bluuebunny/crossref_metadata_2025 huggingface dataset:
Induction of Callus in Leaf Explants of Crinum americanum L. (Amaryllidaceae)
Authors: Copeland Kicia K. P. Gomes- , Santos Izulmé R. I. , Torres Amanda G. , Gomes João V. D. , Almeida Fabrício T. C. de , Fagg Christopher W. , Gomes Sueli M. , Silveira Dâmaris , Simeoni Luiz A.
Published: August 2020

Amaryllidaceae include plant species that present alkaloids with analgesic, anti-cancer, anti-bacterial, anti-viral, anti-fungal and anti-malarial activities. Due to this pharmacological value, several species of this family have been widely studied and among them is White lilly, Crinum americanum. The objective of this work was to induce callogenesis on leaf explants of C. americanum cultivated in vitro for future production of alkaloids. Leaf explants were grown on a culture medium (solid) Murashige and Skoog (1962) supplemented with different concentrations and combinations of plant growth regulators, auxin 2,4-dichlorophenoxyacetic acid and cytokinin 6-benzylaminopurine and their effect on callogenesis assessed for percentage oxidation and explants responsive to callus induction. Callus formation started 10 days after hormone inoculation, and within 30 days after inoculation the best callogenesis and callus biomass growth were observed in medium containing 2.5 mg L-1 of 2,4-dichlorophenoxyacetic acid and 10 mg L-1 of 6-benzylaminopurine. The lowest percentage of oxidation was observed on explants cultivated on medium containing 5 mg L-1 of 6-benzylaminopurine and 2.5 mg L-1 of 2,4-dichlorophenoxyacetic acid. The calli obtained were compact and embryogenic. This work contributes not only to future studies on in vitro callogenesis of this species, but also to a possible protocol for the production of alkaloids of interest from cell suspension cultures produced in vitro. This is the first report of callus formation in Crinum americanum explants.
Effect of Graded Levels of Sulphur on Growth and Yield Attributes of Two Sesame (Sesamum indicum) Varieties at Two Different Soil Sulphur Status
Authors: Venkatakrishnan L. , Backiyavathy M. R. , Meena S. , Kalaiselvi T. , Amirtham D.
Published: August 2020

Field experiment was conducted at farmer’s field in Coimbatore District of Tamil Nadu with two different soil sulphur status i.e., soil with sufficient and deficient levels of sulphur status in order to study the response of graded doses of sulphur on two different sesamum varieties. The experiment was laid out in split plot design with two factors viz., Two sesamum varieties (TMV 7 - Black var., SVPR 1 - White var.) and Six sulphur levels (0, 20, 30, 40, 50, 60 kg S ha-1). Application of 30 and 50 kg S ha-1 at sulphur sufficient and sulphur deficient soils has increased the plant height at VS and FS, at harvest the highest plant height was recorded. Whereas, other important traits like number of branches per plant, number of capsules per plant, length of capsule and dry matter accumulation has registered the highest response for sulphur application at 30 kg ha-1 which was on par with higher doses in sulphur sufficient soil and in case of deficient soil significant response was observed till 50 kg S ha-1. Among the two varieties, TMV 7 has performed better when compared with SVPR 1.
A Case of Cut Throat and Penis
Authors: Thompson Peter Appiah- , Obeng Kwadwo , Maison Patrick , Quansah Kofi , Nortey Michael
Published: August 2020

Aims: The management of cut throat injuries with genital self-mutilation is very challenging due to the complex anatomy of the neck and the need to attain acceptable cosmetic and functional outcomes of penile reconstruction. This report thus seeks to highlight the importance of a multidisciplinary approach to the management of these cases and to raise awareness of the need for early suspicion and diagnosis of mental diseases especially among young people.

Presentation of Case: We present an 18-year old newly diagnosed paranoid schizophrenic man who presented with cut throat and penis after an attempted suicide and was successfully managed at our facility. The family had not suspected any mental disorder. He had repair of the cut throat (involving both trachea and esophagus) after elective tracheostomy, refashioning of the penile stump and psychiatric treatment.

Discussion: Cut throat injuries happen in cases of attempted suicide and among patients with psychiatric problems. Cases of cut throat and genital self-mutilation though rare, have been reported among schizophrenics. Some of such individuals have an impression that destroying their genitals could help them overcome their excessive sexual desires and for others to help them remain righteous. Our patient was diagnosed of schizophrenia after he attempted suicide.

Conclusions: Young people with mental illness must be identified and given appropriate treatment early. Patients presenting with cut throat and penis require a multidisciplinary team approach involving at least an otorhinolaryngologist, a urologist, a general/gastrointestinal surgeon and a psychiatrist for optimum care.
Comparison of the Effectiveness of Portable Ultrasound vs Portable X-Ray as Diagnostic Imaging of Knee Structures in Clinical Medicine
Authors: Wajid Samira Abdul , Ahmed Samith , Devera Vaugn , Dickerson Devin , Thompson Austin , Villela Julia
Published: August 2020

Background: Imaging techniques are providing physicians an opportunity to ensure more accurate diagnostics. While MRI and CT are the gold standard diagnostic tools, they are not utilized as primary diagnostic tools due to their size, immobility, and exposure to radiation. It is not always feasible or practical to wait for a technician in an emergency department to conduct these examinations. As technology continues to advance, diagnostic tools are becoming more portable. X-ray and ultrasound are utilized to diagnose, as well as exclude potential causes of illness. Our objective is to determine which diagnostic tool is more efficient in diagnosing patella/knee injuries.

Methods: This systematic review aims to determine which diagnostic tool is more efficient at diagnosing injuries. By collecting and reviewing existing research to compare the limitations of injury detection, the accessibility and portability of both diagnostic machines, the safety in regards to radiation exposure to patient and physician, the reliability of the diagnoses, and the costs of each machine

Results: Of the reviewed articles, 58% of the articles focusing on knee injuries indicated that ultrasound imaging is a superior diagnostic instrument due to its efficacy and accessibility. When comparing ultrasound to x-ray directly, it was shown that ultrasound is a more precise diagnostic tool.

