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1.
J Environ Manage ; 301: 113880, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34638042

RESUMO

The dairy industry produces vast quantities of dairy processing sludge (DPS), which can be processed further to develop second generation products such as struvite, biochars and ashes (collectively known as STRUBIAS). These bio-based fertilizers have heterogeneous nutrient and metal contents, resulting in a range of possible application rates. To avoid nutrient losses to water or bioaccumulation of metals in soil or crops, it is important that rates applied to land are safe and adhere to the maximum legal application rates similar to inorganic fertilizers. This study collected and analysed nutrient and metal content of all major DPS (n = 84) and DPS-derived STRUBIAS products (n = 10), and created an application calculator in MS Excel™ to provide guidance on maximum legal application rates for ryegrass and spring wheat across plant available phosphorus (P) deficient soil to P-excess soil. The sample analysis showed that raw DPS and DPS-derived STRUBIAS have high P contents ranging from 10.1 to 122 g kg-1. Nitrogen (N) in DPS was high, whereas N concentrations decreased in thermo-chemical STRUBIAS products (chars and ash) due to the high temperatures used in their formation. The heavy metal content of DPS and DPS-derived STRUBIAS was significantly lower than the EU imposed limits. Using the calculator, application rates of DPS and DPS-derived STRUBIAS materials (dry weight) ranged from 0 to 4.0 tonnes ha-1 y-1 for ryegrass and 0-4.5 tonnes ha-1 y-1 for spring wheat. The estimated heavy metal ingestion to soil annually by the application of the DPS and DPS-derived STRUBIAS products was lower than the EU guideline on soil metal accumulation. The calculator is adaptable for any bio-based fertilizer, soil and crop type, and future work should continue to characterise and incorporate new DPS and DPS-derived STRUBIAS products into the database presented in this paper. In addition, safe application rates pertaining to other regulated pollutants or emerging contaminants that may be identified in these products should be included. The fertilizer replacement value of these products, taken from long-term field studies, should be factored into application rates.


Assuntos
Agricultura , Metais Pesados , Fertilizantes/análise , Metais Pesados/análise , Fósforo , Esgotos , Solo
2.
AJNR Am J Neuroradiol ; 42(6): 1080-1086, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33737270

RESUMO

BACKGROUND AND PURPOSE: Despite high interest in machine-learning algorithms for automated segmentation of MRIs of patients with brain tumors, there are few reports on the variability of segmentation results. The purpose of this study was to obtain benchmark measures of repeatability for a widely accessible software program, BraTumIA (Versions 1.2 and 2.0), which uses a machine-learning algorithm to segment tumor features on contrast-enhanced brain MR imaging. MATERIALS AND METHODS: Automatic segmentation of enhancing tumor, tumor edema, nonenhancing tumor, and necrosis was performed on repeat MR imaging scans obtained approximately 2 days apart in 20 patients with recurrent glioblastoma. Measures of repeatability and spatial overlap, including repeatability and Dice coefficients, are reported. RESULTS: Larger volumes of enhancing tumor were obtained on later compared with earlier scans (mean, 26.3 versus 24.2 mL for BraTumIA 1.2; P < .05; and 24.9 versus 22.9 mL for BraTumIA 2.0, P < .01). In terms of percentage change, repeatability coefficients ranged from 31% to 46% for enhancing tumor and edema components and from 87% to 116% for nonenhancing tumor and necrosis. Dice coefficients were highest (>0.7) for enhancing tumor and edema components, intermediate for necrosis, and lowest for nonenhancing tumor and did not differ between software versions. Enhancing tumor and tumor edema were smaller, and necrotic tumor larger using BraTumIA 2.0 rather than 1.2. CONCLUSIONS: Repeatability and overlap metrics varied by segmentation type, with better performance for segmentations of enhancing tumor and tumor edema compared with other components. Incomplete washout of gadolinium contrast agents could account for increasing enhancing tumor volumes on later scans.


