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1.
Br J Nutr ; 123(12): 1406-1414, 2020 06 28.
Artigo em Inglês | MEDLINE | ID: mdl-32131903

RESUMO

Starting university is an important time with respect to dietary changes. This study reports a novel approach to assessing student diet by utilising student-level food transaction data to explore dietary patterns. First-year students living in catered accommodation at the University of Leeds (UK) received pre-credited food cards for use in university catering facilities. Food card transaction data were obtained for semester 1, 2016 and linked with student age and sex. k-Means cluster analysis was applied to the transaction data to identify clusters of food purchasing behaviours. Differences in demographic and behavioural characteristics across clusters were examined using χ2 tests. The semester was divided into three time periods to explore longitudinal changes in purchasing patterns. Seven dietary clusters were identified: 'Vegetarian', 'Omnivores', 'Dieters', 'Dish of the Day', 'Grab-and-Go', 'Carb Lovers' and 'Snackers'. There were statistically significant differences in sex (P < 0·001), with women dominating the Vegetarian and Dieters, age (P = 0·003), with over 20s representing a high proportion of the Omnivores and time of day of transactions (P < 0·001), with Dieters and Snackers purchasing least at breakfast. Many students (n 474, 60·4 %) changed dietary cluster across the semester. This study demonstrates that transactional data present a feasible method for dietary assessment, collecting detailed dietary information over time and at scale, while eliminating participant burden and possible bias from self-selection, observation and attrition. It revealed that student diets are complex and that simplistic measures of diet, focusing on narrow food groups in isolation, are unlikely to adequately capture dietary behaviours.


Assuntos
Inquéritos sobre Dietas/métodos , Dieta/estatística & dados numéricos , Utilização de Instalações e Serviços/estatística & dados numéricos , Serviços de Alimentação/estatística & dados numéricos , Estudantes/estatística & dados numéricos , Adolescente , Comportamento do Consumidor , Comportamento Alimentar , Feminino , Humanos , Estudos Longitudinais , Masculino , Estudos Retrospectivos , Universidades , Adulto Jovem
2.
Vet Pathol ; 54(1): 159-163, 2017 01.
Artigo em Inglês | MEDLINE | ID: mdl-27312368

RESUMO

Five cases of Rhodococcus equi infection in dogs were identified from 2003 to 2014. Three of the dogs had severe, internal lesions attributable to R. equi that have not been previously described: endophthalmitis, endocarditis, and suppurative pleuropneumonia. Isolates from 4 of the dogs were analyzed by polymerase chain reaction for Rhodococcus virulence-associated plasmid (vap) genes. One isolate was vapA-positive, 2 lacked a virulence plasmid, and 1 carried the novel vapN-associated plasmid (pVAPN) recently characterized in bovine isolates. The pVAPN plasmid has not been described in isolates cultured from companion animals. Four of the dogs either were receiving immunosuppressive drugs or had endocrinopathies. R. equi has the potential to cause significant infections in dogs, and immunocompromised animals should be considered at risk for infection.


Assuntos
Infecções por Actinomycetales/veterinária , Doenças do Cão/microbiologia , Rhodococcus equi , Infecções por Actinomycetales/patologia , Animais , Doenças do Cão/patologia , Cães , Feminino , Masculino , Reação em Cadeia da Polimerase/veterinária , Rhodococcus equi/genética , Rhodococcus equi/patogenicidade , Fatores de Virulência/genética
3.
J Comp Pathol ; 155(4): 305-309, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27665042

RESUMO

A 10-year-old, neutered male, Basset hound had a 26 × 21 × 21 cm infiltrative mass on the left abdominal wall that did not extend into the peritoneal cavity based on radiographs and abdominal computed tomography. Cytological examination revealed moderate numbers of neoplastic round cells, which frequently contained numerous round, clear, cytoplasmic vacuoles. Histologically, the tumour was composed of two morphologically distinct cell populations forming a continuum of heterogeneously differentiated cells. The primary spindle-shaped population formed streams with abundant, lightly eosinophilic, alcian blue-positive, myxoid matrix. The second population was arranged in sheets and had a round cell appearance. Scattered within both populations were neoplastic cells containing variably sized, intracytoplasmic, osmium tetroxide-positive vacuoles (lipid). Multifocal large pools of mucin formed pseudocysts, and numerous small capillaries were present throughout the neoplasm. According to the current World Health Organization veterinary classification of liposarcomas, this neoplasm had morphological features of both the myxoid and pleomorphic variants of liposarcoma; however, it was analogous to the recently defined high-grade myxoid liposarcoma in man. Myxoid liposarcoma with round cells has not been described previously in dogs. This case highlights the need to potentially re-evaluate the current classification of liposarcomas in animals.


