bluuebunny/crossref_metadata_2025 huggingface dataset:
Impact of COVID-19 on Liver Enzymes: A Retrospective Study
Authors: Patel Seema , Garima Gini , Anand Nidhi , Pandey Anil Kumar , Kaur Harnam , Pasupuleti Pullaiah
Published: May 2022

Introduction: The global pandemic of novel Coronavirus Disease- 2019 (COVID-19) caused by Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) has spread worldwide crippling the healthcare system. Besides the respiratory system, COVID-19 patients show signs of various degrees of liver damage, the mechanism and implication yet undeciphered. Hence, in this study, we aim to find out pattern and trend of derangement in liver functions in COVID-19 patients. Aim: To depict the pattern and trend of liver enzymes in COVID- 19 admitted patients without history of liver disease. Materials and Methods: The study was conducted as a single centred, retrospective, observational study, from June 2020 till December 2020. It included 1909 admitted COVID-19 positive patients diagnosed via either nasopharyngeal or oropharyngeal swab by Real-Time Reverse Transcription-Polymerase Chain Reaction (RT-PCR). The COVID-19 positive patients were divided into four groups Group I: Age- 15-20 years; 150 patients, Group II: Age 21-40 years; 645 patients, Group III: Age 41-60 years; 560 patients, Group IV: Age >60 years; 554 patients. Abnormality in liver tests was defined as greater than three times of upper limit of normal reference range Alanine Transaminase (ALT) >120 U/L, Aspartate Transaminase (AST) >120 U/L as Hepatocellular injury, greater than two times of upper limit of normal reference range Alkaline Phosphatase (ALP) >250 U/L as Cholestatic Injury (CSI). The statistical analysis was done by the Statistical Package for Social Sciences (SPSS) version 25.0. Results: It was found that with respect to ALT levels, males in age 21-60 years, whereas females in two extremes of age 15-20 and >60 years age were the most affected. Whereas, females in age group <40 years were more affected with higher AST activity. With respect to abnormal ALP levels female in age group <40 years were most affected but males were most affected in age group >40 years. Conclusion: Irrespective of age and gender, abnormality in liver enzymes was observed. Liver Function Tests (LFT) are a part of routine investigations carried out at the time of admission; its abnormalities may guide us in devising strategy to prioritise patient management in correlation with inflammatory markers.
Prevalence of Extended Spectrum Beta-lactamase Encoding Genes in Enterobacteriaceae Isolated from Various Clinical Samples in a Tertiary Care Cancer Centre, Kerala, India
Authors: Samuel Sajani , Neeraja O , Parthiban R , Saravanan M , Sarath KE
Published: May 2022

Introduction: Cancer patients are disproportionately at high risk of developing infections, with a risk of infection about 10 times than that of non cancer patients. Extended Spectrum Beta-Lactamases (ESBLs) enzyme producing gram-negative bacteria have been marked as one of the “serious” threat by the centre for disease control. This enzyme has the ability to hydrolyse the beta-lactam antibiotics and posses a major threat for the immunocompromised and cancer patients. Hence, understanding its prevalence at the grammatical level is important to decide upon the type of drugs to be prescribed. Aim: To study the prevalence of ESBL genes among the Gram- Negative Bacteria (GNB) isolated from cancer patients. Materials and Methods: The present study was a hospital- based cross-sectional study carried out in Microbiology Division, Malabar Cancer Centre, Thalassery, Kerala, India, from January to March 2021. Microbiological identification of the causative agents was done by staining, culturing and biochemical methods. Screening of the isolates for ESBL was done using Double Disc Synergy Test (DDST). Antibiotic susceptibility testing was carried out using modified Kirby-Bauer disc diffusion method. The ESBL producers were genotypically confirmed through Polymerase Chain Reaction (PCR) and typed accordingly. The data were given as average with standard deviation and analysed using Microsoft Excel. Results: Out of 1,310 specimens, 366 (27.9%) were culture positive. Among the GNB, Escherichia coli (50%) followed by Klebsiella pneumoniae (46.07%), Proteus (1.96%) and Enterobacter cloacae (0.98%) were the predominant isolates. Most of these ESBL producers (Escherichia coli, Klebsiella pneumoniae) were Multidrug Resistant (MDR). However, significant isolates of Escherichia coli (98.03%), Klebsiella pneumoniae (36.1%), Proteus, (50%) and Enterobacter cloacae (100%) were sensitive to immunocompromised. Among 102 ESBL producers, prevalence of blaTEM (67.64%) was highest followed by blaCTX-M (10.78%), blaSHV (14.70%) and blaOXA (18.62%). All of the ESBL producers tested showed the presence of one of four beta- lactamase encoding genes by PCR. One of the isolate Proteus mirabilis found to be ESBL producer as confirmed by phenotypic methods but lacked ESBL genes. Conclusion: Higher prevalence of ESBL producing strains warrants stringent measures to tackle the spread of MDR strains. Carbapenems can be considered as the drug of choice against ESBL producers. Highly prevalent blaTEM gene could be considered as potential therapeutical and diagnostic target against the ESBL producing GNBs.
Status of Common Serum Minerals and Trace Elements in COVID-19 Follow-up Cases: A Hospital-based Study from Eastern India
Authors: Maiti Saumyajit , Mukherjee Kheya , Samaddar Debopriyo , Bhattacharjee Debojyoti
Published: May 2022

