Glucocorticoid maturation of mitochondrial respiratory capacity in skeletal muscle before birth.

In adults, glucocorticoids act to match the supply and demand for energy during physiological challenges, partly through actions on tissue mitochondrial oxidative phosphorylation (OXPHOS) capacity. However, little is known about the role of the natural prepartum rise in fetal glucocorticoid concentrations in preparing tissues for the increased postnatal energy demands. This study examined the effect of manipulating cortisol concentrations in fetal sheep during late gestation on mitochondrial OXPHOS capacity of two skeletal muscles with different postnatal locomotive functions. Mitochondrial content, biogenesis markers, respiratory rates and expression of proteins and genes involved in the electron transfer system (ETS) and OXPHOS efficiency were measured in the biceps femoris (BF) and superficial digital flexor (SDF) of fetuses either infused with cortisol before the prepartum rise or adrenalectomised to prevent this increment. Cortisol infusion increased mitochondrial content, biogenesis markers, substrate-specific respiration rates and abundance of ETS complex I and adenine nucleotide translocator (ANT1) in a muscle-specific manner that was more pronounced in the SDF than BF. Adrenalectomy reduced mitochondrial content and expression of PGC1α and ANT1 in both muscles, and ETS complex IV abundance in the SDF near term. Uncoupling protein gene expression was unaffected by cortisol manipulations in both muscles. Gene expression of the myosin heavy chain isoform, MHCIIx, was increased by cortisol infusion and reduced by adrenalectomy in the BF alone. These findings show that cortisol has a muscle-specific role in prepartum maturation of mitochondrial OXPHOS capacity with important implications for the health of neonates born pre-term or after intrauterine glucocorticoid overexposure.

Translatable mitochondria-targeted protection against programmed cardiovascular dysfunction.

The prenatal origins of heart disease in offspring have been established. However, research in species with developmental milestones comparable to humans is lacking, preventing translation of this knowledge to clinical contexts. Using sheep and chickens, two species with similar cardiovascular developmental milestones to humans, we combined in vivo experiments with in vitro studies at organ, cellular, mitochondrial, and molecular levels. We tested mitochondria-targeted antioxidant intervention with MitoQ against cardiovascular dysfunction programmed by developmental hypoxia, a common complication in human pregnancy. Experiments in sheep determined in vivo fetal and adult cardiovascular function through surgical techniques not possible in humans, while those in chicken embryos isolated effects independent of maternal or placental influences. We show that hypoxia generates mitochondria-derived oxidative stress during cardiovascular development, programming endothelial dysfunction and hypertension in adult offspring. MitoQ treatment during hypoxic development protects against this cardiovascular risk via enhanced nitric oxide signaling, offering a plausible intervention strategy.

Development and thyroid hormone dependence of skeletal muscle mitochondrial function towards birth.

KEY POINTS: Skeletal muscle energy requirements increase at birth but little is known regarding the development of mitochondria that provide most of the cellular energy as ATP. Thyroid hormones are known regulators of adult metabolism and are important in driving several aspects of fetal development, including muscle fibre differentiation. Mitochondrial density and the abundance of mitochondrial membrane proteins in skeletal muscle increased during late gestation. However, mitochondrial functional capacity, measured as oxygen consumption rate, increased primarily after birth. Fetal hypothyroidism resulted in significant reductions in mitochondrial function and density in skeletal muscle before birth. Mitochondrial function matures towards birth and is dependent on the presence of thyroid hormones, with potential implications for the health of pre-term and hypothyroid infants. ABSTRACT: Birth is a significant metabolic challenge with exposure to a pro-oxidant environment and the increased energy demands for neonatal survival. This study investigated the development of mitochondrial density and activity in ovine biceps femoris skeletal muscle during the perinatal period and examined the role of thyroid hormones in these processes. Muscle capacity for oxidative phosphorylation increased primarily after birth but was accompanied by prepartum increases in mitochondrial density and the abundance of electron transfer system (ETS) complexes I-IV and ATP-synthase as well as by neonatal upregulation of uncoupling proteins. This temporal disparity between prepartum maturation and neonatal upregulation of mitochondrial oxidative capacity may protect against oxidative stress associated with birth while ensuring energy availability to the neonate. Fetal thyroid hormone deficiency reduced oxidative phosphorylation and prevented the prepartum upregulation of mitochondrial density and ETS proteins in fetal skeletal muscle. Overall, the data show that mitochondrial function matures over the perinatal period and is dependent on thyroid hormones, with potential consequences for neonatal viability and adult metabolic health.