Assuntos
Doenças do Cão/patologia , Lipossarcoma Mixoide/veterinária , Parede Abdominal/patologia , Animais , Cães , Masculino
4.
Meat Sci ; 117: 75-84, 2016 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-26950612

RESUMO

Enhanced pork loin chops, beef longissimus lumborum steaks, semimembranosus steaks (superficial and deep portions), ground beef, and ground turkey were displayed under light emitting diode (LED) and fluorescent (FLS) lighting in two multi-shelf, retail display cases with identical operating parameters. Visual and instrumental color, internal product temperature, case temperature, case cycling, thiobarbituric acid reactive substances (TBARS), and Enterobacteriaceae and aerobic plate counts were evaluated. Under LED, beef products (except the deep portion of beef semimembranosus steaks) showed less (P<0.05) visual discoloration. Pork loin chops had higher (P<0.05) L* values for LED lighting. Other than beef longissimus lumborum steaks, products displayed under LED lights had colder internal temperatures than products under FLS lights (P<0.05). Under LED, pork loin chops, ground turkey, and beef semimembranosus steaks had higher (P<0.05) values for TBARS. LED provides colder case and product temperatures, more case efficiency, and extended color life by at least 0.5d for longissimus and semimembranosus steaks; however, some LED cuts showed increased lipid oxidation.


Assuntos
Armazenamento de Alimentos , Luz , Produtos da Carne/análise , Animais , Bovinos , Enterobacteriaceae/isolamento & purificação , Microbiologia de Alimentos , Conservação de Alimentos , Peroxidação de Lipídeos , Músculo Esquelético , Odorantes , Plantas Comestíveis , Suínos , Temperatura , Substâncias Reativas com Ácido Tiobarbitúrico , Fatores de Tempo , Turquia
5.
Behav Brain Res ; 278: 271-9, 2015 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-25264578

RESUMO

Although the peptide urotensin II (UII) has well studied direct actions on the cardiovascular system, the UII receptor (UIIR) is expressed by neurons of the hindbrain. Specifically, the UIIR is expressed by the cholinergic neurons of the laterodorsal tegmentum (LDTg) and the pedunculopontine tegmentum (PPTg). These neurons send axons to the ventral tegmental area (VTA), for which the PPTg and LDTg are the sole source of acetylcholine. Therefore, it was hypothesized that UIIR activation within the VTA would modulate reward-related behaviors, such as cocaine-induced drug seeking. Intra-VTA microinjections of UII at high concentrations (1 nmole) established conditioned place preference (CPP), but also blocked cocaine-mediated CPP (10 mg/kg). When rats received systemic sub-effectual doses of cocaine (7.5 mg/kg) with intra-VTA injections of 1 or 10 pmole of UII CPP was formed. Furthermore, the second endogenous ligand for the UIIR, urotensin II-related peptide, had the same effect at the 10 pmole dose. The effects of low doses of UII were blocked by pretreatment with the UIIR antagonist SB657510. Furthermore, it was found that intra-VTA UII (10 pmole) further increased cocaine-mediated (7.5 mg/kg) rises in electrically evoked dopamine in the nucleus accumbens. Our study has found that activation of VTA-resident UIIR produces observable behavioral changes in rats, and that UIIR is able to modulate the effects of cocaine. In addition, it was found that UIIR activation within the VTA can potentiate cocaine-mediated neurochemical effects. Therefore, the coincident activation of the UII-system and cocaine administration may increase the liability for drug taking behavior.


Assuntos
Cocaína/farmacologia , Comportamento de Procura de Droga/efeitos dos fármacos , Receptores Acoplados a Proteínas G/efeitos dos fármacos , Urotensinas/farmacologia , Área Tegmentar Ventral/efeitos dos fármacos , Animais , Comportamento Animal/efeitos dos fármacos , Cocaína/administração & dosagem , Condicionamento Psicológico/efeitos dos fármacos , Dopamina/análise , Microinjeções , Vias Neurais/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Núcleo Accumbens/efeitos dos fármacos , Núcleo Accumbens/metabolismo , Hormônios Peptídicos/administração & dosagem , Hormônios Peptídicos/farmacologia , Ratos , Ratos Sprague-Dawley , Receptores Acoplados a Proteínas G/antagonistas & inibidores , Recompensa , Sulfonamidas/administração & dosagem , Sulfonamidas/farmacologia , Urotensinas/administração & dosagem , Área Tegmentar Ventral/citologia , Área Tegmentar Ventral/metabolismo
6.
Neuroscience ; 284: 381-399, 2015 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-25305665

