bluuebunny/crossref_metadata_2025 huggingface dataset:
Susceptibility Pattern of Different Parts of Moringa oleifera against Some Pathogenic Fungi, Isolated from Sputum Samples of HIV Positive Individuals Co-Infected with Pulmonary Tuberculosis
Authors: Onuoha Vivian C. , Enweani Ifeoma B. , Okereke Ogbonnia Ekuma-
Published: August 2020

Background: Opportunistic fungal infection in HIV positive individualshas a high risk of morbidity and mortality as it could be a pointer to systemic infection. Moringa oleifera is one of the 14 species of the family Moringaceae with traditional effectiveness in anti–helminthic, analgesic, anti-ulcerative, anti-microbial, etc.

Aim: This study was designed to determine the susceptibility pattern of different parts of Moringa oleifera against some pathogenic fungi using dried Morringa oleifera seed, bark, pod, and leaf extracts/fractions as agents.

Materials and Methods: A total of 132 subjects (62 males and 70 females) attending outpatient clinics (OPD) at Nnamdi Azikiwe University Teaching Hospital Nnewi, and Holy Rossary Hospital Emekuku Owerri, Nigeria were recruited for this study. The HIV status of the test subjects was confirmed using immuno-chromatographic technique and their sputum screened for Tuberculosis (TB) using Ziehl Nelsen staining technique. The sputum samples were also cultured on Sabouaurd dextrose agar with and without chloramphenicol incubated at 25oC with examination of growth every 3 days for two weeks. Identification of isolates was done using Gram staining methods, chromogenic media and slide culture.700g, 220g, 500g, and 250g each of the different parts of the pulverized Moringa oleifera (seeds, barks, pods, and leaves) were weighed and exctracted using Soxhlet extractor and methanol as solvent.

Results: The prevalence of fungi isolates from test subjects shows that Candida albicans has the highest frequency of 25(26.6%), with the least prevalent being Aspergillus flavus and Phialemonium curvatum at 2(2.1%) respectively. There was also a positive significant correlation between TB and fungi infection in HIV-sero positive subjects (P<0.05).The invitro antifungal activity of these Moringa oleifera extracts/fractions against the test isolates showed no antifungal activity except for ethyl acetate fraction which demonstrated the most prominent activity against A. fumigatus with zones of inhibition of 3-10mm diameter with the least activity against C. krusei with zones of inhibition of 3-6mm diameter. However, the antifungal activities of fluconazole which served as a control in the antifungal susceptibility against the test isolates were more effective than the extract/fractions against the test isolates with a minimum inhibition concentration (MIC) of 50-200mg/ml against 25-100mg/ml for ethylacetate fraction.

Conclusion: The comparable antifungi effect of ethylacetate fraction of M. oleifera leaf with that of fluconazole, suggests that ethylacetate fraction of M. oleifera leaf could aid the treatment of pathogenic fungi due to its potential applications in Pharmaceutical industry, thereby ameliorating the morbidity and mortality rate of HIV positive co-infected TB subjects.
Simulation Study of H1N1 Transmission through Immigrants and Vaccination
Authors: Shah Nita H. , Shukla Nehal , Thakkar Foram , Satia Moksha
Published: August 2020

H1N1 influenza, is a disease caused by the Type A strain of negative-sense single stranded RNA virus which has unique characteristics of reassortment making it different and difficult to control. Due to the unique nature of the virus, the constant globalization of the international population, and the lack of effective immunization, this virus is a great threat to public health. There is only limited data available on impacts of immigrants on H1N1 transmission. So, there is real need to study impacts of immigrants on H1N1 transmission as well as vaccination. We have studied a mathematical model for simulation study of H1N1 transmission through immigrants and vaccination. The class of susceptible people is divided in two types: vaccinated immigrants and non-vaccinated immigrants. The medications and hospitalizations are the remedial steps to get cured. The rate at which people get medicated or hospitalized is analyzed using SEIR model. The system of non-linear ordinary differential equation is formulated for the given model and the reproduction number is then calculated using next generation matrix method which indicates the recovery rate of an individual stability. Numerical simulations are then carried out. Our study showed that increasing the awareness among the general population, immigrants about clinical features and modes of transmission of H1N1 and improving the vaccination rate helps decrease the transmission of H1N1.
Analysis of Influencing Factors of Traffic Accidents in Undersea Tunnel
Authors: Zheng Yang Yong- , Mei Yao
Published: August 2020

This study aims to study the influencing factors of traffic accidents in undersea tunnels and establish a prediction model of undersea tunnel traffic accidents. Through face-to-face inquiry and telephone survey, the relevant information of drivers who had traffic accidents in Jiaozhou Bay undersea tunnel from 2018 to 2019 was obtained, and the relevant information of citizens passing through Jiaozhou Bay undersea tunnel was obtained by posting questionnaires on the Internet. Based on whether there were traffic accidents in undersea tunnel, the drivers with undersea tunnel driving experience were divided into accident group and control group. Through the single factor analysis of the data, it is found that drinking, bad driving habits, having traffic accident experience, speeding and other thirteen factors have an important impact on the occurrence of traffic accidents undersea tunnel. Multivariate analysis was carried out on thirteen factors with significant influence, and a model for predicting the probability of accidents in undersea tunnels was established. Five influencing factors of speeding, the following distance is too small, carelessness, illegal lane change and having traffic accidents experience entered the model, with OR values of 7.11(2.94-18.90), 2.35(1.17-4.08), 3.99(1.23-7.45), 2.68(1.15-6.39) and 3.95(1.78-6.11). The occurrence of accidents in undersea tunnel is related to the length of the tunnel and the traffic flow. For an undersea tunnel with a length of 7 km and an average daily traffic flow of 50000 vehicles, the number of daily traffic accidents is 1.96. The driver’s violations and the driver’s carelessness are the main factors in undersea tunnel traffic accident. In order to improve the traffic safety of undersea tunnel, government departments should step up enforcement against traffic violations. In addition, the undersea tunnel management department needs to consider taking measures to improve the harsh and monotonous environment in the tunnel. This study provides an empirical basis for the prevention of traffic accidents in undersea tunnels.
Maternal Serum Levels of Alpha Tumour Necrotic Factor, Interleukin 10, Interleukin 6 and Interleukin 4 in Malaria Infected Pregnant Women Based on Their Gestational Age in Southeast, Nigeria
Authors: Ifeanyi Obeagu, Emmanuel , Nonyelum Esimai, Bessie , Stella Ekelozie, Ifeoma , Ijego Asogwa Eucharia , Amaeze Amaeze Augustine , Nchekwubedi Chukwu, Stella , Ngozi Amaeze, Florence , Chikodili Ugwuja, Mabel , Kyrian Chukwu, Sunday
Published: August 2020

