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Biomedical subjects

J S Milne

Publications and source records attributed to J S Milne.

At least 19 recordsLinked to original sources

Nutritional modulation of adolescent pregnancy outcome -- a review.

The risks of miscarriage, prematurity and low birth weight are particularly acute in adolescent girls who are still growing at the time of conception. The role of maternal nutrition in mediating pregnancy outcome in this vulnerable group has been examined in sheep models. When singleton bearing adolescent dams are overnourished to promote rapid maternal growth throughout pregnancy, growth of both the placenta and fetus is impaired, and birth occurs prematurely relative to control adolescents of equivalent age. Studies at mid-gestation, prior to alterations in placental mass, suggest that reduced proliferation of the fetal trophectoderm, impaired angiogenesis, and attenuated uteroplacental blood flows are early defects in placental development. By late pregnancy, relative placental mass is reduced by 45% but uteroplacental metabolism and placental glucose transfer capacity remain normal when expressed on a placental weight specific basis. The asymmetrically growth-restricted fetuses are hypoxic, hypoglycemic and have reduced insulin and IGF-1 concentrations. Absolute umbilical nutrient uptakes are attenuated but fetal utilisation of glucose, oxygen and amino acids remains normal on a fetal weight basis. This suggests altered sensitivities to metabolic signals and may have implications for subsequent metabolic health. At the other end of the nutritional spectrum, many girls who become pregnant have inadequate or marginal nutritional status during pregnancy. This situation is replicated in a second model whereby dams are prevented from growing during pregnancy by relatively underfeeding. Limiting maternal intake in this way gradually depletes maternal body reserves leading to a lower transplacental glucose gradient and a modest slowing of fetal growth in late pregnancy. These changes appear to be independent of alterations in placental growth per se. Thus, while the underlying mechanisms differ, maternal intake at both ends of the nutritional spectrum is a powerful determinant of fetal growth in pregnant adolescents.

Adolescent↗

The effects of acute nutrient restriction in the mid-gestational ewe on maternal and fetal nutrient status, the expression of placental growth factors and fetal growth.

This study explores the hypothesis that acute under-nutrition in mid-gestation reduces maternal and fetal nutrient status and affects the expression of specific regulators of placental growth and function. Welsh Mountain ewes were fed a concentrate diet plus wheat straw to provide 100% of their maintenance requirements. The concentrate ration of nutrient restricted (NR) ewes was reduced from day (d) 83 of gestation and withdrawn from d85 to d90. At d90, half the ewes (NR m = 7, control n = 8) were euthanased. The remainder (NR n = 9, control n = 9) were fed their maintenance diet until slaughter at d135. Maternal plasma insulin and IGF-I concentrations decreased during nutrient restriction and NEFA concentrations increased. Fetal IGF-I and insulin concentrations were unaltered by maternal diet. Placental VEGF mRNA expression was reduced at d90 (P < 0.05). IGFBP-3 and IGFBP-2 mRNA expression was reduced at d90 (P < 0.05) and d135 (P < 0.05), respectively. Placental weight was significantly lower in NR ewes at d90 (P < 0.05) and the distribution of placentomes shifted towards the everted phenotype at d135 (P < 0.05). Reduced thoracic girth and uterine fluid volume at d90 (P < 0.05) and decreased fetal lung weight at d90 (P < 0.05) and d135 (P < 0.05) suggest spatial limitation of lung expansion. In summary, acute NR in mid-gestation reduced anabolic drive and mobilised lipid stores in the maternal compartment, whilst fetal nutrient status was maintained. This was accompanied by changes in placental VEGF and IGFBP expression. The growth of the fetal lung appears to have been compromised and this may have adverse consequences for subsequent neonatal respiratory function.

Animals↗

Experimental study of the protein folding landscape: unfolding reactions in cytochrome c.