RESUMO

Rats with lesions of the pedunculopontine tegmental nucleus (PPTg) reliably overconsume high concentration sucrose solution. This effect is thought to be indicative of response-perseveration or loss of behavioral control in conditions of high excitement. While these theories have anatomical and behavioral support, they have never been explicitly tested. Here, we used a contact lickometer to examine the microstructure of drinking behavior to gain insight into the behavioral changes during overconsumption. Rats received either excitotoxic (ibotenic acid) damage to all PPTg neuronal subpopulations or selective depletion of the cholinergic neuronal sub-population (diphtheria toxin-urotensin II (Dtx-UII) lesions). We offered rats a variety of pleasant, neutral and aversive tastants to assess the generalizability and specificity of the overconsumption effect. Ibotenic-lesioned rats consumed significantly more 20% sucrose than sham controls, and did so through licking significantly more times. However, the behavioral microstructure during overconsumption was unaffected by the lesion and showed no indications of response-perseveration. Furthermore, the overconsumption effect did not generalize to highly consumed saccharin. In contrast, while only consuming small amounts of quinine solution, ibotenic-lesioned rats had significantly more licks and bursts for this tastant. Selective depletion of cholinergic PPTg neurons had no effect on consumption of any tastant. We then assessed whether it is the salience of the solution which determines overconsumption by ibotenic-lesioned rats. While maintained on free-food, ibotenic-lesioned rats had normal consumption of sucrose and hypertonic saline. After mild food deprivation ibotenic PPTg-lesioned rats overconsumed 20% sucrose. Subsequently, after dietary-induced sodium deficiency, lesioned rats consumed significantly more saline than controls. These results establish that it is the salience of the solution which is the determining factor leading to overconsumption following excitotoxic PPTg lesion. They also find no support for response-perseveration contributing to this effect. Results are discussed in terms of altered dopamine (DA) and salience signaling.


Assuntos
Comportamento de Ingestão de Líquido/fisiologia , Ingestão de Líquidos/fisiologia , Núcleo Tegmental Pedunculopontino/fisiopatologia , Animais , Colinérgicos/toxicidade , Sacarose Alimentar/administração & dosagem , Toxina Diftérica/toxicidade , Ingestão de Líquidos/efeitos dos fármacos , Comportamento de Ingestão de Líquido/efeitos dos fármacos , Água Potável/administração & dosagem , Agonistas de Aminoácidos Excitatórios/toxicidade , Privação de Alimentos , Ácido Ibotênico/toxicidade , Masculino , Núcleo Tegmental Pedunculopontino/efeitos dos fármacos , Quinina/administração & dosagem , Ratos Sprague-Dawley , Sacarina/administração & dosagem , Sódio na Dieta/administração & dosagem , Urotensinas/toxicidade
7.
J Perinatol ; 34(7): 528-31, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24699219

RESUMO

OBJECTIVE: To compare the effectiveness of 100% oxygen therapy vs oxygen treatment with targeted pulse oximetry in the management of symptomatic small to moderate spontaneous pneumothorax (SP). In total, 100% oxygen treatment for SP has been a common practice in neonatology, albeit there is little evidence to validate its efficacy. STUDY DESIGN: A retrospective chart review of 83 neonatal records with the diagnosis of pneumothorax was conducted. Infants <35 weeks gestation, those with large pneumothoraces requiring chest tube drainage and/or ventilatory support were excluded. Data gathered included demographics, vital signs, treatment information and clinical indicators of resolution of symptoms. RESULT: In total, 45 neonates with SP were included in the study. Groups were similar for gestational age, birth weight, Apgar scores, gravidity, parity, gender, race, pneumothorax size and location. Patients in the 100% oxygen therapy group received a significantly longer oxygen treatment (21.3 vs 8 h, P < 0.001), required longer intravenous fluid treatment (48.6 ± 29.9 vs 31.3 ± 18.8 h, P = 0.03) and were delayed in reaching full feeds (44.1 ± 25.7 vs 29.5 ± 18.8 h, P = 0.03) compared with the oxygen-targeted treatment group. Time to first oral feeding, time to resolution of tachypnea and length of stay were similar in both groups. CONCLUSION: There are no clinically significant advantages to using 100% oxygen in the treatment of symptomatic small to moderate SP. In fact, it may result in longer exposure to unnecessary oxygen treatment and toxicity. Oxygen should be reserved for those who are hypoxic and adjusted to comply with accepted saturation levels in neonates.


