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Biomedical subjects

M P Richards

Publications and source records attributed to M P Richards.

At least 19 recordsLinked to original sources

Genetic regulation of feed intake and energy balance in poultry.

Intensive selection by poultry breeders over many generations for economically important production traits such as growth rate and meat production has been accompanied by significant changes in feed intake and energy balance. For example, the modern commercial broiler, selected for rapid growth and enhanced muscle mass, does not adequately regulate voluntary feed intake to achieve energy balance. When given unrestricted access to feed, broilers exhibit hyperphagia leading to an excessive accumulation of energy (fat) stores, making these birds prone to obesity and other health-related problems. Humoral and neural pathways have been identified and studied in mammals that link appetite and energy balance. A series of highly integrated regulatory mechanisms exists for both of these processes involving complex interactions between peripheral tissues and the central nervous system. Within the central nervous system, the brainstem and the hypothalamus play critical roles in the regulation of feed intake and energy balance. Genes encoding key regulatory factors such as hormones, neuropeptides, receptors, enzymes, transcription factors, and binding/transport proteins constitute the molecular basis for regulatory systems that derive from integrated sensing, signaling, and metabolic pathways. However, we do not yet have a complete understanding of the genetic basis for this regulation in poultry. This review examines what is currently known about the regulation of feed intake and energy balance in poultry. A better understanding of the genes associated with controlling feed intake and energy balance and how their expression is regulated by nutritional and hormonal stimuli will offer new insights into current poultry breeding and management practices.

Animal Husbandry↗

Stable carbon and nitrogen isotope values of bone and teeth reflect weaning age at the Medieval Wharram Percy site, Yorkshire, UK.

We report on the measurements of carbon and nitrogen stable isotopes of both bone and teeth from a single site and population (Medieval Wharram Percy), undertaken to explore variations due to weaning in a past population. There have been a number of recent studies of weaning using delta(15)N values of ribs, and we indicate a number of assumptions that must be met before the results of such studies can be correctly interpreted. We found that rib collagen delta(15)N values decrease to adult levels after age 2 years, indicating that weaning occurred at or before this age. Rib collagen delta(13)C values are also more enriched than adult delta(13)C values before age 2 years, and we argue that this is due to the so-called "carnivore" effect in delta(13)C. We measured teeth and rib delta(15)N values from the same individuals and found that for individuals up to age 11 years, tooth dentine delta(15)N is higher than adult rib delta(15)N values, indicating that the dentine was formed during breast-feeding and that there was almost no turnover of dentine since. We observed some decrease in delta(13)C and delta(15)N rib values, compared to adult rib and teeth values, for the few years after weaning that may relate to a theoretically predicted physiological nitrogen imbalance during this period of rapid growth, but this is more likely due to a childhood diet (up to age 9) which was isotopically different from later diet, possibly consisting of a greater proportion of plant foods.

Adolescent↗

Dietary protein regulates in vitro lipogenesis and lipogenic gene expression in broilers.

The purpose of this experiment was to determine the possible relationship between certain indices of lipid metabolism and specific gene expression in chickens fed graded levels of dietary crude protein. Male, broiler chickens growing from 7 to 28 days of age were fed diets containing 12, 21 or 30% protein ad libitum. In addition, another group of birds was fed on a regimen consisting of a daily change in the dietary protein level (12 or 30%). This latter group was further subdivided such that one-half of the birds received each level of protein on alternating days. Birds were sampled from 28 to 30 days of age. Measurements taken included in vitro lipogenesis, malic enzyme activity the expression of the genes for malic enzyme, fatty acid synthase and acetyl coenzyme carboxylase. In vitro lipogenesis and malic enzyme activity were inversely related to dietary protein levels (12-30%) and to acute changes from 12 to 30%. In contrast, expression of malic enzyme, fatty acid synthase and acetyl CoA carboxylase genes were constant over a dietary protein range of 12-21%, but decreased by feeding a 30% protein diet (acute or chronic feeding). Results of the present study demonstrate a continued role for protein in the regulation of broiler metabolism. It should be pointed out, however, that metabolic regulation at the gene level only occurs when feeding very high levels of dietary protein.