Hydrogen exchange results for cytochrome c have been interpreted in terms of transient hydrogen bond-breaking reactions that include large unfolding reactions and small fluctuational distortions. The differential sensitivity of these opening reactions to denaturant, temperature, and protein stability makes it possible to distinguish the different opening reactions and to characterize their structural and thermodynamic parameters. The partially unfolded forms (PUFs) observed are few and discrete, evidently because they are produced by the reversible unfolding of the protein's several intrinsically cooperative secondary structural elements. The PUFs are robust, evidently because the structural elements do not change over a wide range of conditions. The discrete nature of the PUFs and their small number is as expected for classical folding intermediates but not for theoretically derived folding models apparently because the simplified non-protein models usually analyzed in theoretical studies encompass only a single cooperative unit rather than multiple separable units.

Cytochrome c Group↗

Macrophage-inflammatory protein-1alpha regulates preosteoclast differentiation in vitro.

A validated in vitro system was used to investigate the nature of osteoclast-inducing growth factors (OGF) present in fetal rat calvarial conditioned medium (RCCM). Evidence is presented here that macrophage inflammatory protein-1alpha (MIP-1alpha), a member of the C-C chemokine family, is an essential factor for the induction of osteoclast differentiation in this system. Specific polyclonal antibodies against MIP-1alpha significantly inhibited development of TRAP-positive osteoclast precursors and multinucleated osteoclasts induced by RCCM. Anti-MIP-1alpha antibody treatment was accompanied by an increase in the number of macrophage-like cells, suggesting that bone-derived MIP-1alpha is involved in the direction of preosteoclast formation with an inhibitory action on progenitor cell proliferation. Reverse-phase HPLC of RCCM resolved multiple fractions with OGF activity. OGF fractions separated at low acetonitrile (AcN) concentrations (</=15%) did not bind heparin and were not blocked in their bioactivity by the anti-MIP-1alpha antibody. However, OGF fractions eluted at higher AcN concentrations (30-70%) showed heparin-binding activity and were inhibited in their bioactivity by the anti-MIP-1alpha antibody. Western blotting of RCCM with the anti-MIP-1alpha antibody revealed a distinct band with a molecular mass of around 8-14 kDa corresponding to MIP-1alpha. Recombinant rat MIP-1alpha dose dependently stimulated formation of mononuclear osteoclast precursors with maximum stimulation at 50 ng/ml, though it could not fully mimic RCCM activity. These results identify MIP-1alpha as a candidate responsible for bone-derived OGF bioactivity and confirm that chemokines play an important role in the process of osteoclast recruitment and differentiation.

Animals↗

Handgun safety features: a review for physicians.

OBJECTIVE: Handguns are a ubiquitous consumer product in the United States, which annually cause significant morbidity and mortality. Handgun safety devices are often proposed as potential solutions to this problem. Their effectiveness at reducing handgun injuries and deaths is intensely debated. However, to effectively analyze the potential utility of handgun safety devices, physicians need to be aware of the safety devices available in the consumer market and how they operate. METHODS: A wide variety of safety devices are available in the consumer market, which vary in terms of their ease of operation, cost, and the types of injuries they may prevent. We reviewed several types of handgun safety devices, including loaded chamber indicators, manual thumb safeties, grip safeties, magazine disconnectors, drop safeties, built-in locks, trigger locks, lockboxes, and personalized handguns. Each device is described within the context of reducing unintended discharge and unauthorized use. RESULTS: This review is not exhaustive. There are other types of safety devices that limit access to handguns. Many of these devices, such as barrel locks and chamber locks, work in a similar manner as trigger locks and have the same limitations. The user of any type of safety device should think about the types of injuries the device is designed to prevent and be aware of its limitations. CONCLUSION: Physicians have the potential to reduce the risk of firearm injuries with their patients and communities. Providing accurate information on firearm safety devices and their limitations is important, just as it is for other aspects of health care advice. Armed with accurate information, physicians can hopefully be effective in firearm injury prevention.