Assuntos
Oxigênio/administração & dosagem , Pneumotórax/terapia , Feminino , Humanos , Recém-Nascido , Tempo de Internação/estatística & dados numéricos , Masculino , Ohio , Oximetria , Pneumotórax/fisiopatologia , Estudos Retrospectivos , Resultado do Tratamento
8.
J Neurochem ; 102(1): 112-20, 2007 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-17419804

RESUMO

Urotensin II is a neuropeptide first isolated from fish and later found in mammals: where it has potent cardiovascular, endocrine and behavioral effects. In rat brain the urotensin II receptor (UII-R) is predominately expressed in the cholinergic neurons of the pedunculopontine (PPTg) and laterodorsal tegmental nuclei. Typically, the function of the PPTg has been examined using excitotoxins, destroying both cholinergic and non-cholinergic neurons, which confounds interpretation. We took advantage of UII-R's unique expression profile, by combining UII with diphtheria toxin, to engineer a toxin specific for cholinergic neurons of the PPTg. In vitro, two different toxin constructs were shown to selectively activate UII-R (average EC50 approximately 30 nmol/L; calcium mobility assay) and to be 10,000-fold more toxic to UII-R expressing CHO cells, than wildtype cells (average LD50 approximately 2 nmol/L; cell viability). In vivo, pressure injection into the PPTg of rats, resulted in specific loss of choline transporter and NADPH diaphorase positive neurons known to express the UII-R. The lesions developed over time, resulting in the loss of over 80% of cholinergic neurons at 21 days, with little damage to surrounding neurons. This is the first highly selective molecular tool for the depletion of mesopontine cholinergic neurons. The toxin will help to functionally dissect the pedunculopontine and laterodorsal tegmental nuclei, and advance the understanding of the functions of these structures.


Assuntos
Toxina Diftérica/química , Toxina Diftérica/toxicidade , Neurônios/efeitos dos fármacos , Síndromes Neurotóxicas/patologia , Neurotoxinas/química , Neurotoxinas/toxicidade , Sistema Nervoso Parassimpático/efeitos dos fármacos , Ponte/patologia , Tegmento Mesencefálico/patologia , Urotensinas/química , Urotensinas/toxicidade , Animais , Células CHO , Sobrevivência Celular/efeitos dos fármacos , Cricetinae , Cricetulus , Toxina Diftérica/isolamento & purificação , Eletroforese em Gel de Poliacrilamida , Escherichia coli/genética , Masculino , NADPH Desidrogenase/metabolismo , NADPH Desidrogenase/fisiologia , Neurônios/patologia , Sistema Nervoso Parassimpático/patologia , Plasmídeos/genética , Ratos
9.
Brain Res ; 923(1-2): 120-7, 2001 Dec 27.
Artigo em Inglês | MEDLINE | ID: mdl-11743979

RESUMO

Urotensin II (UII) is a peptide known to be a potent vasoconstrictor. The urotensin II receptor (UII-R) is expressed not only in peripheral tissues but also in the brain of rodents. As a basis for studies of UII central nervous system actions, UII-R localization in the rat brain was analyzed by in situ hybridization and by in situ binding. UII-R mRNA was found in the mesopontine tegmental area colocalizing with choline acetyltransferase. Binding sites were detected throughout the brain with the highest levels found in the pedunculopontine tegmental area, the lateral dorsal tegmental area, and the lateral septal, medial habenular, and interpeduncular nuclei. The majority of these brain nuclei are sites of axonal termination originating from the mesopontine areas, suggesting that UII-R is a presynaptic receptor. This distribution of UII-R in the cholinergic mesopontine area indicates that the UII system may be involved in sensory-motor integration and perhaps in central nervous system blood flow.


