Characteristic transfer of colostrum-derived biologically active substances into cerebrospinal fluid via blood in natural suckling neonatal pigs.

The characteristic transfer of colostral components into cerebrospinal fluid (CSF) via serum after natural suckling has been studied by sodium dodecyl sulphate (SDS) electrophoresis, two-dimensional electrophoresis, immunoblot and enzyme linked immunosorbent assay methods in non-suckling pigs. Total protein concentrations in the serum increased immediately after first suckling, reached a peak value at 12 h, corresponding to a 2.3-fold compared with pre-suckling level. The protein concentration in CSF also increased and reached peak value at 6 h corresponding to 1.6-fold compared with presuckling level. IgG in serum not detected before suckling, increased steeply after suckling, IgG, IgM and IgA transported into the serum were observed in completely intact form by immunoblot method. The IgG transported into serum was quickly transferred into CSF after natural suckling in contrast to the case of bovine IgG. Serum concentration of transferrin was maintained at high level before suckling and was not changed by suckling. Transferrin also detected in CSF was not changed by suckling. Bovine lactoferrin (Lf) administered into the intestinal lumen was transported into serum (0.01%) and also detected in CSF after 6 h as undegraded form (3.1%). Thus, homologous IgG and bovine Lf are transported into CSF, suggesting that the transport of macromolecules into CSF is selective in neonatal pigs.

The effect of casein phosphopeptides on calcium absorption from calcium-fortified milk in growing rats.

The effect of casein phosphopeptides (CPP) prepared from bovine casein by enzymatic hydrolysis (extrinsic CPP) on Ca absorption from Ca-fortified milk was studied in young male rats, in comparison with that produced from casein in the small intestine (intrinsic CPP). The gastrointestinal Ca disappearance (Ca ingested - (gastric Ca + intestinal Ca )) was calculated as an indirect measurement of Ca absorption. After being fasted overnight, the animals were given 2.0 ml Ca-fortified milk (30 g fat, 35 g protein, 2.7 g Ca/kg) without or with 1.0 mg extrinsic CPP/ml, by gastric intubation. The intestinal soluble Ca level after 15 min and the gastrointestinal Ca disappearance after 15 and 30 min in the rats given Ca-fortified milk with 1.0 mg extrinsic CPP/ml were significantly higher than these figures in the rats given Ca-fortified milk without CPP (P < 0.05). When the rats were given unfortified milk (1.35 g Ca/kg) in another reference experiment, no significant effect on intestinal soluble Ca and gastrointestinal Ca disappearance was apparent from the addition of CPP to milk. Ca availability was estimated by measuring 45Ca-deposits in the bones of rats 48 h after being given 2.0 ml Ca-fortified milk labelled with 45Ca (180 kBq/2 ml) with or without 0.25 mg CPP/ml. The levels of 45Ca radioactivity of the femur and tibia from the rats given Ca-fortified milk with extrinsic CPP were significantly higher than those from the control group (P < 0.05). These results suggest that the addition of CPP to Ca-fortified milk could increase Ca absorption by growing rats mainly from CaCO3 added to the milk. The mechanism of CPP related to the interaction of CPP and Ca in the gastrointestinal tract is discussed.

Dietary nucleotides increase the proportion of a TCR gammadelta+ subset of intraepithelial lymphocytes (IEL) and IL-7 production by intestinal epithelial cells (IEC); implications for modification of cellular and molecular cross-talk between IEL and IEC by dietary nucleotides.

We have investigated the effects of dietary nucleotides on intraepithelial lymphocytes (IEL) and intestinal epithelial cells (IEC) in weanling mice. The proportion of T-cell receptor (TCR) gammadelta+ IEL in BALB/c mice fed a diet supplemented with nucleotides (NT(+) diet) was significantly higher than that in mice fed the nucleotide-free diet, while the proportion of TCR alphabeta+ IEL in NT(+) diet-fed mice was significantly decreased. The change of the TCR alphabeta+/TCR gammadelta+ ratio was mainly observed in a CD8 alphaalpha+ subset of IEL. IEC from NT(+) diet-fed mice produced a higher level of IL-7, which is important in the development of TCR gammadelta+ IEL, than those from control diet-fed mice. The expression levels of IL-7 and IL-2 receptors on IEL were not different between the two dietary groups. Our findings suggest that the increased population of a TCR gammadelta+ IEL subset by feeding nucleotides may be caused by the increased production of IL-7 by IEC.