Firearms↗

The availability of extrinsic handgun locking devices in a defined metro area.

Extrinsic safety devices, such as trigger locks, have been central in the recent state discussions on how to reduce firearm injuries. The actual prevalence of handgun locking devices in the consumer market, however, is unknown. This study catalogued the extrinsic safety devices available from handgun dealers and discount retail chains in Milwaukee, WI. We found that all locations studied (n = 13) stock at least 1 type of extrinsic safety device. A total of 21 unique models of safety devices were stocked by the 13 locations, with trigger locks being the most common (n = 9). Other types of devices included lock-boxes (n = 5), cable locks (n = 4), hammer locks (n = 1), barrel locks (n = 1), and a rubber slide strap (n = 1). Handgun owners in the Milwaukee metro area have a selection of extrinsic handgun safety devices available from handgun dealers and discount retailers. However, there does not appear to be a consistent availability on type of device.

Firearms↗

"I didn't know the gun was loaded": an examination of two safety devices that can reduce the risk of unintentional firearm injuries.

Some handguns contain built-in safety devices intended to prevent injuries caused by erroneously believing that a handgun is loaded. A loaded chamber indicator indicates the presence of ammunition in the gun; a magazine safety prevents the gun from being fired when the ammunition magazine is removed, even if one round remains in the firing chamber. In our patent search these devices date back to the turn of the century. But on 1998 pistol models, only 11% contained a loaded chamber indicator and 14% had a magazine safety. In our random-digit-dial telephone survey of U.S. adults, 34.8% of poll respondents (incorrectly) thought that a firearm with its ammunition magazine removed could not be shot, or said that they did not know. Some of the 1100 unintentional gun deaths in the U.S. each year might be prevented if the prevalence of these and other safety devices is increased through legislation, litigation, or voluntary manufacturer action.

Adolescent↗

Determinants of protein hydrogen exchange studied in equine cytochrome c.

The exchange of a large number of amide hydrogens in oxidized equine cytochrome c was measured by NMR and compared with structural parameters. Hydrogens known to exchange through local structural fluctuations and through larger unfolding reactions were separately considered. All hydrogens protected from exchange by factors greater than 10(3) are in defined H-bonds, and almost all H-bonded hydrogens including those at the protein surface were measured to exchange slowly. H-exchange rates do not correlate with H-bond strength (length) or crystallographic B factors. It appears that the transient structural fluctuation necessary to bring an exchangeable hydrogen into H-bonding contact with the H-exchange catalyst (OH(-)-ion) involves a fairly large separation of the H-bond donor and acceptor, several angstroms at least, and therefore depends on the relative resistance to distortion of immediately neighboring structure. Accordingly, H-exchange by way of local fluctuational pathways tends to be very slow for hydrogens that are neighbored by tightly anchored structure and for hydrogens that are well buried. The slowing of buried hydrogens may also reflect the need for additional motions that allow solvent access once the protecting H-bond is separated, although it is noteworthy that burial in a protein like cytochrome c does not exceed 4 angstroms. When local fluctuational pathways are very slow, exchange can become dominated by a different category of larger, cooperative, segmental unfolding reactions reaching up to global unfolding.

Animals↗

Dehydroepiandrosterone (DHEA) and DHEA-S interact with 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) to stimulate human osteoblastic cell differentiation.