Conclusion: Ultrasound imaging is a more effective diagnostic tool than x-ray. It is better able to diagnose bone, soft tissue, and vessel injuries. It is a safer tool for both the patient and the physician because it uses sound rather than radiation to produce an image. It is also more accessible as advances in technology have made portable ultrasounds a protocol for quick assessments of injuries.
Assessment of Atherogenic Indices and Markers of Cardiac Injury in Albino Rats Orally Administered with Tartrazine Azo Dye
Authors: Iroh Geraldine , Weli Benita Oyoburuoma , Adele Uyota Anthony , Briggs Ojoye Ngoye , Waribo Helen Anthony , Elekima Ibioku
Published: August 2020

Aim: Assess the effect of tartrazine azo dye on atherogenic indices and markers of cardiac injury in albino rats.

Study Design: A total number of 63 rats were used for the study. The study was divided into two phases, 1 and 2, which lasted for 30 and 60 days respectively. Phase 1 had 35 rats, 20 as test and 15 as control, while phase 2 had 28 rats, 16 as test and 12 as control. In each phase the test groups were given 7.5mg/kg of tartrazine orally on daily basis over the stipulated period while the control groups were not treated with tartrazine.

Methodology: At the end of the study, 5ml of whole blood was collected from the jugular veins into Lithium Heparin bottles. The sample was spun, plasma collected and analyzed for cardiac Troponin I (cTn-I) and cardiac Troponin T (cTn-T), Total creatinekinase (CK), creatinekinase MM (CK-MM), and creatinekinase MB (CK-MB). Lipid parameters like total cholesterol (TC), High density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C) and Triglyceride (TG) which were also analysed. Atherogenic indices such as atherogenic coefficient (AC), atherogenic index of plasma (AIP), Non High density lipoprotein-cholesterol (nHDL-C), and castelli risk indices 1 and 2 (CRI-1 and CRI-2) were also calculated. In addition, cardiac tissues were collected, fixed in 10% formol saline and examined histologically using Haematoxylin and Eosin stain. Statistical analysis was performed using GraphPad Prism version 8.02.

Results: The results obtained indicate significant increases in nHDL-C, total CK and cTn-T after 30 and 60 days of treatment with tartrazine at ADI doses against controls. Other atherogenic indices such as AIP, AC, CRI-1 and CRI-2 as well as markers of cardiac injury such as cTn-I and CK-MB indicated non-significant increases.

Conclusion: Orally administered tartrazine over a 60 day period induced cardiac injury as shown by the significant increase in the cTn-T and total CK as well as hypertrophied nuclei of cardiomyocytes. This goes to say chronic administration of tartrazine even at the recommended daily dose could pose the risk of cardiovascular disease. This is also supported by an increase in nHDL cholesterol.
Evaluation of Some Cytokines, CD4, Hepcidin, Iron Profile and Some Haematological Parameters of Pulmonary Tuberculosis Patients Coinfected with HIV in Southeast of Nigeria
Authors: Ifeanyi Obeagu, Emmanuel , N. Esimai, Bessie , Uzoma Obeagu, Getrude , O.M.T.B. Ochiabuto, , Ejike Felix Chukwurah , , Stella Ekelozie, Ifeoma , Chinedum Ochei , Kingsley
Published: August 2020

The study was done to determine the levels of interferon-gamma, interleukin 6, interleukin 10, iron status, hepcidin and haematological parameters of patients with pulmonary tuberculosis co-infected with human immunodeficiency virus in Southeast, Nigeria. This study was carried out at the directly observed treatment-short course Tuberculosis (TB DOTS) centre of Federal Medical Centre, Umuahia, located in South-Eastern Nigeria. Therefore, sample size of 240 was used to give room for attrition. A total of two hundred and forty (240) subjects aged 18-60 years were enlisted for this study. Seven milliliters (7ml) of venous blood was collected from each subject and 2.5 ml was dispensed into bottles containing di-potassium salt of ethylenediamine tetra-acetic acid (K2-EDTA) and was used for full blood count, CD4 count and HIV screening. Also, 4.5ml was dispensed into plain tubes. Serum was obtained after clotting by spinning at 3000 RPM for 10 minutes and was used for interferon gamma, interleukin-6, and interleukin-10, iron and hepcidin determination. Data was analysed using statistical package for social science (SPSS) version 20. Student t-test, ANOVA (Analysis of Variance), Pearson Product Moment and Chi-Square were the tools employed. Results were expressed as mean ± standard deviation and are presented in table and significance level was set at P<0.05.The results showed difference that was statistically significant (P<0.05) in IFN-γ (P=0.000), IL-6 (P=0.000) IL-10 (P=0.000), CD4 (P=0.000), hepcidin (P=0.000), Iron (P=0.000), TIBC (P=0.000), %TSA  (P=0.001) ,WBC (P=0.000), Neutrophils (P=0.000), Lymphocyes (P=0.000), Monocytes (P=0.000), Eosinophils (P=0.000), Basophils (P=0.018), RBC (P=0.000), haemoglobin (P=0.000), PCV (P=0.000), MCV (P=0.000), MCH (P=0.000), MCHC (P=0.000), Platelets (P=0.000), ESR (P=0.000) when compared among  control, TB, HIV and TB-HIV subjects respectively. The co infection of HIV on pulmonary TB patients increases the levels of the cytokines. The cytokines and hepcidin can be used as adjunct to prognostic and diagnostic markers as their levels decreased with increased duration of treatment of the patients. The study has shown wide variations in the haemtological indices studied.
MicroRNAs miR-1 and miR-206 Regulate Monocarboxylate Transporter-4 and Vascular Endothelial Growth Factor Gene Expression in Colorectal Cancer
Authors: Khaleel Anas , AlEjielat Rowan , Batarseh Cristina I. , Elbakkoush Abdallah Ahmed , Tarkhan Amneh , Mahdi Aiman , Batarseh Yazan S.
Published: August 2020

Background: Colorectal cancer (CRC) is currently the third most common cancer type in males and the second most occurring in females. The role of microRNA (miRNA) in the development of colorectal cancer is not fully elucidated. Therefore, understanding the mechanistic interaction between miRNA and their target oncogenes may hold great importance as a possible target for interventional anticancer therapy.