Assuntos
Acetilcolina/fisiologia , Ponte/fisiologia , Receptores de Superfície Celular/genética , Receptores Acoplados a Proteínas G , Tegmento Mesencefálico/fisiologia , Animais , Colina O-Acetiltransferase/genética , Expressão Gênica/fisiologia , Habenula/fisiologia , Hibridização In Situ , Radioisótopos do Iodo , Masculino , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley , Receptores Pré-Sinápticos/fisiologia , Núcleos Septais/fisiologia
10.
Protein Expr Purif ; 2(2-3): 151-7, 1991.
Artigo em Inglês | MEDLINE | ID: mdl-1821784

RESUMO

We used site-directed mutagenesis to introduce both a NdeI restriction endonuclease site and an initiator codon at the junction of the leader and structural gene sequences of the metallo-beta-lactamase of Bacillus cereus 5/B/6. This construct allowed us to clone just the beta-lactamase structural gene sequence into an Escherichia coli expression vector. E. coli cells were transformed with the recombinant plasmid, the B. cereus beta-lactamase was expressed, and these E. coli cells were disrupted by sonic oscillation. When the resultant suspensions were clarified by ultracentrifugation, the B. cereus beta-lactamase represented 15% of the total protein in the supernatant. Subsequent gel filtration and ion-exchange chromatography allowed the first reported purification to homogeneity of the B. cereus beta-lactamase from E. coli with an 87% recovery and an overall yield of 17 mg of enzyme per liter of cell culture. The electrophoretic mobilities of the enzyme expressed in and purified from E. coli and the enzyme purified directly from B. cereus were identical in both native and sodium dodecyl sulfate gel electrophoreses. As with the B. cereus enzyme, Km and Vmax (using cephalosporin C as substrate) for the enzyme purified from E. coli were 0.39 mM and 1333 units/mg protein, respectively. Likewise, the Co(II)-reconstituted enzyme purified from E. coli, which retained 29% of the activity of the Zn(II) enzyme, had electronic absorption spectra with maxima at 347, 551, 617, and 646 nm with extinction coefficients of 900, 250, 173, and 150 M-1 cm-1, respectively.


Assuntos
Bacillus cereus/enzimologia , Bacillus cereus/genética , beta-Lactamases/genética , beta-Lactamases/isolamento & purificação , Sequência de Bases , Cromatografia em Gel , Cromatografia por Troca Iônica , Clonagem Molecular , DNA Bacteriano/genética , Escherichia coli/genética , Expressão Gênica , Genes Bacterianos , Cinética , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , beta-Lactamases/metabolismo
11.
Nucleic Acids Res ; 18(14): 4215-21, 1990 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-2198538

RESUMO

Naturally occurring suppressor mutants derived from tRNATrp genes have never been identified in S. cerevisiae. Oligonucleotide-directed mutagenesis was used to generate potential ochre and opal suppressors from a cloned tRNATrp gene. In vitro transcription analyses show the ochre suppressor form of the gene, TRPO, accumulates precursors and tRNA in amounts comparable to the parent. The opal suppressor, TRPOP, accumulates 4-5 fold less tRNA. Both forms of the gene are processed and spliced in vitro to produce tRNAs with the expected base sequences. The altered genes were subcloned into yeast vectors and introduced into yeast strains carrying a variety of amber, ochre, and opal mutations. When introduced on a CEN vector, neither ochre nor opal suppressor forms show suppressor activity. Deletion of the CEN region from the clones increases the copy number to 10-20/cell. The opal suppressor form shows moderate suppressor activity when the gene is introduced on this vector, however, the ochre suppressor form exhibits no detectable biological activity regardless of gene copy number. Northern blot analyses of the steady state levels of tRNATrp in cells containing the high copy-number clones reveal 20-100% increases in the abundance of tRNATrp.


Assuntos
Anticódon , Códon , RNA Mensageiro , RNA de Transferência Aminoácido-Específico/genética , RNA de Transferência de Triptofano/genética , RNA de Transferência , Saccharomyces cerevisiae/genética , Supressão Genética , Sequência de Bases , Northern Blotting , Clonagem Molecular , Genes Fúngicos , Dados de Sequência Molecular , Conformação de Ácido Nucleico , Mapeamento de Nucleotídeos , Splicing de RNA , RNA Fúngico , Transcrição Gênica , Transformação Genética
12.
Stud Fam Plann ; 20(4): 225-34, 1989.
Artigo em Inglês | MEDLINE | ID: mdl-2772996

RESUMO

The Maternal-Child Health/Family Planning (MCH/FP) Extension Project in Bangladesh identifies and examines barriers to implementation of the national MCH/FP program, and determines strategies to overcome them. This study analyzes field-workers' ability to carry out more tasks than they do presently, and how their performance might be improved when additional field-workers are hired. In two experimental subdistricts, researchers observed the work of family welfare assistants (FWAs), the female family planning field-workers, to determine the duration and frequency of their home visits with village women and the content of their exchanges. While many factors influence the FWA's work, researchers found that the preplanned monthly work schedules could be manipulated relatively easily to improve duration and frequency of program contact with village women. With more time available to spend with women, the potential to improve the quality of services is enhanced.