DHEA, an adrenocortical steroid, and its sulfate derivative (DHEA-S), have been implicated in many biological functions, including the regulation of bone mass. In this study, we examined whether DHEA/DHEA-S are capable of directly affecting bone cell proliferation and differentiation, and compared this with the effects of, and interaction with, the established bone cell modulating steroid, 1,25-dihydroxyvitamin D3 (1,25(OH)2D3). Two in vitro models of human osteoblastic cells were used, viz. MG63 osteosarcoma cell line and normal primary osteoblast-like cells (HOB). Our results show that DHEA and DHEA-S failed on their own to exert direct, independent significant effects on the growth and differentiation of human osteoblastic cells, but treating the cells in conjunction with 1,25(OH)2D3 resulted in enhancement of specific A1P activity. Moreover, 1,25(OH)2D3-induced osteocalcin production was potentiated by the adrenal steroids in both cell models. DHEA-S proved in general to be more potent than DHEA. In conclusion, this study shows that the effects of DHEA/DHEA-S on osteoblastic cell growth and differentiation are likely to be mediated via an effect on 1,25(OH)2D3-induced changes in bone cells, suggesting a distinctive role for these steroids in the regulation of bone metabolism.

Calcitriol↗

A novel culture system to generate osteoclasts and bone resorption using porcine bone marrow cells: role of M-CSF.

A novel osteoclast generation and bone resorption assay system is described in which enhanced osteoclastic generation and bone resorption is induced in porcine bone marrow cell cultures cultured in low-serum medium supplemented with fibroblastic cell (L929) conditioned medium (CM). Numerous osteoclasts, which could be identified by TRAP staining and specific labelling with 121F antibody, were generated in a time-dependent and L929-CM concentration-dependent fashion. A specific antibody against murine M-CSF/CSF-1 abolished osteoclast formation indicating that M-CSF is the essential component of the L929-CM driven osteoclast generation. Culturing on devitalized bone slices resulted in extensive osteoclast-mediated resorption as visualized microscopically. After 16 days in culture, practically the entire bone slice surface was excavated by the osteoclastic cells. Bone resorption could be monitored with time using a novel enzyme-linked immunoassay measuring type I collagen N-telopeptides in culture supernatants. Release of collagen fragments from the slices was paralleled by osteoclastic secretion of TRAP. Salmon calcitonin significantly inhibited collagen fragment and TRAP release. 1,25-Dihydroxyvitamin D3 greatly promoted osteoclast generation and subsequent bone resorption, but its presence was not essential for this process to occur.

Acid Phosphatase↗

Thermodynamic parameters from hydrogen exchange measurements.

Just as exchangeable hydrogens that are controlled by global unfolding can be used to measure thermodynamic parameters at a global level, hydrogens that are exposed to exchange by local unfolding reactions may be used to obtain locally resolved energy parameters. Results with the hemoglobin system demonstrate the ability of HX methods to locate functionally important changes in a protein and to measure the energetic contribution of each. These results offer the promise that HX measurements may be used to delineate, in terms of definable bonds and their energies and interactions, the network of interactions that Hb and other proteins use to produce their various functions.

Chromatography, High Pressure Liquid↗

Protein stability parameters measured by hydrogen exchange.

The hydrogen exchange (HX) rates of the slowest peptide group NH hydrogens in oxidized cytochrome c (equine) are controlled by the transient global unfolding equilibrium. These rates can be measured by one-dimensional nuclear magnetic resonance and used to determine the thermodynamic parameters of global unfolding at mild solution conditions well below the melting transition. The free energy for global unfolding measured by hydrogen exchange can differ from values found by standard denaturation methods, most notably due to the slow cis-trans isomerization of the prolyl peptide bond. This difference can be quantitatively calculated from basic principles. Even with these corrections, HX experiments at low denaturant concentration measure a free energy of protein stability that rises above the usual linear extrapolation from denaturation data, as predicted by the denaturant binding model of Tanford.

Amines↗

Effects of dietary acidity on calcium balance and mobilisation, bone morphology and 1,25 dihydroxyvitamin D in prepartal dairy cows.