Introduction: Management of post Coronavirus Disease-2019 (COVID-19) complications are the new challenges nowadays. It has been observed that in post COVID-19 cases the serum levels of several electrolytes and trace elements are deranged. Aim: This study was conducted to estimate the levels of different serum minerals and trace elements in one month post COVID-19 follow-up patients and compared with normal age- sex matched controls. Materials and Methods: This hospital based cross-sectional study was conducted in the Department of Biochemistry in a tertiary care centre, over a period of six months (from January 2021 to June 2021). After fulfilling proper inclusion criteria, 223 COVID-19 follow-up patients of both genders (134 males and 89 females) were included as ‘case’ group. Age-sex matched 250 healthy volunteers were recruited as ‘control’ group. Levels of serum electrolytes (sodium and potassium) and trace elements (zinc, iron, calcium, phosphate, copper, magnesium and selenium) were estimated, analysed and compared against each other. Tables and statistical analysis was done using Statistical Package for the Social Sciences (SPSS) version 20.0. Results: In this study, the levels of serum zinc and iron were found to be higher and serum calcium, phosphate, copper, magnesium and selenium levels were reduced in post COVID-19 one month follow- up cases in comparison to controls. Female cases were in deficient state of iron, calcium, copper and magnesium but had high serum zinc and phosphate when compared to males. No abnormalities were noted in the level of electrolytes in post COVID-19 cases. Conclusion: Assessment and monitoring of levels of the mineral throughout the course of post COVID-19 follow-up is advisable for timely and appropriate measures to combat with post COVID-19 complications.
Seroprevalence of Transfusion Transmitted Infections among Multiple Transfused Patients in Indian Scenario: Step for Safe Transfusion Practices
Authors: Patel Swati , Kesarwani Vishwadeep , Soni Himanshu , Sah Neeharika , Purohit Manju
Published: May 2022

Introduction: Transfusion Transmitted Infections (TTIs) is a worldwide problem both in past and in the present days. The risk of TTIs is increased due to the constant need to receive blood transfusion in patients requiring multiple transfusions. The problem of TTIs runs parallel to the prevalence of the infections in the blood donor group and to the number of units transfused. Aim: To estimate the current prevalence of blood TTIs in patients requiring multiple units of blood transfusions and to determine their association with age, gender, various disease category and number of units transfused. Materials and Methods: This cross-sectional retrospective study was conducted in the Department of Pathology, RD Gardi Medical College, Ujjain, Madhya Pradesh, India from January 2017 to December 2020. Patients of all the age groups and both gender from Outpatient and Inpatient Department, requiring regular transfusions and having a history of 10 or more units of blood transfusion were enrolled. Serological detection of TTIs was done by Enzyme-Linked Immune Sorbent Assay (ELISA) method. Statistically the results were analysed using Statistical Package for Social Sciences (SPSS) version 19.0. Results: In this study, 480 patients were included of which 293 (61.1%) were males and 187 (38.9%) females. Maximum 195 (40.6%) were children in age group of <1-10 years. A total of 63 (13.12%) patients with Hepatitis C virus, 9 (1.87%) patients with Hepatitis B virus and 2 (0.42%) patients with Human Immunodeficiency Virus (HIV) infection were found at the end of study. Highest seropositivity of TTI was seen in patient who had undergone >400 number of transfusions , thus association was established but no association detected with age and sex of the patients. Majority of TTIs were positive in thalassemia patients 164 (34.17%) followed by patient undergoing haemodialysis 151 (31.45%). Conclusion: The results of this study raise an alarm to the existence of a significant risk of TTIs in our society. There is requirement of advanced and vigilant screening of donors with proper strategies to reduce the breach in safe blood transfusion practices.
Comparison of Hand Hygiene Adherence Rate before and after Training in a Multispecialty Hospital, Gujarat, India
Authors: Solanki Rachana Rashesh , Tanna Kruti Jashvant , Halpati Asha Dayabhai , Shah Navin Ishwarbhai
Published: May 2022

Introduction: The most effective way to break the chain of transmission of infection in healthcare facility is Hand Hygiene (HH). However, the HH adherence rate/compliance is varying worldwide. Healthcare facilities should follow comprehensive, systematic approach for assessment of HH adherence rate. Periodic monitoring and necessary feedback is critical to improve HH adherence rate/compliance. Aim: To know the impact of the HH training of Healthcare Workers (HCW) on HH adherence rate. Materials and Methods: This prospective study was conducted at SMS Multispecialty Hospital, Ahmedabad, Gujarat, India. The study was conducted for the duration of six months (October 2019-March 2020). Total 416 opportunities were accessed in two sessions {203 pretraining and 213 post-training} among different HCWs of ICU and NICU. HH adherence rate was measured by direct observation methods by trained, non participating neutral person as per World Health Organisation (WHO) HH guidelines. HH training was given to HCWs (doctors, nursing staff and attendant) of Intensive Care Unit (ICU) and Neonatal Intensive Care Unit (NICU). Pre and post-training HH adherence rate was calculated among HCWs of ICU and NICU in respect to WHO’s five movement of HH. Chi-square test was used to do statistical analysis. Results: Pre and post-training HH adherence rate of HCWs of ICU is 41% and 69% respectively. However, pre and post- training HH adherence rate of HCWs of NICU was 52% and 75%, respectively. Both ICU and NICU post-training HH adherence rate of HCWs was improved which was statistically significant (p-value <0.00001). Both pre and post-training, compared to doctors, nursing staff had better HH adherence rate (45%, 67% in ICU and 58% and 78% in NICU). Hand rub was preferred method of HH (93%). Conclusion: The HH training improved the HH adherence rate of HCWs of ICU and NICU. Induction and periodic training of HCWs improves HH adherence rate of HCWs.
Lipoid Pneumonia and Peritoneal Endometriosis: Rare Incidental Findings on Autopsy
Authors: Chaurasia Zini , Gupta Renu , Agarwal Swapnil
Published: May 2022

