Study of the effect of lactational bone loss on blood lead concentrations in humans.
(9/4708)
Lactation and other clinical states of high bone turnover have been suggested to release lead (Pb) stored in bone into blood and tissues. Previous observations on the influences of lactation have been anecdotal, or at high blood Pb concentrations with varying past exposures, or complicated by postpartum fluid changes. A prospective observational study was performed to investigate possible changes in blood lead concentrations at multiple intervals during lactation for 6 months postpartum and to relate changes in blood lead concentrations to changes in bone density and other variables. Volunteer pregnant subjects (n = 58) were enrolled from a midwifery service at an academic public health hospital. Subjects were mostly Hispanic, recently immigrated, of low economic status, not receiving supplemental calcium, and had low blood Pb concentrations (2.35 +/- 2.05 microg/dl at enrollment). Bone density losses over 6 months for the group averaged -2.46 +/- 6.33% at the vertebral spine and -0.67 +/- 5.21% at the femoral neck. In predicting final bone density, apart from initial bone density only the total number of breast-feedings was a significant independent variable of the variables tested, accounting for an additional 12% of the variability. No changes in blood Pb concentrations were seen over the interval beyond 2 weeks postpartum (minimum detectable change was 0.4 microg/dl). There was no relation between the changes in bone density and changes in blood Pb or the integrated blood Pb over the 2-week to 6-month period. Normal (nonlactating) bone resorption rates contribute a large fraction of the Pb in blood during low-exposure circumstances. However, during lactation the increase in bone resorptive processes is probably relatively small with a larger decrease in deposition accounting for net bone loss, as suggested by other investigations. Thus, concomitant release of Pb from bones of lactating subjects with low blood lead concentrations on this background of high normal resorption was not large enough for detection. (+info)
A new tool for measuring the suckling stimulus during breastfeeding in humans: the orokinetogram and the Fourier series.
(10/4708)
The Fourier series was used to analyse the oral movements recorded by the orokinetogram during breastfeeding in human babies. This is a new method that allows recording of oral movements without introducing any extrinsic element between the nipple and the mouth of the baby. The advantage of displaying suckling activity after fast Fourier transform (FFT) is that this algorithm allows storage, quantification and frequency analysis of the oral movements throughout a suckling bout, which enables the total oral activity to be measured. Two types of oral movements are found: slow high amplitude (SHA) and fast low amplitude (FLA). FLA movements may be derived from peristaltic movements of the tongue that result in tickling stimuli to the mechanoreceptors of the nipple and milk expression. The frequency bandwidth of oral movements is wider (0-8 Hz) than has been described previously (0-3 Hz) and this is due to the presence of the FLA oral movements. An indirect measurement of the energy of oral movements during suckling is obtained by the pattern of energy distribution used in each individual frequency band by oral movements. This pattern changes in relation to the periods of continuous and intermittent suckling activity. SHA and FLA oral movements are more intense during continuous suckling. Statistical analysis showed a correlation between the energy of SHA and FLA waves throughout the suckling bout, and also that the highest level of energy during suckling activity is displayed during the first 2 min. The novel tools described in this paper allow investigation of the role of suckling stimulus in reflex hormone release and other mother-infant interactions. (+info)
Reproductive experience and opioid regulation of luteinizing hormone release in female rats.
