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1.
J Dairy Sci ; 95(1): 83-90, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22192186

RESUMO

Up to 3% of young children develop milk allergy and this may influence the development of immune-mediated diseases in later life. One protein that has been associated with allergic reactions to ruminant milk is α(S1)-casein (CN). Studies suggest that goat milk with low levels of α(S1)-CN may reduce allergenicity of milk, but the dose response to α(S1)-CN has not been confirmed. In this study, we examined the immune response to varying levels of goat α(S1)-CN in a mouse model of gastrointestinal allergy. BALB/c mice (aged 5 wk) were given intraperitoneal injections with α(S1)-CN and aluminum as adjuvant at 1 and 3 wk to sensitize mice to the antigen. In wk 5, groups of fasting mice (n=8/group) were challenged 4 times on alternate days by intragastric gavage with saline or 2, 10, or 20mg of α(S1)-CN. Serum levels of specific IgE, IgG(1), and IgG(2a) antibodies and mouse mast cell protease-I were determined. Interleukin-4, IL-10, and IFN-γ responses to 48-h activation with antigen were measured in cultured splenocytes. We determined that mice sensitized with α(S1)-CN had higher titers of specific IgG(1) and IgE antibodies compared with controls; however, groups challenged with differing doses of α(S1)-CN did not differ. The group challenged with the highest dose of α(S1)-CN had a 10-fold increase in mouse mast cell protease-I compared with the group challenged with saline. Both IL-4 and IL-10 were produced in a dose-dependent manner by cultured splenocytes incubated with α(S1)-CN. Overall, α(S1)-CN stimulated the production of cytokines associated with allergic disease in a dose-dependent manner. Thus, milk with lower levels of α(S1)-CN should contribute to a lesser antigenic burden.


Assuntos
Caseínas/imunologia , Trato Gastrointestinal/imunologia , Hipersensibilidade a Leite/etiologia , Animais , Caseínas/farmacologia , Modelos Animais de Doenças , Relação Dose-Resposta Imunológica , Feminino , Trato Gastrointestinal/efeitos dos fármacos , Cabras , Imunoglobulina E/sangue , Imunoglobulina G/sangue , Interferon gama/sangue , Interleucina-10/sangue , Interleucina-4/sangue , Camundongos , Camundongos Endogâmicos BALB C , Hipersensibilidade a Leite/imunologia
2.
J Dairy Sci ; 93(7): 2857-62, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20630202

RESUMO

Most infant formulas use vegetable oils in place of milk fat to provide an overall fatty acid profile similar to that of breast milk. Vegetable oils have 5 to 20% saturated fatty acids in the sn-2 position of triglycerides unless they are modified by interesterification. Interesterification is increasingly used for the fat for infant formulas to raise the level of saturated fatty acids in the sn-2 position to 40 to 60%. The objective of this study was to verify an alternative approach to providing the appropriate fatty acid profile, including in the sn-2 position, for a goat infant formula. In this method, 55% of total fat was made from goat milk fat and 45% from a mixture of unmodified high oleic sunflower, canola, and sunflower oils in a ratio of 44:30:26. The fatty acid profile was measured by gas-liquid chromatography and the relative percentage of fatty acids in the sn-2 position of triglycerides was measured via partial deacylation with Grignard reagent using trimethylsilyl derivatives of monoacylglycerols. Mixing goat milk fat with vegetable oils produced a formula with a profile of essential fatty acids and a ratio of linoleic:alpha-linolenic fatty acids within the required interval of 5 to 15:1 recommended for infant formula. The proportion of palmitic acid in the sn-2 position was 31%.


Assuntos
Ácidos Graxos/química , Fórmulas Infantis/química , Triglicerídeos/química , Animais , Bovinos , Gorduras/análise , Ácidos Graxos/análise , Cabras , Humanos , Lactente , Leite/química
3.
J Dairy Sci ; 93(10): 4710-21, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20855005