Lipoid pneumonia has been described in as early as 1925, by Laughlen GF, to be a rare variant of pneumonia caused by the inhalation of oil droplets. Around 50% of the patients with lipoid pneumonia are asymptomatic. Here, a case is presented where peritoneal fluid and tissue pieces from various organs of a 12-year-old female were received for examination after autopsy. There was no history of any illness before death. Cytology of fluid revealed features suggestive of endometriosis. Microscopy of lung sections revealed numerous lipid droplets in the alveoli and the interstitium along with a mild inflammatory infiltrate suggestive of lipoid pneumonia. In view of incidental findings in present case report, the occupational history or history of inhalation of oil-based medications becomes very important and should always be investigated. Also, in the absence of any symptoms peritoneal fluid needs to be studied carefully for any pathology.
The Development of Private Digital Currency in China and Its Enlightenment to the Promotion of Legal Digital Currency
Authors: Jun Wang
Published: May 2022

With the rapid development of global financial technology, legal digital currency will become an important form of currency in the future. At present, the central bank is committed to the development of legal digital currency in China, and has carried out experiments on relevant platforms. Legal digital currency is a new thing and lacks successful experience that can be used for reference. Therefore, the analysis and Research on the potential risks of private digital currency and a clear understanding of the shortcomings of China's current response to the risks of private digital currency are important prerequisites for the steady development of China's legal digital currency.

Firstly, it expounds the concept and classification of digital currency, then analyzes the development and supervision status of private digital currency in China and other countries around the world, points out the shortcomings of China's response to the risk of private digital currency, and finally puts forward the enlightenment and suggestions for the issuance of legal digital currency in China.

There are some problems in private digital currency, such as unclear legal attributes and tax policies, non-standard trading platforms, lack of consumer protection, and weak international cooperation. China should pay attention to the above problems when promoting the development of legal digital currency. Specifically, we should clarify the legal attribute of digital currency; Build a complete digital currency supervision system; Improve the digital currency tax policy; Standardize the digital currency trading platform; Building an anti-money laundering mechanism; Strengthen the protection of consumers' rights and interests; Strengthen international digital currency cooperation.
Analysis and Study of Seismic Response of a G+13 Multistoried Building for Different Types of Dampers Using Etabs Software
Authors: Supriya A. , Murali K.
Published: May 2022

Vibration refers to repeated movements that cause fatigue and decreased function of the structure. An earthquake can emit large amounts of energy that can have a detrimental effect on all parts of a building. Therefore, the reduction of vibration or maintenance of buildings such as bridges, dams, roads and buildings is important for the safety of life and the reduction of economic losses. Different types of dampers are currently being developed to reduce vibration in those structures. One of the solutions to reduce vibration vibrations in a building is to use vibration control devices such as dampers and base isolators. The purpose of this study was to investigate the best earthquake response for different types of base isolators and dampers. A 13 storey building is modelled and responses of Storey displacement, story drifts and overturning moments were determined and compared using ETABS software.
Bacterial Isolates and Drug Susceptibility Pattern of Sterile Body Fluids at a Tertiary Care Hospital in Northern India
Authors: Kar Mitra , Chaudhary Radhika , Sahu Chinmoy , Dubey Akanksha , Patel Sangram Singh
Published: May 2022

Background: Body fluids obtained from sterile body sites are expected to be devoid of any pathogenic or commensal microorganisms. The cause of infected body fluids could be any pathological agents or skin contaminants harbored from intensive care units. This study identifies commonly isolated bacteria from the body fluid samples and their antibiotic sensitivity pattern.

Methods: All body fluid samples (except blood, cerebrospinal fluid and contaminated samples) received in Bacteriology section of Department of Microbiology in tertiary care centre in Northern India from November 2019 to May 2020 were included in the study. All microorganisms obtained on bacterial culture were subjected to identification by standard biochemical tests or MALDI-TOF-MS assay and antibiotic sensitivity testing by Kirby Bauer disc diffusion test.

Results: Out of 363 samples of body fluids, 113 (31.12%) showed bacterial growth on culture. Male predominance among patients with liver disease was deemed statistically significant in comparison to those without liver disease. Comorbidities like hypertension and encephalopathy among the patients with liver disease was statistically significant in comparison to those without liver disease. Escherichia coli was the most commonly isolated bacteria (20.35%) which was followed by Pseudomonas spp (15.92%) and Klebsiella pneumoniae (11.50%). Fifty one (45.13%) isolates were multidrug resistant. The isolation of MDR microorganisms from body fluid samples is statistically significant in cancer patients. Among MDR microorganisms, Klebsiella pneumoniae was deemed most resistant with presence of Extended spectrum beta lactamase (ESBL) character and also showed complete resistance to Carbapenems but all isolates were susceptible to Colistin.

Conclusion: This study shows spectrum of bacterial isolates observed from body fluid samples received in our laboratory and helps in empirical treatment of patients based on antibiotic susceptibility patterns. It also highlights importance of adhering to antibiotic sensitivity results and infection control practices to prevent spread of multidrug resistant infection in hospital environment.
Iron Deficiency Anemia among Women of Reproductive Age
Authors: Neshnash Marwa , Usman Muhammad
Published: May 2022

Background:  Iron deficiency anemia is a significant global public health concern that can lead to a decline in mental and physical performance and mortality among women of reproductive age.