Aims: To identify miRNAs that are part of the regulating pathway of Monocarboxylate Transporter-4 (MCT4) and Vascular Endothelial Growth Factor (VEGF) oncogenes.

Study Design: We used publicly available prediction tools (e.g. TargetScan, MicroCosm, PicTar, and DIANA-microT-CDS) to identify the possible miRNA that target the two oncogenes.

Methodology: We used the GeneMania database to visualize the network and verify gene names and remove ambiguity and duplications.  Furthermore, we used miRTarBase database to identify experimentally validated targets which we used to further confirm miRNA-oncogene relationships.  Finally, we utilized miR-Mfold web-tool to further visualize the circular structures and the simulated miR-1 and miR-206 targeting arrangements.

Results: We found two putative miRNA (miR-1 and miR-206) that may downregulate MCT4 coded by SLC16A3 gene and VEGF which is coded by VEGF gene. We found relationships between the validated target genes of miR-1 and miR-206 through GeneMania which we extracted from the literature. And we elucidated the proposed structure of these two miRNAs through miR-Mfold web-tool.

Conclusion: Our results elucidated a novel regulation pathway in CRC cells and may suggest a potential therapeutic approach for CRC therapy. MiR-1 and miR-206 may help cells go to apoptosis and inhibit the angiogenesis of colorectal cancer cells by down-regulation of MCT4 and VEGF proteins in tumor tissues.
Study of Lipid Profile in Metabolic Syndrome Patients
Authors: Karam Pinky , Shanthi B. , Selvi Kalai
Published: August 2020

Background: Metabolic syndrome is a group of metabolic abnormalities in which the chance of developing cardiovascular disease, diabetes mellitus, chronic kidney disease are high.

Aim: It aims at studying the lipid abnormalities in metabolic syndrome patients.

Methods: Total of 100 metabolic syndrome patients were selected for study over a period of 1year. These patients were selected based on the criteria for metabolic syndrome as established by National Cholesterol Education Program (NCEP) adult Treatment Panel III (ATP III). Demographic data were taken and biochemical parameters were estimated by standard guideline.

Results: Total cholesterol is significantly higher in very high risk (272.1 ± 8.591) compared to high risk (241.2 ± 3.901) and moderate risk (231.5 ± 4.498). TGL is significantly higher in very high risk (263.9 ± 13.70) compared to high risk (202.1 ± 6.531) and moderate risk (183.7 ± 7.650). HDL is almost same in very high risk (43.09 ± 1.533), high risk (40.44 ± 0.996) and moderate risk (42.53 ± 1.088). LDL is significantly higher in very high risk (177.9 ± 4.255) and high risk (169.4 ± 3.190) compared to moderate risk (155.7 ± 3.098). VLDL is significantly higher in very high risk (52.78 ± 2.739) compared to high risk (40.43 ± 1.306) and moderate risk (36.73 ± 1.530). CHO: HDL is significantly higher in very high risk (6.648 ± 0.366) compared to moderate risk (5.560 ± 0.207). High risk (6.060 ± 0.156) is not significantly different from very high risk and moderate risk. Thus, TC, TGL, LDL, VLDL, and CHO: HDL is significant as p value < 0.05 while HDL did not have any significance as p value > 0.05.

Conclusion: In this study, high prevalence of dyslipidaemia is seen. So, timely diagnosis and treatment will help in detecting dyslipidaemia patients in future.
Epidemiology of Bacterial Contamination of Inert Hospital Surfaces and Equipment in Critical and Non-critical Care Units: A Brazilian Study
Authors: Rodrigues Dayane Otero , Peixoto Laís da Paixao , Barros Erica Tatiane Mourao , Guimaraes Julianne Rodrigues , Gontijo Bruna Clemente , Almeida Jaisa Leite , Azevedo Lucas Guimaraes de , Lima Julia Cristina Oliveira e , Camara Deyse Silva
Published: August 2020

Aims: The hospital environment is an important reservoir of microorganisms, including multidrug-resistant pathogens, which can cause in-patient contamination and healthcare-related infections.  The objective of this study was to describe the epidemiology of bacterial contamination (contaminated sites, pathogen species and their antimicrobial susceptibility, and identifying of multidrug-resistant microorganisms - MDR) of inert hospital surfaces and medical equipment in two public hospitals in Northern Brazil.

Methods: This was a cross-sectional study with 243 samples (n = 208, from Hospital A; and n = 35, from Hospital B) collected by friction with humidified swabs from inert surfaces and equipment. Sequentially the samples were cultivated and bacterial species were identified by culture-based methods and tested for their susceptibility through agar diffusion assay according to the Clinical and Laboratory Standards Institute (CLSI).

Results: Most inert surfaces and equipment analyzed presented bacterial contamination (95.5%). Staphylococcus aureus was the main pathogen of clinical significance detected both in Hospital A (61.8%) and B (68.6%). Hospital A showed higher rates of isolated MDR bacteria than Hospital B, especially in the Adult Intensive Care Unit, which included methicillin-resistant Staphylococcus aureus (MRSA) (52.7%), Enterobacteria resistant to 4th generation cephalosporins (19.4%), and multidrug-resistant Pseudomonas aeruginosa (2.8%).

