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1.
Pediatr Pulmonol ; 57(2): 498-507, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-34779156

RESUMO

BACKGROUND: The objectives of this study were to analyze the clinical features and laboratory profiles and risk factors associated with critical illness of children with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). METHODS: One hundred and sixty-six coronavirus disease 2019 (COVID-19) Iranian pediatric patients were recruited through a collaborative research network between March and May 2020. Demographics, clinical, laboratory, and radiological results were obtained from patient files. RESULTS: Of 166 patients, 102 (61%) and 64 (39%) were males and females, respectively. Ninety-six (57.8%) and 70 (42.2%), had moderate and severe conditions, respectively. Thirty (18%) of patients died. The common symptoms were fever (73%), cough (54%), and shortness of breath, headache decrease in neutrophil and platelet counts; increase values in lactate dehydrogenase, decrease in the blood pH and HCO3 were significantly associated with the disease severity. 54% and 56% of patients showed abnormal radiographic appearance in Chest X-ray and in chest computed tomography scan, respectively. Sixty-one (36.7%) of patients were referred to intensive care unit (ICU). The coexistence of comorbidity was the main factor associated with ICU admission, shock, arrhythmia, acute kidney injury, acute respiratory distress syndrome, acute cardiac injury, and death. CONCLUSIONS: We describe a higher than previously recognized rate of COVID-19 mortality in Iranian pediatric patients. Epidemiological factors, such as the relatively high case fatality rate in the country and the presence of underlying diseases were the main factors for the high death rate.


Assuntos
COVID-19 , Criança , Criança Hospitalizada , Feminino , Humanos , Irã (Geográfico)/epidemiologia , Laboratórios , Masculino , Estudos Retrospectivos , SARS-CoV-2
2.
Ann Intern Med ; 157(7): 498-511, 2012 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-22893011

RESUMO

BACKGROUND: Debate continues about the comparative benefits and harms of first-generation antipsychotics (FGAs) and second-generation antipsychotics (SGAs) in treating schizophrenia. PURPOSE: To compare the effects of FGAs with those of SGAs in the treatment of adults aged 18 to 64 years with schizophrenia and related psychosis on illness symptoms, diabetes mellitus, mortality,tardive dyskinesia, and a major metabolic syndrome. DATA SOURCES: English-language studies from 10 electronic databases to March 2012, reference lists of relevant articles, and gray literature. STUDY SELECTION: Randomized trials for efficacy and cohort studies at least 2 years in duration for adverse events. DATA EXTRACTION: Two independent reviewers extracted data from 114 studies involving 22 comparisons and graded the strength of evidence for primary outcomes as insufficient, low, moderate, or high using the Grading of Recommendations Assessment, Development and Evaluation approach. DATA SYNTHESIS: Few differences of clinical importance were found for core illness symptoms; lack of precision in effect estimates precluded firm conclusions for many comparisons. Moderate-strength evidence showed a clinically important benefit of haloperidol over olanzapine for improving positive symptoms, but the benefit was scale-dependent: It was seen when the Scale for the Assessment of Positive Symptoms was used but not when the Positive and Negative Syndrome Scale (PANSS) was used. Moderate-strength evidence showed a clinically important benefit of olanzapine over haloperidol in improving negative symptoms when the PANSS and the Scale for the Assessment of Negative Symptoms were used. Low-strength evidence showed no difference in mortality for chlorpromazine verus clozapine or haloperidol versus aripiprazole,increased incidence of the metabolic syndrome for olanzapine versus haloperidol (risk differences, 2% and 22%), and higher incidence of tardive dyskinesia for chlorpromazine versus clozapine (risk differences, 5% and 9%). Evidence was insufficient to draw conclusions for diabetes mellitus. LIMITATIONS: All studies had high or unclear risk of bias. Length of study follow-up was often too brief to adequately measure adverse events. Medication comparisons, dosage, and outcome measurement were heterogenous for head-to-head comparisons. Selective patient populations limit generalizability. CONCLUSION: Clear benefits of FGAs versus SGAs for treating schizophrenia remain inconclusive because of variation in assessing outcomes and lack of clinically important differences for most comparisons. The strength of evidence on safety for major medical events is low or insufficient. PRIMARY FUNDING SOURCE: Agency for Healthcare Research and Quality.