RESUMO

The objectives of the study were to define the sensitivity and specificity of the California Mastitis Test (CMT) in determining the presence of intramammary infection in postpartum dairy goats and to determine whether antibiotic therapy increased bacteriological cure rate and lowered somatic cell count (SCC) compared with untreated controls. A CMT was performed and milk samples were collected for bacteriology from 211 glands of 106 does between 0 and 10 d after kidding. From a population of 3,239 glands from goats in 4 commercial herds, goats with one or both glands with a CMT score of >1 and from which bacteria were isolated were either assigned to be treated with 3 intramammary infusions at 12-h intervals of 75 mg of sodium ampicillin and 250 mg of sodium cloxacillin (n=57 glands) or left as untreated controls (n=49 glands). Milk samples were collected again 14 ± 3 and 21 ± 3 d later for bacteriology and SCC determination. Composite milk yield, goat SCC, length of lactation, and survival data were collected. A partial budget was constructed to assess the cost effectiveness of treatment. At a cut point of greater than trace, the sensitivity, specificity, and positive and negative predictive values of the CMT were 0.74, 0.74, 0.42, and 0.92, respectively. Treatment increased the bacteriological cure rate compared with no treatment [30/57 (53%) vs. 6/49 (12%)], but there was a pathogen by treatment interaction whereby treatment increased cure proportion in glands infected with minor, but not major, pathogens. Treatment reduced the foremilk gland-level SCC [1,595 (95% CI=1,106-2,300) vs. 3,028 (95% CI=2,091-4,385) geometric mean (× 1,000) cells/mL] but not the SCC at goat level [1,596 (95% CI=1,219-2,090) vs. 1,488 (95% CI=1,132-1,955) geometric mean (× 1,000) cells/mL] compared with no treatment. Milk yield, risk of removal from the herd, and length of lactation were not altered by treatment. Treatment resulted in a loss of NZ$20.39/doe. It was concluded that use of the CMT as a screening test resulted in a higher likelihood of finding a gland that would be infected than selecting a gland at random. Treatment increased bacteriological cure rate and reduced SCC at gland level compared with no treatment. However, at goat level, milk yield, SCC, and survival were not altered, resulting in no economic benefit of treatment.


Assuntos
Ampicilina/análogos & derivados , Anti-Infecciosos/uso terapêutico , Cloxacilina/uso terapêutico , Doenças das Cabras/diagnóstico , Doenças das Cabras/tratamento farmacológico , Mastite/veterinária , Ampicilina/uso terapêutico , Animais , Contagem de Células/veterinária , Corynebacterium/isolamento & purificação , Feminino , Cabras , Lactação , Glândulas Mamárias Animais/microbiologia , Mastite/diagnóstico , Mastite/tratamento farmacológico , Leite/citologia , Leite/microbiologia , Sensibilidade e Especificidade , Fatores de Tempo , Resultado do Tratamento
4.
Science ; 213(4509): 787-9, 1981 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-7256280

RESUMO

Intracellular recordings were made from locally thermosensitive preoptic neurons in the green sunfish, Lepomis cyanellus. Stable resting potentials, action potentials, and spontaneous synaptic activity were observed over approximately 4 degrees to 5 degrees C changes in local brain temperature. A small percentage of the warm-sensitive neurons showed exponential firing-rate responses to temperature. These cells discharged rhythmically, lacked visible synaptic input, and showed slowly depolarizing potentials leading to action potentials. Other linear and nonlinear warm-sensitive and cold-sensitive neurons showed spontaneous excitatory and inhibitory synaptic potentials giving rise to action potentials. Cells that appear to be endogenously active may be true thermodetectors, and other thermosensitive neuronal activity may be synaptically mediated.


Assuntos
Regulação da Temperatura Corporal , Hipotálamo/fisiologia , Área Pré-Óptica/fisiologia , Potenciais de Ação , Animais , Temperatura Baixa , Peixes , Temperatura Alta
5.
J Dairy Sci ; 91(6): 2236-46, 2008 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18487646

RESUMO

We have used cDNA microarray analysis to identify genes that play a role in bovine mammary involution. Involution was induced by termination of milking, and alveolar tissue was collected from 48 nonpregnant Friesian cows in mid lactation sacrificed at 0, 6, 12, 18, 24, 36, 72, and 192 h (n = 6/group) postmilking. The most highly upregulated genes were those associated with oxidative stress. Quantitative real-time reverse-transcription PCR analysis confirmed that mRNA expression of spermidine/spermine N(1)-acetyltransferase was increased by 24 h, superoxide dismutase 2 and metallothionein 1A by 36 h, and glutathione peroxidase by 72 h postmilking. The mRNA expression of the host defense proteins lactoferrin and lingual antimicrobial peptide were increased by 192 h postmilking. A dramatic increase in the protein expression of lactoferrin by 192 h postmilking was also detected by Western analysis. Decreased mRNA expression of the milk protein genes alpha(S1)-, beta-, and kappa-casein, and alpha-lactalbumin were early events in the process of involution occurring within 24 to 36 h postmilking, whereas beta-lactoglobulin mRNA was decreased by 192 h postmilking. Decreases in alpha-lactalbumin and beta-lactoglobulin protein levels in alveolar tissue occurred by 24 and 192 h postmilking, respectively, and the cell survival factors beta1-integrin and focal adhesion kinase were decreased by 72 and 192 h postmilking, respectively. The results demonstrate that in the bovine mammary gland, decreased milk protein gene expression and cell survival signaling are associated with multiple protective responses to oxidative stress that occur before the induction of immune responses and mammary epithelial cell apoptosis during involution.