(11/4708)
The objective of the present study was to determine whether reproductive experience that produces shifts in opioid regulation of prolactin secretion and behavioural functions also alters opioid regulation of LH during the oestrous cycle or lactation. In Expt 1 the effect of naloxone administration (i.v.) on LH was compared between age-matched, nulliparous and primiparous, catheterized female rats on dioestrus II. In Expt 2, the effects of multiple reproductive experiences on opiate control of LH were investigated using cyclic, nulliparous and multiparous (three litters) rats. In both experiments, no differences in naloxone-stimulated LH release were found between groups even though multiple reproductive experiences resulted in the prolongation of oestrous cyclicity. In Expt 3, day 8 lactating primiparous rats were administered 2, 5, 10 or 25 mg naloxone kg-1 i.v. The three lowest naloxone doses, but not the 25 mg kg-1 dose, significantly increased LH concentrations. The possible effects of prior reproductive experience on opioid control of LH during lactation were then investigated. Naloxone at 0.5 mg kg-1, but not at 2 mg kg-1 or 10 mg kg-1, stimulated a significantly greater rise in LH in multiparous (two litters) than in primiparous females. Overall, these data indicate that while modest differences were found in naloxone-induced LH responses between multiparous and primiparous animals during lactation, reproductive experience did not significantly alter opioid regulation of LH during subsequent oestrous cycles at the naloxone doses examined. Hence, the effects of reproductive experience on opioid regulation of LH are less pronounced than those previously found for opioid regulation of prolactin and behaviour. (+info)
Demonstration of in vivo mammogenic and lactogenic effects of recombinant ovine placental lactogen and mammogenic effect of recombinant ovine GH in ewes during artificial induction of lactation.
(12/4708)
The present study demonstrates that ovine placental lactogen (oPL) (ovine chorionic somatotrophin) may have an important role in the mammogenesis and/or lactogenesis of the ewe. Its effects were compared with that already described for ovine growth hormone (oGH). In the first experiment, 40 nulliparous ewes were induced to lactate by means of a 7 day (days 1-7) oestro-progestative treatment (E2+P4). The ewes from Group 1 (n=12) received no further treatment, while those of the other groups received either recombinant oGH (roGH, 28 micrograms/kg, i.m., twice daily, Group 2, n=12) or recombinant oPL (roPL, 79 micrograms/kg, i.m., twice daily, Group 3, n=12) from day 11 to 20. All ewes received 25 mg hydrocortisone acetate (HC) twice daily on days 18-20. Control Group 00 (n=2) received no steroid treatment at all, and the control Group 0 (n=2) received only the E2+P4 treatment. Thirteen ewes (three from each experimental group and the two of each control group) were slaughtered at the end of hormone treatments (day 21) before any milking stimulus. The 27 remaining ewes from Groups 1-3 were machine-milked and milk yields recorded daily from day 21 to 76. The E2+P4 treatment enhanced the plasma levels of oPRL, oGH and IGF-I between days 1 and 7 by 1.5, 2. 3 and 2.6 times respectively (P=0.002); roGH treatment induced a highly significant enhancement of IGF-I plasma levels from day 11 to 20, whereas a similar effect appeared for roPL-treated ewes only from day 17 to 20 (P<0.01). Eight weeks after the last exogenous hormone injections, milk yields of both roGH- and roPL-treated groups progressively rose to twice that of unsupplemented groups (P<0.001). The mammary DNA content on day 21 was higher for animals which received either oGH or oPL but, due to individual variations in so few samples (n=3), this difference was not significant. No beta-casein was measured in mammary tissue from control ewes, whereas steroid-treated ewes (E2+P4+HC) had higher casein concentrations regardless of subsequent hormonal treatment on days 11-20 (P<0.001). beta-Casein concentrations in mammary parenchyma of roGH-treated ewes did not differ from that of ewes which received only E2+P4+HC; roPL supplementation clearly enhanced expression of beta-casein (P<0.001). IGF-I stimulation by either roGH or roPL was more precisely examined during a second experiment, in which two twice-daily i.m. doses (58 or 116 micrograms/kg) of either roGH or roPL were administered to four groups of six ewes that were E2+P4 treated as those of Experiment 1. A control group (n=6) received no exogenous hormone from day 11 to 13. On day 13, hourly blood samples were taken from all ewes over 11 h. Both doses of roGH significantly stimulated IGF-I in a dose-dependent manner. The 58 micrograms/kg dose of roPL did not significantly stimulate IGF-I, but although being somewhat less efficient than the 58 micrograms/kg dose of roGH, the 116 micrograms/kg dose of roPL significantly stimulated IGF-I secretion (P<0. 001). These results suggest that mammogenesis and/or lactogenesis in the ewe is in part controlled by somatotrophic hormones such as oGH and oPL and that IGF-I could be one of the mediators of these hormones. (+info)
Changes in properties and neurosteroid regulation of GABAergic synapses in the supraoptic nucleus during the mammalian female reproductive cycle.