Acetyl-CoA Carboxylase↗

A brief review of the archaeological evidence for Palaeolithic and Neolithic subsistence.

Knowledge of our ancestor's diets is becoming increasingly important in evolutionary medicine, as researchers have argued that we have evolved to specific type of 'Palaeolithic' diet, and many modern nutritional disorders relate to the mismatch between the diet to which we have evolved, and the relatively newer agricultural-based 'Neolithic' diets. However, what is the archaeological evidence for pre-agricultural diets and how have they changed over the four million years of hominid evolution? This paper briefly introduces the three lines of evidence we have for Palaeolithic and Neolithic diets; morphological changes, archaeological material evidence, and direct measurement of diet from bone chemistry. The morphological changes, increasing gracilization of the mandible and increasing brain size have been interpreted (based on analogies with living primates) as the move from plants to higher-quality, more digestible, animal meat, although this is debated. The archaeological evidence is especially weak, as many organic materials, especially plants, do not survive well, and are therefore invisible in the archaeological record. Artefacts, such as stone tools which are likely to be used for hunting and animal bones with evidence of human processing and butchering do indicate that hunting did occur at many times in the past, but it is impossible to judge the frequency. Direct evidence from bone chemistry, such as the measurement of the stable isotopes of carbon and nitrogen, do provide direct evidence of past diet, and limited studies on five Neanderthals from three sites, as well as a number of modern Palaeolithic and Mesolithic humans indicates the importance of animal protein in diets. There is a significant change in the archaeological record associated with the introduction of agriculture worldwide, and an associated general decline in health in some areas. However, there is an rapid increase in population associated with domestication of plants, so although in some regions individual health suffers after the Neolithic revolution, as a species humans have greatly expanded their population worldwide.

Agriculture↗

Stable isotope evidence for increasing dietary breadth in the European mid-Upper Paleolithic.

New carbon and nitrogen stable isotope values for human remains dating to the mid-Upper Paleolithic in Europe indicate significant amounts of aquatic (fish, mollusks, and/or birds) foods in some of their diets. Most of this evidence points to exploitation of inland freshwater aquatic resources in particular. By contrast, European Neandertal collagen carbon and nitrogen stable isotope values do not indicate significant use of inland aquatic foods but instead show that they obtained the majority of their protein from terrestrial herbivores. In agreement with recent zooarcheological analyses, the isotope results indicate shifts toward a more broad-spectrum subsistence economy in inland Europe by the mid-Upper Paleolithic period, probably associated with significant population increases.

Animals↗

Sensory nerve conduction deficit in experimental monoclonal gammopathy of undetermined significance (MGUS) neuropathy.

An emerging body of evidence from in vitro studies and in vivo animal models supports a pathogenic role of antibodies in the development of peripheral neuropathy associated with monoclonal gammopathy of undetermined significance (MGUS). Although the assessment of motor and sensory nerve fiber function is of clinical importance, it is seldom applied experimentally. We describe the application of an electrophysiologic method for the evaluation of motor and sensory nerve fiber function using an experimental model of MGUS neuropathy. Supramaximal stimulation of the tibial nerve elicited an early motor response (M-wave, 1.7 +/- 0.1 ms, n = 10) and a late sensory (H-reflex, 7.8 +/- 0.1 ms, n = 10) response that was recorded from the hind foot of anesthetized rats. Intraneural injection of serum antibodies from a MGUS patient with sensorimotor polyneuropathy, but not from an age-matched control subject, produced a marked attenuation of the H-reflex (P < 0.01, n = 10) without affecting the M-wave. Light and electron microscopy of affected nerve showed myelinoaxonal degeneration with sparing of the smaller unmyelinated nerve fibers. The combined electrophysiologic and morphologic findings presented in this study are consistent with a selective sensory conduction deficit in MGUS neuropathy. Selective injury of afferent nerve fibers by this patient's serum antibodies may result from reactivity to neural antigens uniquely expressed by sensory neurons.