Conclusion: The failures in the control of bacterial contamination of inert surfaces and equipment in the two hospitals analyzed reinforce the need for a revised protocol for cleaning and disinfection of the inert surfaces and equipment, and for regulation of antibiotic dispensing, mainly in the AICU of Hospital A, which was found to be a reservoir of MDR pathogens.
Economic Valuation of Santolo Beach Tourism in Garut Regency West Java
Authors: Lestari Viani Puji , Anna Zuzy , Iskandar . , Gumilar Iwang
Published: August 2020

This study aimed to estimate the economic value of Santolo Beach tourism area. This research conducted from December 2019 until January 2020. The method used in this research is survey method with accidental sampling as the data collecting technique. Travel Cost Method and Contingent Valuation Method applied to estimate Santolo Beach's economic value. The factors influenced the number of visits and Willingness to Pay value identified by multiple linear regression. The results of the research showed that the economic value of Santolo Beach with the Travel Cost Method is IDR 114.26 Billion per year for the linear model and IDR 177.79 Billion per year for the semi-log model while the result from the Contingent Valuation Method is IDR 3.65 Billion per year. The factors that influence the number of visits in the linear model are travel costs, income, education, and distance while in the semi-log model is distance. The variables that influence the value of visitors' Willingness to Pay are income and education for both the linear model and the semi-log model.
The inhibition of ribonucleic acid synthesis by the thiol‐oxidizing agent, diamide, in Escherichia coli
Authors: Zehavi-Willner Tova , Wax Richard , Kosower Edward M.
Published: July 2002

The thiol‐oxidizing agent, diamide, has been used to convert glutathione to glutathione disulfide within the cells of a stringent strain of Escherichia coli (CP 78), leading to a cessation of 14C‐leucine incorporation (protein synthesis) and 3H‐uracil incorporation (RNA synthesis). Parallel experiments with an isogenic relaxed strain (CP 79) gave similar results, providing evidence that glutathione is closely linked to RNA synthesis indepently of the link previously shown to protein synthesis.
Activation of the respiratory burst of guinea pig neutrophils by dicyclohexylcarbodiimide
Authors: Aviram Adina , Aviram Irit
Published: July 2002

DCCD activates the respiratory burst in guinea pig peritoneal neutrophils. The onset of the superoxide producing activity is preceeded by a lag, inversely proportional to the dose of the stimulant and to the temperature. Initial rates of superoxide formation exhibit different dependencies on the concentrations of DCCD and on temperature. Activation of NAD(P)H oxidase is inhibited by preincubation of neutrophils with 2‐deoxyglucose and does not require the presence of extra cellular Ca2+.
Differential phosphorylation of ribosomal protein S6 in isolated rat hepatocytes after incubation with insulin and glucagon
Authors: Wettenhall Richard E.H. , Cohen Philip , Caudwell Barry , Holland Ross
Published: July 2002

Glucagon and insulin both stimulated the 32P‐labelling of ribosomal protein S6 in rat hepatocytes that had been incubated with 32Pi. Glucagon selectively enhanced the labelling of the tryptic peptide phosphorylated by cyclic AMP‐dependent protein kinase, demonstrating that 6 S is a physiological substrate for this enzyme. Insulin stimulated the phosphorylation of distinct tryptic peptides, at least one of which appears to be very close in the primary structure to the sites phosphorylated by cyclic AMP‐dependent protein kinase.
Phosphopeptide patterns of the ribosomal protein S6 following stimulation of guineapig parotid glands by secretagogues involving either cAMP or calcium as second messenger
Authors: Padel U. , Kruppa J. , Jahn R. , Söling H.-D.
Published: July 2002

Stimulation of secretion in exocrine cells is associated with the incorporation of up to 3 to 4 phosphates into the ribosomal protein S6. This occurs with secretagogues involving either cAMP or free calcium as second messenger. An analysis of the phosphorylation pattern of S6 from stimulated guineapig parotid glands reveals 3 phosphopeptides (termed A,B,C). The phosphopeptide pattern was identical for cAMP‐ or calcium‐mediated stimulation, whereas phosphorylation of the S6 protein in vitro with catalytic subunit of cAMP‐dependent protein kinase resulted only in the formation of phosphopeptides A and C. Therefore, secretagogue‐mediated phosphorylation is not or not exclusively catalyzed by cAMP‐dependent protein kinase even when cAMP is the second messenger.
A radioimmunoassay for the Hydra head activator
Authors: Bodenmüller Heinz , Zachmann Beate
Published: July 2002

A highly sensitive and specific radioimmunoassay for the head activator has been developed which utilises tritriated head activator. The assay is sensitive in the range of 40–200 fmol. Immunochemical studies showed that the antiserum recognised the intact molecule better than any of the fragments or derivatives produced by enzymatic treatment or chemical synthesis.
A new hemoglobin variant altering the α1β2 contact: Hb Chemilly α2β2 99(G1)Asp→Val
Authors: Rochette J. , Poyart C. , Varet B. , Wajcman H.
Published: July 2002

Hemoglobin Chemilly (α2β2 99(G1)Asp→Val), a high oxygen affinity variant, was uncovered in the red blood cells of a polycythemic patient who reported to the hospital concerning periodic headaches. We describe the molecular abnormality and functional studies of this new abnormal Hb. β 99(G1)Asp, an invariant residue of hemoglobin, is considered a key amino acid for conformational changes between the R⇆T quaternary structures responsible for the allosteric behavior of hemoglobin. Hb Chemilly exhibits a high 02 affinity, very low cooperativity and reduced Bohr effect. Its functional abnormalities are compared to the 5 other Hb variants at site β 99(G1) described up to now of the 7 single base substitutions predictable from the genetic code.
N‐terminal amino acid sequences of chloroform/methanol‐soluble proteins and albumins from endosperms of wheat, barley and related species
Authors: Shewry Peter R. , Lafiandra Domenico , Salcedo Gabriel , Aragoncillo Cipriano , Garcia-Olmedo Francisco , Lew Ellen J.-L. , Dietler Mary D. , Kasarda Donald D.
Published: July 2002

