which was carried at outpatient department (OPD) of obstetrics and gynaecology, Government medical college Srinagar over a period of 3 months. A total of 210 married women in the age group 15-49 years were included in the study. Along with the demographic profile, their knowledge, attitude and practices on contraception were evaluated with the help of self-structured questionnaire.Results: The mean age of the study population was 32.5±6.8 years. Majority of the women were in the age group of 26-35 years (56.2%), literate with formal education above 10th class (42.4%), house-makers (46.2%) and from rural background (65.2%). 82.9% of women had awareness regarding any method of contraception. Knowledge about emergency contraceptive was quiet low (11.0%). The most common source of information on contraception was friends and relatives (64.8%). 41% of women had a negative attitude towards contraception. The most common method of contraceptive used was barriers (21.9%). 43.8% of women had never used any contraceptive.Conclusions: Despite having good knowledge about contraception, utilization of contraceptives were less

Un estudio observacional sobre infecciones bacterianas en pacientes con coronavirus hospitalizados en un centro de tercer nivel de atención

Introduction: The latest global pandemic is caused by SARS-CoV-2 coronavirus. 6% of patients are admitted to the hospital, with 20% of those admitted to the intensive care unit (ICU) due to acute respiratory distress syndrome (ARDS).Aim: to characterise the bacterial infections in patients with coronavirus at an ICU of North Indian Hospital.Method: After receiving ethical approval from the institutional review board, a retrospective study was done on coronavirus subjects admitted to GMC Srinagar's ICUs between January and June 2021. Data on demographics, comorbidities, and microbiology were obtained retrospectively.Results: Overall, “394 COVID-19 patients were admitted to ICU. Median age was 58 years (IQR 51–69) and sex ratio (M/F) was 3. At admission the median SAPS II was 33 (IQR 24–49). Among patients, 232 had at least one co-morbidity and 162 were overweight (body mass index (BMI) = 27.8 kg/m2). In all, 358 patients received antibiotics (244/358 introduced before ICU and 114/358 during ICU stay)”.Conclusion: The current study is the first of its kind in our hospital setting to describe the bacterial persistence in the lungs despite adequate therapy, as well as frequent bloodstream infections possibly associated with bacterial translocations originating from the digestive or oropharyngeal microbiota, in COVID-19 ICU patients.Introducción: La última pandemia mundial está provocada por el coronavirus SARS-CoV-2. El 6 % de los pacientes ingresan en el hospital, con un 20% de los ingresados en la unidad de cuidados intensivos (UCI) debido al síndrome de dificultad respiratoria aguda (SDRA).Objetivo: caracterizar las infecciones bacterianas en pacientes con coronavirus en una UCI del North Indian Hospital.Método: Después de recibir la aprobación ética de la junta de revisión institucional, se realizó un estudio retrospectivo en sujetos con coronavirus admitidos en las UCI de GMC Srinagar entre enero y junio de 2021. Los datos sobre demografía, comorbilidades y microbiología se obtuvieron retrospectivamente.Resultados: en general, “394 pacientes con COVID-19 fueron admitidos en la UCI. La mediana de edad fue de 58 años (RIC 51-69) y la proporción de sexos (M/F) de 3. Al ingreso, la mediana de SAPS II fue de 33 (RIC 24-49). Entre los pacientes, 232 tenían al menos una comorbilidad y 162 tenían sobrepeso (índice de masa corporal (IMC) = 27,8 kg/m2). En total, 358 pacientes recibieron antibióticos (244/358 introducidos antes de la UCI y 114/358 durante la estancia en la UCI)”.Conclusión: El presente estudio es el primero de su tipo en nuestro medio hospitalario en describir la persistencia bacteriana en los pulmones a pesar de una terapia adecuada, así como infecciones frecuentes del torrente sanguíneo posiblemente asociadas con translocaciones bacterianas originadas en la microbiota digestiva u orofaríngea, en COVID-19 pacientes de la UCI

Un estudio observacional sobre infecciones bacterianas en pacientes con coronavirus hospitalizados en un centro de tercer nivel de atención