Assuntos
Neoplasias Encefálicas , Glioblastoma , Algoritmos , Neoplasias Encefálicas/diagnóstico por imagem , Glioblastoma/diagnóstico por imagem , Humanos , Processamento de Imagem Assistida por Computador , Imageamento por Ressonância Magnética , Carga Tumoral
3.
Dis Esophagus ; 28(3): 283-90, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24529029

RESUMO

The association between Barrett's esophagus (BE) and a personal or family history of cancer other than gastroesophageal remains unknown. To evaluate the effect of personal and family history of certain cancers and cancer treatments on the risk of BE, we analyzed data from a Veterans Affairs case-control study that included 264 men with definitive BE (cases) and 1486 men without BE (controls). Patients with history of esophageal or gastric cancer were excluded. Patients underwent elective esophagogastroduodenoscopy or a study esophagogastroduodenoscopy concurrently with screening colonoscopy to determine BE status. Personal and family history of several types of cancer was obtained from self-reported questionnaires, supplemented and verified by electronic medical-record reviews. We estimated the association between personal and family history of cancer or radiation/chemotherapy, and BE. Personal history of oropharyngeal cancer (1.5% vs. 0.4%) or prostate cancer (7.2% vs. 4.4%) was more frequently present in cases than controls. The association between BE and prostate cancer persisted in multivariable analyses (adjusted odds ratio 1.90; 95% confidence interval 1.07-3.38, P = 0.028) while that with oropharyngeal cancer (adjusted odds ratio 3.63; 95% confidence interval 0.92-14.29, P = 0.066) was attenuated after adjusting for retained covariates of age, race, gastroesophageal reflux disease, hiatal hernia, and proton pump inhibitor use. Within the subset of patients with cancer, prior treatment with radiation or chemotherapy was not associated with BE. There were no significant differences between cases and controls in the proportions of subjects with several specific malignancies in first- or second-degree relatives. In conclusion, the risk of BE in men may be elevated with prior personal history of oropharyngeal or prostate cancer. However, prior cancer treatments and family history of cancer were not associated with increased risk of BE. Further studies are needed to elucidate if there is a causative relationship or shared risk factors between prostate cancer and BE.


Assuntos
Esôfago de Barrett/etiologia , Neoplasias/complicações , Idoso , Esôfago de Barrett/genética , Estudos de Casos e Controles , Colonoscopia , Endoscopia do Sistema Digestório , Família , Predisposição Genética para Doença , Nível de Saúde , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias/genética , Neoplasias Orofaríngeas/complicações , Neoplasias Orofaríngeas/genética , Neoplasias da Próstata/complicações , Neoplasias da Próstata/genética , Fatores de Risco , Autorrelato , Estados Unidos , United States Department of Veterans Affairs
4.
J Med Virol ; 86(2): 224-34, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24166351

RESUMO

Hepatitis C virus subgenotypes 1a and 1b are found worldwide and cause 60% of all hepatitis C cases. It has been reported recently that viral genetic variations have a critical impact on the patient treatment outcome. In particular, polymorphisms of the HCV core protein have been linked to poor treatment response. However, most of these studies were conducted on Asian populations, Japanese in particular who are infected with HCV subgenotype 1b. Hence, we aimed in this study to examine the core protein polymorphisms in Saudi patients who are infected with chronic HCV genotype 1 (1a and 1b subtypes) and its association with treatment outcome. Direct sequencing of full-length core protein and data mining analyses were utilized. Results have shown that the response to treatment is dependent on subgenotypes. Indeed, HCV-1b showed different point mutations that are associated with treatment outcome where the point mutations at positions 70 (Arg(70) Gln) and 75 (Thr(75) Ala) in HCV-1b are significantly associated with PEG-IFN/RBV treatment response. In contrast, HCV-1a showed no significant association between core protein mutations and response to treatment. In addition, analyses of HCV-1a core protein sequences revealed a highly conserved region especially in the responder group. This study provides a new insight in the genetic variability of full-length core protein in HCV genotype 1 in Saudi infected patients.


Assuntos
Antivirais/uso terapêutico , Variação Genética , Hepacivirus/efeitos dos fármacos , Hepatite C Crônica/tratamento farmacológico , Interferons/uso terapêutico , Ribavirina/uso terapêutico , Proteínas do Core Viral/genética , Adulto , Biologia Computacional , Feminino , Genótipo , Hepacivirus/genética , Hepatite C Crônica/virologia , Humanos , Masculino , Pessoa de Meia-Idade , Mutação Puntual , Arábia Saudita , Análise de Sequência de DNA , Resultado do Tratamento
5.
Psychol Rep ; 111(1): 47-63, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23045847