Assuntos
Antipsicóticos/uso terapêutico , Esquizofrenia/tratamento farmacológico , Antipsicóticos/efeitos adversos , Antipsicóticos/classificação , Pesquisa Comparativa da Efetividade , Humanos , Adesão à Medicação , Recidiva , Indução de Remissão , Resultado do Tratamento
3.
Pediatrics ; 129(3): e771-84, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22351885

RESUMO

BACKGROUND AND OBJECTIVE: Despite increasing on-label and off-label use of antipsychotics, prescribing antipsychotics to children remains controversial due to uncertainty of their relative benefits and safety. We systematically reviewed the effectiveness and safety of first- (FGA) and second-generation antipsychotics (SGA) for patients aged ≤24 years with psychiatric and behavioral conditions. METHODS: We searched 10 databases from January 1987 to February 2011, gray literature, trial registries, and reference lists. Two reviewers independently selected studies, assessed methodologic quality, and graded the evidence. One reviewer extracted, and a second verified, data. We summarized findings qualitatively and conducted meta-analyses when appropriate. RESULTS: Sixty-four trials and 17 cohort studies were included. Most trials had a high risk of bias; cohort studies had moderate quality. All comparisons of FGAs versus SGAs, FGAs versus FGAs, and FGAs versus placebo had low or insufficient strength of evidence. There was moderate strength of evidence for the following comparisons. Olanzapine caused more dyslipidemia and weight gain, but fewer prolactin-related events, than risperidone. Olanzapine caused more weight gain than quetiapine. Compared with placebo, SGAs improved clinical global impressions (schizophrenia, bipolar and disruptive behavior disorders) and diminished positive and negative symptoms (schizophrenia), behavior symptoms (disruptive behavior disorders), and tics (Tourette syndrome). CONCLUSIONS: This is the first comprehensive review comparing the effectiveness and safety across the range of antipsychotics for children and young adults. The evidence on the comparative benefits and harms of antipsychotics is limited. Some SGAs have a better side effect profile than other SGAs. Additional studies using head-to-head comparisons are needed.


Assuntos
Antipsicóticos/uso terapêutico , Transtornos Mentais/diagnóstico , Transtornos Mentais/tratamento farmacológico , Adolescente , Fatores Etários , Antipsicóticos/efeitos adversos , Transtorno do Deficit de Atenção com Hiperatividade/diagnóstico , Transtorno do Deficit de Atenção com Hiperatividade/tratamento farmacológico , Estudos de Casos e Controles , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Transtornos do Humor/diagnóstico , Transtornos do Humor/tratamento farmacológico , Prognóstico , Ensaios Clínicos Controlados Aleatórios como Assunto , Medição de Risco , Esquizofrenia/diagnóstico , Esquizofrenia/tratamento farmacológico , Fatores Sexuais , Resultado do Tratamento , Adulto Jovem
4.
Ann Intern Med ; 155(4): 234-45, 2011 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-21844549

RESUMO

BACKGROUND: Pain management is integral to the management of hip fracture. PURPOSE: To review the benefits and harms of pharmacologic and nonpharmacologic interventions for managing pain after hip fracture. DATA SOURCES: 25 electronic databases (January 1990 to December 2010), gray literature, trial registries, and reference lists, with no language restrictions. STUDY SELECTION: Multiple reviewers independently and in duplicate screened 9357 citations to identify randomized, controlled trials (RCTs); nonrandomized, controlled trials (non-RCTs); and cohort studies of pain management techniques in older adults after acute hip fracture. DATA EXTRACTION: Independent, duplicate data extraction and quality assessment were conducted, with discrepancies resolved by consensus or a third reviewer. Data extracted included study characteristics, inclusion and exclusion criteria, participant characteristics, interventions, and outcomes. DATA SYNTHESIS: 83 unique studies (64 RCTs, 5 non-RCTs, and 14 cohort studies) were included that addressed nerve blockade (n = 32), spinal anesthesia (n = 30), systemic analgesia (n = 3), traction (n = 11), multimodal pain management (n = 2), neurostimulation (n = 2), rehabilitation (n = 1), and complementary and alternative medicine (n = 2). Overall, moderate evidence suggests that nerve blockades are effective for relieving acute pain and reducing delirium. Low-level evidence suggests that preoperative traction does not reduce acute pain. Evidence was insufficient on the benefits and harms of most interventions, including spinal anesthesia, systemic analgesia, multimodal pain management, acupressure, relaxation therapy, transcutaneous electrical neurostimulation, and physical therapy regimens, in managing acute pain. LIMITATIONS: No studies evaluated outcomes of chronic pain or exclusively examined participants from nursing homes or with cognitive impairment. Systemic analgesics (narcotics, nonsteroidal anti-inflammatory drugs) were understudied during the search period. CONCLUSION: Nerve blockade seems to be effective in reducing acute pain after hip fracture. Sparse data preclude firm conclusions about the relative benefits or harms of many other pain management interventions for patients with hip fracture. PRIMARY FUNDING SOURCE: Agency for Healthcare Research and Quality.