Introduction: The latest global pandemic is caused by SARS-CoV-2 coronavirus. 6% of patients are admitted to the hospital, with 20% of those admitted to the intensive care unit (ICU) due to acute respiratory distress syndrome (ARDS).Aim: to characterise the bacterial infections in patients with coronavirus at an ICU of North Indian Hospital.Method: After receiving ethical approval from the institutional review board, a retrospective study was done on coronavirus subjects admitted to GMC Srinagar's ICUs between January and June 2021. Data on demographics, comorbidities, and microbiology were obtained retrospectively.Results: Overall, “394 COVID-19 patients were admitted to ICU. Median age was 58 years (IQR 51–69) and sex ratio (M/F) was 3. At admission the median SAPS II was 33 (IQR 24–49). Among patients, 232 had at least one co-morbidity and 162 were overweight (body mass index (BMI) = 27.8 kg/m2). In all, 358 patients received antibiotics (244/358 introduced before ICU and 114/358 during ICU stay)”.Conclusion: The current study is the first of its kind in our hospital setting to describe the bacterial persistence in the lungs despite adequate therapy, as well as frequent bloodstream infections possibly associated with bacterial translocations originating from the digestive or oropharyngeal microbiota, in COVID-19 ICU patients.Introducción: La última pandemia mundial está provocada por el coronavirus SARS-CoV-2. El 6 % de los pacientes ingresan en el hospital, con un 20% de los ingresados en la unidad de cuidados intensivos (UCI) debido al síndrome de dificultad respiratoria aguda (SDRA).Objetivo: caracterizar las infecciones bacterianas en pacientes con coronavirus en una UCI del North Indian Hospital.Método: Después de recibir la aprobación ética de la junta de revisión institucional, se realizó un estudio retrospectivo en sujetos con coronavirus admitidos en las UCI de GMC Srinagar entre enero y junio de 2021. Los datos sobre demografía, comorbilidades y microbiología se obtuvieron retrospectivamente.Resultados: en general, “394 pacientes con COVID-19 fueron admitidos en la UCI. La mediana de edad fue de 58 años (RIC 51-69) y la proporción de sexos (M/F) de 3. Al ingreso, la mediana de SAPS II fue de 33 (RIC 24-49). Entre los pacientes, 232 tenían al menos una comorbilidad y 162 tenían sobrepeso (índice de masa corporal (IMC) = 27,8 kg/m2). En total, 358 pacientes recibieron antibióticos (244/358 introducidos antes de la UCI y 114/358 durante la estancia en la UCI)”.Conclusión: El presente estudio es el primero de su tipo en nuestro medio hospitalario en describir la persistencia bacteriana en los pulmones a pesar de una terapia adecuada, así como infecciones frecuentes del torrente sanguíneo posiblemente asociadas con translocaciones bacterianas originadas en la microbiota digestiva u orofaríngea, en COVID-19 pacientes de la UCI

Contiguous Patches of Translational Hydration Dynamics on the Surface of K-Ras

Proteins involved in signaling pathways represent an interesting target for experimental analysis by ODNP (Overhauser Dynamic Nuclear Polarization), which determines the translational mobility at the surface of proteins. They also represent a challenge, since the hydration dynamics at all sites remains relatively rapid, requiring sensitive measurements capable of drawing finder distinctions. Targeting the protein K-Ras, we find ODNP cross-relaxivity values that appear consistent within similar regions of 3D space, regardless of the specific residue where the spin probe used to select the location has been attached

Un estudio observacional sobre infecciones bacterianas en pacientes con coronavirus hospitalizados en un centro de tercer nivel de atención

Introduction: The latest global pandemic is caused by SARS-CoV-2 coronavirus. 6% of patients are admitted to the hospital, with 20% of those admitted to the intensive care unit (ICU) due to acute respiratory distress syndrome (ARDS).Aim: to characterise the bacterial infections in patients with coronavirus at an ICU of North Indian Hospital.Method: After receiving ethical approval from the institutional review board, a retrospective study was done on coronavirus subjects admitted to GMC Srinagar's ICUs between January and June 2021. Data on demographics, comorbidities, and microbiology were obtained retrospectively.Results: Overall, “394 COVID-19 patients were admitted to ICU. Median age was 58 years (IQR 51–69) and sex ratio (M/F) was 3. At admission the median SAPS II was 33 (IQR 24–49). Among patients, 232 had at least one co-morbidity and 162 were overweight (body mass index (BMI) = 27.8 kg/m2). In all, 358 patients received antibiotics (244/358 introduced before ICU and 114/358 during ICU stay)”.Conclusion: The current study is the first of its kind in our hospital setting to describe the bacterial persistence in the lungs despite adequate therapy, as well as frequent bloodstream infections possibly associated with bacterial translocations originating from the digestive or oropharyngeal microbiota, in COVID-19 ICU patients.Introducción: La última pandemia mundial está provocada por el coronavirus SARS-CoV-2. El 6 % de los pacientes ingresan en el hospital, con un 20% de los ingresados en la unidad de cuidados intensivos (UCI) debido al síndrome de dificultad respiratoria aguda (SDRA).Objetivo: caracterizar las infecciones bacterianas en pacientes con coronavirus en una UCI del North Indian Hospital.Método: Después de recibir la aprobación ética de la junta de revisión institucional, se realizó un estudio retrospectivo en sujetos con coronavirus admitidos en las UCI de GMC Srinagar entre enero y junio de 2021. Los datos sobre demografía, comorbilidades y microbiología se obtuvieron retrospectivamente.Resultados: en general, “394 pacientes con COVID-19 fueron admitidos en la UCI. La mediana de edad fue de 58 años (RIC 51-69) y la proporción de sexos (M/F) de 3. Al ingreso, la mediana de SAPS II fue de 33 (RIC 24-49). Entre los pacientes, 232 tenían al menos una comorbilidad y 162 tenían sobrepeso (índice de masa corporal (IMC) = 27,8 kg/m2). En total, 358 pacientes recibieron antibióticos (244/358 introducidos antes de la UCI y 114/358 durante la estancia en la UCI)”.Conclusión: El presente estudio es el primero de su tipo en nuestro medio hospitalario en describir la persistencia bacteriana en los pulmones a pesar de una terapia adecuada, así como infecciones frecuentes del torrente sanguíneo posiblemente asociadas con translocaciones bacterianas originadas en la microbiota digestiva u orofaríngea, en COVID-19 pacientes de la UCI