The N‐terminal amino acid sequences of two chloroform/methanol soluble globulins from barley and one form wheat are reported. They are homologous with N‐terminal sequences previously reported for α‐amylase and trypsin inhibitors from cereals and 2 S storage proteins from castor bean and rape. Three albumins were also purified from Aegilops squarrosa and Triticum monococcum. These had N‐terminal amino acid sequences most closely related to the α‐amylase and trypsin inhibitors. The relationships of this superfamily of seed proteins are discussed.
The negative surface charge density of plasmalemma vesicles from wheat and oat roots
Authors: Møller Ian M. , Lundborg Tomas , Bérczi Alajos
Published: July 2002

Plasmalemma vesicles were isolated in a sucrose‐containing medium from wheat and oat roots and the net negative surface charge density was determined with 9‐aminoacridine fluorescence [Chow, W.S. and Barber, J. (1980) J. Biochem. Biophys. Methods 3, 173‐185]. The outer surface of the vesicles (measured in the presence of sucrose) had densities of − 16 to − 20 mC·m−2 and −29 mC·m−2 for wheat and oat roots, respectively. The inner surface ‐ presumed to be the cytoplasmic side and calculated from the values measured in the presence and absence of sucrose ‐ was more negative, and its size depended on the salt status of the roots.
The competitive equilibrium between aluminium and ferric ions for the binding sites of transferrin
Authors: Cochran M. , Coates J. , Neoh S.
Published: July 2002

Human transferrin is shown to bind 2 mol of aluminium per mol of protein using spectrophotometric titration. Competitive equilibrium between aluminium and ferric ions for transferrin binding sites is observed, and a value of 2.5 (± 0.4) × 1015 M−1 is found for the apparent binding constant under physiological conditions.
Phosphorylation of elongation factor 1 in polyribosome fraction of rabbit reticulocytes
Authors: Davydova E.K. , Sitikov A.S. , Ovchinnikov L.P.
Published: July 2002

A single protein, M
r, ~ 50000, is shown to be phosphorylated during incubation of a mono‐ and polyribosome fraction of rabbit reticulocytes with [γ‐32P]ATP at a low ionic strength. This protein has been identified as the elongation factor 1α (EF‐1α). The phosphorylated EF‐1α, in contrast to the unmodified factor, is not detected in complexes with mono‐ and polyribosomes. It is suggested that the phosphorylation of EF‐lα can result in its decompartmentation from polyribosomes and thus affect the rate of protein synthesis.
A restriction endonuclease SuaI from the thermoacidophilic archaebacterium Sulfolobus acidocaldarius
Authors: Prangishvili D.A. , Vashakidze R.P. , Chelidze M.G. , Gabriadze I.Yu.
Published: July 2002

A type II restriction endonuclease (SuaI) has been isolated from the thermoacidophilic archaebacterium Sulfolobus
acidocaldarius. The enzyme is an isoschizomer of BspRI. It does not cut S. acidocaldarius DNA, as the recognition sequence GGCC in this DNA contains modified nucleotide(s). The enzyme is most active at 60–70°C and is highly thermostable.
Activity and action pattern of Bacillus licheniformis α‐amylase in aqueous ethanol
Authors: Blakeney Anthony B. , Stone Bruce A.
Published: July 2002

A purified B. licheniformis α‐amylase in a mixture of ethanol‐aqueous buffer (1:1, ) retains half the activity shown in water alone. In ethanol‐aqueous buffer (7:3, ) about 20% of the activity is retained. The pattern of oligosaccharides produced from amylose changed with ethanol concentration; in aqueous buffer the products are: DP 1 and 2, 33.7%; DP 3, 28.5%; DP 4, 4.4% and DP 5, 33.4%. Whereas in ethanolaqueous buffer (7:3, ) the products are DP 1 and 2, 66.8%; DP 3, 17.3%; DP 4, 4.1 % and DP 5, 11.8%. These results suggest that a change in substrate affinity at the active centre subsites is induced in the ethanolaqueous buffer medium.
3β,17β‐Hydroxysteroid dehydrogenase of Pseudomonas testosteroni
Authors: Minard Philippe , Legoy Marie-Dominique , Thomas Daniel
Published: July 2002

3β,17β‐Hydroxysteroid dehydrogenase (3β17/βHSDH) is an NAD‐dependent dehydrogenase which has a double specificity for the 3‐ and 17‐positions on the steroid skeleton. When dehydroepiandrosterone (DHEA) is used as steroid substrate, and the assay coupled with ketosteroid‐isomerase, the two reactions occur alternately and each reaction on the 3‐position produces a chromophoric molecule. These two reactions can follow one another without dissociation of the coenzyme from the enzyme binding site. This is confirmed by competition experiments with another dehydrogenase.
Thiophilic adsorption ‐ a new method for protein fractionation
Authors: Porath Jerker , Maisano Federico , Belew Makonnen
Published: July 2002

Divinylsulfone
Immunoglobulin
Affinity chromatography
Hydrophobic interaction (salting‐out)
Serum fractionation
Elevated chondroitin 6‐sulfotransferase activity in fetal bovine serum
Authors: Sugahara Kazuyuki , Shibamoto Sayumi , Yamashina Ikuo
Published: July 2002

Sulfation
Chondroitin sulfate
Sulfotransferase
Purification and composition of photosystem I reaction center of Prochloron sp., an oxygen‐evolving prokaryote containing chlorophyll b
Authors: Schuster Gadi , Nechushtai Rachel , Nelson Nathan , Ohad Itzhak
Published: July 2002