Assuntos
Apoptose , Bovinos/fisiologia , Lactação/metabolismo , Glândulas Mamárias Animais/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos/veterinária , RNA Mensageiro/genética , Regulação para Cima , Animais , Antioxidantes/metabolismo , Apoptose/genética , Western Blotting/veterinária , Bovinos/genética , Feminino , Lactoferrina/genética , Lactoferrina/imunologia , Lactoferrina/metabolismo , Glândulas Mamárias Animais/imunologia , Proteínas do Leite/genética , Proteínas do Leite/imunologia , Proteínas do Leite/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Estresse Oxidativo/genética , Estresse Oxidativo/fisiologia , RNA Mensageiro/análise , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Fatores de Tempo
6.
Mar Pollut Bull ; 126: 610-621, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29129320

RESUMO

Chemical analyses and toxicity testing using six marine species were used to characterize the hazard of produced waters (PW) to marine life from twelve Australian offshore platforms. Hazard data were used in conjunction with platform-specific plume discharge dilution and species sensitivity distribution modeling to estimate cumulative risks by calculating the multiple substance potentially affected fraction of species in the local marine environment. Results provided two independent lines of evidence demonstrating that cumulative risks to marine life from these discharges meet intended 95% species protection goals at the edge of the mixing zone. A limited number of PW constituents (hydrocarbons, sulphide and ammonia) appeared to dictate risk thereby informing management and providing a rationale for more targeted analyses in future monitoring studies. Based on these findings a tiered framework is proposed to foster consistent screening and potential refinement of cumulative risk evaluations for PW discharges.


Assuntos
Resíduos Industriais , Indústria de Petróleo e Gás , Poluentes Químicos da Água/análise , Amônia/análise , Austrália , Monitoramento Ambiental , Hidrocarbonetos/análise , Medição de Risco , Sulfetos/análise
7.
Vet Immunol Immunopathol ; 115(3-4): 205-15, 2007 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-17173979

RESUMO

The bovine mammary gland requires lymphocytes for immune protection of the gland from foreign pathogens and, in addition, to transfer immune protection to the neonate via colostrum and milk. The process of homing primed lymphocytes to tissues is mediated by the interaction of cell-adhesion molecules displayed on the surface of lymphocytes and counter receptors displayed on the vascular endothelium. This study was conducted to identify the cell-adhesion molecules involved in homing lymphocytes to the bovine mammary gland at four different physiological stages; pregnant, colostral, lactation and involution. The expression and distribution of adhesion molecules in alveolar tissues and supramammary lymph nodes from the mammary glands of healthy cows was determined in situ by immunohistochemical analysis and compared with bovine Peyer's patch, used as a typical mucosal-associated lymphoid tissue and positive control. The mucosal addressin molecule, MAdCAM-1, was not detected in bovine mammary tissues at any of the four different physiological stages. Absence of MAdCAM-1 expression was verified by quantitative real-time RT-PCR analysis. Transcription levels of MAdCAM-1 mRNA were found to be more then 5 x 10(3)-fold lower in mammary alveolar tissues compared with bovine Peyer's patch tissues. In contrast to MAdCAM-1, phase-dependent protein expression of VCAM-1 was detected in both mammary alveolar tissues and the supramammary lymph nodes, with the highest expression observed in colostral phase cows. The protein expression in mammary alveolar tissues was limited to larger venules, although in colostral phase cows, VCAM-1 was also detected around the alveoli perimeter. In the supramammary lymph node, VCAM-1 protein was observed on both small and large venules. PNAd was detected in supramammary lymph nodes at all physiological stages of the mammary gland; however, it was not found in mammary alveolar tissues. Lymphocytes expressing beta7 were not detected in mammary tissues and lymphocytes expressing CD62L were only observed in the supramammary lymph nodes. Overall the data suggest that MAdCAM-1 and VCAM-1 are not involved in homing lymphocytes to the bovine mammary gland; whereas, VCAM-1 and PNAd may have this role in the supramammary lymph node.