Assuntos
Fraturas do Quadril/complicações , Manejo da Dor , Acupressão , Analgésicos/uso terapêutico , Raquianestesia , Terapia Combinada , Pesquisa Comparativa da Efetividade , Delírio/etiologia , Delírio/prevenção & controle , Humanos , Bloqueio Nervoso , Dor/tratamento farmacológico , Dor/etiologia , Terapia de Relaxamento , Tração , Estimulação Elétrica Nervosa Transcutânea
5.
Neotrop Entomol ; 40(6): 653-60, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-23939271

RESUMO

The tribe Epiponini comprehends the swarm-founding Neotropical wasps, with several species endemic to Brazil, which are extremely important in studies of social evolution of wasps. The Epiponini diverge in several ways from the definitions of high eusociality, since caste syndromes range from species without morphological caste differentiation to those with complete caste dimorphism, and all species are polygynous. Frequently, indirect studies based on morphometry and physiology are the only solutions to collect data regarding the natural history and caste system in this tribe, since most species are extremely aggressive and build enveloped nests, usually in places of difficult access. We analyzed morphological parameters in seven colonies of the Epiponini species Polybia (Trichothorax) sericea Olivier in different phases of colonial development. Nine body variables were taken and females were classified according to their ovary development and spermathecal contents. The results showed that caste differences in this species are based on a contrast among variables: queens have larger mesosoma and abdomen, but are smaller in head width and wing length. These results suggest that morphological caste differentiation in this species is based mainly on body shape. We considered this combination of characters as being adaptive. We also showed that caste differences varied according to the colony cycle, with more conspicuous differences when queen number is reduced.


Assuntos
Comportamento Animal , Vespas/classificação , Animais , Feminino , Masculino , Vespas/anatomia & histologia
6.
Ann Intern Med ; 153(4): 246-55, 2010 Aug 17.
Artigo em Inglês | MEDLINE | ID: mdl-20621893

RESUMO

BACKGROUND: Many approaches exist for managing rotator cuff tears. PURPOSE: To compare the benefits and harms of nonoperative and operative interventions on clinically important outcomes in adults with rotator cuff tears. DATA SOURCES: 12 electronic databases (1990 to September 2009), gray literature, trial registries, and reference lists were searched. STUDY SELECTION: Controlled and uncontrolled studies that assessed nonoperative or operative treatments or postoperative rehabilitation for adults with confirmed rotator cuff tears were included. Operative studies in English-language publications and nonoperative and postoperative rehabilitation studies in English, French, or German were considered. Studies were assessed in duplicate. DATA EXTRACTION: 2 reviewers assessed risk for bias by using the Cochrane Risk of Bias tool and the Newcastle-Ottawa Scale. One reviewer rated the evidence by using a modified GRADE (Grading of Recommendations Assessment, Development, and Evaluation) approach. Data were extracted by one reviewer and verified by another. DATA SYNTHESIS: 137 studies met eligibility criteria. All trials had high risk for bias. Cohort and uncontrolled studies were of moderate quality. Reported functional outcomes did not differ between open versus mini-open repair, mini-open versus arthroscopic repair, arthroscopic repair with versus without acromioplasty, or single-row versus double-row fixation. Earlier return to work was reported for mini-open repair versus open repair and for continuous passive motion with physical therapy versus physical therapy alone. Open repairs showed greater improvement in function than did arthroscopic debridement. Complication rates were low across all interventions. LIMITATIONS: Limited evidence, which was often of low quality, precluded conclusions for most comparisons. Language restrictions may have excluded some relevant studies, and selective outcome reporting may have introduced bias. CONCLUSION: Evidence on the comparative effectiveness and harms of various operative and nonoperative treatments for rotator cuff tears is limited and inconclusive. PRIMARY FUNDING SOURCE: Agency for Healthcare Research and Quality.