The photosystem I reaction center complex of the photosynthetic prokaryote Prochloron, present as a symbiont in ascidians from the Red Sea at Eilat (Israel), was isolated and characterized. The complex consists of 4 polypeptide subunits, and therefore is similar to that of cyanophytes and green algae. Their apparent molecular masses (in kDa) are about 70 (subunit I), 16 (subunit II), 10 (subunit III), and 8 (subunit IV). The purified reaction center contains about 40 chlorophyll molecules per P‐700 as compared to about 800 in the intact thylakoids. Subunit I has the same apparent electrophoretic mobility and appears as a double polypeptide band as reported for this subunit in higher plants and algae. Immunological cross‐reactivity was detected among subunits I and II of Prochloron and photosystem I reaction centers of higher plants and algae.
A protein‐tyrosine kinase in the nuclear matrix from rat liver
Authors: Ohmura Yoshihisa , Teraoka Hirobumi , Tsukada Kinji
Published: July 2002

Protein kinase activity in isolated nuclei from rat liver was detected in situ after electrophoresis on SDS‐polyacrylamide gel containing no exogenous protein substrate. After renaturation of polypeptides, the gel was incubated with [γ‐32P]ATP and divalent cations. Among five major protein kinase activities observed as radioactive bands by autoradiography, a protein kinase autophosphorylating on tyrosine (M
r 30 000) was identified and found to be localized in the nucleus, particularly in the nuclear matrix. The intensity of the activity band representing the level of the protein‐tyrosine kinase in rat liver nuclei did not appreciably change during 3–24 h after partial hepatectomy.
The gene for the 10 kDa phosphoprotein of photosystem II is located in chloroplast DNA
Authors: Hird Sean M. , Dyer Tristan A. , Gray John C.
Published: July 2002

Nucleotide sequencing of a region of wheat chloroplast DNA between the genes for the 47 kDa chlorophyll a‐binding protein of photosystem II (psbB) and cytochrome b‐563 (petB) has revealed an open reading frame of 73 codons. This open reading frame has been identified as the gene (psbH) for the 10 kDa phosphoprotein of photosystem II by comparison with the published N‐terminal amino acid sequence and amino acid composition of the purified spinach protein. The predicted sequence of the protein shows some homology with the N‐terminal region of the light‐harvesting chlorophyll a/b‐binding protein of photosystem II (LHCII).
Diphtheria toxin and its mutant crm 197 differ in their interaction with lipids
Authors: Papini Emanuele , Colonna Raffaele , Schiavo Giampietro , Cusinato Federico , Tomasi Maurizio , Rappuoli Rino , Montecucco Cesare
Published: July 2002

The interaction of diphtheria toxin and its enzymatically deficient mutants crm 176 and crm 197 with liposomes has been studied by turbidity measurement and hydrophobic photolabelling with photoactivatable phosphatidylcholines. Diphtheria toxin and crm 176 at neutral pH bind to the surface of lipid bilayers while crm 197 also appears to interact with the fatty acid chains of phospholipids. All proteins undergo a change in conformation over the same range of acidic pH and become able to insert in the lipid bilayer. The tighter lipid interaction of crm 197 may account for its higher cell association constant. The possibility is discussed that the binding of diphtheria toxin to cells is mediated by both a protein receptor and an interaction with the head group of phospholipids.
Phosphorylation of high‐ and low‐molecular‐mass atrial natriuretic peptide analogs by cyclic AMP‐dependent protein kinase
Authors: Olins Gillian M. , Mehta Pramod P. , Blehm Delores J. , Patton Dennis R. , Zupec Mark E. , Whipple Deborah E. , Tjoeng Foe S. , Adams Steven P. , Olins Peter O. , Gierse James K.
Published: July 2002

Synthetic high‐ and low‐molecular‐mass atrial peptides were phosphorylated in vitro by cyclic AMP‐dependent protein kinase and [32P]ATP. From a series of atrial peptide analogs, it was deduced that the amino acid sequence, Arg101–Ser104 of atriopeptin was required for optimal phosphorylation. Phosphorylated AP(99–126) was less potent than the parent atriopeptin in vasorelaxant activity and receptor‐binding properties. These results indicate that the presence of a phosphate group at the N‐terminus of AP(99–126) decreases the interaction of the peptide with its receptor and, as a consequence, decreases bioactivity. These observations are in contrast to those of Rittenhouse et al. [(1986) J. Biol. Chem. 261, 7607–7610] who reported that phosphorylation of AP(101–126) enhanced the stimulation of Na/K/Cl cotransport in cultured vascular smooth muscle cells.
Proteolytic activation of protein kinase C in the extracts of cells treated for a short time with phorbol ester
Authors: Buday László , Seprödi János , Farkas Gyöngyi , Mészáros György , Romhányi Tibor , Bánhegyi Gábor , Mandl József , Antoni Ferenc , Faragó Anna
Published: July 2002

A 10 min treatment of human neutrophils with phorbol 12‐myristate 13‐acetate (PMA) has been reported to induce accumulation of the proteolytically activated Ca2 +/phospholipid‐independent catalytic fragment of protein kinase C in the cytosol of intact cells [(1986) J. Biol. Chem. 261, 4101‐4105]. We investigated the proteolytic conversion of protein kinase C to the Ca2 +/phospholipid‐independent form in the cytosol and membrane fractions of pig neutrophils. The activity of protein kinase C was measured with its specific oligopeptide substrate Ala‐Ala‐Ala‐Ser‐Phe‐Lys‐Ala‐Lys‐Lys‐amide designed previously. In our experiments the short‐term treatment of neutrophils with PMA did not induce the accumulation of the proteolytically activated form of protein kinase C in the cytosol of intact cells. However, treatment of cells with PMA enhanced the limited proteolysis of protein kinase C during the preparation of cell extracts.
Evidence for a novel voltage‐activated channel in the outer mitochondrial membrane
Authors: Kinnally K.W. , Tedeschi H. , Mannella C.A.
Published: July 2002