Assuntos
Antígenos de Superfície/biossíntese , Bovinos/imunologia , Glândulas Mamárias Animais/imunologia , Proteínas de Membrana/biossíntese , Mucoproteínas/biossíntese , Molécula 1 de Adesão de Célula Vascular/biossíntese , Animais , Antígenos de Superfície/genética , Antígenos de Superfície/imunologia , Feminino , Imuno-Histoquímica/veterinária , Glândulas Mamárias Animais/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana/imunologia , Mucoproteínas/genética , Mucoproteínas/imunologia , Nódulos Linfáticos Agregados/imunologia , Período Pós-Parto , Gravidez , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Receptores de Retorno de Linfócitos/imunologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Molécula 1 de Adesão de Célula Vascular/genética , Molécula 1 de Adesão de Célula Vascular/imunologia
8.
J Dairy Sci ; 89(12): 4520-6, 2006 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17106082

RESUMO

Goat milk and cow milk are commonly used in infant formula preparations and, as such, understanding the nutritional characteristics of infant formulas made from these milks is important. In this study, a goat milk infant formula was compared with an adapted (whey-enhanced) cow milk infant formula with respect to mineral absorption and deposition using the 3-wk-old piglet as a model for the 3-mo-old infant. Equal numbers of piglets (n = 8) were fed either the goat milk formula or the cow milk formula. The mineral composition of the prepared goat milk formula was higher than that of the prepared cow milk formula for most minerals, including calcium (75.1 vs. 56.7 mg/100 mL) but excluding iron, which was higher in the prepared cow milk formula (0.92 vs. 0.74 mg/100 mL). The amounts of calcium, phosphorus, and manganese absorbed by the piglets were significantly higher for the goat milk formula, whereas the amounts of zinc, iron, and magnesium absorbed were significantly higher for the cow milk formula. Apparent mineral absorption, relative to intake, was statistically higher in the cow milk formula for calcium and phosphorus, although the actual differences were very small (less than 1.3%). For copper, zinc, iron, and magnesium there was no significant difference between treatments in apparent mineral absorption, whereas for manganese, absorption was higher for the goat milk infant formula. The absolute mineral deposition was higher in piglets fed the goat milk formula for calcium, phosphorus, and manganese, whereas iron deposition was higher in the piglets fed cow milk formula. For all other minerals tested, there were no significant differences between treatments. The goat milk infant formula provided a pattern of mineral retention in the 3-wk-old piglet very similar to that of the adapted cow milk infant formula. The minor differences observed between the 2 appeared to be due to the different mineral contents of the 2 formulas.


Assuntos
Fórmulas Infantis/química , Leite/química , Minerais/farmacocinética , Absorção , Animais , Disponibilidade Biológica , Osso e Ossos/química , Cálcio/análise , Bovinos , Creatinina/urina , Fezes/química , Cabras , Masculino , Minerais/análise , Minerais/urina , Modelos Animais , Valor Nutritivo , Suínos
9.
J Dairy Sci ; 89(7): 2408-13, 2006 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-16772556

RESUMO

Goat milk is used as an alternative to cow milk for the production of infant formulas. However, little is known about the protein quality and, specifically, about the digestible AA pattern of goat milk formulas compared with their cow milk counterparts. In this study, the true ileal AA digestibility of a goat milk infant formula was compared with a premium cow milk infant formula. The 3-wk-old piglet was used as a model for the 3-mo-old infant. Both milk formulas were prepared as described by the manufacturer, with titanium dioxide added as an indigestible marker. The formulas were fed to the piglets over a 2-wk trial period. Digesta from the terminal ileum were collected post euthanasia and analyzed for AA content, along with samples of the formulas. True AA digestibility was determined after correcting for endogenous AA loss at the terminal ileum of pigs fed an enzyme-hydrolyzed casein-based diet, followed by ultrafiltration (5,000 Da) of the digesta. Total urine and feces collection was also undertaken to determine the nitrogen retention from the diets. The true ileal AA digestibility was similar between the goat and cow milk infant formulas for all AA except Gly and Trp. There was no significant difference in the nitrogen retention of piglets fed the two different formulas. The goat milk infant formula and the premium cow milk infant formula were similar in terms of protein quality.


Assuntos
Aminoácidos/metabolismo , Bovinos , Digestão , Cabras , Fórmulas Infantis/química , Leite/química , Aminoácidos/administração & dosagem , Animais , Proteínas Alimentares/administração & dosagem , Proteínas Alimentares/análise , Proteínas Alimentares/metabolismo , Fezes/química , Íleo/metabolismo , Masculino , Nitrogênio/análise , Nitrogênio/urina , Suínos
10.
Biochim Biophys Acta ; 613(1): 130-9, 1980.
Artigo em Inglês | MEDLINE | ID: mdl-7378414

RESUMO

An improved method is described for the preparation of milligram quantities of substrate-modified sulphatase A. The latter has the same molecular weight and the same ability to form a tetramer as has native sulphatase A. It has been shown that the modified enzyme prepared with nitrocatechol [35S]sulphate as substrate contains 1 mol 35SO24- per mol enzyme and that any treatment which causes reversion of the modified enzyme to native enzyme is accompanied by the loss of the bound SO24-. Dialysis of the 35S-modified enzyme against a solution containing SO24- causes a loss of 35SO24- with no change in the amount of modified enzyme in the preparation. It has been shown that the activation of the substrate-modified enzyme by SO24- does not lead to the formation of a third stable form of sulphatase A.