Objective: The goal of this literature review is to review and integrate information from the literature on the prevalence, etiology, consequences, and treatment options associated with iron deficiency anemia.

Review: Prevalence of iron deficiency anemia is high among women of reproductive age especially in low-income countries such as African and Asian countries compared to high-income countries. This could be due to differences in topographical, dietary, and cultural factors. Additionally, iron deficiency can be caused by poor absorption of iron or decreased iron intake. Poor absorption can occur as a side effect of different medications, while decreased iron intake could be the result of a diet high in legumes but low in protein and fruits. Moreover, iron deficiency anemia can be caused by persistent diseases, infections, increased demand during pregnancy, and obesity. If left untreated, the consequences of iron deficiency anemia include low quality of life, poor mental and physical health, a decline in work performance, and increased comorbidities. The current mainstay of treatment for iron deficiency anemia includes oral iron administration, parenteral iron administration, and blood transfusion.

Conclusion: Iron deficiency anemia is one of the most common health care issues among women of reproductive age. Different factors like blood loss, distant health care facilities, and poor diet can all influence the iron status in women of reproductive age. If left untreated, anemia can cause a variety of negative consequences, which can be avoided by implementing timely, cost-effective therapies.
High Resolution Computed Tomography (HRCT) for Evaluation of Pulmonary Manifestations in Patients with Autoimmune Diseases
Authors: Habib Marina Sherif Fouad , Khadra Rania Sobhy Mohamed Abo , Younis Rasha Lotfy , Sherif Mohamed Fouad
Published: May 2022

Background: The systemic immune system afflictions can reason very a couple of pneumonic parenchymal, vascular, aviation route, and pleural anomalies. The highlight thoracic signs of those afflictions are incited through the basic immune system prepare.

Aim of the Work: Evaluation of the radiological findings on HRCT of the lung in patients with autoimmune diseases was the aim of this research

Methods: This retrospective cross- sectional research was conducted on 25 cases (18 female and 7 male) ranging from 20-80 years old with autoimmune diseases proved by laboratory and clinical finding attending Tanta university Hospitals. Our patients were referred from chest department to Radiology department (CT units) at Tanta University Hospitals. The period of this research was from May 2020 to May 2021.

Results: Chest complications are common among autoimmune patients, the most frequent chest manifestations were fatigue followed by dyspnea representing 72% and 60% respectively,  40% of patients were presented with cough, The presence of clubbing and weight loss in autoimmune individuals with chest manifestations is frequent representing about (24% and 16%) respectively.

Conclusion: For evaluation of patients with connective tissue diseases of chest manifestations (pain, shortness of breath, cough), HRCT is the imaging technique of choice), According to HRCT, ground glass opacities was the most prevalent chest findings representing 56%, followed by consolidation and bronchiectasis representing 28% and lastly pericardial effusion represents 4%.
Thoracic Involvement in Polyangiitis Granulomatosis: A Moroccan Series of 21 Case
Authors: Benjelloun H. , Anniche H. , Jalloul W. , Zaghba N. , Chaanoun K. , Yassine N.
Published: May 2022

Granulomatosis with polyangiitis, formerly known as Wegener's disease, is a severe systemic necrotizing vasculitis, associating an inflammation of the vascular wall with a peri and extravascular granulomatosis. It is a rare disease that affects small vessels.it is classically translated by an otorhinolaryngological, respiratory and renal involvement. Other systemic manifestations of vasculitis may also be present.of slow evolution, the disease is often misleading and difficult to diagnose.The aim of our work is the analysis of the clinico-radiological thoracic and extra-thoracic manifestations, as well as the therapeutic and evolutionary profile in a retrospective study of a series of 22 cases.
Outcome of Minimally Invasive Plate Osteosynthesis for Comminuted Diaphyseal Humeral Fractures
Authors: Shamy Ahmad Al , Forse El Sayed El , Mahmoud Ashraf , Amin Osama
Published: May 2022

Aims: To examine the clinical outcomes of the minimally invasive plate osteosynthesis technique for comminuted diaphyseal humeral fractures based on clinical and radiologic records.

Study Design:  Prospective study.

Place and Duration of Study: This study was carried on twenty-one adult patients with comminuted humeral shaft fracture treated with MIPO between February 2020 to February 2021 were included in this study.

Methods: Patients with pathological fracture, Gustilo and Anderson type 2 or 3, intra-articular fracture and preoperative radial nerve palsy were excluded. The surgery time, blood loss and time of union were noted. Clinical outcome was assessed by Q-DASH score.

Results: We had thirteen cases of 12-B type and eight cases of 12-C type of fracture. According to Q-DASH score, eighteen cases had satisfactory results. The mean radiological fracture union time was 13.85 weeks. Mean elbow flexion was 1250, mean extension range was -2.140. out of all cases, two cases had superficial wound infection. One patient had iatrogenic radial nerve palsy. One patient had non-union.