Patch‐clamp studies of the outer mitochondrial membrane indicate a voltage‐dependent increase in conductance for potentials positive relative to the exterior of the mitochondrion. The time course of the conductance changes is consistent with an activation of channels. Voltage pulse experiments suggest that the activation phenomenon corresponds to assembly of the channels from subunits with disassembly occurring after recovery of the original conductance. Effects of temperature and concanavalin A on the voltage‐induced conductances are also consistent with a channel assembly model.
Fluorimetric pH measurement in whole cells of dark aerobic and anaerobic cyanobacteria
Authors: Hinterstoisser Barbara , Peschek Günter A.
Published: July 2002

9‐Aminoacridine and atebrin fluorescence quenching by dark aerobic and anaerobic suspensions of Anacystis nidulans, Plectonema boryanum and Gloeobacter violaceus was determined at external pH 7–9. Individual pH values in cytosol and thylakoid compartments were calculated from the simultaneously different intracellular enrichment factors of the monovalent and the divalent base. Concomitantly, at external pH 4–7 the cytosolic pH was measured with fluorescein diacetate which is taken up and decomposed in the cell by cytosolic hydrolases. The pH‐dependent fluorescence of the free fluorescein, which remains trapped in the cell, monitors the cytosolic pH. The fact that the latter was higher in aerobic than in anaerobic cells, and insensitive to saturating concentrations of dicyclohexylcarbodiimide aerobically, was taken to support the concept of a proton‐translocating respiratory chain in the plasma membrane.
Intracellular stimulation of mast cells with guanine nucleotides mimic antigenic stimulation
Authors: Fernandez J.M. , Lindau M. , Eckstein F.
Published: July 2002

Exocytosis was followed in single rat peritoneal mast cells, by measuring the cell membrane capacitance using circuit analysis and patch‐clamp techniques. After antigenic stimulation or intracellular perfusion with guanine nucleotides, exocytosis followed a time course characterized by a lag period d, area expansion factor A, and a time constant τ. We suggest that A depends entirely on the cell's morphology, d reflects the properties of a GTP‐binding regulatory protein that appears to rate limit the response and τ is due to an independent and yet unknown process. In contrast, cells stimulated by compound 48/80 can respond without a measurable delay and degranulate within 2 s, suggesting that this compound acts at a site after the GTP‐binding regulatory protein.
The chymotrypsin‐like activity of human prostate‐specific antigen, γ‐seminoprotein
Authors: Akiyama Kazuko , Nakamura Takanori , Iwanaga Sadaaki , Hara Mitsuwo
Published: July 2002

γ‐Seminoprotein (γ‐Sm) is a human prostate‐specific antigen and a serine protease judging from the complete amino acid sequence which shows extensive homology with the kallikrein family. The enzymatic activity of γ‐Sm was defined as a chymotypsin‐like activity using reduced and S‐3‐(trimethylated amino)propylated lysozyme and insulin‐oxidized A and B chains as substrates. The ‐LeuSer‐ peptide bond of lysozyme was rapidly hydrolyzed by γ‐Sm. γ‐Sm also hydrolyzed the ‐PheGlu‐ of lysozyme and the ‐LeuCys(SO3H) ‐of insulin B chain. Insulin A chain and arginyl‐ or lysyl‐linkage of these proteins were not hydrolyzed by γ‐Sm at all.
Effect of adrenergic agonists on phosphoinositide breakdown in rat skeletal muscle preparations
Authors: Schadewaldt Peter , Stapper Norbert J. , Staib Wolfgang
Published: July 2002

Adrenergic regulation of phosphoinositide breakdown in rat skeletal muscle was investigated in 30‐min incubations with 10 mM LiCl. In rat hemidiaphragms, prelabelled with D‐myo‐[2‐3H]inositol, addition of α‐agonists (epinephrine, norepinephrine, phenylephrine) induced a 5‐8‐fold increase of [3H]inositol monophosphate accumulation. This could be prevented by inclusion of α‐antagonists (phentolamine, prazosin). β‐Agonists and/or β‐antagonists had no effect. Similar experiments with isolated flexor digitorum brevis muscle fibers yielded confirmatory results. Functional integrity of β‐receptor mediated processes was suggested by the β‐agonist‐induced increase of glucose 6‐phosphate in hemidiaphragms and cAMP in fiber preparations. The results indicate that phosphoinositide breakdown in differentiated rat skeletal muscle is, at least in part, under α‐adrenergic control.
The antimycin‐sensitive electrogenesis in Rhodopseudomonas sphaeroides chromatophores
Authors: Drachev Lel A. , Mamedov Mahir D. , Semenov Alexey Yu.
Published: July 2002

Electrogenesis linked to cyclic electron transfer through the ubiquinol‐cytochrome 5c
2 oxidoreductase complex in Rhodopseudomonas sphaeroides chromatophores was investigated by a direct electrometric method. Rapid kinetic measurements of laser flash‐induced ΔΨ generated in chromatophores associated with a phospholipid‐impregnated collodion film revealed an exponential rise ofΔΨ with a τ of ~ 700 μs. This exponential electrogenic phase contributes about 11% of the total photoelectric response and is sensitive to antimycin.
Amino acid sequence of rat liver cathepsin L
Authors: Towatari Takae , Katunuma Nobuhiko
Published: July 2002