Assuntos
Arilsulfatases/metabolismo , Fígado/enzimologia , Sulfatases/metabolismo , Animais , Arilsulfatases/isolamento & purificação , Catecóis/metabolismo , Bovinos , Fenômenos Químicos , Química , Estabilidade de Medicamentos , Ativação Enzimática/efeitos dos fármacos , Cinética , Peso Molecular , Espectrofotometria Ultravioleta , Sulfatos/farmacologia
11.
Biochim Biophys Acta ; 687(2): 303-9, 1982 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-7093261

RESUMO

The effects of thermal acclimation of goldfish upon the bulk fluidity of synaptic, mitochondrial and myelin membrane fractions of brain was determined using steady-state and differential polarised phase fluorimetry. Membrane fluidity decreased in the order, mitochondria greater than synaptic membranes greater than myelin. in each case membranes from cold-acclimated goldfish were more fluid than the corresponding membranes of warm-acclimated goldfish, though the adjustment of fluidity in each case was insufficient to compensate for the direct effects of the temperature difference. The extent of fluidity compensation was greatest in the mitochondrial fraction and least in the myelin fraction, indicating heterogeneous responses in different membrane-types. Steady-state and dynamic fluorimetric techniques provided identical estimates of the homeoviscous responses, indicating that despite its short-comings, the steady-state technique provided as good a measure of adaptive responses as the more complex and sophisticated technique.


Assuntos
Encéfalo/fisiologia , Membranas Intracelulares/fisiologia , Membranas Sinápticas/fisiologia , Animais , Carpa Dourada , Fluidez de Membrana , Mitocôndrias/fisiologia , Bainha de Mielina/fisiologia , Espectrometria de Fluorescência , Temperatura
12.
Biochim Biophys Acta ; 599(2): 341-58, 1980 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-7407099

RESUMO

The liver mitochondrial and microsomal membranes of green sunfish and rat were examined by steady state polarisation and differential polarised phase fluorimetry to determine the effects of seasonal adaptation of membrane dynamic structure to temperature. Steady state polarisation studies indicated that the liver mitochondria of green sunfish acclimated to different temperatures showed a greater partial compensation of membrane fluidity for the fatty acid composition of both membrane preparations generally became more unsaturated at lower acclimation temperatures, though the differences between 5 degrees C and 25 degrees C acclimated fish were more pronounced in the mitochondrial fraction than in the microsomal fraction. Differential polarised phase fluorimetric studies indicated that the rotations of diphynylhexatriene in mitochondrial and microsomal membranes were highly hindered, though the hindrance offered by membranes of 25 degrees C acclimated green sunfish was far greater than that offered by the membranes of 5 degrees C acclimated fish, thus supporting the concept of homeoviscous adaptation. The absolute rotational rate was not consistently affected by acclimation treatment.


Assuntos
Aclimatação , Membranas Intracelulares/ultraestrutura , Lipídeos de Membrana/análise , Microssomos Hepáticos/ultraestrutura , Mitocôndrias Hepáticas/ultraestrutura , Fosfolipídeos/análise , Animais , Ácidos Graxos/análise , Peixes , Ratos , Estações do Ano , Especificidade da Espécie , Espectrometria de Fluorescência
13.
Biochim Biophys Acta ; 511(3): 442-52, 1978 Aug 17.
Artigo em Inglês | MEDLINE | ID: mdl-687623

RESUMO

Temperature adaptation of biological membranes was examined by comparing the fragmented sarcoplasmic reticulum preparation of goldfish acclimated to different temperatures. Membrane fluidity was estimated using the fluorescence polarization technique. There was considerable variation between preparations, but no consistent differences in fluidity were observed between 5- and 25 degrees C-acclimated goldfish, fish species adapted over an evolutionary period to arctic or desert temperatures, and rat. The fatty acid composition of the sarcoplamic reticulum preparations of differently acclimated goldfish showed differences in the proportion of mono- and polyunsaturated fatty acids while the proportion of saturated fatty acids remained relatively constant. However, the fatty acid composition of sarcoplasmic reticulum phosphoglycerides became more unsaturated in the order rat, desert pupfish, arctic sculpin, which correlates with their respective environmental or body temperature. It is concluded that differences in membrane components other than fatty acids are important in determining membrane dynamic structure. The inability to demonstrate homeoviscous adaptation in sarcoplasmic reticulum is supported by other evidence suggesting that functions of the sarcoplasmic reticulum that are measured in vitro are not affected by such modifications of their phosphoglyceride fatty acid composition as occur during thermal acclimation.


