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[Protein metabolism in the newborn lamb. IV. Consequences of amino acid and lactose ingestion].

A study was made on protein metabolism and hormonal changes following birth in newborn lambs fed amino acids alone or in combination with lactose. Eight newborn lambs taken from their mother immediately after birth were fed hourly for 8 h, either with a solution of peptides and free amino acids obtained by mild hydrolysis of whey proteins (4 lambs; diet AP) or with the same solution + lactose (4 lambs; diet APL). L-[4,5-3H] leucine was continuously perfused into a jugular vein for 6 h when the lambs were 2 h 30 min old. Plasma glucose and insulin levels increased after birth in APL lambs whereas they decreased in the AP; these differences were significantly different. Plasma cortisol levels remained unchanged throughout the experiment. Free essential amino acid levels did not vary when lambs were older than 4.5 h; they depended on the corresponding amino acid intakes. Plasma free threonine, valine, isoleucine, leucine, tyrosine and lysine were lower in APL than in AP lambs. The plasma leucine irreversible loss and leucine oxidation were higher in AP than in APL lambs. The plasma flux of leucine from whole body protein breakdown was lower in APL than in AP lambs inasmuch as the plasma flux of dietary leucine may be estimated by the amounts of leucine ingested in both cases. No significant difference was found for the fractional synthesis rates of tissue proteins such as liver, skin, skeletal muscle, lung, brain and whole body. These rates for skin, muscle and whole body were close to those previously measured in colostrum fed lambs. The increase in whole body protein accretion resulting from lactose feeding in combination with amino acids seemed to result from a decreased protein breakdown that could be mediated by the insulin response.

Amino Acids↗

Intracloacal Salmonella typhimurium infection of broiler chickens: reduction of colonization with anaerobic organisms and dietary lactose.

The combined effect of treatments with dietary lactose plus anaerobic organisms on cecal colonization of broiler chicks by Salmonella typhimurium was evaluated. Chickens treated with a combination of anaerobic organisms and 7% dietary lactose were resistant to cecal colonization by S. typhimurium. The number of recoverable S. typhimurium cells per gram of cecal contents taken on days 10 and 15 after infection was significantly reduced. Treatment with anaerobes without the addition of lactose did not effectively control cecal colonization. Intracloacal inoculations with bacterial concentrations that varied by 10,000-fold resulted in roughly similar levels of colonization. The treatments resulted in reduced cecal pH and elevated levels of undissociated volatile fatty acids. Statistically significant correlations (P less than 0.01) were observed between the S. typhimurium concentrations in cecal material and the concentrations of undissociated fatty acids (r = -0.79, and between the bacterial counts and pH (r = 0.72).

Animals↗

Cross-linking of the components of lactose synthetase with dimethylpimelimidate.

The cross-linking of the two components of lactose synthetase, alpha-lactalbumin and a galactosyltransferase, with dimethylpimelimidate was examined. The extent of the cross-linking at pH 8.1 was found to be dependent upon the presence of substrates or inhibitors for the galactosyltransferase. N-acetylglucosamine and mixtures of either N-acetylglucosamine, Mn-2+ and UDP, or UDP-galactose and Mn-2+ promoted the formation of cross-linked species. Glucose or a mixture of UDP and Mn-2+ were much less effective in promoting cross-linking. Two types of intermolecularly cross-linked species of alpha-lactalbumin and the galactosyltransferase were obtained. Each was a 1:1 cross-linked complex of alpha-lactalbumin and either of the two forms of the transferase with molecular weights of about 42,000 and 48,000, respectively. Cross-linked complexes were not observed with more than 1 molecule each of alpha-lactalbumin and the transferase. The cross-linked complexes were obtained in homogeneous form by gel filtration on Sephadex and absorption of uncross-linked enzyme by affinity chromatography on alpha-lactalbumin-Sepharose in the presence of N-acetylglucosamine. They migrated on gel electrophoresis in sodium dodecyl sulfate with mobilities in accord with their predicted molecular weights as 1:1 complexes of alpha-lactalbumin and the transferase. The amino acid composition of the cross-linked complex was in reasonable agreement with the expected composition of a 1:1 mixture of alpha-lactalbumin and galactosyltransferase. The enzymic properties of the cross-linked and uncross-linked enzymes were compared. The cross-linked complex had a much higher intrinsic lactose synthetase activity than did uncross-linked enzyme although only about 1% of the potential activity of uncross-linked enzyme in the presence of optimal concentrations of alpha-lactalbumin. The lactose synthetase activity of the cross-linked complex, however, was unaffected by exogenous alpha-lactalbumin. In addition, the complex readily catalyzed the transfer of galactose from UDP-galactose to xylose in the absence of exogenous alpha-lactalbumin. The N-acetyllactosamine synthetase activity of the complex was low compared to its activity with other monosaccharides. Ovalbumin, which is a good acceptor for the uncross-linked transferase, was not an acceptor for the cross-linked complex. Kinetic studies of the complex suggest that its modified catalytic activity is not the result of the modification by dimethylpimelimidate but reflects the expected effects of is provided, and that