Conclusion: Although technically demanding, MIPO technically should be considered one of the management options in the treatment of complex humeral diaphyseal fractures as it adheres to biological fixation principles with minimal soft tissue dissection, preserving fracture hematoma and periosteal blood supply.
Effect of Seed Priming of Two Stored Sudangrass Genotypes on Seed Germination, Seedling Vigor, Growth and Forage Yield and Its Component
Authors: El-Gaafarey Tamer G. , EL-Nahrawy Shereen M. , Shahein Alaa M. E. A.
Published: May 2022

Seed priming is a technique, which could improve the germination and establishment on the seeds, which were stored for 1.5 years under normal conditions. Therefore, this study was designed to evaluate the effect of different seed priming techniques, through un-soaked seed (control), Hydro-priming (soaked with distill water), 25 ppm of salicylic acid, 1.5% of Calcium chloride (CaCl2) and 3% of Potassium dihydrogen phosphate (KH2PO4) for 10 hours on seed germination%, seedling vigor and forage yield of two sudangrass genotypes (Piper black and Giza1). Two experiments (laboratory and field) were conducted at Sakha Agric. Res. Station, ARC, Egypt, during the two successive summer seasons 2019 and 2020.The results obtained indicated that 25 ppm of salicylic acid followed by 1.5% of CaCl2 and 3% of KH2PO4 treatments significantly affected on seed germination percentage, shoot and root length, fresh and dry seedling weight, seed vigor index, seedling vigor index, electrical conductivity and forage yield. Piper black genotypes were the highest in seed germination percentage, seed vigor index and seedling dry weight, but it was the lowest of electrical conductivity. It could be concluded that seed priming may serve as an appropriate treatment for accelerating the emergence so improve cutting and forage yield of sudangrass genotypes under study. Significant differences for fresh yield ton /hectare and piper black had higher fresh, dry and total yield ton /hectare than Giza1. Seed priming had highly significant differences at all cuts in the two seasons and SA25 ppm Salsilc acid treatment had superior and the highest dry yield in the two seasons, while control treatment was the lowest dry yield. Technique of seed priming led to improving germination, seedling characters, and forage yield under experiment conditions in comparison to non-primed seed (dry seed) in sudangrass seed stored for 1.5 years. Salicylic acid with 25 ppm had the highest values followed by1.5% of CaCl2 and 3% of KH2PO4 in the field conditions. Piper black genotype was the best as compared with Giza1 genotype, which gave the highest seed germination %, seedling vigor and viability. May be recommended for hydropriming seeds for 10 hours in water, which led to an increase in the total fresh yield by 30%, the highest seed germination percentage (94.00 and 92.50%), seed vigor index (20.79 and 19.84) and seedling vigor index (2194.11 and 2236.47) as practical seeds about 10 hours treatment.
Increased protein tyro sine‐phosphorylation in primary T‐cells transduced with Tax l of Human T‐cell leukemia virus type I
Authors: Akagi Tsuyoshi , Ono Hiroaki , Sasaki Terukatsu , Shimotohno Kunitada
Published: July 2002

Protein tyrosine‐phosphorylation in primary human T‐cells transduced with Taxi of Human T ‐cell leukemia virus type I was investigated. In comparison with control T‐cells, the level of protein tyrosine‐phosphorylation after stimulation with anti‐CD3 antibody increased significantly in Tax1‐transduced T‐cells. This enhancement in tyrosine‐phosphorylation possibly accounted for the augmented proliferative response of these cells, which has been reported previously
Prolactin induces growth inhibition and promotes differentiation of CHO cells stably transfected with prolactin receptor complementary DNA
Authors: Bignon Christophe , Daniel Nathalie , Kermabon Anne-Yvonne , Djiane Jean
Published: July 2002

We have characterized a stable and functional transfectant of the rabbit prolactin receptor in Chinese hamster ovary cells, and investigated the action of prolactin (PRL) on the growth and differentiation of this transfectant (clone E32). PRL induced a significant inhibition of E32 cell proliferation. Growth inhibition correlated with gene induction of the molecular marker of ovarian differentiation cholesterol side chain cleavage P450 (P450scc). Both effects were inversely proportional to cell confluence. The limits and potential development of such transfected cellular systems are discussed.
An 8.5‐kDa ribonuclease from the extreme thermophilic archaebacterium Sulfolobus solfataricus
Authors: Fusi Paola , Grisa Margareth , Tedeschi Gabriella , Negri Armando , Guerritore Andrea , Tortora Paolo
Published: July 2002

Protein p3, a ribonuclease we previously isolated from the archaebacterium Sulfolobus solfataricus [P. Fusi et al. (1993) Eur. J. Biochem. 211, 305–310], was subjected to complete amino acid sequencing. It consisted of 75 residues, with a calculated M
r of 8582, a pI of 10.1, and had some degree of monomethylation at Lys‐4 and Lys‐6. p2, a previously sequenced, 62‐residue ribonuclease from the same organism, had an identical sequence for 57 consecutive residues starting from the N‐terminus. p2 and p3 also showed a striking similarity to five other proteins previously isolated from Sulfolobus strains and identified as DNA‐binding proteins. However, the C‐terminus, 10 residue region of p3 did not show any similarity to these proteins; in contrast, it was significantly similar to stretches in three eubacterial ribonucleases from Bacillus strains. No difference between p2 and p3 has so far been detected as regards their catalytic properties. Available data suggest that these molecules have a narrow substrate specificity and probably play specific roles in RNA processing.
Wild type and constitutively activated forms of the Drosophila Toll receptor have different patterns of N‐linked glycosylation
Authors: Kubota Ken , Keith Fionna J. , Gay Nicholas J.
Published: July 2002