Assuntos
Lesões do Manguito Rotador , Anti-Inflamatórios/uso terapêutico , Dexametasona/uso terapêutico , Humanos , Ácido Hialurônico/uso terapêutico , Procedimentos Ortopédicos/métodos , Modalidades de Fisioterapia , Complicações Pós-Operatórias , Amplitude de Movimento Articular , Projetos de Pesquisa/normas , Manguito Rotador/fisiologia , Manguito Rotador/cirurgia , Esteroides/uso terapêutico , Fatores de Tempo , Resultado do Tratamento , Viscossuplementos/uso terapêutico , Ferimentos e Lesões/cirurgia , Ferimentos e Lesões/terapia
7.
Biochem Soc Trans ; 32(Pt 2): 269-72, 2004 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-15046586

RESUMO

Methanopyrus kandleri is a hyperthermophilic methanogenic archaeon, which grows on H(2) and CO(2) as its sole energy source. Its growth temperature optimum is 98 degrees C. One of the interesting characteristics of this archaeon is its high intracellular salt content. The organism has been reported to contain the trianionic cDPG (cyclic 2,3-diphosphoglycerate) and K+ at concentrations of 1.1 and 3 M, respectively. Reflecting the high cellular salt concentration, the enzymes in this organism are adapted not only to high temperature but also to high salt concentrations. The formyltransferase from M. kandleri was characterized extensively with respect to thermo- and halophilicity. The crystal structure of the formyltransferase at 1.73 A shows the enzyme to be composed of four identical subunits of molecular mass 32 kDa. The formyltransferase is thermostable and active only at relatively high concentrations of potassium phosphate (1 M) or other salts with strongly hydrated anions (strong salting-out salts). Potassium phosphate and potassium cDPG were found to be equivalent in activating and stabilizing the enzyme. At low concentrations of these salts, the enzyme is inactive and thermolabile. It was shown by equilibrium sedimentation analysis that the enzyme is in a monomer/dimer/tetramer equilibrium, the equilibrium constant being dependent on the concentration of salts: the higher oligomeric species increase with increasing salt concentrations. Evidence was provided that the monomer is both inactive and thermolabile. Experiments using a mutation which is directed to break surface ion pairs between two dimers indicated that dimerization is required for activity and tetramerization leads to thermostability.


Assuntos
Archaea/enzimologia , Hidroximetil e Formil Transferases/química , Dióxido de Carbono , Divisão Celular , Cristalografia por Raios X , Dimerização , Relação Dose-Resposta a Droga , Hidroximetil e Formil Transferases/metabolismo , Modelos Moleculares , Mutação , Fosfatos/química , Fosfatos/farmacologia , Potássio/química , Compostos de Potássio/química , Compostos de Potássio/farmacologia , Estrutura Secundária de Proteína , Sais/farmacologia , Temperatura
8.
EMBO J ; 20(23): 6561-9, 2001 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-11726492

RESUMO

Cofactor F420 is a 5'-deazaflavin derivative first discovered in methanogenic archaea but later found also to be present in some bacteria. As a coenzyme, it is involved in hydride transfer reactions and as a prosthetic group in the DNA photolyase reaction. We report here for the first time on the crystal structure of an F420-dependent oxidoreductase bound with F420. The structure of F420H2:NADP+ oxidoreductase resolved to 1.65 A contains two domains: an N-terminal domain characteristic of a dinucleotide-binding Rossmann fold and a smaller C-terminal domain. The nicotinamide and the deazaflavin part of the two coenzymes are bound in the cleft between the domains such that the Si-faces of both face each other at a distance of 3.1 A, which is optimal for hydride transfer. Comparison of the structures bound with and without substrates reveals that of the two substrates NADP has to bind first, the binding being associated with an induced fit.


Assuntos
NADH NADPH Oxirredutases/química , NADP/química , Riboflavina/análogos & derivados , Riboflavina/química , Sítios de Ligação , Catálise , Domínio Catalítico , Desoxirribodipirimidina Fotoliase/metabolismo , Dimerização , Modelos Químicos , Modelos Moleculares , NADP/metabolismo , Ligação Proteica , Conformação Proteica , Dobramento de Proteína , Estrutura Terciária de Proteína , Especificidade por Substrato
9.
J Mol Biol ; 309(1): 315-30, 2001 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-11491299