Amino Acids↗

[Incidence of lactose malabsorption in the population 6-18 years of age].

Breath test was performed in 664 school-aged children in order to measure lactose malabsorption. The first screening showed that 23.4% of the children evidenced malabsorption of milk sugar. In these children further stool examination for Giardia lamblia infection, and saccharose breath test was performed to identify more complex absorption problems. The remaining 146 children were tested again after a period of 3-9 month and 45.8% of this population showed lactose malabsorption. In conclusion the authors determined that 10.1% of school aged children were permanently hypo- or alactasic. During the examination, they measured the approximate consumption of milk considering the quantity of milk intake showed that the consumption of milk and lactose malabsorption were not closely related normal and abnormal absorption among children who would not normally consume milk.

Adolescent↗

[The biological value of the proteins of dairy products with varying contents of lactose].

In experiments on growing Wistar male rats biological value of proteins was estimated in 5 milk products containing proteins: "Detolact", "Humana O", "Novolact MM", lactalbumin and "Dialact" containing from 17 to 58% of lactose. The final level of lactose in the ration (7% of protein by mass) varied from 4.4 to 30.6%. The parameters of protein biological value in the tested products did not significantly differ from those of the control protein (casein) that has proved the validity of using rats for investigation of biological value of proteins in the rations containing up to 30% of lactose.

Animals↗

Characterization and sequencing of the lac Y54-41 "uncoupled" mutant of the lactose permease.

The Escherichia coli strain carrying the lac Y54-41 gene encodes a mutant lactose permease which carries out normal downhill transport of galactosides but is defective in uphill accumulation. In this study, the mutant lac Y54-41 gene was cloned onto the multicopy vector pUR270. As expected, the cloned gene was shown to express normal downhill transport activity but was markedly defective in the uphill transport of methyl-beta-D-thiogalactopyranoside. Direct measurements of H+ transport revealed that the mutant permease can transport H+ with methyl-beta-D-thiogalactopyranoside but at a significantly reduced capacity compared to the wild-type strain. However, under conditions where the mutant and wild-type strains both transport lactose at similar rates, no detectable H+ transport was observed in the mutant strain. The entire cloned lac Y54-41 gene was subjected to DNA sequencing, and a single base substitution was found which replaces glycine 262 in the protein with a cysteine residue. Inhibition experiments showed that the mutant permease is dramatically more sensitive to three different sulfhydryl reagents: N-ethylmaleimide, p-hydroxymericuribenzoate, and p-hydroxymercuriphenylsulfonic acid. However, the lactose analogue, thiodigalactoside, was only marginally effective at protecting against inhibition in the mutant strain. The results are consistent with the idea that the sulfhydryl reagents are inhibiting the mutant permease activity by reacting with cysteine 262.

Base Sequence↗

Ion chromatographic determination of lactose, galactose, and dextrose in grated cheese using pulsed amperometric detection.

A method is presented to determine lactose, galactose, and dextrose in grated cheese using ion chromatography with pulsed amperometric detection (IC/PAD). The method is specific, sensitive, rapid, and precise. Sugar levels as low as 0.01% are detectable, and recoveries ranged from 114% at the 0.1% level to 93.3% at the 2.0% level. The precision expressed as coefficient of variation (CV) was 2.3% for lactose, 2.1% for galactose, and 3.1% for dextrose. Lactose, galactose, and dextrose levels are given for 45 authentic hard cheeses and 26 authentic soft cheeses.