(13/4708)
1. GABAA receptor-mediated synaptic innervation of oxytocin neurones in the supraoptic nucleus (SON) was analysed in adult female rats going through their first reproductive cycle by recording the spontaneous inhibitory postsynaptic currents (sIPSCs) at six stages of female reproduction. 2. During pregnancy we observed a reduction in the interval between monoquantal sIPSCs. The synaptic current amplitude, current decay and neurosteroid sensitivity of postsynaptic GABAA receptors observed at this stage were not distinguishable from those measured in virgin stage SON. 3. Upon parturition an increase in monoquantal synaptic current decay occurred, whereas potentiation by the progesterone metabolite allopregnanolone (3alpha-OH-DHP) was suppressed. 4. Throughout a substantial part of the lactation period the decay of synaptic currents remained attenuated, whilst the potentiation by 3alpha-OH-DHP remained suppressed. 5. Several weeks after the end of lactation sIPSC intervals, their current decay velocity as well as the potentiation by 3alpha-OH-DHP were restored to pre-pregnancy levels, which is indicative of the cyclical nature of synaptic plasticity in the adult SON. 6. Competitive polymerase chain reaction (PCR) analysis showed that virgin animals expressed alpha1 and alpha2 GABAA receptor subunit mRNA at a relative ratio of 2 : 1 compared with beta-actin. After pregnancy both alpha1 and alpha2 subunit mRNA levels were transiently increased, although at a relative ratio of 1 : 4, in line with the hypothesis that alpha2 plays a large role in postsynaptic receptor functioning. During post-lactation both alpha subunits were downregulated. 7. We propose that synaptic remodelling in the SON during pregnancy includes changes in the putative number of GABA release sites per neurone. At parturition, and during the two consecutive weeks of lactation, a subtype of postsynaptic GABAA receptors was observed, distinct from the one being expressed before and during pregnancy. Synaptic current densities, calculated in order to compare the impact of synaptic inhibition, showed that, in particular, the differences in 3alpha-OH-DHP potentiation of these two distinct GABAA receptor subtypes produce robust shifts in the impact of synaptic inhibition of oxytocin neurones at the different stages of female reproduction. (+info)
Guinea-pig milk-protein synthesis. Isolation and characterization of messenger ribonucleic acids from lactating mammary gland and identification of caseins and pre-alpha-lactalbumin as translation products in heterologous cell-free systems.
(14/4708)
1. The major milk proteins synthesized by the lactating mammary gland of the guinea pig were identified and designated as caseins A, B and C and alpha-lactalbumin, with estimated mol.wts. of 28000, 25500, 20500 and 14500 respectively. 2. Antisera to the total casein fraction and to alpha-lactalbumin were prepared from rabbits. The milk proteins were also iodinated with either 131I or 125I. 3. A poly(A)-rich RNA fraction was isolated from lactating guinea-pig mammary glands. Isolation was by affinity chromatography on oligo(dT)-cellulose. 4. Examination of this RNA fraction by electrophoresis on polyacrylamide gels containing formamide indicated three major species 1, 2 and 3, with estimated wol.wts. of 5.4 X 10(5) and 3.3 X 10(5), and the apparent absence of rRNA species. 5. The poly(A)-rich RNA stimulated protein synthesis in heterologous cell-free systems based on wheat germ, Krebs II ascites-tumour cells, and the latter supplemented with an initiation factor-3 fraction from rabbit reticulocyte ribosomes. 6. Between 80 and 90% of the protein synthesis directed by the mRNA was for milk proteins. 7. Analysis of the proteins immunoprecipitated by the alpha-lactalbumin antiserum showed in the wheat-germ system that the product was a protein with a molecular weight greater than that of alpha-lactalbumin, whereas in the ascites-tumour-cell systems both this protein and alpha-lactalbumin were found. When the larger protein was treated with CNBr and the resulting peptides were examined, it was shown that the extra peptide was at the N-terminus. This and other evidence is adduced for the initial translation product of alpha-lactalbumin being a precursor with an addition of about ten amino acids at the N-terminus. 8. Similar analysis of the casein immlnospecific proteins produced under the direction of mRNA indicated that the products had a molecular weight that was apparently a littel smaller than that of the caseins synthesized in vivo. This was not consistent with higher-molecular weight casein precursors. 9. Possible explanations for the results obtained are discussed, especially in terms of the physiological significance of the pre-alpha-lactalbumin as a secretory protein. (+info)
Redistribution of protein kinase C isoforms in rat pancreatic acini during lactation and weaning.