A survey of northern Victorian dairy farmers to investigate dairy calf management: colostrum feeding and management.

OBJECTIVES: To describe colostrum management practices carried out in northern Victorian dairy herds and to identify weaknesses in these areas that may affect calf health and welfare by comparing the results with the current industry recommendations METHODS: A questionnaire to obtain information about colostrum management and calf-rearing practices was sent to commercial dairy farming clients of Rochester Veterinary Practice between June and September 2013. The questionnaire consisted of a general herd overview and colostrum harvesting practices. RESULTS: The response rate was 39% (58/150). Many dairy producers were not meeting the current industry recommendations in the following areas: (1) time of removal calf from the dam, (2) relying on calf suckling colostrum from the dam to achieve adequate passive transfer, (3) failing to supplement calves with colostrum, (4) feeding inadequate volumes of colostrum, (5) delayed colostrum harvesting, (6) pooling of colostrum, (7) failing to objectively assess colostrum quality or relying on visual assessment and (8) storing colostrum for a prolonged periods of time at ambient temperatures. CONCLUSION: The results from this survey highlight the need for greater awareness of industry standards for colostrum management and feeding hygiene.

A survey of northern Victorian dairy farmers to investigate dairy calf management: calf-rearing practices.

OBJECTIVES: To describe the calf-rearing practices carried out in northern Victorian dairy herds and to identify weaknesses that may affect calf health and welfare by comparing the results with current industry recommendations. METHODS: Survey of dairy farms from Rochester and the surrounding farming area. RESULTS: The response rate was 39% (58/150). Many dairy producers were not meeting the current industry recommendations in the following areas: (1) delayed access to pellets and roughage, (2) failing to provide access to water from birth, (3) delayed disbudding of calves, (4) delayed timing of booster vaccinations, (5) weaning based on age alone, (6) failing to isolate sick calves and (7) early sale age of excess calves. CONCLUSION: The results from this survey highlight the need for greater awareness of industry standards for calf husbandry and weaning.

Factors associated with colostrum immunoglobulin G concentration in northern-Victorian dairy cows.

OBJECTIVES: To determine the proportion of first-milking colostrum samples produced on four northern-Victorian dairy farms that meet industry standards in terms of immunoglobulin G (IgG) concentration and to identify risk factors that affect colostrum quality. METHODS: Colostrum IgG concentrations from 442 dairy cows on four farms were estimated using a Brix refractometer and risk factors for colostrum IgG concentration were determined using multivariable logistic regression. RESULTS: Only 39% of samples met the definition of high quality. The strongest predictor for colostrum quality was the interval from calving to colostrum harvesting. Colostrum harvested from cows within 12 h of calving was 6-fold more likely to be high quality compared with colostrum harvested later. Colostrum from cows in ≥ 4th lactation was nearly twice as likely to be high quality compared with cows entering their 1st lactation. If the calf was not allowed to suckle from the dam prior to colostrum harvesting, the odds of producing high-quality colostrum were nearly 4-fold greater. If the cow had not leaked colostrum prior to harvesting, it was more than 3-fold more likely to produce high-quality colostrum. CONCLUSIONS: The majority of samples assessed were below industry standard. Herd, lactation number, calf suckling or cow leaking colostrum prior to harvesting and time between calving and colostrum harvesting were factors that influenced colostrum IgG concentration. The results support current industry recommendations of harvesting colostrum shortly after parturition (ideally within 12 h of calving) and testing the quality of all colostrum prior to feeding to dairy calves.