Cheese↗

Is lactose malabsorption a risk factor in fractures of the elderly?

The prevalence of lactose malabsorption (LM) was determined by lactose tolerance tests in 18 women with spinal fragility fractures and 28 women with hip fractures and compared with 35 female controls of the same ethnic background. The association of LM with bone density in the spine, bone mineral density in the distal radius and histomorphometric values of iliac crest biopsies was also evaluated. The prevalence of LM did not differ between the fracture groups and the control subjects. Patients with LM did not differ from those with normal lactose tolerance with regard to bone densitometry or histomorphometric findings in iliac crest biopsies. These results suggest that in Finland LM is not a risk factor for osteopenic fractures in the elderly.

Adult↗

Characteristics of lactose-fermenting Salmonella strains from Poland.

In this study 184 lactose-fermenting Salmonella strains, collected in the National Salmonella Centre from the northern and central parts of Ponad were examined. Epidemiological, serological and biochemical investigations were carried out. Apart from this, chemotherapeutic resistance and male-phage sensitivity were determined. Most of strains belonged to S. agona serotype (S. typhimurium and S. oranienburg were also presented) which apart from the lactose-fermenting ability retained all the remaining biochemical features typical of Salmonella bacilli, were male-phage M13 resistant and showed a high resistance to a wide spectrum of chemotherapeutics. In order to establish the way of the acquiring lac+ property by Salmonella bacilli P22 phage transduction and conjugation experiments, with E. coli F'lac and Hfr H as donors, were performed. S. agona lac- strains were shown to acquire the lactose-fermenting ability by mating with E. coli.

Adult↗

[Effect of lactose ingestion on body development. Study in rats].

Dams (rats) and their offsprings were fed on diets containing casein (Cas) or rice and beans (RB) as protein source. The presence of lactose improved the whole animal as well as the intestinal development in Cas diet but not from RB ones. It is suggested that lactose must be maintained permanently in diet but in low concentration. In this investigation it was also observed that the small intestinal development occurs initially by enhancement of length and weight, while after weaning only weight continues to increase independently of the presence or not of lactose in diet.

Animals↗

Site-specific alteration of cysteine 176 and cysteine 234 in the lactose carrier of Escherichia coli.

In the present study, Cys-176 and Cys-234 in the lactose carrier have been modified to serine residues via site-specific mutagenesis. The resultant mutants have been characterized with regard to galactoside transport activity and sulfhydryl reagent sensitivity. The mutant proteins (in which Cys-176 or Cys-234 had been replaced with serine) are able to effectively transport galactosides, although the transport rates for lactose and methyl-beta-D-galactopyranoside are slightly reduced compared to the normal lactose carrier. In addition, both mutants are less sensitive than the wild-type to high concentrations of two different sulfhydryl reagents, N-ethylmaleimide and p-hydroxymercuribenzoate. Overall, the data are consistent with the idea that Cys-176 and Cys-234 are close to the substrate recognition site. However, neither residue appears to be essential for galactoside transport by providing an ionizable group near the active site or by forming a disulfide bond.

Cysteine↗

Characterization of lactose carrier mutants which transport maltose.

Brooker and Wilson (Brooker, R. J., and Wilson, T. H. (1985) Proc. Natl. Acad. Sci. U. S. A. 82, 3959-3963) previously isolated lactose carrier mutants which were able to transport maltose. All of the mutants were found to be single amino acid substitutions for alanine 177 or for tyrosine 236. In the present study, we have examined the ability of these mutants to transport maltose, lactose, o-nitrophenyl-beta-D-galactopyranoside, methyl-beta-D-thiogalactopyranoside, and H+. Both the position 177 and 236 mutants have enhanced rates of maltose transport and exhibit apparent Km values for maltose which are substantially less than that of the wild-type strain. The position 177 mutants transport lactose and other galactosides at a normal rate and with normal affinity during downhill transport and show counterflow transport rates which are faster than the wild-type strain. Interestingly, these mutants are markedly defective in accumulating substrates against a concentration gradient, yet retain a normal H+:galactoside stoichiometry. The position 236 mutants appear to be defective in the downhill, uphill, and counterflow transport of galactosides but exhibit a normal H+:galactoside stoichiometry.