RESUMO

The purpose of this study was to examine the psychometric properties of the adapted Irrational Beliefs Inventory (IBI-34) and thus begin the process of assessing its adequacy for use in an Arab culture. The scale was translated and then administered to two samples of undergraduate students from the United Arab Emirates University. Data from 384 students were used in the main analysis, and data from 251 students were used for cross-validation. Principal components analysis (PCA) with varimax rotation followed by PCA with oblimin rotation yielded the same five components in both the main sample and the validation sample, thus consistent with the original Dutch study. Only 34 of the original 50 items were adequate to represent the five constructs. Cronbach's alpha coefficient for the overall scale was .76 and for the subscales ranged between .71 and .76, except for the Rigidity subscale, which was .54. The adapted IBI-34 correlated significantly and negatively with the General Health Questionnaire and Beck Depression Inventory, providing support for concurrent validity. Due to the non-significant differences between male and female participants on the total score of the IBI-34, the scale can be used for both sexes by summing across all items to give a total score that can be used as a general indicator of the irrational thinking.


Assuntos
Árabes/psicologia , Comparação Transcultural , Cultura , Inventário de Personalidade/estatística & dados numéricos , Teste de Realidade , Percepção Social , Adolescente , Adulto , Mecanismos de Defesa , Feminino , Humanos , Individualidade , Masculino , Psicometria/estatística & dados numéricos , Reprodutibilidade dos Testes , Fatores Sexuais , Estudantes/psicologia , Emirados Árabes Unidos , Adulto Jovem
6.
J Surg Educ ; 69(4): 544-9, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22677595

RESUMO

BACKGROUND: Literature suggests declining interest in General Surgery (GS) and other surgical specialties, with fewer Canadian medical residency applicants identifying a surgical specialty as their first choice. Although perceptions of surgical careers may begin before enrollment in clerkship, clerkship itself provides the most concentrated environment for perceptions to evolve. Most students develop perceptions about specialties during their clinical clerkships. This study examines the immediate impact of GS clerkship on student attitudes toward GS as a career, and on preferences towards GS compared with other specialties. METHODS: A pre-post design involved 61 McMaster clinical clerks. Two instruments were used to collect data from students over the course of clerkship (2008-2009). Paired comparison (PC) compared ranking of career choices before and after clerkship. Semantic differential (SD) measured attitudes toward GS and variables that may have affected attitudes before and after clerkship. Analyses used SPSS 16.0 (SPSS Inc., Chicago, IL). RESULTS: Clerks ranked preferences for GS changed substantially after clerkship, moving from the 10th to the 5th position compared with other specialties. Ranks of surgical subspecialties also changed, though GS demonstrated the largest improvement. SD results were consistent with PC, showing improved attitudes after rotation, with differences both statistically and practically significant (t = 3.81, p < 0.000, effect size = 0.23). Results indicated that attitudes toward all areas related to GS clerkship (attending physicians, surgical residents, ward nurses, scrub nurses, workload, knowledge achieved, technical skills acquired) improved significantly except attitude toward technical skills acquired. CONCLUSIONS: Clinical clerkship at McMaster was a positive experience and significantly enhanced preferences towards GS and attitudes towards GS as a career. Medical schools should foster positive interaction between clinical clerks and staff (including attending surgeons and nurses), ensure that teaching hospital staff provide a positive experience for clerks, and should provide opportunities to learn basic technical skills during GS clerkship.


Assuntos
Estágio Clínico/organização & administração , Competência Clínica , Educação de Graduação em Medicina/organização & administração , Cirurgia Geral/educação , Adulto , Atitude do Pessoal de Saúde , Escolha da Profissão , Avaliação Educacional , Feminino , Previsões , Hospitais Universitários , Humanos , Masculino , Ontário , Desenvolvimento de Programas , Avaliação de Programas e Projetos de Saúde , Autoavaliação (Psicologia) , Estudantes de Medicina/psicologia , Estudantes de Medicina/estatística & dados numéricos , Adulto Jovem
7.
Genet Mol Res ; 11(2): 855-62, 2012 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-22576913