Knowledge of emergency contraception among women of childbearing age at a teaching hospital of Karachi

OBJECTIVES: To assess knowledge and attitudes about Emergency Contraception among women of childbearing age in Karachi, Pakistan. METHODS: A questionnaire based survey was conducted on 400 married women, attending the family practice clinics at a teaching hospital in Karachi, Pakistan from July to December 2006. Questionnaire was administered to women at the family practice clinic-seeking level of knowledge of emergency contraception (EC) and attitudes towards its use, Ethical requirements of informed consent and confidentiality were ensured Data was entered into Epi data and analyzed in SPSS.RESULTS: Eighty-eight percent of women were not aware of EC. 83% were housewives. Only a small number (11.5%) ever used EC to prevent pregnancy, out of those, the correct timing of effectiveness of post-coital pill was known to only 40% of women while none of these women were aware of the existence of Intra Uterine Contraceptive Device (IUCD) insertion as an option for EC About 50% of women identified general practitioners or family medicine clinics as their main sources of knowledge about EC. Increased advertising was considered desirable by 72% while 37% considered over the counter availability of EC pill desirable and only 36% of women were uncomfortable to use EC because of religious reasons. CONCLUSION: EC has a potential to offer women an important option for fertility control. Lack of women\u27s knowledge about EC use and availability may account in part for its limited use. There is a need to improve women\u27s education about EC. The primary health care providers can play a major role in informing their patients about emergency contraception

The Behavior Of Profit Of Musharakah Mutanaqisah Homeownership Partnership By The Case Of Abandoned Housing Project In Malaysia

Amalan projek perumahan di Malaysia adalah membimbangkan kerana sistem pembiayaan perumahan melaksanakan kontrak jual kemudian bina. Hal ini dipercayai menjadi faktor utama yang membawa kepada projek perumahan terbengkalai. Sistem ini membolehkan jualan dibuat daripada pemaju kepada pembeli sebelum projek perumahan selesai. Isu utama daripada projek perumahan terbengkalai adalah apabila ia melibatkan sistem pembiayaan Islam terutamanya pada kemudahan Bai 'Bithaman Ajil (BBA). Daripada kajian kes di Malaysia, sebahagian besar pembeli rumah dalam pembinaan kecewa dengan sistem perbankan Islam kerana projek perumahan terbengkalai tiba-tiba berlaku dan mereka perlu menanggung kerugian dan menghadapi lebih banyak kesukaran untuk menyelesaikan masalah ini secara sah. Bank Islam tidak boleh membiarkan penghutang untuk menghadapi kerugian semata-mata kerana setiap pihak bertanggungjawab untuk menjual hak yang gagal menyiapkan dalam tempoh. Tambahan pula, bank Islam dipercayai oleh masyarakat untuk membawa keadilan dan belas kasihan kepada rakyat kerana ia menyediakan amalan yang sah dan beretika mengikut Syariah. Ulama memperkenalkan Musharakah Mutanaqisah Homeownership (MMP) sebagai kemudahan pembiayaan perumahan alternatif menggantikan BBA. The practice of housing project in Malaysia is worrisome because housing financing system implement the sell-then-built contract. This is believed to be a major factor that leads to the abandoned housing project. The system let the selling made from the developer to the purchaser before the housing project is complete. The main issue cause from the abandoned housing project is when it involving the Islamic financing system particularly on Bai’ Bithaman Ajil(BBA) facility. From the cases study in Malaysia, most purchasers of houses under construction were disappointed with the Islamic banking system because of the abandoned housing project is suddenly occurred and they have to suffer the losses and facing more difficulties to solve the problem legally. The Islamic bank should not let the debtors to face the losses alone because each of the parties responsible for selling the rights which failed to complete in duration. Furthermore, the Islamic bank is trusted by the society to bring justice and compassion for the people as it provides legitimate and ethical practice following the Shariah. Scholars introduce Musharakah Mutanaqisah Homeownership (MMP) as an alternative home financing facility replacing the BBA