Assuntos
Adaptação Biológica , Membranas/metabolismo , Retículo Sarcoplasmático/metabolismo , Animais , Ácidos Graxos/análise , Peixes , Polarização de Fluorescência , Carpa Dourada , Fluidez de Membrana , Fosfolipídeos/metabolismo , Polienos/metabolismo , Ratos , Sinaptossomos/metabolismo , Temperatura , Termodinâmica , Viscosidade
14.
Endocrinology ; 119(1): 91-6, 1986 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-3522215

RESUMO

Epithelial cells were isolated from mammary glands of mice in various physiological states, and the rate of carrier-mediated glucose transport was determined with 3-O-methylglucose. The basal rate (in the absence of exogenous insulin) increases about 40-fold as the animal of origin progresses from the virgin to the midlactating state and declines precipitously during involution. Insulin does not acutely stimulate carrier-mediated transport by cells isolated from virgin or lactating mice and evokes only about a 50% enhancement of this transport rate in the cells derived from pregnant and early postlactational animals. However, culture of mammary explants from pregnant mice in the presence of insulin, cortisol, and PRL before isolation of the epithelial cells results in a 400% increase in the basal rate of carrier-mediated glucose transport in the absence of exogenous hormones; this effect requires 3 days of incubation. The enhanced rate is equivalent to that in cells freshly isolated from animals in early lactation. Insulin-like growth factor I can mimic the chronic effect of insulin, but multiplication-stimulating activity, epidermal growth factor, and 20% fetal calf serum do not supplant insulin in this system. The maximum velocity, but not the Km, of 3-O-methylglucose transport is markedly increased during ontogeny; this is compatible with an increase in the number of functional transporters during development. The results suggest that insulin and/or insulin like growth factor I are implicated in development of the high basal rate of carrier-mediated glucose transport in mammary cells from lactating mice.


Assuntos
Fator de Crescimento Insulin-Like I/farmacologia , Insulina/farmacologia , Glândulas Mamárias Animais/metabolismo , Metilglucosídeos/metabolismo , Metilglicosídeos/metabolismo , Proteínas de Transporte de Monossacarídeos/metabolismo , Somatomedinas/farmacologia , 3-O-Metilglucose , Animais , Transporte Biológico/efeitos dos fármacos , Células Cultivadas , Fator de Crescimento Epidérmico/farmacologia , Células Epiteliais , Feminino , Hidrocortisona/farmacologia , Fator de Crescimento Insulin-Like II/farmacologia , Cinética , Lactação , Glândulas Mamárias Animais/efeitos dos fármacos , Glândulas Mamárias Animais/crescimento & desenvolvimento , Camundongos , Camundongos Endogâmicos C3H , Gravidez , Prolactina/farmacologia
15.
Endocrinology ; 120(4): 1411-6, 1987 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-3549260

RESUMO

The concentration-activity profiles for insulin and insulin-like growth factor I (IGF-I; in the presence of and insulin-like growth factor I (IGF-I; in the presence of hydrocortisone and PRL) have been compared in terms of the accumulation of beta-casein mRNA, total casein synthesis, and alpha-lactalbumin and basal carrier-mediated glucose transport activities in mammary epithelial cells from midpregnant mice. For the accumulation of the casein mRNA and the induction of casein synthesis and alpha-lactalbumin activity, the insulin ED50 is 1-2 ng/ml, while that for IGF-I is 10- to 20-fold greater. The effects of insulin and IGF-I are not additive in these instances. For the induction of basal carrier-mediated glucose transport, the insulin ED50 is 8 ng/ml, and that for IGF-I is 16 ng/ml. Either factor can induce transport activity up to the level present in the cells from 2-day lactating mice. In this instance the effects are additive; insulin and IGF-I together can induce the transport up to the 10-day lactating level.


Assuntos
Caseínas/biossíntese , Glucose/metabolismo , Fator de Crescimento Insulin-Like I/farmacologia , Insulina/farmacologia , Lactalbumina/metabolismo , Glândulas Mamárias Animais/metabolismo , Somatomedinas/farmacologia , 3-O-Metilglucose , Animais , Transporte Biológico/efeitos dos fármacos , Caseínas/genética , Epitélio/metabolismo , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Glândulas Mamárias Animais/efeitos dos fármacos , Metilglucosídeos/metabolismo , Camundongos , Gravidez , RNA Mensageiro/metabolismo
16.
J Clin Endocrinol Metab ; 67(1): 25-9, 1988 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-3259954