Enhancing effect of dietary oil emulsions on immune responses to protein antigens fed to mice.

Repeated intragastric administration of beta-lactoglobulin (beta-Lg) with emulsified soybean oil elicited an antigen-specific, systemic humoral immune response in different strains of mice. The antibody response was enhanced as the dose of oil was increased and the particle size of emulsions was decreased. Feeding of aqueous beta-Lg could induce the antibody response only when emulsified oil was fed almost simultaneously. However, the emulsion-driven humoral immune response was not observed when mice were treated with anti-CD40 ligand antibody or in athymic mice. It is likely that the intestinal coexistence of emulsified oil with dietary antigens modulates the immune system to crucially support B cell response. A practical application of the present results to the prevention of cow's milk protein sensitization in infants is proposed.

Characteristic transfer of colostral components into cerebrospinal fluid via serum in neonatal pigs.

In order to evaluate the possibility of modification of brain function by colostral suckling, the characteristic transfer of colostral components into serum and cerebrospinal fluid (CSF) has been studied by SDS electrophoresis, immunoblot and ELISA methods in nonsuckling pigs. Total protein concentrations in the serum increased immediately after oral administration of bovine colostrum, reaching a peak value (7.0 +/- 0.7 g/dl) at 24 h after administration, corresponding to a 3-fold increase compared to preinfusion levels. IgG and other macromolecular components (MW 19, 000-58,000) were recognized in serum by electrophoretic and ELISA analysis. Total protein concentrations in the CSF collected from the cisterna magna also increased steeply after colostral administration, reaching a maximal value (54.1 +/- 5.0 mg/dl) at 4 h, corresponding to a 4-fold increase compared to preinfusion levels. Two colostral components (MW 19,000 and 31,000) in serum were confirmed to be present in the CSF by electrophoresis. The component of MW 19,000 was identified by immunoblot as beta-lactoglobulin. IgG in serum transferred from colostrum could not be detected in the CSF by ELISA. Lactoferrin administered into the intestine was also detected in the CSF via serum. These results indicate that some components of colostrum can be transported into the CSF via the serum, suggesting the possibility of modification of immature brain functions by colostral suckling in neonatal pigs.

Effect of the cholesterol content of a formula on the lipid compositions of plasma lipoproteins and red blood cell membranes in early infancy.

We investigated whether or not a regular formula for full-term infants supplemented with cholesterol (cholesterol-fortified) would increase the plasma cholesterol concentration and alter the red blood cell (RBC) membrane lipid composition in healthy full-term infants compared with their breast-fed counterparts. At 1 mo of age, total plasma cholesterol and low-density-lipoprotein (LDL) cholesterol were significantly higher in the breast-fed infants than in the cholesterol-unfortified, formula-fed infants. At 3 mo of age, total cholesterol and LDL cholesterol were significantly higher in the breast-fed infants than in the two formula-fed infant groups. These significant differences had disappeared by 6 mo of age. Although the cholesterol-unfortified, formula-fed infants had lower proportions of docosahexaenoic acid (DHA, 22:6n-3) and eicosapentaenoic acid (EPA, 20:5n-3) in the RBC membranes compared with the breast-fed group at 6 mo, DHA and EPA concentrations in the cholesterol-fortified, formula-fed infants were not significantly different. The results of the present study suggest that the plasma cholesterol concentration and fatty acid pattern of the RBC membranes in infants fed a cholesterol-fortified formula may be much closer to those in breast-fed infants than in infants fed a cholesterol-unfortified formula.

Dietary casein phosphopeptides prevent bone loss in aged ovariectomized rats.