RESUMO

Methyl-coenzyme M reductase (MCR) catalyzes the final reaction of the energy conserving pathway of methanogenic archaea in which methylcoenzyme M and coenzyme B are converted to methane and the heterodisulfide CoM-S-S-CoB. It operates under strictly anaerobic conditions and contains the nickel porphinoid F430 which is present in the nickel (I) oxidation state in the active enzyme. The known crystal structures of the inactive nickel (II) enzyme in complex with coenzyme M and coenzyme B (MCR-ox1-silent) and in complex with the heterodisulfide CoM-S-S-CoB (MCR-silent) were now refined at 1.16 A and 1.8 A resolution, respectively. The atomic resolution structure of MCR-ox1-silent describes the exact geometry of the cofactor F430, of the active site residues and of the modified amino acid residues. Moreover, the observation of 18 Mg2+ and 9 Na+ ions at the protein surface of the 300 kDa enzyme specifies typical constituents of binding sites for either ion. The MCR-silent and MCR-ox1-silent structures differed in the occupancy of bound water molecules near the active site indicating that a water chain is involved in the replenishment of the active site with water molecules. The structure of the novel enzyme state MCR-red1-silent at 1.8 A resolution revealed an active site only partially occupied by coenzyme M and coenzyme B. Increased flexibility and distinct alternate conformations were observed near the active site and the substrate channel. The electron density of the MCR-red1-silent state aerobically co-crystallized with coenzyme M displayed a fully occupied coenzyme M-binding site with no alternate conformations. Therefore, the structure was very similar to the MCR-ox1-silent state. As a consequence, the binding of coenzyme M induced specific conformational changes that postulate a molecular mechanism by which the enzyme ensures that methylcoenzyme M enters the substrate channel prior to coenzyme B as required by the active-site geometry. The three different enzymatically inactive enzyme states are discussed with respect to their enzymatically active precursors and with respect to the catalytic mechanism.


Assuntos
Metano/metabolismo , Methanobacterium/enzimologia , Oxirredutases/química , Oxirredutases/metabolismo , Sítios de Ligação , Catálise , Cloretos/metabolismo , Coenzimas/metabolismo , Cristalografia por Raios X , Íons/metabolismo , Ligantes , Magnésio/metabolismo , Modelos Moleculares , Oxirredução , Peptídeos/metabolismo , Maleabilidade , Ligação Proteica , Conformação Proteica , Subunidades Proteicas , Sódio/metabolismo , Solventes , Especificidade por Substrato , Temperatura , Zinco/metabolismo
10.
Appl Environ Microbiol ; 67(7): 3041-5, 2001 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-11425719

RESUMO

Recently it was reported that methanogens of the genus Methanobrevibacter exhibit catalase activity. This was surprising, since Methanobrevibacter species belong to the order Methanobacteriales, which are known not to contain cytochromes and to lack the ability to synthesize heme. We report here that Methanobrevibacter arboriphilus strains AZ and DH1 contained catalase activity only when the growth medium was supplemented with hemin. The heme catalase was purified and characterized, and the encoding gene was cloned. The amino acid sequence of the catalase from the methanogens is most similar to that of Methanosarcina barkeri.


Assuntos
Catalase/metabolismo , Heme/metabolismo , Methanobacteriaceae/enzimologia , Sequência de Aminoácidos , Catalase/química , Catalase/genética , Catalase/isolamento & purificação , Dados de Sequência Molecular , Análise de Sequência de DNA
12.
FEBS Lett ; 485(2-3): 200-4, 2000 Nov 24.
Artigo em Inglês | MEDLINE | ID: mdl-11094167

RESUMO

The hmd gene, which encodes the metal-free hydrogenase in methanogenic archaea, was heterologously expressed in Escherichia coli. The overproduced enzyme was completely inactive. High activity could, however, be induced by the addition of ultrafiltrate from active enzyme denatured in 8 M urea. The active fraction in the ultrafiltrate was heat-labile and migrated on gel filtration columns with an apparent molecular mass well below 1000 Da.


Assuntos
Euryarchaeota/enzimologia , Hidrogenase/química , Cromatografia em Gel , Estabilidade Enzimática , Escherichia coli/genética , Expressão Gênica , Temperatura Alta , Hidrogenase/genética , Hidrogenase/metabolismo , Peso Molecular , Desnaturação Proteica , Renaturação Proteica , Ultrafiltração , Ureia
13.
J Mol Biol ; 303(2): 329-44, 2000 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-11023796

RESUMO

The nickel enzyme methyl-coenzyme M reductase (MCR) catalyzes the terminal step of methane formation in the energy metabolism of all methanogenic archaea. In this reaction methyl-coenzyme M and coenzyme B are converted to methane and the heterodisulfide of coenzyme M and coenzyme B. The crystal structures of methyl-coenzyme M reductase from Methanosarcina barkeri (growth temperature optimum, 37 degrees C) and Methanopyrus kandleri (growth temperature optimum, 98 degrees C) were determined and compared with the known structure of MCR from Methanobacterium thermoautotrophicum (growth temperature optimum, 65 degrees C). The active sites of MCR from M. barkeri and M. kandleri were almost identical to that of M. thermoautotrophicum and predominantly occupied by coenzyme M and coenzyme B. The electron density at 1.6 A resolution of the M. barkeri enzyme revealed that four of the five modified amino acid residues of MCR from M. thermoautotrophicum, namely a thiopeptide, an S-methylcysteine, a 1-N-methylhistidine and a 5-methylarginine were also present. Analysis of the environment of the unusual amino acid residues near the active site indicates that some of the modifications may be required for the enzyme to be catalytically effective. In M. thermoautotrophicum and M. kandleri high temperature adaptation is coupled with increasing intracellular concentrations of lyotropic salts. This was reflected in a higher fraction of glutamate residues at the protein surface of the thermophilic enzymes adapted to high intracellular salt concentrations.