Malaria has been reported as a condition caused by infestation with Plasmodium parasite species, is a major public health problem globally especially in developing countries like Nigeria. This study was carried out in Federal Medical Centre Umuahia in Abia State, Nigeria. A study was done to determine the maternal serumlevels of alpha tumour necrotic factor, interleukin 10, interleukin 6and interleukin 4 in malaria infected pregnant women based on their gestational age in Southeast, Nigeria. A total of 150 subjects between the ages of 18-45 years were recruited for the study comprising of fifty (50) subjects each of the 3 trimesters. Commercial ELISA Kit by MELSIN Medical Co Limited was used to measure all the cytokines. The results of Table 1 showed no significant difference of TNF-α (p=0.346), IL-10 (p=0.059), IL-6 (p=0.811) and IL-4 (p=0.257) of malaria infected pregnant women at first trimester and second trimester respectively. The results of Table 2 showed no significant difference of TNF-α (p=0.642), IL-10 (p=0.678), IL-6 (p=0.551) and IL-4 (p=0.280) of malaria infected pregnant women at first trimester and third trimester respectively. The results of Table 2 showed no significant difference of TNF-α (p=0.062), IL-10 (p=0.016), IL-6 (p=0.352) and IL-4 (p=0.914) of malaria infected pregnant women at first trimester and third trimester respectively. The study showed no changes in the cytokines studied among the malaria infected pregnant women based on gestational ages except when IL-10 was compared between the subjects on second trimester and third trimester. This study shows that malaria infection does not changes these cytokines in pregnant women based on gestational ages except the il-10 when compared at second trimester and third trimester but changes when compared at other trimesters.
Bacterial Contamination of Chicken Eggs from Poultry Farms and Retail Markets in the New Juaben Municipality, Ghana
Authors: Addo Matthew Glover , Raissa Manomba-Asamoah , Badu Kingsley , Agordzo Samuel
Published: August 2020

This cross-sectional study seeks to investigate the bacterial contamination of poultry products (eggs) produced from selected farms and sold within the major markets in the New Juaben Municipality in the Eastern region of Ghana. A total of 100 eggs were sampled randomly from October to December, 2018 from five poultry farms and five retail markets in the municipality for microbial evaluation. The collected swabs of eggshells and its contents collected at various times of the day were cultured on suitable media and standard microbiological test were performed to identify the isolated organisms. The entire eggshell samples were positive for bacteria with only 13 (13%) having growth in the egg content. The genera of bacteria recorded are Staphylococcus, Escherichia, Klebsiella, Proteus and Citrobacter. The mean total viable count of bacteria from both shell and content sampled from poultry farms and retail markets were 13.2×107cfu/ml and 8.0 ×107cfu/ml (p>0.05) respectively. These were higher than the acceptable value of 10×105 gm/cfu. The study also showed that the level of microbial contamination of the eggs was highest in the afternoon as compared to the morning and evening. The bacteria isolates and the sheer numbers from both the eggshells and the egg content confirm the poor hygienic conditions under which both the poultry farms and markets were kept. Even though serious human pathogens were not prevalent in this study, it is recommended that strict hygienic protocols be adhered to on the poultry farms and in the markets.
Fungal Infections in Plateau State, Nigeria
Authors: Enurah L. U. , Shaibu S. J. , Bitrus A. A. , Zekeri A. H. Momoh- , Lombin L. H. , Barde I. J.
Published: August 2020

This paper describes a study carried out on people of different ages with various skin diseases who submitted themselves for examination and diagnoses at the Dermatophylosis Research Centre of the National Veterinary Research Institute, Vom, Plateau State, Nigeria. Samples of skin scrapping were collected and cultured on Sabouraud Dextrose Agar (SDA) while some were examined under the microscope for possible morphological identification of the fungi. The results showed that out of the thirty samples analyzed, nine genera of fungi were isolated comprising of Trichophyton 10 (33.3%), Aspergillus 9 (30%), Penicillum 4 (13.32%), Microsporum 2 (6.70%), Epidermorphyton 1 (3.33%), Trichosporum   1 (3.33%), Cryptococcus 1 (3.33%), Mucor 1 (3.33%), and Cephalosporium  1 (3.33).