Mitochondrial - Endoplasmic reticulum interactions in the trophoblast: Stress and senescence.

Placental stress has been implicated in the pathophysiology of complications of pregnancy, including growth restriction and pre-eclampsia. Initially, attention focused on oxidative stress, but recently mitochondrial and endoplasmic reticulum stress have been identified. Complex molecular interactions exist among these different forms of stress, making it unlikely that any occurs in isolation. In part, this is due to close physiological connections between the two organelles principally involved, mitochondria and the endoplasmic reticulum (ER), mediated through Ca2+ signalling. Here, we review the involvement of the mitochondria-ER unit in the generation of stress within the trophoblast, and consider consequences for obstetric outcome. Mild stress may induce adaptive responses, including upregulation of antioxidant defences and autophagy, while moderate levels may affect stem cell behaviour and reduce cell proliferation, contributing to the growth-restricted phenotype. High levels of stress can stimulate release of pro-inflammatory cytokines and anti-angiogenic factors, increasing the risk of pre-eclampsia. In addition, chronic stress may promote senescence of the trophoblast, which in other cell types leads to a pro-inflammatory senescence-associated secretory phenotype. Evidence from rodents suggests that a degree of trophoblastic stress develops with increasing gestational age in normal pregnancies. The increase in maternal concentrations of soluble fms-like tyrosine kinase-1 (sFlt-1) and reduction in placental growth factor (PlGF) suggest the same may occur in the human, starting around 30 weeks of pregnancy. Placental malperfusion, or co-existing maternal conditions, such as diabetes, will exacerbate that stress. Amelioration of trophoblastic stress should remain a research priority, but will be difficult due to the complexity of the molecular pathways involved.

Survey of bovine colostrum quality and hygiene on northern Victorian dairy farms.

One of the major challenges for dairy producers is to produce, harvest, and store high-quality colostrum and feed it to their replacement heifer calves. Limited published data are available in Australia regarding the relationship between colostrum management, hygiene, and quality. The objectives of this study were to investigate (1) the colostrum storage and handling practices carried out on farm; (2) the immunoglobulin concentration and bacterial composition of colostrum being fed to replacement dairy heifer calves; (3) the percentage of colostrum being fed to replacement dairy heifer calves that meet industry recommendations; and (4) risk factors for bacterial contamination of colostrum. The study was carried out on 24 dairy farms located near Rochester, Victoria, Australia. Two hundred forty colostrum samples were collected (10 samples per farm). Each farm harvested and stored first-milking colostrum under normal farm conditions. A 10-mL sample of the colostrum was collected in a sterile container immediately before feeding, and a Brix refractometer reading was taken. The samples were then frozen at -4°C and submitted for bacterial concentration analysis. Fifty-eight percent of colostrum samples met the recommended industry standard of a total plate count (TPC) of <100,000cfu/mL, and 94% of colostrum samples met the recommended industry standard of total coliform count (TCC) of 10,000cfu/mL. However, when all the current industry recommendations for TPC, TCC, and Brix refractometer percentage for colostrum quality were considered, only 23% of the samples met all standards. These findings demonstrate that a large number of calves were at risk of receiving colostrum of poor quality, with high bacterial loads that may have interfered with the acquisition of transfer of passive immunity and affected calf health. Further investigation is required to identify the farm-specific factors that may influence the level of bacterial contamination of colostrum. Recommendations as a result of this study include refrigeration of excess colostrum shortly (within 1h) after collection and thorough disinfection of the calf feeding apparatus before use.

Electron- and photon-impact ionization of furfural.