The effect of dietary Ca-bound casein phosphopeptides (CaCPP) on the bones of aged ovariectomized (OVX) rats was studied as a model for post-menopausal bone loss. Three groups of ovariectomized rats were fed a control diet or one of two experimental diets, and one group of sham-operated rats (SHAM) was fed the control diet. The experimental diets contained 0.5% Ca and 0.4% P. In one diet, CaCPP was the sole source of calcium and provided 62.5% of dietary phosphorus (CaCPP diet). In the other, Ca-free CPP provided 100% of dietary phosphorus (Ca-free CPP diet). In the control diet, CaCO3 and KH2PO4 were used. During a 17-wk feeding period, there was little change in femoral bone mineral densities (BMD) of ovariectomized rats fed CaCPP and Ca-free CPP, or in the SHAM rats fed the control diet, whereas the bone mineral densities in the control ovariectomized rats decreased with time. Some of the segmental bone mineral densities of the excised femurs from the rats fed CaCPP were significantly higher than those from the control ovariectomized rats, but the values of the Ca-free CPP group were similar to those of the control ovariectomized rats. In the Ca-free CPP group, the discrepancy in bone mineral densities obtained between in vivo results and excised specimens might have been the result of a loss in bone mass due to their significant loss in body weight. There were no significant differences in serum inorganic phosphorus, alkaline phosphatase activity, osteocalcin or 1 alpha-25-dihydroxycholecalciferol concentrations among the ovariectomized groups. In the CaCPP and Ca-free CPP groups, urinary phosphorus excretion decreased and urinary calcium excretion increased significantly with time. The inhibitory effect on bone loss in aged ovariectomized rats could be due to the effects of dietary CaCPP on phosphorus and calcium metabolism.

Trace lipid from whey-mineral complex enhances calcium availability in young ovariectomized rats.

The effects of the trace lipids (L1 and L2) extracted from two kinds of whey-mineral complex on bone metabolism were studied in young ovariectomized (OVX) rats. The concentrations of oestradiol in the L1 and L2 oils were 305 and 1313 pg/ml respectively, while the concentrations of progesterone were 34.3 and 36.9 ng/ml respectively. Trace amounts of 1 alpha,25-dihydroxycholecalciferol (1 alpha,25-(OH)2D) were detected: 56 pg/ml in the L1 oil and 132 pg/ml in the L2 oil. The OVX rats were orally given 0.4 ml each test oil twice weekly. Other animals were given with only a vehicle (control) or an oil mixture with similar levels of oestradiol and progesterone to those in L2 oil (PE oil) for comparison. The Ca absorption rate, bone density of the humerus and femoral Ca content in the rats given L2 were significantly higher than those in the control rats (P < 0.05). Serum calcitonin and 1 alpha,25-(OH)2D from rats given L2 were both significantly higher than those from the control rats, while serum progesterone in the L2 group was slightly lower. It is suggested that the bone metabolism of rats given L2 reflected the compound effect of 1,25-(OH)2D and oestradiol in the L2 oil. In contrast, the relatively low level of these hormones might have been associated with the mediocre bone characteristics of the L1 group. It is suggested from these results that the trace level of hormones could be one of the reasons why whey-mineral complex showed good Ca availability.

The effect of casein phosphopeptides on calcium utilization in young ovariectomized rats.

The effect of casein phosphopeptides (CPPs) on Ca utilization in ovariectomized (OVX) rats was studied. A mixture of CPPs corresponding to the amino acid sequences 1-25 and 1-28 in the beta-casein was isolated from the tryptic digest of beta-casein (beta CPP). After being fed a low Ca diet for 30 days, OVX rats were fed experimental diets of which the Ca level was 0.1%, 0.3% or 0.5% with or without 0.15% beta CPP for 28 days. During days 1-3 of the Ca refeeding period, rats fed beta CPP with 0.5% Ca showed a higher Ca absorption than control rats not supplemented with beta CPP. During days 7-9 and 26-28, there were no significant differences in Ca and P balances between the beta CPP group and the control group for each dietary Ca level. Femoral Ca and P contents from rats fed beta CPP tended to be higher than those from control rats. These results suggest that beta CPP supplementation could have an effect on Ca absorption at a certain degree of Ca deficiency.