The study revealed high incidence of human fungal diseases, a situation that calls for good hygiene practices in the study area to mitigate and possibly eradicate the prevalence of human skin diseases in the rural area of Plateau State.
Nursing discharge plan in hospitals: an experience report
Authors: Carneiro Jayanne Moreira , Jesus Larissa Oliveira de , Silva Caroline Santos , Santiago Ainara dos Santos , Santos Anderson Alves Lima , Marques Patrícia Figueiredo
Published: August 2020

Objetivo: descrever o processo de implantação de Planos de Alta na Clínica Cirúrgica de um hospital de grande porte de Salvador (BA). Método: relato de experiência que descreve aspectos relacionados a elaboração, atualização e implantação de cinco planos de alta hospitalar de enfermagem para pacientes atendidos nessa clínica cirúrgica. Foram direcionados aos seguintes procedimentos cirúrgicos: hernioplastias, colecistectomia, histerectomia, amputação e prostatectomia. Resultados: foram atualizados cinco planos de alta, os quais foram analisados pela coordenadora de enfermagem, médico cirurgião geral e supervisora do componente obrigatório; após sugestões e alterações realizadas, foram encaminhados para a Comissão de Padronização desse Hospital para serem implantados nas Unidades de Clínica Cirúrgica. Conclusão: a experiência possibilitou a afirmação dos instrumentos como um importante material didático para os pacientes sobre as condutas pós-cirúrgicas e para o enfermeiro, que através desse material consegue direcionar as orientações necessárias.
Health-related quality of life of patients bearing diabetes mellitus
Authors: Marques Jennifer Vieira Paschoalin , Teixeira Carla Regina de Souza , Zanetti Maria Lúcia , Kusumota Luciana , Becker Tânia Alves Canata , Hodniki Paula Parisi
Published: August 2020

Objective: The purpose of this study was to investigate the health-related quality of life and clinical parameters of patients with type 2 diabetes mellitus. Methods: This is a cross-sectional study with a quantitative approach, which was undertaken with 73 patients with type 2 diabetes mellitus in a health unit of the Sistema Único de Saúde (SUS) [Brazilian Unified Health System] in Ribeirão Preto city, São Paulo State, Brazil, in 2016. The DQOL-Brazil questionnaire, which covers the domains of satisfaction, impact, social and/or vocational concerns, and concerns about diabetes mellitus, was used for data collection. Results: Most of the participants were female, married, and retired with an average age of 62.7 years old. The domain of vocational and/or social concerns had the best score for quality of life while satisfaction had the worst. The quality of life of patients with altered glycated hemoglobin levels was worse than that of patients without such alterations. For those with unchanged hemoglobin glycated levels, the domain of social and/or vocational concerns had the best quality of life scores while the domain of satisfaction had the worst. Conclusion: The results can help the development of intervention studies and strategic plans in health services.
Female sexuality after bariatric surgery according to the Merleau-Ponty phenomenology: integrative Review
Authors: Delatore Silvana , Dell'Agnolo Cátia Millene , Marcon Sonia Silva , Pelloso Sandra Marisa
Published: August 2020

Objetivo:Analisar as alterações na sexualidade das mulheres submetidas à cirurgia bariátrica, por meio de revisão integrativa da literatura. Método:A busca ocorreu nas bases Medical Literature Analysis and Retrieval System Online, Literatura Latino-Americana e do Caribe em Ciências da Saúde, Enfermagem, Scientific Electronic Library Onlinee Portal de Periódicos Coordenação de Aperfeiçoamento de Pessoal de Nível Superior. Resultados: Foram identificados 158 publicações, sendo incluídos estudos originais, quantitativos e qualitativos, publicados em inglês e português, com as palavras-chave sexualidade feminina e cirurgia bariátrica, restando 7 artigos, os quais foram analisados segundo a fenomenologia de Merleau-Ponty. Foram identificadas 4 categorias temáticas: Alterações positivas, negativas, questões médicas e fatores psicossociais. Conclusão:A essência do fenômeno se mostrou multifatorial de modo que a representação do corpo físico em alguns casos gerou melhora psicossocial, conjugal, cultural, e na sexualidade, porém não na sua plenitude de prazer e satisfação e em outros repercutiu em alterações negativas.
Nurse managers’ moral suffering in a university hospital
Authors: Drago Laila Crespo , Ramos Flávia Regina Souza , Brehmer Laura Cavalcante de Farias , Silveira Luciana Ramos , Brito Maria José Menezes
Published: August 2020

Objetivo: Descrever situações geradoras e elementos envolvidos no processo de sofrimento moral na experiência de enfermeiros gerentes de um hospital público do sul do Brasil. Método: Estudo descritivo e exploratório de abordagem qualitativa, realizado durante o período de março a setembro de 2014, cujas informações foram coletadas mediante entrevista semiestruturada. Participaram da pesquisa 17 enfermeiros gerentes. Os dados foram organizados no software Atlas. Ti© e analisados segundo a técnica de análise textual discursiva. Resultados: Emergiram três categorias: Organização e Condições de trabalho, Gestão de Pessoas, e O Ser Gerente. Conclusão: Constatou-se que os gerentes vivenciam o sofrimento moral decorrente de situações conflituosas relacionadas às condições de trabalho, profissionais insubordinados, falta de autonomia, assédio moral e as múltiplas faces da função gerencial. Assim, buscou-se contribuir para área da saúde, nos aspectos de sensibilizar moralmente os profissionais para as atividades que desempenham e no tocante ao sofrimento moral.
Mapping of the main types of neoplasms among children and adolescents across the regional health agencies from the Rio de Janeiro state, Brazil
Authors: Valente Rayssa Goulart , Silva Aline Cerqueira Santos Santana da , Knupp Virginia Maria de Azevedo Oliveira , Silva Bianka Queiroz da , Barcia Leila Leontina do Couto , Neto José Antônio de Sá
Published: August 2020