The He(i) photoelectron spectrum of furfural has been investigated, with its vibrational structure assigned for the first time. The ground and excited ionized states are assigned through ab initio calculations performed at the outer-valence Green's function level. Triple differential cross sections (TDCSs) for electron-impact ionization of the unresolved combination of the 4a″ + 21a' highest and next-highest occupied molecular orbitals have also been obtained. Experimental TDCSs are recorded in a combination of asymmetric coplanar and doubly symmetric coplanar kinematics. The experimental TDCSs are compared to theoretical calculations, obtained within a molecular 3-body distorted wave framework that employed either an orientation average or proper TDCS average. The proper average calculations suggest that they may resolve some of the discrepancies regarding the angular distributions of the TDCS, when compared to calculations employing the orbital average.

Influence of speed of sample processing on placental energetics and signalling pathways: implications for tissue collection.

INTRODUCTION: The placenta is metabolically highly active due to extensive endocrine and active transport functions. Hence, placental tissues soon become ischaemic after separation from the maternal blood supply. Ischaemia rapidly depletes intracellular ATP, and leads to activation of stress-response pathways aimed at reducing metabolic demands and conserving energy resources for vital functions. Therefore, this study aimed to elucidate the effects of ischaemia ex vivo as may occur during tissue collection on phosphorylation of placental proteins and kinases involved in growth and cell survival, and on mitochondrial complexes. METHODS: Eight term placentas obtained from normotensive non-laboured elective caesarean sections were kept at room-temperature and sampled at 10, 20, 30 and 45 min after delivery. Samples were analyzed by Western blotting. RESULTS: Between 10 and 45 min the survival signalling pathway intermediates, P-AKT, P-GSK3α and ß, P-4E-BP1 and P-p70S6K were reduced by 30-65%. Stress signalling intermediates, P-eIF2α increased almost 3 fold after 45 min. However, other endoplasmic reticulum stress markers and the Heat Shock Proteins, HSP27, HSP70 and HSP90, did not change. Phosphorylation of AMPK, an energy sensor, was elevated 2 fold after 45 min. Contemporaneously, there was an ∼25% reduction in mitochondrial complex IV subunit I. DISCUSSION AND CONCLUSIONS: These results suggest that for placental signalling studies, samples should be taken and processed within 10 min of caesarean delivery to minimize the impact of ischaemia on protein phosphorylation.

Oxygen delivery and fetal-placental growth: beyond a question of supply and demand?

Towards the end of the first trimester, blood flow and oxygenation rise within the placenta, supporting an increased capacity for mitochondrial oxidative metabolism in both the placenta and developing fetus. In this regard, the placenta acts uniquely as both a conduit of oxygen to the fetal circulation and a significant consumer of oxygen in order to support its own energy demands for the processes of nutrient transport and protein synthesis for hormone production and growth. When the supply of oxygen becomes restricted, for example during chronic exposure to hypobaric hypoxia at high altitude, placental and fetal tissues respond in order to optimise the allocation of oxygen between competing demands. In this case, the placenta appears to remodel its metabolism to decrease oxygen consumption, probably by increasing ATP production via glycolysis. This process can maintain oxygen supply to the fetus but is still associated with growth restriction. Oxidative stress, a feature of pre-eclampsia, might elicit similar metabolic changes in the absence of hypoxia. This review considers what is known about the metabolic response of the placenta and fetal tissues to hypoxia and oxidative stress, and suggests possible mechanisms that might underlie such metabolic remodelling using lessons from other tissues and organ systems. Aspects of the hypoxia response that remain to be addressed are highlighted and future studies suggested. Much remains unknown about the coordinated metabolic response of the fetal-placental unit to chronic hypoxia and oxidative stress, but it would appear to be more than a simple question of supply and demand.

Cryopreservation of placental biopsies for mitochondrial respiratory analysis.

Mitochondrial function is required to support energetically-demanding processes in the placenta. As such, a compromise in mitochondrial function could severely impact fetal growth and development. Respirometry is a highly useful method for studying mitochondrial function, but is not possible in freeze-thawed mitochondria, which become uncoupled. We have developed a novel method that permits respiratory analysis of cryopreserved placental tissue. We studied mitochondrial function in 7 normal human placentas, analysing both fresh and cryopreserved samples. We found no impairments in respiration following cryopreservation in the delivery suite, with enhanced coupling, as indicated by higher respiratory control ratios, than in fresh placental samples transported to the laboratory on ice.