Assuntos
Serviços de Saúde Materna/organização & administração , Qualidade da Assistência à Saúde , Saúde da População Rural , Trabalho , Bangladesh , Agentes Comunitários de Saúde , Serviços de Planejamento Familiar , Feminino , Humanos , Centros de Saúde Materno-Infantil
13.
J Clin Invest ; 80(5): 1280-8, 1987 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-2824558

RESUMO

The regulation of the expression of interstitial collagenase and tissue inhibitor of metalloproteinases (TIMP) was examined in response to both retinoid compounds and glucocorticoids. Effective retinoids induced a dose-dependent, specific increase in the production of TIMP of approximately two- to threefold by monolayer cultures of human fibroblasts derived from various tissues, while simultaneously causing a decrease in collagenase secretion of similar magnitude. These effects were apparent by 8-12 h in culture and disappeared within 24 h after the withdrawal of retinoid compounds. The retinoid effect on TIMP production was mediated via an increased biosynthesis of new inhibitor protein. Similarly, increased steady state levels of TIMP messenger RNA (mRNA) accompanied by decreased quantities of collagenase mRNA were demonstrated, suggesting transcriptional control of the retinoid action. The data suggest that retinoids co-regulate the expression of collagenase and TIMP, and do so in an inverse manner. Dexamethasone caused a dose-dependent, specific decrease in collagenase production without altering the biosynthesis of TIMP. These findings were paralleled by a marked reduction in collagenase mRNA, without any accompanying change in TIMP mRNA. Therefore, TIMP and collagenase expression appear to be independently modulated by glucocorticoids.


Assuntos
Fibroblastos/metabolismo , Glucocorticoides/farmacologia , Metaloendopeptidases/antagonistas & inibidores , Colagenase Microbiana/genética , Inibidores de Proteases/genética , Retinoides/farmacologia , Linhagem Celular , Dexametasona/farmacologia , Fibroblastos/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Técnicas de Imunoadsorção , Cinética , Hibridização de Ácido Nucleico , RNA Mensageiro/metabolismo , Tretinoína/farmacologia
14.
Proc Natl Acad Sci U S A ; 83(11): 3756-60, 1986 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-3012533

RESUMO

Human skin fibroblasts secrete collagenase as two proenzyme forms (57 and 52 kDa). The minor (57-kDa) proenzyme form is the result of a partial posttranslational modification of the major (52-kDa) proenzyme through the addition of N-linked complex oligosaccharides. Human endothelial cells as well as fibroblasts from human colon, cornea, gingiva, and lung also secrete collagenase in two forms indistinguishable from those of the skin fibroblast enzyme. In vitro tissue culture studies have shown that the level of constitutive synthesis of this fibroblast-type interstitial collagenase is tissue specific, varies widely, and correlates with the steady-state level of a single collagenase-specific mRNA of 2.5 kilobases. The tumor promoter, phorbol 12-myristate 13-acetate, apparently blocks the control of collagenase synthesis resulting in a similarly high level of collagenase expression (approximately equal to 3-7 micrograms of collagenase per 10(6) cells per 24 hr) in all examined cells. The constitutive level of synthesis of a 28-kDa collagenase inhibitor does not correlate with that of the enzyme. Phorbol 12-myristate 13-acetate stimulates the production of this inhibitor that in turn modulates the activity of collagenase in the conditioned media. As a result, the apparent activity of the enzyme present in the medium does not accurately reflect the rate of its synthesis and secretion.


Assuntos
Colagenase Microbiana/metabolismo , Endotélio/enzimologia , Fibroblastos , Regulação da Expressão Gênica/efeitos dos fármacos , Glicoproteínas/metabolismo , Humanos , Colagenase Microbiana/genética , Processamento de Proteína Pós-Traducional , RNA Mensageiro/genética , Pele/enzimologia , Acetato de Tetradecanoilforbol/farmacologia , Distribuição Tecidual , Tunicamicina/farmacologia
15.
Arch Biochem Biophys ; 241(1): 36-44, 1985 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-2992392