Assuntos
Adaptação Fisiológica , Substituição de Aminoácidos , Sequência Conservada , Cisteína/análogos & derivados , Euryarchaeota/enzimologia , Oxirredutases/química , Oxirredutases/metabolismo , Filogenia , Arginina/análogos & derivados , Arginina/metabolismo , Sítios de Ligação , Catálise , Cristalografia por Raios X , Cisteína/metabolismo , Meio Ambiente , Evolução Molecular , Glutamina/análogos & derivados , Glutamina/metabolismo , Glicina/metabolismo , Temperatura Alta , Ligação de Hidrogênio , Methanobacterium/enzimologia , Methanosarcina barkeri/enzimologia , Metilistidinas/metabolismo , Modelos Moleculares , Concentração Osmolar , Oxirredutases/genética , Oxirredutases/isolamento & purificação , Conformação Proteica , Dobramento de Proteína , Subunidades Proteicas , Solventes , Eletricidade Estática
14.
Arch Microbiol ; 174(3): 213-6, 2000 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-11041352

RESUMO

Methanosarcina barkeri is a methanogenic archaeon that can only grow under strictly anoxic conditions but which can survive oxidative stress. We have recently reported that the organism contains a monofunctional catalase. We describe here that it also possesses an active iron superoxide dismutase. The enzyme was purified in three steps over 130-fold in a 14% yield to a specific activity of 1500 U/mg. SDS-PAGE revealed the presence of only one band, at an apparent molecular mass of 25 kDa. The primary structure determined from the cloned and sequenced gene revealed similarity to iron- and manganese superoxide dismutases. The highest similarity was to the iron superoxide dismutase from Methanobacterium thermoautotrophicum. The enzyme from M. barkeri was found to contain, per mol, 1 mol iron, but no manganese in agreement with the general observation that anaerobically growing organisms only contain iron superoxide dismutase. The enzyme was not inhibited by cyanide (10 mM), which is a property shared by all iron- and manganese superoxide dismutases. The presence of superoxide dismutase in M. barkeri is noteworthy since a gene encoding superoxide dismutase (sod) has not been found in Archaeoglobus fulgidus, a sulfate-reducing archaeon most closely related to the Methanosarcinaceae.


Assuntos
Methanosarcina barkeri/enzimologia , Estresse Oxidativo/fisiologia , Superóxido Dismutase/isolamento & purificação , Superóxido Dismutase/metabolismo , Sequência de Aminoácidos , Clonagem Molecular , Meios de Cultura , Metanol/metabolismo , Methanosarcina barkeri/fisiologia , Dados de Sequência Molecular , Análise de Sequência de DNA , Superóxido Dismutase/química , Superóxido Dismutase/genética
15.
Eur J Biochem ; 267(22): 6619-23, 2000 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-11054114

RESUMO

Formyltransferase from Methanopyrus kandleri is composed of only one type of subunit of molecular mass 32 kDa. The enzyme is in a monomer/dimer/tetramer association equilibrium, the association constant being affected by lyotropic salts. Oligomerization is required for enzyme activity and thermostability. We report here on a subunit interface mutation (R261E) which affects the dimer/tetramer part of the association equilibrium of formyltransferase. With the mutant protein it was shown that tetramerization is not required for activity but is necessary for high thermostability.


Assuntos
Hidroximetil e Formil Transferases/química , Hidroximetil e Formil Transferases/metabolismo , Substituição de Aminoácidos , Archaea/enzimologia , Arginina , Dimerização , Estabilidade Enzimática , Ácido Glutâmico , Hidroximetil e Formil Transferases/genética , Cinética , Modelos Moleculares , Mutagênese Sítio-Dirigida , Conformação Proteica , Subunidades Proteicas , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Termodinâmica
16.
J Mol Biol ; 300(4): 935-50, 2000 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-10891279