RESUMO

Besides the host immune response, genetic and environmental factors play crucial roles in the manifestation of hepatitis B virus (HBV) infection. "Regulated on activation normal T-cell expressed and secreted" factor (RANTES) plays a vital role in CD4(+), CD8(+) T-lymphocyte and dendritic cell activation and proliferation in inflammation. Single nucleotide polymorphisms (SNPs) in the RANTES gene are associated with several viral and non-viral diseases. Association studies have invariably indicated a lack of association between RANTES gene SNPs and HBV infection in ethnic populations, even though RANTES gene SNPs exhibit distinct ethnic distributions. Despite the high prevalence of HBV infections in Saudi Arabia, no studies have been made concerning a possible relationship between RANTES gene polymorphisms and susceptibility to and progression of HBV infection. We examined -403G>A and -28C>G RANTES gene variants in 473 healthy controls and 484 HBV patients in ethnic Saudi populations. Significant differences were found in the genotype and allele distributions of the SNPs between the controls and the HBV patients. Both SNPs were significantly linked to viral clearance in these subjects. Our data demonstrate for the first time in a Saudi population, a relationship between the RANTES gene polymorphisms and the clinical course of HBV infection and underscore the importance of evaluating the genetic background of the affected individual to determine how it may affect disease progression.


Assuntos
Quimiocina CCL5/genética , Hepatite B/genética , Polimorfismo de Nucleotídeo Único , Sequência de Bases , Primers do DNA , Feminino , Genética Populacional , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Arábia Saudita
8.
J Egypt Soc Parasitol ; 33(3): 679-86, 2003 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-14708845

RESUMO

Anchitrema sanguineum and Prosthodendrium (Prosthodendrium) urna are two digenean trematodes infecting many species of bats in Egypt. The surface topography of them was studied by scanning electron microscopy. Examination of A. sanguineum revealed the presence of posteriorly directed broadbase spines allover the body. The oral sucker is bordered by several small sucker-like papillae and few irregularly distributed pores. The ventral sucker is smaller than the oral one and surrounded by several papillae. In P. (P.) urna the body is covered with simple, spines posteriorly directed. The oral sucker has few papillae and its tegumental rim slightly elevated. The ventral sucker is slightly larger than the oral one.


Assuntos
Quirópteros/parasitologia , Trematódeos/ultraestrutura , Infecções por Trematódeos/veterinária , Animais , Egito , Microscopia Eletrônica de Varredura/métodos , Microscopia Eletrônica de Varredura/veterinária , Trematódeos/anatomia & histologia , Infecções por Trematódeos/parasitologia
9.
Acta Neurochir (Wien) ; 140(11): 1121-5, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-9870056

RESUMO

In this study, the authors attempt to provide an account of the factors that affect the outcome of hydrocephaly in 26 foetuses. The hydrocephalus was related to a myelomeningocele in 35% of patients. Sixty-two percent of foetuses showed intra-uterine progression of their hydrocephalus and 50% were shunted postnatally. At a mean follow up of 2 years, the outcome was considered "fair" in 54% of patients. Our findings demonstrate that in foetal hydrocephaly a more favourable outcome is expected in patients with hydrocephalus which does not progress in utero, in whom the labour is not induced before 36 weeks of gestation, who are delivered vaginally weighing more than 2.5 kg and have a head circumference below the 95th centile and a CT cortical mantle thickness of 2 cm and more and who are treated by CSF shunting. The diagnosis of the foetal hydrocephaly at or before 28 weeks of gestation and the presence of a myelomeningocele did not affect the outcome significantly. Neurosurgeons are reminded to keep an open mind for infants with foetal hydrocephaly and to offer active treatment to patients with a potentially favourable outcome.


Assuntos
Hidrocefalia/diagnóstico , Peso ao Nascer , Pré-Escolar , Progressão da Doença , Feminino , Seguimentos , Idade Gestacional , Humanos , Hidrocefalia/etiologia , Hidrocefalia/cirurgia , Lactente , Recém-Nascido , Masculino , Meningomielocele/complicações , Meningomielocele/diagnóstico , Meningomielocele/cirurgia , Gravidez , Tomografia Computadorizada por Raios X , Resultado do Tratamento , Ultrassonografia Pré-Natal , Derivação Ventriculoperitoneal
10.
J Biol Chem ; 268(11): 7646-9, 1993 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-8385115