RESUMO

The concentration of immunoreactive insulin-like growth factor I (IGF-I) in human mammary secretions, assayed after acid-ethanol extraction, was high [mean, 4.1 +/- 0.5 (+/- SE) nmol/L; n = 13] for several weeks prepartum. It then decreased during the first 3 days postpartum to 1.3 +/- 0.1 nmol/L (n = 28), in parallel with changes in epidermal growth factor (EGF) and protein concentrations. However, between the first and sixth weeks postpartum, the IGF-I concentration increased to 2.5 +/- 0.2 nmol/L (n = 18), while levels of EGF and protein decreased further. Given that the volume of milk produced increases during this period, the total IGF-I output rose by up to 4-fold, while EGF output remained constant. The increase in IGF-I and decrease in EGF in milk suggest that different regulatory mechanisms control the output of different growth factors by the mammary gland.


Assuntos
Fator de Crescimento Insulin-Like I/análise , Lactação/metabolismo , Leite Humano/análise , Somatomedinas/análise , Fator de Crescimento Epidérmico/análise , Feminino , Humanos , Proteínas do Leite/análise , Período Pós-Parto/metabolismo , Gravidez , Terceiro Trimestre da Gravidez , Radioimunoensaio , Fatores de Tempo
17.
J Immunol Methods ; 177(1-2): 175-84, 1994 Dec 28.
Artigo em Inglês | MEDLINE | ID: mdl-7822824

RESUMO

The extraction and purification of serum-derived immunoglobulin fraction in the egg yolk of hens by the combined treatment of the raw egg yolk with caprylic (octanoic) acid and ammonium sulphate is described. This simple two-step method proved to be both rapid, reproducible and suitable for batch processing of pooled egg yolk. The method recovered in excess of 130 mg of immunoglobulin per egg yolk. Two chickens were inoculated at two weekly intervals with 100 micrograms each of ovine alpha-lactalbumin over a ten week period. The alpha-lactalbumin antigen was purified by a hydrophobic-interaction chromatographic procedure and further purified by a gel excision-elution process. No precipitating antibodies could be demonstrated in gel diffusion techniques with this antibody. The specificity and specific activity of the antibody were monitored by western blotting and demonstrated the presence of highly specific antibodies to ovine alpha-lactalbumin in the treated egg yolk. The extraction procedure had no adverse effects on antibody titre. We concluded, and confirmed previous reports, that the use of chickens for the production of highly specific antibodies to mammalian proteins with particular reference to milk proteins presented numerous advantages over conventional procedures.


Assuntos
Anticorpos/isolamento & purificação , Caprilatos/química , Gema de Ovo/imunologia , Imunoglobulinas/isolamento & purificação , Lactalbumina/imunologia , Animais , Galinhas , Imunização , Imunoeletroforese , Peso Molecular , Ovinos
18.
J Endocrinol ; 150(1): 121-7, 1996 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-8708553

RESUMO

Plasma clearance of insulin-like growth factors-I and -II (IGF-I and -II) and insulin-like growth factor-binding protein-2 (IGFBP-2) from lactating goats (n = 4) was determined following a single intravenous injection of the corresponding 125I-labelled human protein. Transfer of these proteins out of the vascular space was monitored by their subsequent appearance in mammary-derived lymph and milk. Clearance of 125I-IGFBP-2 from circulation was 0.37 +/- 0.06 ml/min/kg, which is markedly greater than that of 125I-IGF-I or -II (0.11 +/- 0.01 and 0.12 +/- 0.01 ml/min/kg respectively). This was also reflected in longer elimination half-lives for IGF-I (353 +/- 6 min) and -II (254 +/- 8 min) compared with IGFBP-2 (110 +/- 9 min). Three hours after injection of the 125I-labelled protein, the plasma:lymph ratio of trichloroacetic acid-precipitable radioactivity was 1.54 +/- 0.04, 3.3 +/- 0.6 and 4.1 +/- 0.4 for IGFBP-2, IGF-I and -II respectively. The form of 125I-IGFBP-2 in lymph was not different from that of plasma. Elevation of plasma concentrations of IGFBP-2 by its intravenous infusion significantly decreased plasma half-life of both IGF-I and -II (251 +/- 8 and 198 +/- 7 min respectively). Although the amount and rate of transfer of IGF into mammary-derived lymph was decreased slightly by IGFBP-2, concentrations eventually obtained were not different from control. However, secretion of IGFs into milk was significantly reduced by IGFBP-2, particularly in the case of IGF-I. These results are consistent with the ability of all three compounds to cross the vascular endothelium intact and of IGFBP-2 to decrease the uptake of IGF by mammary epithelium and subsequent secretion into milk. IGFBP-2 may well have acted to target plasma IGF towards non-mammary tissues, thus explaining the more rapid plasma clearance of IGFs in the presence of elevated IGFBP-2.