Escherichia coli↗

Roentgenographic observation of gas-fluid levels in the colon of children with abdominal pain and malabsorption of lactose.

Gas-fluid levels in the colon observed in radiographs are abnormal and usually indicate serious gastrointestinal disease. Colonic gas-fluid levels associated with concurrent abdominal pain and malabsorption of lactose, documented by lactose breath hydrogen testing, were observed in five children. Incomplete lactose absorption is a relatively benign condition that can be added to the differential diagnosis of gas-fluid levels in the colon and may account for some cases of spontaneous resolution of clinical and radiologic signs in children presenting with acute recurrent abdominal pain.

Child↗

Isolation, purification & properties of lactose binding agglutination factor from rabbit skeletal muscle.

The lactose binding agglutination factor from rabbit skeletal muscle is isolated through the use of lactose and urea. The factor is purified with DEAE cellulose and Sepharose 4B chromatography. The molecular weight of the factor is determined with Sephadex G-75 chromatography and found to be 28,000 Daltons. The subunit's molecular weight is determined by SDS gel electrophoresis, and found to be 14,000 Daltons. It was found that this binding factor can agglutinate trypsin-treated rabbit erythrocytes. Moreover, this agglutination is inhibited by EDTA and lactose.

Animals↗

Milk insulin, GH and TSH: relationship to changes in milk lactose, glucose and protein during lactogenesis in women.

Changes in concentration of insulin, growth hormone (GH) and thyroid stimulating hormone (TSH) in the whey fraction of mammary secretion of women during late pregnancy and lactogenesis were determined by radioimmunoassay. The milk hormone changes were compared to changes in the concentration of milk lactose, glucose and proteins which reflect the transition from colostrum to milk production during lactogenesis. During late pregnancy the average concentrations of insulin, GH and TSH in the colostrum of 2 women were 114.6 microU ml-1, 21.7 microU ml-1 and 14.1 microU ml-1, respectively. The concentrations of these hormones fell from high to low levels between day 1 and day 2 post partum concomitantly with the changes in the concentration of milk lactose, glucose and protein. On day 5 post partum the average milk concentrations of insulin, GH and TSH were 21.0 microU ml-1, 4.0 microU ml-1, and 5.0 microU ml-1, respectively. Similar milk hormone changes occurred in non breast feeding women during the first 11 days post partum. During 2 to 13 months of lactation one woman, the average concentration of milk insulin was 12 microU ml-1. In another woman, from 0 to 22 days after termination of breast feeding, the concentration of insulin increased from 19 to 56 microU ml-1. There were significant positive correlations between hormones and total protein and negative correlations between hormones and lactose, and hormones and glucose in women during lactogenesis and involution. The results showed that the changes in insulin, GH and TSH in milk were closely related to changes in secretory activity and permeability of the breast.

Colostrum↗

[Method of determining the activity of beta-galactosidase with lactose as substrate].

A method for estimating beta-galactosidase activity (beta-D-galactosidase-galactohydrolase, EC 3.2.1.23) with lactose as substrate was developed. The glucose formed as a result of lactose hydrolysis was determined by glucoseoxidase method with potassium ferrocyanide as a substrate-dye. The method is convenient, simple and reproducible, as well as gives an opportunity to characterize the enzyme due to its ability to hydrolyze lactose that is of great value when putting the enzyme into practice.

Buffers↗

Mechanism of regulation of the lactose permease by the phosphotransferase system in Escherichia coli: evidence for protein-protein interaction.

Binding of enzyme IIIglc to membranes was demonstrated in vitro, using membrane fragments from an E. coli strain which produces elevated levels of the lactose permease. Lactose and other substrates of the lactose permease enhanced the binding, but phosphoenolpyruvate decreased it in the presence of enzyme I and HPr. HPr also bound to the membranes under some conditions. The results support a model of permease regulation involving allosteric protein-protein interactions.

Allosteric Regulation↗