RESUMO

Phorbol myristate acetate (PMA), a tumor promotor known to stimulate collagenase production in fibroblasts and endothelial cells, was examined with regard to its ability to regulate the expression of the collagenase inhibitor secreted by human skin fibroblasts. Confluent human skin fibroblasts were incubated with concentrations of PMA ranging from 10(-11) to 10(-7) M, and the conditioned medium was analyzed by enzyme-linked immunosorbent assay for both immunoreactive collagenase and collagenase inhibitor. PMA stimulated the production of both collagenase and collagenase inhibitor in several cell lines to maximal rates that were very similar, 300 to 350 vs 230 to 330 pmol 10 micrograms DNA-1 48 h-1, respectively. Due to differences in the basal levels of expression of these proteins, such rates reflected a two- to sevenfold stimulation in collagenase production, in comparison to a more uniform two- to threefold enhancement in inhibitor synthesis. Production of inhibitor was 50% of maximal at 7 X 10(-9) M and maximal at 10(-7) M phorbol. This concentration-dependent effect was very similar to that observed for collagenase expression. Total protein synthesis by the phorbol-conditioned cells, as studied by incorporation of [3H]leucine into newly synthesized protein, was not significantly increased, nor was cellular DNA content. The onset of the effect of PMA on inhibitor production occurred between 4 and 8 h, was maximal by 8 h, and continued undiminished for at least another 64 h. After the first 8 h, inhibitor production continued at a roughly constant rate of approximately 10 pmol 10 micrograms DNA-1 h-1. Interestingly, following the removal of phorbol from culture medium, such fibroblasts continued to produce increased quantities of inhibitor protein for at least 72 h. Metabolic labeling studies in which fibroblasts were exposed to [3H]leucine followed by immunoprecipitation using inhibitor-specific antibody suggested that stimulation of inhibitor production by PMA was mediated via an increased synthesis of new inhibitor protein. Therefore, in response to the tumor promoter, PMA collagenase and collagenase inhibitor expression by human skin fibroblasts appear to be coregulated.


Assuntos
Inibidores Enzimáticos/metabolismo , Colagenase Microbiana/metabolismo , Forbóis/farmacologia , Pele/enzimologia , Acetato de Tetradecanoilforbol/farmacologia , Compartimento Celular , Células Cultivadas , Meios de Cultura , DNA/biossíntese , Relação Dose-Resposta a Droga , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Biossíntese de Proteínas , Pele/efeitos dos fármacos , Fatores de Tempo , Inibidores Teciduais de Metaloproteinases
16.
Fam Plann Perspect ; 16(2): 77-82, 1984.
Artigo em Inglês | MEDLINE | ID: mdl-6723943

RESUMO

Eighty-seven percent of black adolescent males attending an inner-city junior high school and senior high school in Baltimore report that they have had sexual intercourse. Eighty percent of the sexually experienced teenagers have used a contraceptive method--and 60 percent used one at last intercourse. Contraceptive practice is characterized by heavy reliance on male methods: Of those using a method at last intercourse, 41 percent relied on the condom; 15 percent, on withdrawal; and 14 percent, on a combination of male and female methods. Use of female methods alone (mostly the pill) accounted for 28 percent of contraceptive practice at last intercourse. Nine-tenths of the teenagers have heard of the pill and condom, but far smaller majorities have heard of the IUD and diaphragm (about three-fifths each). In general, this population regards the condom as comparable in effectiveness to the pill. More than two-fifths of those recognizing these methods believe the pill--and the condom--to be "very good" at preventing pregnancy, and one-fifth believe that about the IUD and diaphragm. Nearly four in 10 mistakenly believe that they need a parent's permission to attend a birth control clinic, while slightly more than half think they need permission to buy nonprescription contraceptives in a drugstore . Although nearly nine in 10 respondents recognize that boys share a responsibility for preventing pregnancy when they have sex, more than half are willing to tolerate unprotected coitus. Most wish to delay parenthood until their early 20s, but many of these adolescents appear to hold attitudes conducive to out-of- wedlock conception.(ABSTRACT TRUNCATED AT 250 WORDS)


Assuntos
Atitude , Negro ou Afro-Americano/psicologia , Comportamento Contraceptivo , Comportamento Sexual , Adolescente , Anticoncepção/métodos , Feminino , Conhecimentos, Atitudes e Prática em Saúde , Humanos , Masculino , Maryland , Pais , Inquéritos e Questionários
17.
Hybridoma ; 2(2): 201-9, 1983.
Artigo em Inglês | MEDLINE | ID: mdl-6205977