RESUMO

Coenzyme F(420)-dependent methylenetetrahydromethanopterin reductase (Mer) is an enzyme of the Cl metabolism in methanogenic and sulfate reducing archaea. It is composed of identical 35-40 kDa subunits and lacks a prosthetic group. The crystal structure of Mer from Methanopyrus kandleri (kMer) revealed in one crystal form a dimeric and in another a tetrameric oligomerisation state and that from Methanobacterium thermoautotrophicum (tMer) a dimeric state. Each monomer is primarily composed of a TIM-barrel fold enlarged by three insertion regions. Insertion regions 1 and 2 contribute to intersubunit interactions. Insertion regions 2 and 3 together with the C-terminal end of the TIM-barrel core form a cleft where the binding sites of coenzyme F(420) and methylene-tetrahydromethanopterin are postulated. Close to the coenzyme F(420)-binding site lies a rarely observed non-prolyl cis-peptide bond. It is surprising that Mer is structurally most similar to a bacterial FMN-dependent luciferase which contains a non-prolyl cis-peptide bond at the equivalent position. The structure of Mer is also related to that of NADP-dependent FAD-harbouring methylenetetrahydrofolate reductase (MetF). However, Mer and MetF do not show sequence similarities although they bind related substrates and catalyze an analogous reaction.


Assuntos
Euryarchaeota/enzimologia , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/química , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/metabolismo , Riboflavina/análogos & derivados , Riboflavina/metabolismo , Adaptação Fisiológica , Sequência de Aminoácidos , Sítios de Ligação , Cristalografia por Raios X , Dimerização , Meio Ambiente , Flavina-Adenina Dinucleotídeo/química , Flavina-Adenina Dinucleotídeo/metabolismo , Flavinas/química , Flavinas/metabolismo , Methanobacterium/enzimologia , Modelos Moleculares , Dados de Sequência Molecular , Estrutura Quaternária de Proteína , Estrutura Secundária de Proteína , Riboflavina/química , Alinhamento de Sequência
17.
J Biol Chem ; 275(6): 3755-60, 2000 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-10660523

RESUMO

The global production of the greenhouse gas methane by methanogenic archaea reaches 1 billion tons per annum. The final reaction releasing methane is catalyzed by the enzyme methyl-coenzyme M reductase. The crystal structure of methyl-coenzyme M reductase from Methanobacterium thermoautotrophicum revealed the presence of five modified amino acids within the alpha-subunit and near the active site region. Four of these modifications were C-, N-, and S-methylations, two of which, 2-(S)-methylglutamine and 5-(S)-methylarginine, have never been encountered before. We have now confirmed these modifications by mass spectrometry of chymotryptic peptides. With methyl-coenzyme M reductase purified from cells grown in the presence of L-[methyl-D(3)]methionine, it was shown that the methyl groups of the modified amino acids are derived from the methyl group of methionine rather than from methyl-coenzyme M, an intermediate in methane formation. The D(3) labeling pattern was found to be qualitatively and quantitatively the same as in the two methyl groups of the methanogenic coenzyme F(430), which are known to be introduced via S-adenosylmethionine. From the results, it is concluded that the methyl groups of the modified amino acids in methyl-coenzyme M reductase are biosynthetically introduced by an S-adenosylmethionine-dependent post-translational modification. A mechanism for the methylation of glutamine at C-2 and of arginine at C-5 is discussed.


Assuntos
Aminoácidos/biossíntese , Methanobacterium/enzimologia , Oxirredutases/química , Sequência de Aminoácidos , Aminoácidos/química , Sítios de Ligação , Cromatografia Líquida de Alta Pressão , Quimotripsina , Glutamina/metabolismo , Espectrometria de Massas , Metaloporfirinas/química , Metano/metabolismo , Metionina/metabolismo , Metilação , Dados de Sequência Molecular , Estrutura Molecular , Fragmentos de Peptídeos/química , Processamento de Proteína Pós-Traducional , S-Adenosilmetionina/metabolismo , Análise de Sequência
18.
Surg Laparosc Endosc Percutan Tech ; 10(6): 404-8, 2000 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11147919

RESUMO

The treatment of hepatocellular carcinoma associated with liver cirrhosis necessitates local therapy in some patients because of severe hepatic dysfunction. Percutaneous ethanol injection therapy, the local therapy for such cancer of the liver, and percutaneous microwave coagulation therapy are detailed. The significant disadvantages of these procedures is their inability to evaluate precisely whether the tumor will develop complete necrosis after treatment because the cancer tissue cannot be excised with use of these procedures. Conversely, laparoscopic hepatectomy, which is minimally invasive surgery, has a disadvantage, that is, its difficulty in complex maneuvers, including hemostasis, ligation, and suture. The authors developed laparoscopic-assisted hepatectomy, which is hepatectomy by small incision during laparotomy with the use of laparoscopic observation. This report describes laparoscopic-assisted hepatectomy, which may allow the solving of problems with percutaneous ethanol injection therapy, percutaneous microwave coagulation therapy, and laparoscopic hepatectomy.