Aged↗

The ontogeny of leptin mRNA expression in growing broilers and its relationship to metabolic body weight.

The polypeptide hormone leptin is produced by both adipose tissue and the liver and has been shown to induce satiety in chickens. In this study we have investigated the developmental regulation of leptin mRNA expression in growing broiler chickens. Leptin expression generally increases in all tissues from 1-12 weeks of age. In the subcutaneous fat depot there is an apparent pattern of increased leptin mRNA expression occurring at 2, 6, and 10 weeks post-hatch. This pattern was not evident in the other tissues surveyed and may relate to the cycle of loading and unloading of adipocytes with lipid. No consistent gender differences in leptin expression patterns were detected in the tissues surveyed, as is often observed in mammals. Positive correlations between metabolic body weight and adipose leptin expression levels were observed. Leptin expression by the liver was highly correlated with metabolic body weight from 1-6 weeks of age, and uncorrelated from 6-12 weeks of age. This pattern of increasing liver leptin expression with increasing body weight during the early rapid growth phase of the bird may be due to limited fat storage during this period, which is followed by rapid body fat accumulation from 6-12 weeks. The characterization and tissue specific distribution of leptin mRNA expression in the growing broiler indicate similar patterns of leptin production to that of growing mammals. Leptin may be involved in lipid flux through the adipocyte as well as the shift in lipid metabolism to increased storage during pre-puberty.

Adipose Tissue↗

Comparison of gonadal hormone levels in turkey embryos incubated in long-term shell-less culture and in ovo.

Changes in concentrations of 17beta-estradiol (E2) and androgenic hormones were measured in turkey embryos incubated in long-term, shell-less culture (ex ovo) and in ovo. Blood samples were obtained from both sets of embryos on Days 14, 16, 18, 20, and 22 and from embryos incubated in ovo on Days 24, 26, and 28. Ex ovo and in ovo embryos showed no differences in either hormone within sexes, with one exception. On Day 14 of incubation, the ex ovo females had higher (P < 0.05) E2 levels (55.6+/-5.1 pg/mL) than the in ovo females (32.2+/-2.3 pg/mL); however, this result might have been due to the small sample size (n = 3) for ex ovo females. No significant differences were found in androgen concentrations between sexes in ovo on Days 24, 26, and 28 of incubation. However, on Days 24, 26, and 28, in ovo females showed highly significant differences (P < 0.01) in E2 compared with males of the same age. These results indicate a similar developmental pattern for the endocrine system in ovo and ex ovo through Day 22 of incubation. Further, there were sex differences in E2 that are likely to be critical for sexual differentiation that emerges late in embryonic development.

Age Factors↗

Design and application of a polyclonal peptide antiserum for the universal detection of leptin protein.

An epitope-specific polyclonal antiserum was produced in rabbits immunized against a synthetic 15 amino acid peptide (QRVTGLDFIPGLHPV) derived from the coding sequence reported for the porcine leptin gene (GenBank Accession No. U59894). This peptide contains a core sequence comprised of eight amino acids (GLDFIPGL) that is totally conserved in all leptin proteins studied to date. Purified recombinant human, mouse, rat, pig, and chicken leptin proteins were separated by polyacrylamide gel electrophoresis (SDS-PAGE) and electro-blotted onto PVDF membranes. Western blots were developed employing the leptin-specific peptide antiserum with an alkaline-phosphatase-conjugated anti-rabbit IgG second antibody chromogenic system. The peptide antiserum was found to be highly specific for leptin which exhibited an estimated molecular weight of about 16 kDa for all species analyzed. The sensitivity of the Western blot assay was not sufficient to permit the direct detection of leptin in chicken serum or plasma. However, with this assay we were able to detect native leptin protein in an enriched fraction prepared from chicken plasma using a combination of gel filtration and ion exchange column chromatography. Slot blots indicated a potential application of the immunostaining technique for quantitative analysis of leptin protein. Finally, the peptide antiserum was successfully employed to localize leptin protein by immunohistochemical staining of thin sections prepared from adipose (chicken and pig) and liver (chicken) tissue samples. This study is the first to report a polyclonal peptide antiserum that apparently recognizes intact leptin protein, both native and recombinant, regardless of the species of origin.