Objetivo: Analisar os principais tipos de neoplasia malignas em pacientes de 0 a 19 anos de idade nas regionais de saúde do estado do Rio de Janeiro. Método: Estudo ecológico que analisou as neoplasias mais frequentes através dos dados contidos no Sistema de Informação Hospitalar em 2017. Os dados foram analisados através do cálculo da frequência relativa e medidas de tendências central. O mapeamento foi realizado por meio do TabWin. Resultados: foram observados 2.662 casos de neoplasias malignas na população de 0-19 anos de idade residentes nas regionais de saúde do Estado do Rio de Janeiro, sendo a Metropolitana I a região de maior proporção e a leucemia o tipo de câncer infanto-juvenil mais frequente. Conclusão: apropriação desses dados torna possível concretizar estratégias para a construção de políticas públicas, visando medidas de prevenção, diagnosticas e tratamento vislumbrando maior sobrevida, melhor qualidade de vida e redução da taxa de mortalidade infanto-juvenil.
Experimental evidence against the applicability of the Saffman‐Delbrück model to the translational diffusion of lipids in phosphatidylcholine bilayer membranes
Authors: Vaz Winchil L.C. , Dieter Hallmann
Published: July 2002

The translational diffusion coefficient (D
t) of the fluorescent lipid derivative, N‐(4‐nitrobenz‐2‐oxa‐1,3‐diazole)‐phosphatidylethanolamine (NBD‐PE), with acyl chains of 6, 12, and 18 carbon atoms was measured in multibilayers of 1‐palmitoyl‐2‐oleoyl‐, dilauroyl‐, and distearoyl‐phosphatidylcholines using the fluorescence recovery after photobleaching technique. In a given bilayer, in the liquid crystalline phase, D
t was found to be independent of the height (length in a direction parallel to the bilayer normal) of the diffusing species. This is in disagreement with fluid hydrodynamic models for diffusion in bilayers which predict an almost linear dependence of D
t upon the height of the diffusing species in the bilayer.
24R,25‐Dihydroxyvitamin D stimulates creatine kinase BB activity in chick cartilage cells in culture
Authors: Sömjen Dalia , Kaye Alvin M. , Binderman Itzhak
Published: July 2002

In chick limb‐bud cartilage cell cultures 24R,25‐dihydroxycholecalciferol (24R,25(OH)2D3), but not 24S,25(OH)2D3, 1α,25(OH)2D3 or 25(OH)D3, stimulates the activity of the brain type (BB) isozyme of creatine kinase (EC 2.7.3.2), the ‘estrogen‐induced protein’ first identified in rat uterus. Cultures treated with bromodeoxyuridine, in which cartilage formation in inhibited, show no stimulation of creatine kinase BB by 24R,25(OH)2D3.
γ‐Hexachlorocyclohexane activation of alveolar macrophage phosphatidylinositol cycle, calcium mobilization and O−2 production
Authors: Holian Andrij , Marchiarullo Michael A. , Stickle Douglas F.
Published: July 2002

The influence of γ‐hexachlorocyclohexane (HCC) on phosphatidyl inositol (PI) cycle activity was investigated in the guinea pig alveolar macrophage. Similar to stimulation by the chemotactic peptide N‐formyl‐nle‐leu‐phe (FNLLP), 125μM HCC was found to stimulate PI cycle activity, calcium mobilization (45Ca2+ efflux and cytosolic [Ca2+] elevation) and O−
2 production, although the action of HCC was prolonged in comparison. HCC treatment did not block subsequent stimulation by FNLLP. HCC is proposed to act at an early stage in the same activation sequence as does FNLLP.
Molecular cloning of unintegrated closed circular DNA of porcine retrovirus
Authors: Suzuka Iwao , Shimizu Nobuaki , Sekiguchi Kiichi , Hoshino Hiroo , Kodama Michi , Shimotohno Kunitada
Published: July 2002

Viral DNA of unintegrated closed circular form was isolated from a swine kidney cell line (SKL) which was infected with a porcine retrovirus Tsukuba‐1 (PRetV) produced from a swine malignant lymphoma‐derived cell line, Shimozuma‐1 and cloned using a λ phage vector; Charon 21 A. One of ten independent clones contained the 8.3 kb DNA fragment as an insert, which was thought to be a full length of viral DNA molecule carrying a long terminal repeat (LTR) sequence. We have analyzed this insert by mapping the recognition sites of some restriction endonucleases by Southern blot hybridization with appropriate probes.
Differential response of the human hepatoma‐derived cell line HA22T/VGH to polypeptide mitogens
Authors: Clementi Massimo , Testa Ivano , Festa Antonella , Bagnarelli Patrizia , Chang Chungming , Carloni Guido
Published: July 2002

Several human cell lines derived from primary cancer of the liver are able to grow under serum‐free conditions and produce spreading and growth factors which are released into the culture medium. Since this autocrine growth under hormone‐free conditions might play a basic role in malignant transformation, we studied the effect on cell replication and the presence of specific membrane receptors of epidermal growth factor (EGF) and insulin on a dedifferentiated human hepatoma cell line, named HA22T/VGH. Our results point to a similar inhibitory effect on cell replication in the presence of both EGF and insulin, in spite of detecting different affinities of binding.
Time‐resolved fluorescence studies of flavodoxin Demonstration of picosecond fluorescence lifetimes of FMN in Desulfovibrio flavodoxins
Authors: Visser Antonie J.W.G. , van Hoek Arie , Kulinski Tadeusz , Le Gall Jean
Published: July 2002