The effect of an acid or alkali diet was investigated in 14 mature dairy cows during the last 28 days of pregnancy. The acid diet reduced the incidence of parturient hypocalcaemia compared with the alkali diet and was associated with higher blood ionised calcium and plasma chloride concentrations and lower blood pH and acid-base excess before parturition. Plasma 1,25-dihydroxyvitamin D concentrations before parturition were increased by the acid diet but the concentrations of parathyroid hormone and intact 1-86 parathyroid-hormone-related peptide were unaffected. The estimated fractional calcium absorption and calcium mobilisation rate during an ethylenediamine tetraacetic acid infusion were increased by the acid diet 14 days before parturition. Cortical bone remodelling occurred in all the animals during late pregnancy but was particularly evident in the cows given the acid diet. The data suggest that an acid diet ameliorates parturient hypocalcaemia by enhancing calcium mobilisation before parturition by increasing calcium absorption and bone resorption, these increases possibly being mediated by increases in the plasma concentration of 1,25-dihydroxyvitamin D.

Animal Feed↗

Primary structure effects on peptide group hydrogen exchange.

The rate of exchange of peptide group NH hydrogens with the hydrogens of aqueous solvent is sensitive to neighboring side chains. To evaluate the effects of protein side chains, all 20 naturally occurring amino acids were studied using dipeptide models. Both inductive and steric blocking effects are apparent. The additivity of nearest-neighbor blocking and inductive effects was tested in oligo- and polypeptides and, surprisingly, confirmed. Reference rates for alanine-containing peptides were determined and effects of temperature considered. These results provide the information necessary to evaluate measured protein NH to ND exchange rates by comparing them with rates to be expected for the same amino acid sequence is unstructured oligo- and polypeptides. The application of this approach to protein studies is discussed.

Alanine↗

Urease (EC 3.5.1.5) inhibition in the sheep rumen and its effect on urea and nitrogen metabolism.

The urease (EC 3.5.1.5) inhibitor, phenylphosphoryldiamidate (PPDA), was given by continuous infusion into the rumen of two sheep nourished by intragastric infusion and into either the rumen or abomasum of four sheep given a pelleted diet containing 119 g crude protein (nitrogen x 6.25)/kg dry matter. PPDA was given at 1 g/d in infusion sheep and 1.5 g/d in the normally-fed sheep. Measurements of urea kinetics were made using single injections of [14C]urea. Urease inhibition was complete within 24 h of starting PPDA infusions to the rumen; in this time-period, urea concentration in rumen contents reached equilibrium with that in plasma and this situation persisted until infusions were terminated. Relative to the control periods, plasma urea and rumen ammonia concentrations were unchanged but urea irreversible loss rate decreased by 26% in infusion sheep and 33% in fed sheep when PPDA was given. Urinary urea excretion was not affected, hence urea degradation, measured by difference, decreased by 77 and 58% respectively in response to urease inhibition. Administration of PPDA to the abomasum resulted in a reduction in rumen urease activity to about 40% of control values but had no effect on urea metabolism. Differences between treatments in daily nitrogen retention were not significant, indicating that under the dietary conditions imposed in these experiments, even substantial changes in urea recycling had only minor effects on the overall N economy of the animal.

Abomasum↗

Urea degradation in sheep nourished by intragastric infusion: effects of level and nature of energy inputs.

Four female sheep nourished wholly by infusions of volatile fatty acids (VFA), buffer and minerals to the rumen and casein to the abomasum were given in addition infusions of supplementary energy calculated to increase the energy input to the rumen by 30%. The design was a Latin square and the supplements given were (a) nil, (b) a standard VFA mixture similar to the basal infusion, (c) butyric acid alone and (d) glucose. Measurements were made of nitrogen retention and rumen fermentation characteristics and the kinetics of urea metabolism were measured over 24 h by means of a single injection of [14C]urea. An active microbial fermentation was established in the rumen in response to the infusion of glucose and estimates of microbial protein synthesis derived from urinary purine excretion agreed well with those calculated from stoichiometric principles. The presence of a microbial population in the rumen resulted in a decrease in urinary urea excretion and reductions in plasma urea concentration, urea pool size and rumen ammonia (NH3) concentration. Infusion of the mixed VFA or butyric acid supplements had no effect on these indices of urea metabolism. Measurements of urea irreversible loss rate showed high variability and the mean values did not differ significantly between the four treatments. Urea degradation in the gastrointestinal tract was also highly variable but increased, on average, by 2.4 g day-1 on the high-energy treatments. Examination of regression relationships between these variables also indicated a difference between the glucose treatment and the others in the metabolic fate of the NH3 derived from urea hydrolysis. It is concluded that urea degradation increased in response to additions of energy but did not differ according to the nature of the supplements supplied. In the glucose-supplemented group, the NH3 arising from degraded urea was incorporated into microbial protein and so removed from the urea-NH3 cycle; when additions of mixed VFA or butyric acid were given, the NH3 arising in hydrolysis appeared simply to be reabsorbed as NH3 and to contribute anew to urea formation.