(15/4708)
Freshly enzymatically isolated pancreatic acini from lactating and weaning Wistar rats were used to investigate the role of protein kinase C (PKC) isoforms during these physiologically relevant pancreatic secretory and growth processes. The combination of immunoblot and immunohistochemical analysis shows that the PKC isoforms alpha, delta, and epsilon are present in pancreatic acini from control, lactating and weaning rats. A vesicular distribution of PKC-alpha, -delta, and -epsilon was detected by immunohistochemical analysis in the pancreatic acini from all the experimental groups. PKC-delta showed the strongest PKC immunoreactivity (PKC-IR). In this vesicular distribution, PKC-IR was located at the apical region of the acinar cells. No differences were observed between control, lactating and weaning rats. However, the immunoblot analysis of pancreatic PKC isoforms during lactation and weaning showed a significant translocation of PKC-delta from the cytosol to the membrane fraction when compared with control animals. Translocation of PKC isoforms (alpha, delta and epsilon) in response to 12-O-tetradecanoyl phorbol 13-acetate (TPA) 1 microM (15 min, 37 degrees C) was comparable in pancreatic acini from control, lactating and weaning rats. In the control group, a significant translocation of all the isoforms (alpha, delta and epsilon) from the cytosol to the membrane was observed. The PKC isoform most translocated by TPA was PKC-delta. In contrast, no statistically significant increase in PKC-delta translocation was detected in pancreatic acini isolated from lactating or weaning rats. These results suggest that the PKC isoforms are already translocated to the surface of the acinar cells from lactating or weaning rats. In addition, they suggest that isoform specific spatial PKC distribution and translocation occur in association with the growth response previously described in the rat exocrine pancreas during lactation and weaning. (+info)
Use of doxycycline-controlled gene expression to reversibly alter milk-protein composition in transgenic mice.
(16/4708)
A reverse tetracycline transactivator-encoding cDNA under the control of the mammary specific beta-lactoglobulin promoter was linked to a bovine alpha-lactalbumin transcription unit driven by a reverse tetracycline-controlled transactivator/doxycycline-inducible human cytomegalovirus promoter. The construct was microinjected into eggs from alpha-lactalbumin-deficient mice. These mice produce a highly viscous lactose-free milk and have a shortened lactation period. Mice from three out of the nine transgenic lines investigated expressed reverse tetracycline-controlled transactivator mRNA in their lactating mammary glands at levels detectable by Northern analysis. Following doxycycline addition to the drinking water, lactation was fully restored in animals from the three lines. Doxycycline removal resulted in a reversal of phenotype. The observed mammary-specific and high expression of the doxycycline inducible reporter gene (up to 5.2 mg of recombinant alpha-lactalbumin.mL-1 of milk, i.e. up to 13-fold induction) opens up exciting prospects to use the tetracycline system to study the development and functioning of the mammary gland, and to control the production level of active pharmaceutical proteins in the milk of transgenic animals. (+info)