RESUMO
Macrophages are essential components of the innate and adaptive immune responses, playing a decisive role in atherosclerosis, asthma, obesity, and cancer. The differential gene expression resulting from adhesion of macrophages to the extra-cellular matrix (ECM) has been studied in the J774A1 murine macrophage cell line using quantitative polymerase chain reaction (qPCR). The goal of this study was to identify housekeeping genes (HKGs) that remain stable and unaltered under normal culture conditions and in the presence of laminin after a time lapse of 6 and 24 h. The expression stabilities of eight commonly used reference genes were analyzed by determining the comparative threshold cycle ((ΔΔ)Ct) values, and using the BestKeeper, NormFinder, and geNorm algorithms. BestKeeper analysis revealed that the glyceraldehyde-3-phosphate dehydrogenase (GAPDH), peptidylprolyl isomerase A (PPIA), and ribosomal protein L13a (RPL13A) genes were highly stable, confirming the results of the (ΔΔ)Ct analysis. On the other hand, NormFinder proposed RPL13A and beta-glucuronidase (GUSB) to be the most suitable combination, and geNorm adjudged RPL13A, PPIA, and GUSB to be the most stable across all culture conditions. All programs discarded the use of actin beta and beta-2-microglobulin for normalization. The collected data indicated that RPL13A, PPIA, GAPDH, and GUSB as highly suitable as reference genes for qPCR analysis of murine macrophages under normal and ECM-simulated culture conditions. This study also emphasizes the importance of evaluating HKGs used for normalization to ensure the accuracy of qPCR data.
Assuntos
Expressão Gênica , Genes Essenciais/genética , Macrófagos/metabolismo , Reação em Cadeia da Polimerase em Tempo Real/normas , Animais , Linhagem Celular , Confiabilidade dos Dados , Camundongos , RNA Mensageiro , Reação em Cadeia da Polimerase em Tempo Real/métodos , Padrões de ReferênciaRESUMO
Bowman-Birk inhibitors (BBIs) are cysteine-rich and highly cross-linked small proteins that function as specific pseudosubstrates for digestive proteinases. They typically display a "double-headed" structure containing an independent proteinase-binding loop that can bind and inhibit trypsin, chymotrypsin and elastase. In the present study, we used computational biology to study the structural characteristics and dynamics of the inhibition mechanism of the small BBI loop expressing a 35-amino acid polypeptide (ChyTB2 inhibitor) which has coding region for the mutated chymotrypsin-inhibitory site of the soybean BBI. We found that in the BBI-trypsin inhibition complex, the most important interactions are salt bridges and hydrogen bonds, whereas in the BBI-chymotrypsin inhibition complex, the most important interactions are hydrophobic. At the same time, ChyTB2 mutant structure maintained the individual functional domain structure and excellent binding/inhibiting capacities for trypsin and chymotrypsin at the same time. These results were confirmed by enzyme-linked immunosorbend assay experiments. The results showed that modeling combined with molecular dynamics is an efficient method to describe, predict and then obtain new proteinase inhibitors. For such study, however, it is necessary to start from the sequence and structure of the mutant interacting relatively strongly with both trypsin and chymotrypsin for designing the small BBI-type inhibitor against proteinases.
Assuntos
Endopeptidases/metabolismo , Inibidor da Tripsina de Soja de Bowman-Birk/química , Inibidor da Tripsina de Soja de Bowman-Birk/farmacologia , Sequência de Aminoácidos , Animais , Bovinos , Quimotripsina/antagonistas & inibidores , Análise por Conglomerados , Desenho de Fármacos , Modelos Moleculares , Dados de Sequência Molecular , Estrutura Secundária de Proteína , Software , Propriedades de Superfície , Inibidores da Tripsina/químicaRESUMO
Bowman-Birk inhibitors (BBIs) are cysteine-rich and highly cross-linked small proteins that function as specific pseudosubstrates for digestive proteinases. They typically display a "double-headed" structure containing an independent proteinase-binding loop that can bind and inhibit trypsin, chymotrypsin and elastase. In the present study, we used computational biology to study the structural characteristics and dynamics of the inhibition mechanism of the small BBI loop expressing a 35-amino acid polypeptide (ChyTB2 inhibitor) which has coding region for the mutated chymotrypsin-inhibitory site of the soybean BBI. We found that in the BBI-trypsin inhibition complex, the most important interactions are salt bridges and hydrogen bonds, whereas in the BBI-chymotrypsin inhibition complex, the most important interactions are hydrophobic. At the same time, ChyTB2 mutant structure maintained the individual functional domain structure and excellent binding/inhibiting capacities for trypsin and chymotrypsin at the same time. These results were confirmed by enzyme-linked immunosorbend assay experiments. The results showed that modeling combined with molecular dynamics is an efficient method to describe, predict and then obtain new proteinase inhibitors. For such study, however, it is necessary to start from the sequence and structure of the mutant interacting relatively strongly with both trypsin and chymotrypsin for designing the small BBI-type inhibitor against proteinases.