Picosecond‐resolved fluorescence spectroscopy on FMN bound in flavodoxin from Desulfovibrio vulgaris and Desulfovibrio gigas revealed fluorescence lifetimes of about 30 ps. The lifetime shortening can be explained by charge transfer interaction in the flavin excited state with aromatic amino acid residues. In addition to the short lifetime component, nanosecond lifetimes were also observed in both flavodoxins. The latter fluorescence lifetime in D. vulgaris flavodoxin (5.6 ns) is even longer than that of free FMN (4.7 ns A fixed lifetime analysis of the results from a diluted D. vulgaris flavodoxin solution yielded the dissociation constant of the FMN‐apoflavodoxin complex: K
d = 9.4 ∓ 1.4 nM at 20 °C and pH 7.0.
Sequence of an intestinal cDNA encoding human motilin precursor
Authors: Seino Y. , Tanaka K. , Takeda J. , Takahashi H. , Mitani T. , Kurono M. , Kayano T. , Koh G. , Fukumoto H. , Yano H. , Fujita J. , Inagaki N. , Yamada Y. , Imura H.
Published: July 2002

A cDNA clone encoding the human motilin precursor was isolated from an intestinal library using synthetic oligonucleotide probes. The predicted amino acid sequence indicates that the motilin precursor consists of 115 amino acids and includes a 25‐residue N‐terminal signal peptide followed by the 22‐amino‐acid motilin sequence and a long, 68‐residue C‐terminal peptide. The amino acid sequence of human motilin predicted from the cDNA sequence is indentical to its porcine counterpart, which has been determined by protein sequencing. Proteolytic processing of promotilin to motilin occurs at the sequence, Lys‐Lys, this being the first reported instance of processing occurring at a pair of Lys residues. In other precursors it occurs at Lys‐Arg, Arg‐Arg, Arg, or very rarely Lys.
Calcium‐activated neutral protease and its endogenous inhibitor Activation at the cell membrane and biological function
Authors: Suzuki Koichi , Imajoh Shinobu , Emori Yasufumi , Kawasaki Hiroshi , Minami Yasufumi , Ohno Shigeo
Published: July 2002

The structures of calcium‐activated neutral protease (CANP) and its endogenous inhibitor elucidated recently have revealed novel features with respect to their structure‐function relationship and enzyme activity regulation. The protease is regarded as a proenzyme which can be activated at the cell membrane in the presence of Ca2+ and phospholipid, and presumably regulates the functions of proteins, especially membrane‐associated proteins, by limited proteolysis. Protein kinase C is hydrolysed and activated by CANP at the cell membrane to a cofactor‐independent form. These results are reviewed and the possible involvement of CANP in signal transduction is discussed.
The γ‐subunit of ATP synthase from spinach chloroplasts Primary structure deduced from the cloned cDNA sequence
Authors: Miki Junji , Maeda Masatomo , Mukohata Yasuo , Futai Masamitsu
Published: July 2002

cDNA clones encoding the γ‐subunit of chloroplast ATP synthase were isolated from a spinach library using synthetic oligonucleotide probes. The predicted amino acid sequence indicated that the mature chloroplast γ‐subunit consists of 323 amino acid residues and is highly homologous (55% identical residues) with the sequence of the cyanobacterial subunit. The positions of the four cysteine residues were identified. The carboxyl‐terminal region of the choloroplast γ‐subunit is highly homologous with those of the γ‐subunits from six other sources (bacteria and mitochondria) sequenced thus far.
Autoantibodies to the core proteins of hnRNPs
Authors: Dangli Amalia , Guialis Apostolia , Vretou Evangelia , Sekeris Constantine E.
Published: July 2002

A novel autoantibody reacting with the core polypeptides of hnRNP particles has been detected in the serum of a patient with systemic lupus eythematosus (SLE) and Sjögren's syndrome manifestations. Immunoblot analysis, using either rat liver or HeLa nuclear extracts as the antigen source, demonstrated that the autoantibody interacts with a specific subgroup of the core polypeptides of hnRNP particles, namely A2, B1 and B2, but not with A1, C1 and C2.
Rapid preparation of the nicotinic acetylcholine receptor for crystallization in detergent solution
Authors: Hertling-Jaweed Sigrid , Bandini Giampiero , Müller-Fahrnow Anke , Dommes Veronika , Hucho Ferdinand
Published: July 2002

A novel rapid purification method for the nicotinic acetylcholine receptor from Torpedo electric tissue was developed. It allows preparation of 10 mg quantities of pure and stabile receptor protein within 2 days. This protein is used for crystallization attempts. Conditions are described which reproducibly yield crystals.
The γ subunit of F1 and the PVP protein of Fo (FoI) are components of the gate of the mitochondrial FoF1 H+‐ATP synthase
Authors: Papa S. , Guerrieri F. , Zanotti F. , Fiermonte M. , Capozza G. , Jirillo E.
Published: July 2002

The γ subunit of the F1 moiety of the bovine mitochondrial H+‐ATP synthase is shown to function as a component of the gate. Addition of purified γ subunit to Fo‐liposomes inhibits transmembrane proton conduction. This inhibition can be removed by the bifunctional thiol reagent diamide. Immunoblot analysis shows that the diamide effect is likely due to disulphide bridging of the γ subunit with the PVP protein of the Fo sector.
The deazaguanine‐derivative, queuine, affects cell proliferation, protein phosphorylation and the expression of the proto oncogenes c‐fos and c‐myc in HeLa cells
Authors: Langgut W. , Kersten H.
Published: July 2002

In higher eukaryotes the hypermodified guanine analogue, queuine (7‐(5‐(((1S,4S,5R)‐4,5‐dihydroxy‐2‐cyclopentene‐1‐yl)amino)‐methyl)‐7‐deazaguanine), occurs free or as modified nucleoside (Q) in the anticodon of specific tRNAs. Fast proliferating tissues and tumors contain considerable amounts of free queuine and Q‐deficient tRNAs. Here we show that HeLa cells can be grown in the absence or presence of queuine. In response to queuine, and under appropriate conditions, (i) the proliferation of HeLa cells was stimulated, (ii) the steady‐state level of c‐fos mRNA was reduced, contrary that of c‐myc mRNA was elevated, and (iii) in cytosolic extracts protein phosphorylation especially for a 42 kDa protein was significantly increased. The results suggest that queuine substitutes for growth factors in a signal transduction pathway.
Human c‐myc gene contains a regulatory site similar to consensus of interferon response sequence (IRS)
Authors: Alexandrova N.M. , Itkes A.V. , Imamova L.R. , Chernov B.K. , Tulchinsky E.M. , Ulyanov N.B. , Kisselev L.L.
Published: July 2002