RESUMO

The molecular events that lead from the interaction of insulin with its receptor to the activation of protein serine/threonine kinases are still unknown. In this study, we have examined the role of GTP-binding proteins in this signaling pathway using differentiated 3T3-L1 adipocytes permeabilized with alpha-toxin from Staphylococcus aureus. Addition of GTP gamma S (guanosine 5'-O-(3-thiotriphosphate)) or insulin to such permeabilized cells markedly increases protein kinase activities in cell lysates using the microtubule-associated protein-2 kinase substrate peptide KRELVE-PLTPSGEAPNQALLR, which contains the threonine 669 phosphorylation site on the epidermal growth factor receptor. Similar stimulations of protein kinase activity by these agents are observed using the peptide KRRRLASLAA, which is selectively phosphorylated by ribosomal protein S6 kinases. The effects of insulin and GTP gamma S are not additive. Importantly, the GTP-binding protein antagonist GDP beta S (guanosine 5'-O-(2-thiodiphosphate)) inhibits the activation of the protein kinase activities by insulin in permeabilized 3T3-L1 adipocytes. These data are consistent with the hypothesis that activation of Ras or other GTP-binding proteins is a key element of the signaling mechanism whereby insulin receptor tyrosine kinase activates the microtubule-associated protein-2 kinase cascade.


Assuntos
Tecido Adiposo/enzimologia , Guanosina Trifosfato/análogos & derivados , Guanosina Trifosfato/farmacologia , Insulina/farmacologia , Proteínas Quinases/metabolismo , Células 3T3 , Sequência de Aminoácidos , Animais , Proteínas Quinases Dependentes de Cálcio-Calmodulina , Permeabilidade da Membrana Celular , Enterotoxinas , Ativação Enzimática , Guanosina 5'-O-(3-Tiotrifosfato)/farmacologia , Guanosina Difosfato/análogos & derivados , Guanosina Difosfato/farmacologia , Guanilil Imidodifosfato/farmacologia , Cinética , Camundongos , Dados de Sequência Molecular , Peptídeos/metabolismo , Staphylococcus aureus , Especificidade por Substrato , Tionucleotídeos/farmacologia
11.
J Biol Chem ; 266(7): 4052-5, 1991 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-1847913

RESUMO

Insulin action leads to the rapid stimulation of a cytosolic Kemptide (Leu-Arg-Arg-Ala-Ser-Leu-Gly) kinase (KIK) that has been recently purified to near homogeneity (Klarlund, J. K., Bradford, A. P., Milla, M. G., and Czech, M. P. (1990) J. Biol. Chem. 265, 227-234). To examine its activation mechanism, purified KIK was treated with purified protein phosphatases. The catalytic subunit of phosphatase 2A inhibited the activity of control KIK by about 50% and abolished the 5-fold elevation in KIK activity due to insulin action. The catalytic subunit of phosphatase 1 with equivalent activity based on dephosphorylation of 32P-labeled phosphorylase alpha had no effect on either control or insulin-stimulated KIK activity. The deactivation of insulin-stimulated KIK by phosphatase 2A was time- and concentration-dependent and was blocked by phosphatase inhibitors. The purified native complexes of phosphatase 2A, phosphatase 2A1, and phosphatase 2A2 similarly deactivated KIK. Analyis of control or insulin-stimulated KIK with two antiphosphotyrosine antibodies by immunoblotting and immunoprecipitation failed to detect the presence of phosphotyrosine in the kinase. These results indicate that KIK is activated by phosphorylation as part of a kinase cascade emanating from insulin receptor stimulation.


Assuntos
Fígado/enzimologia , Oligopeptídeos/metabolismo , Fosfoproteínas Fosfatases/metabolismo , Proteínas Quinases/metabolismo , Sequência de Aminoácidos , Animais , Ativação Enzimática/efeitos dos fármacos , Insulina/farmacologia , Cinética , Masculino , Dados de Sequência Molecular , Oligopeptídeos/química , Fosfoproteínas/metabolismo , Proteína Fosfatase 1 , Proteína Fosfatase 2 , Ratos , Ratos Endogâmicos
12.
Biochem J ; 268(3): 539-45, 1990 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-2114095

RESUMO

Purified rat liver ATP citrate-lyase is phosphorylated on serine residues by an insulin-stimulated cytosolic kinase activity partially purified from rat adipocytes [Yu, Khalaf & Czech (1987) J. Biol. Chem. 262, 16677-16685]. The Km for lyase phosphorylation by this hormone-sensitive kinase activity is approx. 3 microM. Two-dimensional tryptic-peptide mapping of the 32P-labelled lyase reveals that the kinase-catalysed phosphorylation occurs primarily on a specific peptide. In intact 32P-labelled adipocytes, insulin enhances the serine phosphorylation of ATP citrate-lyase by 2-3-fold. Tryptic digestion of the 32P-labelled lyase immunopurified from insulin-treated adipocytes also yields one major phosphopeptide. 32P-labelled lyase tryptic peptides derived from labelling experiments in vitro and in vivo exhibit identical electrophoretic and chromatographic migration profiles. Furthermore, radio-sequencing of the phosphopeptide from lyase 32P-labelled in vitro indicates that serine-3 from the N-terminus is phosphorylated by the insulin-stimulated cytosolic kinase, in agreement with previous studies on the position of the phosphoserine residue in ATP citrate-lyase isolated from insulin-treated cells. Taken together, the similarity in site-specific phosphorylation of ATP citrate-lyase from insulin-treated adipocytes to that catalysed by the hormone-activated cytosolic kinase in vitro strongly suggests that this kinase mediates insulin action on lyase phosphorylation in intact cells.