Assuntos
Animais , Endopeptidases/metabolismo , Inibidor da Tripsina de Soja de Bowman-Birk/química , Modelos Moleculares , Sequência de Aminoácidos , Bovinos , Análise por Conglomerados , Desenho de Fármacos , Inibidor da Tripsina de Soja de Bowman-Birk/farmacologia , Inibidores da Tripsina/química , Quimotripsina/antagonistas & inibidoresRESUMO
Twenty five newborns weighting less than 1501 g were followed after electively installing a percutaneous catheter with subcutaneous tunneling; they were used for parenteral nutrition through the basilic, cephalic or external jugular veins, lasting an average of 10.3 +/- 2.08 days. Complications encountered were obstruction, accidental extraction, edema and phlebitis. There were no complications at the puncture site or subcutaneous tunnel. The arms were the first choice site of installation and were occluded with sterile gauze and adhesive tape in a bracelet form. We concluded that this technique is an appropriate alternative for parenteral nutrition in the low birth weight newborn.
Assuntos
Cateterismo Venoso Central/métodos , Recém-Nascido de Baixo Peso , Nutrição Parenteral , Cateterismo Venoso Central/efeitos adversos , Feminino , Humanos , Recém-Nascido , Masculino , Estudos ProspectivosRESUMO
We present a case-report of a six weeks old premature baby who developed acute acalculous gangrenous cholecystitis related to septicemia due to Citrobacter freundii. We analyze other etiological factors and remark the importance of physical examination and ultrasonographic exploration of the biliary tract. Early cholecystectomy is suggested.
Assuntos
Colecistite/etiologia , Doenças do Prematuro , Doença Aguda , Colecistectomia , Colecistite/diagnóstico , Colecistite/cirurgia , Citrobacter , Infecções por Enterobacteriaceae/complicações , Humanos , Lactente , Recém-Nascido , Perfuração Intestinal , Masculino , Necrose , Sepse/complicações , UltrassonografiaRESUMO
We discuss the case of a premature baby weighing 1,200 g who developed right diaphragmatic paralysis when a venous cutdown was performed on his internal jugular vein. Acquired unilateral diaphragmatic paralysis has been associated with obstetric trauma of the cervical plexus and surgical procedures of the neck and thorax, when the phrenic nerve is injured. We suggest fluoroscopy as the best diagnostic method; early diaphragmatic plication reduces the morbimortality of these patients.
Assuntos
Sangria/efeitos adversos , Doenças do Prematuro , Veias Jugulares , Nervo Frênico/lesões , Paralisia Respiratória/etiologia , Enterocolite Pseudomembranosa/complicações , Enterocolite Pseudomembranosa/terapia , Humanos , Recém-Nascido de Baixo Peso , Recém-Nascido , Masculino , Nutrição ParenteralRESUMO
PIP: Tetanus is an infectious disease that can occur at any age and is highly lethal. In the present study, a review was made of all cases of tetanus between January 1979 and June 1985 at the University Hospital "Dr. Eleuterio Gonzalez" in Monterrey, Nuevo Leon, Mexico. Out of a total of 76 cases, 10 were newborns. Of these, 5 were female and 5 male. The average age at admission was 8.5 days and the period of Collis was 32.3 hours. All had been born in a septic environment at term and with an average birthweight of 3120 kgs. Using Jandra's classification, 3 cases were considered mild, 5 moderate, and 2 severe. The laboratory tests were not relevant. 9 patients were given Phenobarbital and Diazepam as sedatives; 6 received total parenteral alimentation, and 5 newborns were given mechanical ventilation during an average of 26.6 days. The most common complications were respiratory problems and septicemia. The average length of hospitalization was 26.3 days with a mortality rate of 50%. (author's)^ieng
Assuntos
Doença , Pesquisa sobre Serviços de Saúde , Hospitais , Mortalidade Infantil , Lactente , Mortalidade , Tétano , Terapêutica , Adolescente , Fatores Etários , América , América Central , Atenção à Saúde , Demografia , Países Desenvolvidos , Países em Desenvolvimento , Saúde , Instalações de Saúde , Infecções , América Latina , México , América do Norte , Organização e Administração , População , Características da População , Dinâmica Populacional , Avaliação de Programas e Projetos de Saúde , PesquisaRESUMO
This study aims at comparing performance among subjects along a series of tests dealing with verbal comprehension, verbal fluency, and reasoning. The subjects were, approximately, the same age. However theirs were different educational levels. The samples drawn were 180 male subjects, approximately 22-years old, distributed into 3 groups according to their education level, i.e. primary school, high school, and college. Three subtests, Verbal Meaning, Word Fluency, and Reasoning (SRA Primary Mental Abilities Test Ages 11 to 17-Form AH) with a high saturation of the above mentioned factors were administered. The results of the simple variation analysis on each subtest scores showed a significant principal effect on education although a strict covariation among scores and the subjects' education level were not detected.
Assuntos
Escolaridade , Testes de Linguagem , Resolução de Problemas , Comportamento Verbal , Adulto , Análise de Variância , Humanos , MasculinoRESUMO
This study aims at comparing performance among subjects along a series of tests dealing with verbal comprehension, verbal fluency, and reasoning. The subjects were, approximately, the same age. However theirs were different educational levels. The samples drawn were 180 male subjects, approximately 22-years old, distributed into 3 groups according to their education level, i.e. primary school, high school, and college. Three subtests, Verbal Meaning, Word Fluency, and Reasoning (SRA Primary Mental Abilities Test Ages 11 to 17-Form AH) with a high saturation of the above mentioned factors were administered. The results of the simple variation analysis on each subtest scores showed a significant principal effect on education although a strict covariation among scores and the subjects education level were not detected.