Differential cross sections for the ionization of oriented H2 molecules by electron impact.

A nonperturbative close-coupling technique is used to calculate differential cross sections for the electron-impact ionization of H2 at an energy of 35.4 eV. Our approach allows cross sections for any orientation of the molecule with respect to the incident electron beam to be analyzed. New features in the resulting cross sections are found compared with the case where the molecular orientation is averaged, and also with cross sections for He at equivalent electron kinematics. When averaged over all possible molecular orientations, good agreement is found with recent experimental results.

Rosiglitazone treatment improves cardiac efficiency in hearts from diabetic mice.

Isolated perfused hearts from type 2 diabetic (db/db) mice show impaired ventricular function, as well as altered cardiac metabolism. Assessment of the relationship between myocardial oxygen consumption (MVO(2)) and ventricular pressure-volume area (PVA) has also demonstrated reduced cardiac efficiency in db/db hearts. We hypothesized that lowering the plasma fatty acid supply and subsequent normalization of altered cardiac metabolism by chronic treatment with a peroxisome proliferator-activated receptor-gamma (PPARgamma) agonist will improve cardiac efficiency in db/db hearts. Rosiglitazone (23 mg/kg body weight/day) was administered as a food admixture to db/db mice for five weeks. Ventricular function and PVA were assessed using a miniaturized (1.4 Fr) pressure-volume catheter; MVO(2) was measured using a fibre-optic oxygen sensor. Chronic rosiglitazone treatment of db/db mice normalized plasma glucose and lipid concentrations, restored rates of cardiac glucose and fatty acid oxidation, and improved cardiac efficiency. The improved cardiac efficiency was due to a significant decrease in unloaded MVO(2), while contractile efficiency was unchanged. Rosiglitazone treatment also improved functional recovery after low-flow ischemia. In conclusion, the present study demonstrates that in vivo PPARgamma-treatment restores cardiac efficiency and improves ventricular function in perfused hearts from type 2 diabetic mice.

A case of multiple cutaneous schwannomas; schwannomatosis or neurofibromatosis type 2?

A 54 year old man presented with numerous cutaneous schwannomas, cranial nerve lesions, and spinal cord lesions, but no evidence of vestibular nerve involvement. There was no family history of neurocutaneous lesions. To help discriminate between the various possible diagnoses in this patient, molecular analysis of two cutaneous schwannomas was undertaken. An identical point mutation in the NF2 gene in the two anatomically distinct tumours was found, confirming this as a case of NF2 mosaicism.

A study of metabolic compartmentation in the rat heart and cardiac mitochondria using high-resolution magic angle spinning 1H NMR spectroscopy.

High-resolution magic angle spinning (MAS) (1)H nuclear magnetic resonance (NMR) spectroscopy is increasingly being used to monitor metabolic abnormalities within cells and intact tissues. Many toxicological insults and metabolic diseases affect subcellular organelles, particularly mitochondria. In this study high-resolution (1)H NMR spectroscopy was used to examine metabolic compartmentation between the cytosol and mitochondria in the rat heart to investigate whether biomarkers of mitochondrial dysfunction could be identified and further define the mitochondrial environment. High-resolution MAS spectra of mitochondria revealed NMR signals from lactate, alanine, taurine, choline, phosphocholine, creatine, glycine and lipids. However, spectra from mitochondrial extracts contained additional well-resolved resonances from valine, methionine, glutamine, acetoacetate, succinate, and aspartate, suggesting that a number of metabolites bound within the mitochondrial membranes occur in 'NMR invisible' environments. This effect was further investigated using diffusion-weighted measurements of water and NMR spectroscopy during state 2 and state 3 respiration. State 3 respiration caused a decrease in the resonance intensity of endogenous succinate compared with state 2 respiration, suggesting that coupled respiration may also modulate the NMR detection of metabolites within mitochondria.