[Effects of the cervical spinal cord stimulation on persistent vegetative syndrome: experimental and clinical study].

Influence of spinal cord stimulation (SCS) on electroencephalograms (EEGs) and intracerebral neurotransmitters were studied in cats, and clinical usefulness of the stimulation was reviewed in patients with vegetative syndrome. Cat spinal cords were transdurally stimulated at the second cervical spine for two hours with a pulse width of 1.0 milliseconds, a frequency of 200 Hz and an amplitude ranging from 10 to 15 volts. EEGs were recorded at the frontal epidural space and neurotransmitters in the frontal cortex, such as acetylcholine (ACh), 5-hydroxyindoleacetic acid (5-HIAA), aspartate, glutamate and gamma-aminobutyric acid (GABA), were measured by an in vivo microdialysis method. The background activities of the EEGs were increased by 2 Hz after the SCS. The ACh content increased to 320% three hours after initiation of the SCS, although the other four neurotransmitters did not show any change. Epidural SCS at the upper cervical level was applied to 15 vegetative patients for the amelioration of their vegetative state. Eleven of the 15 patients were treated later than three months after the onset of vegetative state and the other four within three months. Three of the 11 patients treated late and one of the four treated early escaped from the vegetative state (good group). Some clinical improvements were observed in four cases (fair group), two of whom were treated late and the other treated early. The other seven patients showed no improvement (no change group).(ABSTRACT TRUNCATED AT 250 WORDS)

Bioavailabilities of calcium, phosphorus and magnesium from whey mineral complex in growing male rats.

The bioavailabilities of Ca, P and Mg from whey mineral complex (WMC) were studied from the viewpoint of the mineral balance and bone properties in growing male rats and compared with bovine bone ash (BBA) and CaCO3. Ca source showed no significant effect on feed intake, body weight gain or feed efficiency (p < 0.05). When the dietary Ca level was 0.3%, the dry weight of the femur in rats fed WMC was significantly higher than that in rats fed BBA. The femur P content of rats fed 0.3% Ca as WMC was significantly higher than that of rats fed BBA or CaCo3. The breaking energy of the femur from rats fed WMC was significantly higher than that from rats fed BBA at a Ca level of 0.2% or 0.3%. There was the same tendency in bone densities as was observed in breaking properties. There was no significant difference in quantitative values for Ca balance among three groups of rats, whereas those rats fed WMC had a significantly higher P retention than other groups. Ca bioavailability from WMC and the effect on the utilization of P and Mg were discussed.

[Effects and indications of spinal cord stimulation on the vegetative syndrome].

The effects of spinal cord stimulation (SCS) on the vegetative syndrome were studied in six patients. Factors affecting the results were mentioned with a view to establishing indications as to whether or not the SCS should be performed. "Persistent vegetative states" were thought to be identical with Ohta's "vegetative syndrome" which consists of eleven signs. Six of these signs--polyphasic cycle of waking and sleeping, urinary incontinence, being bedridden and being tube fed etc--were important criteria of the vegetative syndrome. SCS was thought to be effective if one or more of the 6 signs disappeared after SCS. SCS was performed at level from C2 to C4 with a frequency of 25 to 120 Hz, an intensity of 2.5 to 6 volts, a pulse duration of 0.3 to 0.5 msec. and a duration of 3 to 11 hours per day. Neurological signs, ABR, CT/MRI, EEG and the grade of the vegetative syndrome were estimated before and after SCS. In the course of SCS, 2 of the 6 patients recovered from the vegetative syndrome. Both had a localized lesion in the brain stem without a cerebral lesion on CT/MRI, with bilateral appearance of the fifth peak with prolonged latency and decreased amplitude of main peaks on ABR. The other 4 patients showed little or no improvement. They all had diffuse cerebral atrophy or low density areas on CT and almost normal ABR. One of these patients, who suffered a cerebral contusion leading to transtentorial herniation with unilateral cerebral contusion on CT and unilateral disappearance of the fifth peak on ABR, showed no recovery from the vegetative syndrome.(ABSTRACT TRUNCATED AT 250 WORDS)