Amino Acid Sequence↗

Relationship between functional evaluation measures and self-assessment in nonacute low back pain.

STUDY DESIGN: The correlations between objective biomechanical indicators of function and self-assessment scores were examined retrospectively for 91 subjects with nonacute low back pain. OBJECTIVES: To examine the correlation between self-assessment, trunk range of motion (ROM), velocity, and complex mechanical coordination patterns of the spine in nonacute low back pain. SUMMARY OF BACKGROUND DATA: In low back pain, there is often little concordance between pain, physical impairment, and disability. Use of range of motion and velocity to enhance objectivity in impairment evaluations has been ineffectual. In this study, two hypotheses were examined: range of motion and velocity are controllable and inherently correlated with self-assessment; complex spinal coordination patterns such as range of lordosis cannot be controlled and are independent of self-assessment. METHODS: Self-assessment questionnaires were administered, and indexes of spinal motion and coordination were measured through skin marker kinematics. The correlation between self-assessments and biomechanical measures was determined. RESULTS: Self-assessments of function were significantly correlated with parameters prone to regulation: range of motion, velocity, and load lifted. In contrast, little correlation was found with measures of complex spinal coordination less susceptible to conscious or affective regulation, namely, range of lordosis and estimated segmental mobility. This effect was magnified with increased load. Self-assessment scores were significantly poorer among insurance referrals, regardless of functional status. CONCLUSIONS: Simple parameters of the functional examination, such as range of motion and velocity, are strongly correlated with cognitive state, and thus the information they supply is less than ideal. Complex spinal coordination is a better indicator of the degree of spinal dysfunction and enhances the process of differentiating between pain, disability, and functional impairment.

Adult↗

Neanderthal diet at Vindija and Neanderthal predation: the evidence from stable isotopes.

Archeological analysis of faunal remains and of lithic and bone tools has suggested that hunting of medium to large mammals was a major element of Neanderthal subsistence. Plant foods are almost invisible in the archeological record, and it is impossible to estimate accurately their dietary importance. However, stable isotope (delta(13)C and delta(15)N) analysis of mammal bone collagen provides a direct measure of diet and has been applied to two Neanderthals and various faunal species from Vindija Cave, Croatia. The isotope evidence overwhelmingly points to the Neanderthals behaving as top-level carnivores, obtaining almost all of their dietary protein from animal sources. Earlier Neanderthals in France and Belgium have yielded similar results, and a pattern of European Neanderthal adaptation as carnivores is emerging. These data reinforce current taphonomic assessments of associated faunal elements and make it unlikely that the Neanderthals were acquiring animal protein principally through scavenging. Instead, these findings portray them as effective predators.

Animals↗

Quantitative analysis of leptin mRNA using competitive reverse transcription polymerase chain reaction and capillary electrophoresis with laser-induced fluorescence detection.

Leptin, the protein hormone product of the obese (ob) gene, functions in the regulation of appetite, energy expenditure, and reproduction in animals and humans. Since changes in the level of circulating leptin can have marked physiological consequences, it is important to be able to accurately quantify leptin gene expression. Toward this goal, we have constructed a chicken leptin RNA competitor and successfully employed it as an internal standard in the development of a quantitative-competitive reverse transcription polymerase chain reaction (QC-RT-PCR) assay for leptin mRNA. Capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) was utilized for the separation and analysis of chicken leptin target (261 bp) and competitor (234 bp) dsDNA products from QC-RT-PCR assay samples. Leptin amplicons were separated using a DB-1 coated capillary (27 cm x 100 microm ID) at a field strength of 300 V/cm in a replaceable sieving matrix consisting of 0.5% hydroxypropylmethyl cellulose (HPMC) in 1 x TBE (89 mM Tris-base, 89 mM boric acid, 2 mM EDTA, pH 8.3) buffer with 0.5 microg/mL EnhanCE fluorescent intercalating dye. Samples were diluted 1:100 with deionized water and introduced into the capillary by electrokinetic injection. QC-RT-PCR/CE-LIF was used to quantify leptin mRNA in liver and adipose tissue from 8-week-old male and female broiler chickens. This study is the first report of quantitative analysis of leptin gene expression using QC-RT-PCR/CE-LIF.