Assuntos
Carcinoma Hepatocelular/cirurgia , Hepatectomia/métodos , Laparoscopia/métodos , Neoplasias Hepáticas/cirurgia , Recidiva Local de Neoplasia/cirurgia , Idoso , Carcinoma Hepatocelular/complicações , Carcinoma Hepatocelular/patologia , Hepatectomia/efeitos adversos , Hepatectomia/instrumentação , Humanos , Laparoscopia/efeitos adversos , Cirrose Hepática/complicações , Neoplasias Hepáticas/complicações , Neoplasias Hepáticas/patologia , Masculino , Recidiva Local de Neoplasia/complicações , Recidiva Local de Neoplasia/patologia , Seleção de Pacientes , Escleroterapia , Tomografia Computadorizada por Raios X , Resultado do Tratamento
19.
Structure ; 7(10): 1257-68, 1999 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-10545331

RESUMO

BACKGROUND: The reduction of carbon dioxide to methane in methanogenic archaea involves the tetrahydrofolate analogue tetrahydromethanopterin (H(4)MPT) as a C(1) unit carrier. In the third step of this reaction sequence, N(5)-formyl-H(4)MPT is converted to methenyl-H(4)MPT(+) by the enzyme methenyltetrahydromethanopterin cyclohydrolase. The cyclohydrolase from the hyperthermophilic archaeon Methanopyrus kandleri (Mch) is extremely thermostable and adapted to a high intracellular concentration of lyotropic salts. RESULTS: Mch was crystallized and its structure solved at 2.0 A resolution using a combination of the single isomorphous replacement (SIR) and multiple anomalous dispersion (MAD) techniques. The structure of the homotrimeric enzyme reveals a new alpha/beta fold that is composed of two domains forming a large sequence-conserved pocket between them. Two phosphate ions were found in and adjacent to this pocket, respectively; the latter is displaced by the phosphate moiety of the substrate formyl-H(4)MPT according to a hypothetical model of the substrate binding. CONCLUSIONS: Although the exact position of the substrate is not yet known, the residues lining the active site of Mch could be tentatively assigned. Comparison of Mch with the tetrahydrofolate-specific cyclohydrolase/dehydrogenase reveals similarities in domain arrangement and in some active-site residues, whereas the fold appears to be different. The adaptation of Mch to high salt concentrations and high temperatures is reflected by the excess of acidic residues at the trimer surface and by the higher oligomerization state of Mch compared with its mesophtic counterparts.


Assuntos
Aminoidrolases/química , Euryarchaeota/enzimologia , Sequência de Aminoácidos , Aminoidrolases/genética , Aminoidrolases/metabolismo , Domínio Catalítico/genética , Cristalografia por Raios X , Euryarchaeota/genética , Temperatura Alta , Modelos Moleculares , Dados de Sequência Molecular , Conformação Proteica , Estrutura Quaternária de Proteína , Estrutura Terciária de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Homologia de Sequência de Aminoácidos , Eletricidade Estática
20.
J Affect Disord ; 54(1-2): 161-9, 1999 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-10403159

RESUMO

BACKGROUND: The present study investigated the structure of depressive symptoms in the perinatal period. METHOD: The Zung Self-Rating Depression Scale (SDS) was administered to a total of 1329 women in early, middle and late pregnancy and 5 days, 1 month, 6 months, 12 months and 18 months after the delivery. RESULTS: A number of somatic items and the suicidal ideation item of the SDS made low contributions to the evaluation of the severity of depression, and as a consequence these were excluded in the principal component analysis. Three factors were interpretable as "Cognitive", "Affective insomnia" and "Attentional" emerged at all eight assessment points. The goodness-of-fit index (GFI) generated by confirmatory factor analyses (LISREL 7.20) proved sufficiently high on all eight occasions. LIMITATION: The present study investigated only one self-rating scale and the sample comprised Japanese mothers only. CONCLUSION: The three-factor model of the SDS in the perinatal period was derived from exploratory and confirmatory factor analyses. It is noteworthy that the same three-factor structure emerged at all eight collection points in the present study.


Assuntos
Depressão Pós-Parto/psicologia , Período Pós-Parto/psicologia , Adolescente , Adulto , Análise Fatorial , Feminino , Seguimentos , Humanos , Gravidez , Autoavaliação (Psicologia) , Inquéritos e Questionários , Fatores de Tempo
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