Assuntos
Cabras/metabolismo , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/farmacocinética , Linfa/metabolismo , Leite/metabolismo , Somatomedinas/farmacocinética , Animais , Feminino , Meia-Vida , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/sangue , Fator de Crescimento Insulin-Like I/análise , Fator de Crescimento Insulin-Like I/farmacocinética , Fator de Crescimento Insulin-Like II/análise , Fator de Crescimento Insulin-Like II/farmacocinética , Radioisótopos do Iodo , Linfa/química , Taxa de Depuração Metabólica , Leite/química , Somatomedinas/análise
19.
J Endocrinol ; 135(2): 311-6, 1992 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-1474338

RESUMO

The milk yield and mammary blood flow responses to close-arterial, intra-mammary infusion of IGF-I were investigated in five Saanen goats milked frequently or normally the day before. Animals were infused for 6 h with recombinant human IGF-I (1.3 nmol/min) and milked hourly following i.v. injection of oxytocin beginning 2 h before infusion and then every 2 h. On one occasion animals were milked five times (after i.v. injection of oxytocin) on the day before infusion and on the other they were milked twice, without oxytocin. The ratio of milk yield from the infused to that from non-infused gland increased by 17 +/- 4% (mean +/- S.E.M.) in goats milked twice the day before infusion and by 6 +/- 2% when the infusion was preceded by frequent milking. Maximal responses were obtained 4 h after the start of the infusion and differed significantly (P < 0.05), according to pretreatment milking. Blood flow through the infused gland rose in parallel to the milk yield response. At 5 h, when maximal levels were achieved, blood flow was 182 +/- 23% of the pre-infusion flow rate following twice-daily milking and 139 +/- 3% of the pre-infusion flow rate following more frequent milk removal. Thus, more frequent milk removal on the day before close-arterial infusion of IGF-I attenuated both the milk yield and mammary blood-flow response to the infusion of IGF-I.


Assuntos
Cabras/fisiologia , Fator de Crescimento Insulin-Like I/farmacologia , Glândulas Mamárias Animais/fisiologia , Leite/metabolismo , Estimulação Física , Animais , Glicemia/metabolismo , Feminino , Cabras/sangue , Insulina/sangue , Fator de Crescimento Insulin-Like I/metabolismo , Lactação , Glândulas Mamárias Animais/irrigação sanguínea , Glândulas Mamárias Animais/efeitos dos fármacos , Ocitocina/farmacologia , Fluxo Sanguíneo Regional/fisiologia
20.
J Endocrinol ; 128(3): 457-63, 1991 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-2013749

RESUMO

Lactating goats exhibiting widely divergent responses to short-term (4 days) treatment with bovine GH (bGH) were retrospectively divided into two groups based on the magnitude of this response. There was no difference between groups in terms of the pretreatment milk yield, but by day 4 of treatment milk secretion had increased by 4.99 +/- 2.5 (S.E.M.) ml/h (P greater than 0.05 compared with pretreatment) for group 1 and 22.9 +/- 2.4 ml/h (P less than 0.001) for group 2. Plasma GH increased in both groups, but concentrations were significantly higher both before and during treatment in group 1 compared with group 2. Plasma concentrations of insulin-like growth factor-I (IGF-I) increased significantly during bGH treatment for both groups and there was no significant difference between the two until day 4 of treatment when levels of IGF-I in group 1 began to decline, whereas those from group 2 were maintained. Concentrations of IGF-I in milk from goats in group 1 were not significantly altered by GH administration, whereas those in goats in group 2 were increased by 40% (P less than 0.01 compared with pretreatment). Levels of IGF-I in mammary secretory tissue from four animals from group 1 were not altered by bGH (2.8 +/- 0.2 and 2.77 +/- 0.08 nmol/kg tissue before and after treatment respectively), but were significantly (P less than 0.05) increased in four animals from group 2 (2.80 +/- 0.2 and 9.9 +/- 1.1 nmol/kg tissue).(ABSTRACT TRUNCATED AT 250 WORDS)


Assuntos
Hormônio do Crescimento/farmacologia , Fator de Crescimento Insulin-Like I/metabolismo , Glândulas Mamárias Animais/metabolismo , Leite/metabolismo , Animais , Feminino , Cabras , Insulina/metabolismo , Lactação , Glândulas Mamárias Animais/efeitos dos fármacos , Glândulas Mamárias Animais/fisiologia , Leite/efeitos dos fármacos , Radioimunoensaio , Taxa Secretória/efeitos dos fármacos
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