Animal Nutritional Physiological Phenomena↗

The effect of a rumen microbial fermentation on urea and nitrogen metabolism of sheep nourished by intragastric infusion.

Four sheep were maintained by infusion of volatile fatty acid (VFA), mineral and buffer solutions to the rumen and casein to the abomasum. After a 3 week period in which control measurements were made, glucose was introduced gradually to the rumen infusion mixture until glucose replaced about 65% of VFA energy. Further measurements were made in the fourth week of glucose additions after which the control VFA infusions were re-established and measurements repeated. Urea kinetics were measured with [14C]urea. Rumen ATP concentrations were used as an index of microbial growth and increased from 1.5 nmol ml-1 on control treatments to 46 nmol ml-1 when glucose was given. The presence of a rumen fermentation resulted in a 40% decrease in plasma urea concentration but was without effect on rumen ammonia concentration. Urea irreversible loss rate decreased from 18.0 to 12.8 g urea day-1 when glucose was given. This, however, was matched by a decrease in urinary urea excretion, with the result that urea degradation, obtained by difference, remained constant throughout the three treatment periods. Daily nitrogen (N) retention increased from 1.0 to 2.7 g when a rumen microbial population was present. A model of urea and NH3 transactions for the two dietary situations is presented. Calculations indicated that 0.40 of the casein N supply was reutilized as urea by the rumen micro-organisms. It is suggested, however, that the improvement in N retention resulted from a change in protein:energy ratio of the infused nutrients rather than from an enhanced supply of protein to the intestine. The use of endogenous urea clearance as an index of epithelial permeability to urea is questioned.

Animal Nutritional Physiological Phenomena↗

Urea metabolism in sheep given conventional feeds or nourished by intragastric infusion.

A comparison has been made of the kinetics of urea metabolism in sheep given conventional feeds or maintained wholly by intragastric infusions of volatile fatty acids, buffer, mineral and casein solutions. Daily nitrogen supply was 13.2 g/day in 'fed' sheep and 6.0 g/day in 'infusion' sheep. On each feeding system measurements were made at the basal (maintenance) level of intake and when the basal level was supplemented with infusions of urea into the abomasum (125 mmol/day) or into the rumen (300 mmol/day). Additional measurements were made when ammonium carbonate (100 mmol/day) was infused into the rumen, when cassava (150 g/day) was added to the diet of 'fed' sheep or when the input of casein to 'infusion' sheep was reduced by half. Urea kinetics were measured by means of a single intravenous injection of [14C]urea. Urea irreversible loss rate, urea pool size, urinary urea excretion, plasma urea concentrations and rumen ammonia concentrations were all significantly higher in sheep given conventional feeds than in those nourished by infusion but urea degradation in the gastrointestinal tract did not differ between the two methods of feeding. Regression analysis indicated only minor differences between the two feeding systems in the relationships between the various indices of urea metabolism. It is concluded that most of these findings can be attributed to differences in the quantity and nature of the nitrogen supplied in the basal diets and that the sheep nourished by infusion would be a suitable model for the study of factors involved in the control of urea recycling.

Ammonia↗