Expression of c‐myc proto oncogene is regulated by multiple mechanisms. Here, we report that the consensus of the regulatory region of interferondependent genes, GGAAAN1−3 GAAA, was found after computer search in the 5'‐terminal flank of human c‐myc gene in position (−76:−67). In vitro transcription of c‐myc gene fragments showed that the consensus region competes with oligonucleotide GGGAAAATGAAACT for binding to specific protein(s). This oligonucleotide was shown to bind selectively the interferon‐dependent positive transcription factor [1]. Transcription of c‐myc fragments lacking 5'‐terminal region up to positions −101 or +71 was initiated at two sites located in the first intron. These sites did not coincide with P1 in vivo RNA cap‐site. Binding of the protein factor(s) to the regulatory region of c‐myc gene −76:−67 blocked the in vitro transcription initiated in the first intron.
Genetically tailored atrial natriuretic factor‐dependent guanylate cyclase Immunological and functional identity with 180 kDa membrane guanylate cyclase and ATP signaling site
Authors: Marala Ravi , Duda Teresa , Goraczniak Rafal M. , Sharma Rameshwar K.
Published: July 2002

Biochemical and immunological studies have established that one of the signal transducers of atrial natriuretic factor (ANF) is a 180 kDa membrane guanylate cyclase (180 kDa mGC), which is also an ANF receptor; obligatory in the transduction process is an intervening ATP‐regulated step, but its mechanism is not known. GCα is a newly discovered member of the guanylate cyclase family whose activity is independent of the known natriuretic peptides, and the enzyme is not an ANF receptor. The genetically tailored GCα, GCα‐DmutGln338 Leu364, however, is not only a guanylate cyclase but also an ANF receptor and is structurally and functionally identical to the cloned wild‐type ANF receptor guanylate cyclase, GC‐A. We now report that the ANF‐dependent guanylate cyclase activity in the particulate fractions of cells transfected with GCα‐DmutGln338 Leu364 was inhibited by the 180 kDa mGC polyclonal antibody, and with this antibody probe it was possible to purify the 130 kDa expressed receptor; the hormone‐dependent cyclase activity of this receptor was exclusively dependent upon ATP; and through site‐directed mutational studies with GCα mutants, the signaling sequence that defines ATP binding site was identified. We thus conclude that 180 kDa mGC and the mutant protein are immunologically similar; both proteins are linked to the ANF signal in the generation of cyclic GMP synthesis; and in both the ligand binding and catalytic activities are bridged through a defined ATP binding module.
Antisem to carcinoembryonic antigen recognizes a phosphotyrosine‐containing protein in human colon cancer cell lines
Authors: Skubitz Keith M. , Ducker Thomas P. , Skubitz Amy P.M. , Goueli Said A.
Published: July 2002

Members of the carcinoembryonic antigen (CEA) family include CEA, non‐specific cross reacting antigen (NCA), and biliary glycoprotein (BGP), and appear to function as cell adhesion molecules. Immunoprecipitation and subsequent gel electrophoresis of proteins from several colon cancer cell lines labeled with [γ‐32P]ATP, under conditions designed to detect ecto‐kinase‐catalyzed phosphorylation of cellular proteins, revealed that polyclonal anti‐CEA antiserum recognized a 175–190 kDa phosphoprotein on the surface of colon cancer cells. The ability to detect this phosphoprotein did not correlate with CEA production, and immunoprecipitation studies suggested that the phosphoprotein is BGP. Phosphoamino acid analysis of the 175–190 kDa protein showed that it contained predominantly phosphotyrosine.
In vivo biosynthesis of cholesterol in the rat retina
Authors: Fliesler Steven J. , Florman Richard , Rapp Laurence M. , Pittler Steven J. , Keller R.Kennedy
Published: July 2002

Previous reports have suggested that the rate of de novo cholesterol synthesis in the adult vertebrate retina is extremely slow. We investigated cholesterol biosynthesis in the adult rat retina in vivo, following intravitreal injection of [3H]acetate. HPLC analysis of retinal non‐saponifiable lipid extracts revealed co‐elution of radioactivity with endogenous cholesterol mass within 4.5 h post‐injection. Incorporation of [3H]acetate into cholesterol was markedly reduced by co‐injection of known inhibitors of the cholesterol pathway. In contrast to previous results with retinas from other species, no radiolabel or mass corresponded to squalene, except in lipid extracts from retinas treated with NB‐598, a squalene epoxidase inhibitor. These results demonstrate, for the first time, the capacity of the adult vertebrate retina to rapidly synthesize cholesterol de novo.
Molecular cloning of a novel putative G protein‐coupled receptor (GPCR21) which is expressed predominantly in mouse central nervous system
Authors: Saeki Yoshinaga , Ueno Satoshi , Mizuno Ryuzo , Nishimura Tomoya , Fujimura Harutoshi , Nagai Yoshitaka , Yanagihara Takehiko
Published: July 2002