Toll is a Drosophila membrane protein related in sequence to the mammalian platelet glycoprotein 1B and to the interleukin‐1 receptor. It mediates a signal transduction pathway leading to the development of dorsoventral polarity in the Drosophila embryo. In this paper we show that a constitutively activated mutant receptor, Toll10B, is processed into a distinct isoform of slower electrophoretic mobility when compared with the wild type molecule in both cell lines and the embryo. The wild type protein can also be processed into this form if over‐expressed but in the embryo is present as the smaller species. We show that the decrease in the mobility of Toll10B and over‐expressed wild type receptors is caused by altered patterns of N‐linked glycosylation and that both forms are secreted to the cell surface. On the basis of these results, we propose that the Toll10B receptor is unable to associate with a limiting co‐factor which when bound directly or indirectly masks supplementary N‐linked glycosylation sites.
Subunit‐specific inhibition of inward‐rectifier K+ channels by quinidine
Authors: Doi T. , Fakler B. , Schultz J.H. , Ehmke H. , Brändle U. , Zenner H.-P. , Süßbrich H. , Lang F. , Ruppersberg J.P. , Busch A.E.
Published: July 2002

Distinct inward‐rectifier K+ channel subunits were expressed in Xenopus oocytes and tested for their sensitivity to the channel blocker quinidine. The ‘strong’ inward‐rectifier K+ channel IRK1 was inhibited by quinidine with an EC50 of 0.7 mM, while the ‘weak’ reactifier channel ROMK1 was only moderately inhibited. ROMK1(N171D)‐IRKIC‐term chimeric channels, which carry both sites for strong rectification of IRK1 channels (the negatively charged D171 in the second transmembrane domain and the IRK1‐C‐terminus including E224), displayed strong rectification like IRK1, but showed weak sensitivity to quinidine‐like ROMK1, suggesting independence of quinidine binding and rectification mechanisms. Moreover, BIR10 and BIR11, two strong rectifier subunits originally cloned from rat brain, exerted subunit‐specific sensitivity to quinidine, being much higher for BIR11. Quinidine blockade of IRK1 was not voltage‐dependent, but strongly dependent on the pH in the superfusate. These results strongly suggest a subunit‐specific interaction of inward‐rectifier K+ channels with neutral quinidine within membrane lipid bilayers.
Dynamic light scattering study of the two‐domain structure of Humicola insolens endoglucanase V
Authors: Boisset Claire , Borsali Redouane , Schülein Martin , Henrissat Bernard
Published: July 2002

Endoglucanase V (EG V) of Humicola insolens is composed of a catalytic domain and of a cellulose‐binding domain linked by a 33 amino acid long peptide rich in Ser, Thr and Pro residues. This work describes the dynamic behavior of the two‐domain structure of EG V as revealed by quasi‐elastic light scattering experiments. For both the full‐length and the isolated catalytic domain, the autocorrelation function is essentially described by a single relaxation mode. The equivalent hydrodynamic radius of the catalytic domain was found to correspond precisely to the dimensions measured from the previously determined three‐dimensional structure. The results obtained with the full‐length protein allow a description of the two domain structure of EG V similar to that resulting from earlier studies using small angle X‐ray scattering on cellulases from Trichoderma reesei. The hydrodynamic dimensions of the entire enzyme can be approximated as an ellipsoid with dimensions of .
The effect of regular exercise on women receiving danazol for treatment of endometriosis
Authors: Carpenter S.E. , Tjaden B. , Rock J.A. , Kimball A.
Published: July 2002

Objective: To determine the effects of regular exercise on women receiving danazol for the treatment of endometriosis. Methods: Thirty‐nine patients were randomized to a danazol‐only or a danazol/exercise regimen in a prospective clinical trial carried out at tertiary care institutions. Patients in the danazol/exercise group were instructed to exercise four times per week, for 40 min per session, at an intensity of 20 metabolic units. Side effect profiles, pelvic symptoms, aerobic fitness, strength and hormone levels were compared for all subjects. The number of side effects of danazol was analyzed by the method of generalized estimating equations. Results: The number of side effects reported during a 4‐week period was 1.09–2.17 times greater for the danazol‐only than for the danazol/exercise group. All patients had improvement of symptoms during treatment. The danazol/exercise group had significantly lower testosterone levels during treatment. The time to recurrence of endometriosis was not different between groups. Conclusions: Exercise during danazol therapy reduces the number of androgenic side effects. Relief of pain and time to recurrence are unaffected.
ATP‐induced activation of expressed RyR3 at low free calcium
Authors: Manunta Maria , Rossi Daniela , Simeoni Ilenia , Butelli Eugenio , Romanin Christoph , Sorrentino Vincenzo , Schindler Hansgeorg
Published: July 2002

Ca2+ channel properties of the mink ryanodine receptor type 3 (RyR3), expressed in HEK293 cells, were studied in planar lipid bilayers to which RyR3 rich membrane fragments had been fused. RyR3 channels were not active at resting levels of Ca2+
free but were gated by an additional 1 mM ATP, exhibiting long open times. The second major finding was the absence of channel inactivation at millimolar Ca2+
free. Insertion of a myc tag at the N‐terminus of RyR3 did not affect the channel properties. As to skeletal muscle, the observed type 3 channel properties appear physiologically meaningful by assisting type 1 channels in calcium release.
The catalytic RNA of RNase P from Escherichia coli cleaves Drosophila 2S ribosomal RNA in vitro: a new type of naturally occurring substrate for the ribozyme
Authors: Hori Yoshiaki , Tanaka Terumichi , Kikuchi Yo
Published: July 2002