Assuntos
ATP Citrato (pro-S)-Liase/metabolismo , Insulina/farmacologia , Proteínas Quinases/metabolismo , Tecido Adiposo/citologia , Tecido Adiposo/enzimologia , Animais , Epididimo/enzimologia , Técnicas In Vitro , Masculino , Fragmentos de Peptídeos/metabolismo , Mapeamento de Peptídeos , Fosforilação , Ratos , Ratos Endogâmicos , Serina/metabolismo
13.
Arzneimittelforschung ; 39(5): 612-4, 1989 May.
Artigo em Alemão | MEDLINE | ID: mdl-2667525

RESUMO

The effects of three oral doses of nifedipine 20 mg or nifedipine 20 mg/dihydroergotoxin 2 mg (Pontuc, HN 85) during three subsequent days on an oral glucose load (100 g) were compared to placebo. Neither drug altered the glucose load or inhibited the secretion of insulin or C-peptide. The fall of serum potassium was also identical to controls. Basal plasma norepinephrine concentrations were lower following nifedipine/dihydroergotoxine than after nifedipine alone (2 p less than 0.05). A decrease of the serum sodium concentration by 2 mmol/l was observed with nifedipine but not with the combined drugs.


Assuntos
Di-Hidroergotoxina/farmacologia , Eletrólitos/metabolismo , Glucose/metabolismo , Nifedipino/farmacologia , Adulto , Peptídeo C/sangue , Di-Hidroergotoxina/administração & dosagem , Teste de Tolerância a Glucose , Frequência Cardíaca/efeitos dos fármacos , Humanos , Insulina/sangue , Masculino , Nifedipino/administração & dosagem , Norepinefrina/sangue , Renina/sangue
14.
J Biol Chem ; 262(34): 16677-85, 1987 Dec 05.
Artigo em Inglês | MEDLINE | ID: mdl-2960679

RESUMO

The cytosolic fraction of insulin-treated adipocytes exhibits a 2-fold increase in protein kinase activity when Kemptide is used as a substrate. The detection of insulin-stimulated kinase activity is critically dependent on the presence of phosphatase inhibitors such as fluoride and vanadate in the cell homogenization buffer. The cytosolic protein kinase activity exhibits high sensitivity (ED50 = 2 X 10(-10) M) and a rapid response (maximal after 2 min) to insulin. Kinetic analyses of the cytosolic kinase indicate that insulin increases the Vmax of Kemptide phosphorylation and ATP utilization without affecting the affinities of this enzyme toward the substrate or nucleotide. Upon chromatography on anion-exchange and gel filtration columns, the insulin-stimulated cytosolic kinase activity is resolved from the cAMP-dependent protein kinase and migrates as a single peak with an apparent Mr = 50,000-60,000. The partially purified kinase preferentially utilizes histones, Kemptide, multifunctional calmodulin-dependent protein kinase substrate peptide, ATP citrate-lyase, and acetyl-coenzyme A carboxylase as substrates but does not catalyze phosphorylation of ribosomal protein S6, casein, phosvitin, phosphorylase b, glycogen synthase, inhibitor II, and substrate peptides for casein kinase II, protein kinase C, and cGMP-dependent protein kinase. Phosphoamino acid analyses of the 32P-labeled substrates reveal that the insulin-stimulated cytosolic kinase is primarily serine-specific. The insulin-activated cytosolic kinase prefers Mn2+ to Mg2+ and is independent of Ca2+. Unlike ribosomal protein S6 kinase and protease-activated kinase II, the insulin-sensitive cytosolic kinase is fluoride-insensitive. Taken together, these results indicate that a novel cytosolic protein kinase activity is activated by insulin.