RESUMO

LE cell formation is one feature of systemic lupus erythematosus exhibited by virtually all mice of the NZB/NZW strain and is the result of accumulation of antibodies directed against components of cell nuclei. A hybrid cell line which produces antibodies capable of inducing LE cell formation in vitro has been isolated in a hybridoma fusion using the splenocytes of unimmunized NZB/NZW mice. These monoclonal autoantibodies provide an intense staining of the chromatin in cells of a number of divergent species and tissues. They bind strongly to the histone rich (2 M NaCl) fraction of extracted, isolated nuclei. Further analyses using the antibodies in immune precipitations and in antibody labeling of capillary blots on nitrocellulose sheets of calf thymus histone demonstrate that the antibodies are directed against histones and are capable of reacting with H1, H2a, H2b, H3, and H4 histones individually. In contrast to human autoantibodies with histone specificity, no cross-reactivity of this monoclonal autoantibody with the lymphocyte surface could be detected by either immunofluorescence or immunoelectron microscopy.


Assuntos
Anticorpos Monoclonais/imunologia , Autoanticorpos/imunologia , Histonas/imunologia , Animais , Hibridomas/imunologia , Lúpus Eritematoso Sistêmico/imunologia , Camundongos , Camundongos Endogâmicos NZB , Neutrófilos , Linfócitos T/imunologia
18.
Fam Plann Perspect ; 15(1): 25-9, 1983.
Artigo em Inglês | MEDLINE | ID: mdl-6680700

RESUMO

PIP: This study explores the reasons why teenagers coming for professional birth control help for the 1st time choose the particular clinic in which they enrolled and the clinic characteristics which may encourage the young patients to come for contraceptive help early or contribute to a delay in seeking services. 1234 never pregnant teenagers in 31 family planning clinics of various types in 8 cities answered a questionnaire in the spring of 1980. About 44% of the respondents were white, 48% black and the rest from other racial groups. Just over 14% came to the clinic before 1st coitus, 36% came only because they suspected they were pregnant, and the rest enrolled after they began intercourse but did not suspect pregnancy. The interval between intercourse and enrollment ranged from a few weeks to 2 years with a median interval of 11.5 months. There were 15 possible responses to the question, "Why did you choose this clinic?" The 5 principal reasons given in order were: It doesn't tell their parents; the people there care about teens; it is the closest, their friends come to it; and it is the only 1 they know of. Also significant was, for blacks only, an adult, usually the mother, chose the clinic. Additional information on responses to this question and others is given in charts. The questionnaire also showed a consistent but weak association between the reasons for selecting a particular facility and the characteristics of the clinic such as cost of services, special teen hours, teen rap sessions, and outreach program. The responses indicated that many factors including age, race, and the time when the teenager 1st seeks clinic services affect her choice. The conclusion drawn is that a network of facilities with a variety of services is needed with the common characteristics of open caring atmosphere, women clinicians available and absolute confidentiality so that the proportion of young women who appear only after they are pregnant might be greatly reduced.^ieng


Assuntos
Comportamento do Adolescente , Serviços de Planejamento Familiar , Adolescente , Instituições de Assistência Ambulatorial , Comportamento de Escolha , Confidencialidade , Anticoncepção , Humanos
20.
Aviat Space Environ Med ; 53(7): 666-9, 1982 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-7115255

RESUMO

In order to examine the effects of chronic low G acceleration on blood flow distribution and cardiac output, chickens (N=10) were centrifuged at +2Gz for 30-61 d. Controls (N=12) were not centrifuged. The animals were anesthetized with sodium pentobarbital after removal from the centrifuge and surgically prepared in order to measure cardiac output and regional blood flows by the reference sample method with 85Sr labeled microspheres (15 +/- 5 mum diam.). Both brachial arteries were cannulated to withdraw timed, paired blood samples at a known rate. The chest was opened and a cannula inserted into the left ventricle for administration of microspheres. Tissue samples were taken after completion of experimental procedures and their radioactivity was determined. The cardiac outputs in the two groups were not significantly different. Regional blood flows to the kidney, eyes, and skeletal muscle were significantly increased in the animals subjected to chronic +2Gz. While the mechanism by which these increases in blood flow occurred is not known, results indicate that chronic exposure to hyperdynamic gravitational fields can alter circulatory dynamics. We conclude that the cardiovascular system is directly involved in the process of adaptation to chronic positive acceleration.


Assuntos
Aceleração/efeitos adversos , Fluxo Sanguíneo Regional , Adaptação Fisiológica , Animais , Débito Cardíaco , Galinhas/sangue , Olho/irrigação sanguínea , Feminino , Gravitação , Masculino , Microesferas , Músculos/irrigação sanguínea , Circulação Renal , Radioisótopos de Estrôncio , Fatores de Tempo
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