We have found that the catalytic RNA of RNase P of Escherichia coli (M1 RNA) can cleave 2S ribosomal RNA (2S rRNA) of Drosophila melanogaster at specific positions in vitro. The cleavage mainly occurred at two sites between nucleotides 11 and 12, and between 16 and 17 of 2S rRNA. Kinetic analyses of the reaction revealed that a dimer caused by intermolecular interaction of 2S rRNA may be the substrate for the cleavage between 11 and 12, while a simple monomer is the substrate for the cleavage between 16 and 17. Substrate recognition by M1 RNA is also discussed.
Heat induced expression of CD95 and its correlation with the activation of apoptosis upon heat shock in rat histiocytic tumor cells
Authors: Sreedhar A.S , Pardhasaradhi B.V.V , Khar Ashok , Srinivas Usha K
Published: July 2002

The heat shock response is a universal phenomenon and is among the most highly conserved cellular responses. However, BC‐8, a rat histiocytoma, fails to mount a heat shock response unlike all other eukaryotic cells. In the absence of induction of heat shock proteins, apoptotic cell death is activated in BC‐8 tumor cells upon heat shock. We demonstrate here that stable transformants of BC‐8 tumor cells transfected with hsp70 cDNA constitutively express hsp70 protein and are transiently protected from heat induced apoptosis for 6–8 h. In addition heat stress induces CD95 gene expression in these tumor cells. There is a delay in CD95 expression in hsp70 transfected cells suggesting a correlation between the cell surface expression of CD95 and the time of induction of apoptosis in this tumor cell line. Also expression of CD95 antigen appears to inhibit the interaction between heat shock factors and heat shock elements in these cells resulting in the lack of heat shock response.
Ribonucleotide reductase is regulated via the R2 subunit during the life cycle of Trypanosoma brucei
Authors: Breidbach Tanja , Krauth-Siegel R.Luise , Steverding Dietmar
Published: July 2002

We have examined the occurrence of the R1 and R2 subunits of ribonucleotide reductase during the life cycle of Trypanosoma brucei. Whereas the R1 protein is present throughout the life cycle, the R2 protein is not found in cell cycle‐arrested short stumpy trypanosomes. RT‐PCR/hybridization analysis revealed almost equal amounts of the R1 and R2 mRNAs in all life cycle stages of the parasite. The data indicate that ribonucleotide reductase of African trypanosomes is developmentally controlled by post‐transcriptional regulation of the R2 subunit.
Synergistic activation of the mkp‐1 gene by protein kinase A signaling and USF, but not c‐Myc
Authors: Sommer Anette , Burkhardt H. , Keyse Steven M. , Lüscher Bernhard
Published: July 2002

Mitogen‐activated protein kinase phosphatase 1 (MKP‐1) is negatively regulating mitogen‐activated protein kinases and is therefore involved in early signaling processes. The expression of the mkp‐1 gene is induced by growth factors and stress. The promoters of the human and murine mkp‐1 genes contain several conserved DNA binding elements, including two cAMP response elements and an E box. We observed that the upstream stimulatory factor (USF), but not c‐Myc, activated mkp‐1. USF synergized with protein kinase A, thus providing evidence for a role of the E box, during signal‐regulated stimulation of mkp‐1.
Successful mimicry of a complex viral antigen by multiple peptide insertions in a carrier protein
Authors: Feliu Jordi X , Carbonell Xavier , Villaverde Antonio
Published: July 2002

The antigenic properties of a viral peptide from the surface of foot‐and‐mouth disease virus particles have been successfully mimicked by multiple insertion in solvent‐exposed regions of Escherichia coli β‐galactosidase. By increasing the number of viral peptides per enzyme monomer, the average IC50 of hybrid proteins in a competitive enzyme‐linked immunosorbent assay) have decreased to values close to that presented by natural virions. Moreover, the antigenic diversity of these new recombinant enzymes when measured with different anti‐virus antibodies has also been largely reduced, indicating a better presentation of the epitopes located in the viral peptide. Although bivalent antibody binding could have been favoured by multiple presentation, conformational modifications of the viral peptide, due to the presence of other insertions or a cooperative antibody binding cannot be excluded. In addition, a multidimensional antigenic analysis have grouped together the multiple‐inserted proteins with the native virus, suggesting that increasing the number of insertions could be a good strategy to reproduce the antigenic properties of an immunoreactive peptide in a natural multimeric disposition.
Fas‐triggered phosphatidylserine exposure is modulated by intracellular ATP
Authors: Gleiss Bettina , Gogvadze Vladimir , Orrenius Sten , Fadeel Bengt
Published: July 2002

Recognition signals are displayed on the cell surface during apoptosis that enable macrophages to engulf and dispose of the dying cell. A common signal is the externalization of phosphatidylserine (PS). Studies in erythrocytes and platelets have suggested that PS exposure requires the concomitant activation of a phospholipid scramblase (PLS) and inhibition of an adenosine triphosphate (ATP)‐dependent aminophospholipid translocase. However, the molecular mechanism underlying PS exposure during apoptosis remains poorly understood. In this study, we provide evidence that expression of PLS is neither necessary nor sufficient for PS exposure during Fas‐triggered apoptosis. On the other hand, egress of PS is shown to correlate with a decline in intracellular ATP and inhibition of aminophospholipid translocase activity upon Fas stimulation. Moreover, suppression of intracellular ATP levels by the glucose anti‐metabolite, 2‐deoxyglucose, alone or in combination with glucose‐free medium, potentiates Fas‐induced PS exposure in the PLS‐expressing Jurkat cell line and enables PLS‐defective Raji cells to externalize PS in response to Fas ligation. These studies suggest that intracellular ATP levels can modulate the externalization of PS during apoptosis, and implicate the ATP‐dependent aminophospholipid translocase in this process.
Dependence of Apparent Viscosity on Mycelial Morphology of Streptomyces fradiae Culture in Various Nitrogen Sources
Authors: Choi Du Bok , Park Enoch Y. , Okabe Mitsuyasu
Published: July 2002