Assuntos
Tecido Adiposo/enzimologia , Insulina/farmacologia , Manganês/metabolismo , Proteínas Quinases/metabolismo , Animais , Cromatografia por Troca Iônica , Citosol/enzimologia , Ditiotreitol/farmacologia , Relação Dose-Resposta a Droga , Heparina/farmacologia , Concentração de Íons de Hidrogênio , Anticorpos Anti-Insulina , Masculino , Oligopeptídeos/metabolismo , Proteínas Serina-Treonina Quinases , Ratos , Ratos Endogâmicos , Especificidade por Substrato , Temperatura
15.
J Biol Chem ; 262(16): 7865-73, 1987 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-2438282

RESUMO

The action of insulin on tyrosine phosphorylation of plasma membrane-associated proteins in rat adipocytes was investigated. Incubation of plasma membranes from insulin-treated adipocytes with [gamma-32P] ATP results in a marked increase in tyrosine phosphorylation of Mr = 160,000 (P160) and Mr = 92,000 proteins when compared to controls. Based on the immunoreactivities of these two proteins with anti-insulin receptor antibodies, the Mr = 92,000 species is identified as the insulin receptor beta subunit while P160 is unrelated to the receptor structure. P160 appears to be a glycoprotein as evidenced by its adsorption to wheat germ agglutinin-agarose. The tyrosine phosphorylation of P160 exhibits a rapid response to insulin (maximal within 2 min at 37 degrees C) and is readily reversed following removal of the free hormone by anti-insulin serum. The time courses of insulin-stimulated phosphorylation as well as the dephosphorylation of P160 coincide with those of the activation and deactivation of the insulin receptor kinase in the same plasma membrane preparation. Concanavalin A and hydrogen peroxide mimic insulin stimulation of the insulin receptor kinase and enhance the tyrosine phosphorylation of P160. Isoproterenol, epidermal growth factor, and phorbol diester are without effects. Analysis of the insulin dose-response relationship between P160 tyrosine phosphorylation and insulin receptor kinase activity reveals that maximal phosphorylation of P160 occurs when only a fraction (25%) of the receptor kinase is activated by the hormone. A similar relationship between these two parameters is observed for the insulinomimetic agent hydrogen peroxide. The close correlation between the level of P160 phosphorylation and insulin receptor kinase activity suggests that P160 may be tyrosine phosphorylated by the receptor kinase following receptor kinase activation by the hormone or insulin-like agents. This hypothesis is further supported by the finding that the insulin receptor kinase is the only insulin-sensitive tyrosine kinase detectable in adipocyte plasma membranes under the conditions of our experiments.


Assuntos
Tecido Adiposo/metabolismo , Glicoproteínas/metabolismo , Insulina/farmacologia , Proteínas de Membrana/metabolismo , Tecido Adiposo/efeitos dos fármacos , Animais , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Técnicas In Vitro , Cinética , Masculino , Peso Molecular , Fosforilação , Fosfotirosina , Proteínas Tirosina Quinases/metabolismo , Ratos , Receptor de Insulina , Tirosina/análogos & derivados , Tirosina/análise
16.
Proc Natl Acad Sci U S A ; 84(12): 3972-6, 1987 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-2438690

RESUMO

Triton X-100-solubilized high-density microsomes from insulin-treated rat adipocytes exhibit a marked increase in serine/threonine and tyrosine kinase activities toward exogenous histone when compared to controls. The insulin-dependent activation of microsomal histone kinase activities occurs within the physiological range of hormone concentrations (ED50 = 0.6 nM). The hormone-enhanced histone phosphorylation by the high-density microsomes appears to be catalyzed by two distinct kinases, based on their differential interaction with wheat germ agglutinin-agarose. The insulin-sensitive serine/threonine kinase is not retained by The insulin-sensitive serine/threonine kinase is not retained by the lectin column, whereas the tyrosine kinase appears to be a glycoprotein as evidenced by its adsorption to the immobilized lectin. The insulin-stimulated serine/threonine kinase exhibits preferential phosphorylation of histone and Kemptide (synthetic Leu-Arg-Arg-Ala-Ser-Leu-Gly) compared to a number of other peptide substrates. The substrate specificity of this serine/threonine kinase shows that it is distinct from the kinases that phosphorylate ribosomal protein S6, casein, phosvitin, ATP citrate lyase, and glycogen synthase and from multifunctional calmodulin-dependent, cAMP- and cGMP-dependent, and Ca2+/phospholipid-dependent protein kinases. Furthermore, 22% of the insulin-sensitive serine/threonine kinase activity can be adsorbed by monoclonal anti-phosphotyrosine antibodies immobilized on agarose. Its adsorption is specifically inhibited by excess free phosphotyrosine but not phosphoserine or phosphothreonine. The data suggest that this insulin-stimulated serine/threonine kinase in adipocyte high-density microsomes is tyrosine-phosphorylated, consistent with the hypothesis that the stimulatory action of insulin on this kinase may be mediated by tyrosine phosphorylation.