A novel cDNA clone encoding a putative G protein‐coupled receptor (named GPCR21) was isolated from a mouse brain cDNA library along with its homologue, GPCR01 (the mouse counterpart of previously reported rat receptor R334 [(1991) FEBS Lett. 292, 243‐248]) by the polymerase chain reaction using degenerate oligonucleotide primers. Northern blotting and reverse transcription‐polymerase chain reaction analyses showed predominant expression of these two receptors in the central nervous system. In situ hybridization analysis revealed their prominent expression in the limbic system and further demonstrated the differential distribution of their mRNAs in mouse brain. Although the ligands for these receptors are yet to be identified, the significant sequence homology between these receptors suggests that they constitute a new receptor subfamily and they possibly represent different receptor subtypes for an unknown neurotransmitter.
Human amnion epithelial cells assemble tenascins and three fibronectin isoforms in the extracellular matrix
Authors: Linnala Auli , Balzac Enrica , Zardi Luciano , Virtanen Ismo
Published: July 2002

Monoclonal antibodies (MAb) were used to show that cultured human amnion epithelial (HuA) cells produce tenascins (Tn) and isoforms of cellular fibronectin (cFn). Tn polypeptides of Mr
280,000 and 190,000, assembled into extracellular matrix (ECM) but not secreted into the culture medium by HuA cells, were electrophoretically similar to those produced by human fibroblasts as revealed with domain‐specific MAbs. The results suggested that most Fn produced by HuA cells contained the extradomain (ED) A and an oncofetal domain but only a minor fraction EDB. In immunofluorescence Tn and Fn were seen in different cytoplasmie granules upon monensin‐induced intracellular accumulation. Tn appeared to be deposited in the ECM in colocalization with Fn but distinctly slower. The present results show that cultured normal human epithelial cells synthesize Tn and three isoforms of cFn and secrete them by using different cytoplasmie pathways.
KEKE motifs
Authors: Realini Claudio , Rogers Scott W. , Rechsteiner Martin
Published: July 2002

A stretch of 28 ‘alternating’ lysine (K) and glutamate (E) residues is found in an activator of the multicatalytic protease. Such ‘KEKE sequences’ are also present in subunits of the multicatalytic protease, in subunits of the 26S protease and in a variety of chaperonins. We propose that KEKE regions promote association between protein complexes. Furthermore, they may contribute to the selection of peptides presented on MHC Class I receptors.
Involvement of cell wall β‐glucan in the action of HM‐1 killer toxin
Authors: Kasahara Shin , Inoue Shunsuke Ben , Mio Toshiyuki , Yamada Toshiko , Nakajima Tasuku , Ichishima Eiji , Furuichi Yasuhiro , Yamada Hisafumi
Published: July 2002

HM‐1 killer toxin secreted from Hansenula mrakii inhibits the growth of Saccharomyces cerevisiae cells by interfering with β‐1,3‐glucan synthesis. We found that HM‐1 killer toxin killed intact cells but not protoplasts. In addition, cells lacking the functional KRE 6 allele (kre6Δ) became resistant to higher concentration of HM‐1 killer toxin. As reported by Roemer and Bussey [(1991) Proc. Natl. Acad. Sci. 88 11295–11299], cells lacking functional KPE6 had a reduced level of the cell wall β‐1,6‐glucan compared to that in cells harboring the normal KRE6. These results suggest that the cell wall ⨿‐glucan is involved in the action of HM‐1 killer toxin. Addition of HM‐1 killer toxin with several kinds of oligosaccharides revealed that either ⨿‐1,3‐ or β‐1,6‐glucan blocked the cytocidal action of HM‐1 killer toxin whereas α‐1,4‐glucan and chitin did not. Mannan also interfered with HM‐1 killer toxin action, but this inhibitory effect was much weaker than that observed with β‐1,3‐ or β‐1,6‐glucans. Thus, it appears that the cell wall β‐glucan interacts with HM‐1 killer toxin, and that this toxin‐β‐glucan commitment is required for the action of HM‐1 killer toxin.
Distribution and localization of a G protein‐coupled inwardly rectifying K+ channel in the rat
Authors: Karschin Christine , Schreibmayer Wolfgang , Dascal Nathan , Lester Henry , Davidson Norman , Karschin Andreas
Published: July 2002

The cellular distribution of the mRNA of the inwardly rectifying K+ channel KGA (GIRK1) was investigated in rat tissue by in situ hybridization. KGA was originally cloned from the heart and represents the first G protein‐activated K+ channel identified. It is expressed in peripheral tissue solely in the atrium, but not in the ventricle, skeletal muscle, lung and kidney. In the central nervous system KGA is most prominently expressed in the Ammon's horn and dentate gyrus of the hippocampus, neocortical layers II–VI, cerebellar granular layer, olfactory bulb, anterior pituitary, thalamic nuclei and several distinct nuclei of the lower brainstem. The abundant expression of KGA in many CNS neurons support its important role as a major target channel for G protein mediated receptor function.
Removal of Mg2+ inhibition of cardiac ryanodine receptor by palmitoyl coenzyme A
Authors: Connelly Timothy , Ahern Christopher , Sukhareva Manana , Coronado Roberto
Published: July 2002

45Ca2+ fluxes and planar bilayer recordings indicated that the fatty acid metabolise palmitoyl coenzyme A, but not free coenzyme A or palmitic acid, stimulated the cardiac ryanodine receptor channel of pig heart sarcoplasmic reticulum. Palmitoyl CoA reactivated channels inhibited by concentrations of cytoplasmic free Mg2+ in the physiological range. Reactivation by palmitoyl CoA in the presence of Mg2+ was stimulated by myoplasmic free Ca2+ in the micromolar range. Acyl coenzyme A derivatives may be utilized by cardiac muscle cells to compensate for the severe Mg2+ inhibition of ryanodine receptors which would otherwise leave Ca2+ stores unresponsive to Ca2+ and to other cytosolic ligands involved in signal transduction.
Fish glucocorticoid receptor with splicing variants in the DNA binding domain
Authors: Takeo Jiro , Hata Jun-ichiro , Segawa Chisako , Toyohara Haruhiko , Yamashita Shinya
Published: July 2002