To examine what causes increased viscosity in culture broth in Streptomyces fradiae culture, various natural nitrogen sources were investigated. Extracellular protease activity increased with culture time and decomposed the natural nitrogen source into amino acids. In the case of gluten meal, after a culture time of 5 d, concentrations of glutamic acid and aspartic acid had increased to 600 and 200 mg/L, respectively, which were about 3‐ and 2‐fold as high as levels in cultures under similar conditions using Pharmamedia. For various amino acids tested, the addition of glutamic acid or aspartic acid mixture to the culture medium raised the apparent viscosity to its highest demonstrated value, 260 mPa·s after 5 d of culture, which was 3‐fold higher than without amino acids. Consumption of the decomposed glutamic acid and aspartic acid was dependent on the activities of glutamate dehydrogenase and aspartate aminotransferase, respectively. When ammonium ion was used as the nitrogen source, cell concentration reached 1.75 g/L measured as an intracellular nucleic acid concentration,which was about 2.3‐fold higher than that with any other natural nitrogen source. However, apparent viscosity was only 75 mPa·s, a value one‐third that of the amino acid mixture, and 70% of the pellets were bigger than 1.2 × 104 μm2. In the case of gluten meal or the amino acid mixture, pellets bigger than 1.2 × 104 μm2 comprised only 8%. This demonstrates that consumption of some amino acids affected the formation of filamentous morphology, which caused an increase in the apparent viscosity of the culture broth, and the apparent viscosity was not caused by the mycelial concentration but the mycelial morphology.
Monitoring Embryo Development in Chicken Eggs using Acoustic Resonance Analysis
Authors: Coucke Peter M. , Room Gwendolien M. , Decuypere Eddy M. , De Baerdemaeker Josse G.
Published: July 2002

Early embryonic development was monitored using acoustic resonance frequency analysis. Based on the course of the resonance frequency (RF) of a chicken egg during incubation, detection of viable and infertile eggs or eggs in which the embryo died during the first days of incubation was possible at the beginning of day 5. A sudden decrease of the RF of viable eggs was noticed after 96–104 h of incubation, whereas infertile eggs or eggs in which the embryo died during the first days of incubation showed a smooth, continuous increase of RF during the entire period of observation. Furthermore, the interruption of the embryonic development by cold storage after 24 or 84 h of incubation delayed the moment of decreasing RF. This proves that the sudden decrease of RF corresponds to a certain stage of development. This application of acoustic resonance frequency analysis offers the potential of early, automatic detection of inviable embryo's in eggs during incubation.
Introducing allergists to food chemistry
Authors: Steinhart H. , Wigotzki M. , Zunker K.
Published: July 2002

A report of common analytical methods for the identification and characterization of food allergens is presented. The different electrophoretic and immunologic methods describe the present analytical tools to characterize allergens. With respect to the lack of knowledge on the structure and the properties of food allergens, additional chemical techniques such as chromatography, NMR and MALDI‐TOF are of particular importance.
Divergent solid‐phase synthesis and candidacidal activity of MUC7 D1, a 51‐residue histidine‐rich N‐terminal domain of human salivary mucin MUC7
Authors: Satyanarayana J. , Situ H. , Narasimhamurthy S. , Bhayani N. , Bobek L.A. , Levine M.J.
Published: July 2002

Abstract: Domain 1 of the low‐molecular‐weight human salivary mucin, designated MUC7 D1, spans the 51 N‐terminal amino acid residues. This domain contains a 15‐residue basic histidine‐rich subdomain (R3‐Q17) which has 53% sequence similarity to histatin 5 (Hsn‐5), a salivary molecule known to exert potent in vitro cidal activity against Candida albicans and many other medically important fungi. The MUC7 D1‐15mer and its derivatives have previously been synthesized in our laboratory and their candidacidal activities have been found to be inferior to that of Hsn‐5. We were therefore intrigued to explore the candidacidal potency of the full‐length MUC7 D1 (51‐mer). Linear solid‐phase synthesis of this domain has been accomplished following standard Fmoc chemistry. The problems of partial coupling, owing to the peptide chain length, at several stages of the solid‐phase step‐by‐step synthesis were circumvented either by double‐coupling techniques or efficient coupling procedures. The MUC7 D1 peptide was purified to homogeneity by conventional reverse‐phase HPLC using two columns connected in series. Secondary structure of the purified peptide was assessed by circular dichroism (CD) spectroscopy in phosphate buffer and trifluoroethanol and compared to that of MUC7 D1‐15mer and Hsn‐5. The MUC7 D1 candidacidal activity was assessed against azole‐sensitive and azole‐resistant C. albicans strains and was found, unlike that of the MUC7 D1‐15mer, to be comparable with that of Hsn‐5, indicating that in addition to Hsn‐5, MUC7 D1 could provide an attractive alternative to the classical antifungal agents. The candidacidal potency of MUC7 D1, like that of MUC7 D1‐15mer, and of Hsn‐5, appears to be largely dependent on peptide charge, irrespective of α‐helical structure.