The complete amino acid sequences of the heavy and light chains of rat liver cathepsin L (EC 3.4.22.15) were determined at the protein level. The heavy and light chains consisted of 175 and 44 amino acid residues, respectively, and their M
r values without glycosyl groups calculated from these sequences were 18941 and 5056, respectively. The amino acid sequence was also determined from the N‐terminal sequences of the heavy and light chains, and the sequences of cleavage fragments of the heavy chain with lysylendopeptidase and cyanogen bromide. The fragments were aligned by comparison with the amino acid sequence deduced from the sequence of cDNA of rat preprocathepsin L. The sequence of rat liver cathepsin L determined at the protein level was identical with that deduced from the cDNA sequence except that in the heavy chain, residues 176–177 (Asp‐Ser) were not present at the C‐terminus and alanine was replaced by proline at residue 125. Asn‐108 in the heavy chain is modified with carbohydrate.
The modification of chlorophyll fluorescence of Chlamydomonas reinhardtii by photoinhibition and chloramphenicol addition suggests a form of photosystem II less susceptible to degradation
Authors: Briantais Jean-Marie , Cornic Gabriel , Hodges Michael
Published: July 2002

The effect of photoinhibition and chloramphenicol addition on room‐temperature chlorophyll fluorescence induction parameters exhibited by Chlamydomonas reinhardtii has been studied. The observed changes induced by these different treatments suggest the presence of a photosystem II (PS II) reaction centre having an abnormal turnover rate and which gives rise to the fast initital variable fluorescence rise (F
i−F
o) in the absence of DCMU. It is proposed that a pool of PS II with a modified D1 protein exists which is less susceptible to degradation. Furthermore, it is shown that the fluorescence quenching associated with photoinhibition is not directly linked to PS II reaction centre degradation.
Light‐regulated expression of the psbA transcript in the cyanobacterium Anacystis nidulans
Authors: Lönneborg A. , Kalla S.R. , Samuelsson G. , Gustafsson P. , Öquist G.
Published: July 2002

The level of photoinhibition as well as the rate of recovery of photosynthesis increases with increasing light intensities in Anacystis nidulans. In addition, the recovery from photoinhibition is insensitive to rifampicin. We found that the amount of psbA‐specific mRNAs, coding for the D1 protein, was higher in cells grown in high rather than in low photon flux densities of white light. The results support the hypothesis that the rate of synthesis of the D1 protein plays a role both in the susceptibility of photosynthesis to photoinhibition and during recovery from photoinhibition. In addition, we found that a majority of the mRNAs specific for the psbA gene could be detected intact 90 min after the addition of rifampicin.
Is the 9 kDa thylakoid membrane phosphoprotein functionally and structurally analogous to the ‘H’ subunit of bacterial reaction centres?
Authors: Packham Nigel K.
Published: July 2002

Although the amino acid sequence of the 9 kDa (phospho)protein of chloroplasts has been determined, the function of this thylakoid membrane protein in photosynthetic electron transport and the reason for its physiological control remains unclear. In this paper, I briefly review the evidence which indicates that the phosphorylation of the 9 kDa protein results in a partial inhibition of photosynthetic oxygen evolution by increasing the stability of the semiquinone bound to QA the primary, plastoquinone‐binding site of photosystem II (PS II). I propose that in its dephosphorylated state, the 9 kDa thylakoid membrane protein may serve PS II to ensure efficient photochemical charge separation by aiding the transfer of reducing equivalents out of the reaction centre to the attendant plastoquinone pool. This function is analogous to that proposed for the H‐subunit of the reaction centre of photosynthetic eubacteria. Whether these two proteins have evolved from a common ancestral reaction centre protein is discussed in the light of a comparison of their amino acid sequences and predicted secondary structures.
Core histone acetylation during lymphocyte activation
Authors: Profumo Aldo , Querzola Flavia , Vidali Giorgio
Published: July 2002

Histone acetylation has been followed in cultures of human lymphocytes, in PHA‐stimulated lymphocytes and in mixed lymphocytes obtained from identical twins and from unrelated donors. A computer assisted analysis of two‐dimensional gels and autoradiograms revealed that in cultured lymphocytes only H3 and H4 core histones incorporate labeled acetate and that two H3 variants greatly differ in their rate of acetate uptake.
Identification by flow cytometry of two distinct rhodamine‐123‐stained mitochondrial populations in rat liver
Authors: Lopez-Mediavilla C. , Orfao A. , Gonzalez M. , Medina J.M.
Published: July 2002

Isolated rat liver mitochondria were split into three fractions of increasing density when applied to a Percoll gradient. NADH‐ubiquinone oxidoreductase, succinate dehydrogenase and cytochrome‐c oxidase but not F1‐ATPase activities increased with density as well as respiratory rate in state 3 and the respiratory control index. Flow cytometry of mitochondrial density fractions stained with rhodamine‐123 revealed the occurrence in each density fraction of two distinct mitochondrial populations with different fluorescence intensity. The high fluorescence population was minor and its proportion decreased with density. The extent of high fluorescence population staining depended on the deenergized state of the mitochondria suggesting that this population represents an immature form of the mitochondria which may develop into a fully functional organelle by the incorporation of structural and/or functional proteins.
Formylmethanofuran dehydrogenase activity in cell extracts of Methanobacterium thermoautotrophicum and of Methanosarcina barkeri
Authors: Börner G. , Karrasch M. , Thauer R.K.
Published: July 2002

Cell extracts of Methanobacterium thermoautotrophicum catalyzed the reduction of methyl viologen (apparent K
m=0.1 mM) with formylmethanofuran (apparent K
m=10 μM) at a specific rate of 4 μmol · min−1 · mg protein−1. Coenzyme F420 (apparent K
m=25 μM) rather than NAD, NADP, FAD, or FMN was used as physiological electron acceptor. With coenzyme F420 the specific activity was 0.4 μmol · min−1 · mg−1. More than 60% of the formylmethanofuran dehydrogenase activity was associated with the pellet fraction, which was obtained by centrifugation at 160 000 × g. An enzyme system with very similar properties was also found to be present in cell extracts of Methanosarcina barkeri grown on methanol. In both organisms the formylmethanofuran dehydrogenase activity was rapidly inactivated by cyanide.