Here we describe the isolation of a rainbow trout cDNA containing an entire GR coding region. Although the encoded protein is highly homologous to other GRs, especially in its DNA binding domain, it contains a nine amino acid insertion between the two zinc fingers. This novel form is found in all rainbow trout tissues examined; however, the testis also contains a splice variant lacking this insert, making it completely continuous to other GRs. In transient transfection assays of cultured cells, the two rainbow trout GR variants activated transcription from the glucocorticoid‐responsive mouse mammary tumor virus promoter to comparable levels.
The emergence of major cellular processes in evolution
Authors: Ouzounis Christos , Kyrpides Nikos
Published: July 2002

The phylogenetic distribution of divergently related protein families into the three domains of life (archaea, bacteria and eukaryotes) can signify the presence or absence of entire cellular processes in these domains and their ancestors. We can thus study the emergence of the major transitions during cellular evolution, and resolve some of the controversies surrounding the evolutionary status of archaea and the origins of the eukaryotic cell. In view of the ongoing projects that sequence the complete genomes of several Archaea, this work forms a testable prediction when the genome sequences become available. Using the presence of the protein families as taxonomic traits, and linking them to biochemical pathways, we are able to reason about the presence of the corresponding cellular processes in the last universal ancestor of contemporary cells. The analysis shows that metabolism was already a complex network of reactions which included amino acid, nucleotide, fatty acid, sugar and coenzyme metabolism. In addition, genetic processes such as translation are conserved and close to the original form. However, other processes such as DNA replication and repair or transcription are exceptional and seem to be associated with the structural changes that drove eukaryotes and bacteria away from their common ancestor. There are two major hypotheses in the present work: first, that archaea are probably closer to the last universal ancestor than any other extant life form, and second, that the major cellular processes were in place before the major splitting. The last universal ancestor had metabolism and translation very similar to the contemporary ones, while having an operonic genome organization and archaean‐like transcription. Evidently, all cells today contain remnants of the primordial genome of the last universal ancestor.
Convenient fluorometric assay for matrix metalloproteinase activity and its application in biological media
Authors: Beekman Bob , Drijfhout Jan Wouter , Bloemhoff Willem , Ronday H.Karel , Tak Paul Peter , te Koppele Johan M.
Published: July 2002

Matrix metalloproteinases (MMPs) are involved in physiological tissue remodeling and pathological conditions like tumour metastasis and joint destruction. Until now, no convenient and sensitive MMP‐activity assay in crude media like synovial fluid has been available. Therefore, the highly soluble fluorogenic substrate TNO211 (Dabcyl‐Gaba‐Pro‐Gln‐Gly‐Leu‐Glu(EDANS)‐Ala‐Lys‐NH2), containing the MMP cleavable Gly‐Leu bond and EDANS/Dabcyl as fluorophore/quencer combination, was synthesized and characterized as an MMP specific substrate. We show that the fluorogenic assay using TNO211 is sensitive and can detect MMP activity in culture medium from endothelial cells and untreated synovial fluid (SF) from RA and OA patients, and control subjects. MMP activity in SF significantly increased in the order C < OA < RA, thus the frequent use of OA samples as control in studies on RA is debatable.
Mitochondria and programmed cell death: back to the future
Authors: Petit Patrice X. , Susin Santos-Antonio , Zamzami Naoufal , Mignotte Bernard , Kroemer Guido
Published: July 2002

Programmed cell death, or apoptosis, has in the past few years undoubtedly become one of the most intensively investigated biological processes. However, fundamental questions concerning the molecular and biochemical mechanisms remain to be elucidated. The central question concerns the biochemical steps shared by the numerous death induction pathways elicited by different stimuli. Heterogeneous death signals precede a common effector phase during which cells pass a threshold of ‘no return’ and are engaged in a degradation phase where they acquire the typical onset of late apoptosis. Alterations in mitochondrial permeability transition linked to membrane potential disruption precede nuclear and plasma membrane changes. In vitro induction of permeability transition in isolated mitochondria provokes the release of a protein factor capable of inducing nuclear chromatin condensation and fragmentation. This permeability transition is regulated by multiple endogenous effectors, including members of the bcl‐2 gene family. Inhibition of these effects prevents apoptosis.
Labor scheduling using NPV estimates of the marginal benefit of additional labor capacity
Authors: Thompson Garry M.
Published: July 2002

An extensive literature exists on the problems of daily (shift) and weekly (tour) labor scheduling. In representing requirements for employees in these problems, researchers have used formulations based either on the model of Dantzig (1954) or on the model of Keith (1979). We show that both formulations have weakness in environments where management knows, or can attempt to identify, how different levels of customer servve affect profits. These weaknesses results in lower‐than‐necessary profits.

This paper presents a New Formulation of the daily and weekly Labor Scheduling Problems (NFLSP) designed to overcome the limitations of earlier models. NFLSP incorporates information on how changing the number of employees working in each planning period affects profits. NFLP uses this information during the development of the schedule to identify the number of employees who, ideally, should be working in each period. In an extensive simulation of 1,152 service environments, NFLSP outperformed the formulations of Dantzig (1954) and Keith (1979) at a level of significance of 0.001. Assuming year‐round operations and an hourly wage, including benefits, of $6.00, NFLSP's schedules were $96,046 (2.2%) and $24,648 (0.6%) more profitable, on average, than schedules developed using the formulations of Danzig (1954) and Keith (1979), respectively. Although the average percentage gain over Keith's model was fairly small, it could be much larger in some real cases with different parameters. In 73 and 100 percent of the cases wa simulated NFLSP yielded a higheprofit than the models of Keith (1979) and Danzig (1954), respectively.