Assuntos
Tecido Adiposo/enzimologia , Insulina/farmacologia , Microssomos/enzimologia , Proteínas Quinases/metabolismo , Tirosina , Tirosina/análogos & derivados , Animais , Anticorpos , Complexo Antígeno-Anticorpo/análise , Cinética , Masculino , Fosforilação , Fosfotirosina , Proteínas Serina-Treonina Quinases , Ratos , Tirosina/análise
17.
Farmaco Sci ; 42(6): 457-63, 1987 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-3115826

RESUMO

The synthesis of a series of 2-chloro-4-nitrobenzoylamino acid methyl esters (II-VIII), corresponding hydrazides (IX-XV), dipeptide methyl esters (XVI-XXII) and dipeptide hydrazides (XXIII-XXIX) was achieved employing the carbodiimide and azide methods. The derivatives containing the residues of Phe and Tyr were found to be active against several microorganisms.


Assuntos
Antibacterianos/síntese química , Dipeptídeos/síntese química , Nitrobenzoatos/síntese química , Amidas/síntese química , Amidas/farmacologia , Bacillus subtilis/efeitos dos fármacos , Fenômenos Químicos , Química , Dipeptídeos/farmacologia , Escherichia coli/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Nitrobenzoatos/farmacologia
19.
J Anim Sci ; 62(1): 77-85, 1986 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-3957811

RESUMO

Previous research in this laboratory demonstrated the existence of a major gene (hg), inherited as a homozygous recessive, which increases postweaning growth by 40 to 50% in C57Bl/6 mice (Ch) compared to the same genetic stock without the major gene (CH). Although its effect has not been previously evaluated, this single recessive allele is also in a line of mice selected for rapid postweaning gain for over 70 generations. Gh represents that line of mice with the major gene for growth (hg) in the growth-selected background and GH the growth-selected background without the major gene. Total body weight, daily weight gain, feed consumption and gain/feed, measured daily from 21 to 42 d of age, were all significantly greater (p less than .01) in the two lines with the hg/hg genotype (Ch and Gh) compared with their respective control lines (CH and GH). Differences in body composition at 42 d of age between CH compared with Ch and GH compared with Gh were accounted for by difference in body weight. Gross and net energetic efficiency, calculated assuming a similar maintenance energy requirement, were improved (P less than .01) in Ch and Gh compared to CH and GH, respectively. The results demonstrated that hg influences growth in growth-neutral and growth-selected backgrounds. The gene also alters energy metabolism by increasing energetic efficiency of growth and(or) decreasing maintenance energy requirement.


Assuntos
Composição Corporal , Genes Recessivos , Homozigoto , Camundongos Endogâmicos C57BL/genética , Fatores Etários , Animais , Ingestão de Alimentos , Metabolismo Energético , Camundongos , Camundongos Endogâmicos C57BL/crescimento & desenvolvimento , Desmame
20.
Farmaco Sci ; 39(2): 154-61, 1984 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-6714410

RESUMO

The synthesis of a series of tetrachlorophthaloylamino acids and some of the corresponding methyl esters is described. Coupling of tetrachlorophthaloylamino acids with amino acid methyl esters by the N,N'-dicyclohexylcarbodiimide method yielded the desired tetrachlorophthaloyldipeptide methyl esters. All of the synthesized tetrachlorophthaloylamino acids, esters and some dipeptide methyl esters were found to be active against a number of microorganisms.


Assuntos
Antibacterianos/síntese química , Dipeptídeos/síntese química , Ácidos Ftálicos/síntese química , Bactérias/efeitos dos fármacos , Fenômenos Químicos , Química , Dipeptídeos/farmacologia , Espectroscopia de Ressonância Magnética , Testes de Sensibilidade Microbiana , Ácidos Ftálicos/farmacologia , Espectrofotometria Infravermelho , Espectrofotometria Ultravioleta
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