Adipose Tissue↗

Effect of pH on lipid oxidation using trout hemolysate as a catalyst: a possible role for deoxyhemoglobin.

Hemoglobin-mediated lipid oxidation was studied by adding hemolysate to washed cod muscle. Three pH values were examined (pH 7.6, 7.2, and 6.0). The lag time prior to rancidity and thiobarbituric acid reactive substance development decreased greatly as the pH was reduced (p < 0.01). Formation of methemoglobin due to autoxidation of the heme pigment was found to occur more rapidly at reduced pH. Also, the level of deoxyhemoglobin was found to sharply increase with pH reduction in the range of pH 7.6-6.0. This suggested a potential role for deoxyhemoglobin as a catalyst. ATP lowered hemoglobin oxygenation at pH 7.2. Peroxidation of linoleic acid by oxy/deoxyhemoglobin and methemoglobin was investigated at two levels of preformed lipid hydroperoxides. At a reduced level of preformed lipid hydroperoxides, oxy/deoxyhemoglobin stimulated peroxidation of linoleic acid, whereas methemoglobin did not. At the higher level of preformed lipid hydroperoxides, both oxy/deoxyhemoglobin and methemoglobin were active. This investigation suggests that reduced hemoglobins played an important role in lipid oxidation processes.

Animals↗

Analysis of leptin gene expression in chickens using reverse transcription polymerase chain reaction and capillary electrophoresis with laser-induced fluorescence detection.

Leptin is a peptide hormone product of the obese (ob) gene that functions in the regulation of appetite, energy expenditure and reproduction in animals and humans. We have developed a technique using capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) for the analysis of chicken leptin (261 base pairs, bp) and beta-actin (612 bp) double-stranded DNA products from reverse transcription polymerase chain reaction (RT-PCR) assays. Amplicons were separated using a DB-1 coated capillary (27 cm x 100 microns I.D.) at a field strength of 300 V/cm in a replaceable sieving matrix consisting of 0.5% hydroxypropylmethylcellulose (HPMC) in 1X TBE (89 mM Tris-base, 89 mM boric acid, 2 mM EDTA, pH 8.3) buffer with 0.5 microgram/ml EnhanCE fluorescent intercalating dye. RT-PCR samples (1-2 microliters) were diluted 1:100 with deionized water and introduced into the capillary by electrokinetic injection. Separations were completed in less than 6 min and the total time required per sample, including capillary conditioning, was 8 min. We have applied RT-PCR-CE-LIF to determine the effects of insulin and estrogen treatment on leptin gene expression relative to that of beta-actin in chicken liver and adipose tissue. In addition, we have constructed a chicken leptin mRNA competitor (234 bp amplicon) and evaluated it for use as an internal standard in the development of a quantitative-competitive RT-PCR assay. Our findings represent the first reported application of capillary electrophoresis to the analysis of leptin gene expression by RT-PCR.

Animals↗

Improving the evaluation of benign low back pain.

STUDY DESIGN: A prospective, blind study was conducted to investigate the factors underlying the decisions of expert clinicians in diagnosis of acute, benign low back pain, compared with results obtained with an automated physical examination by machine. From the results, a strategy to significantly improve clinical diagnosis in cases of discordance was determined. OBJECTIVES: To identify factors in the clinical assessment of low back pain that indicate when independent diagnostic testing would be useful. SUMMARY OF BACKGROUND DATA: The clinical evaluation of low back pain is often dominated by subjective reports of pain. Published medical literature has underscored several inherent weaknesses of the clinical examination, and concerns have been raised about its effectiveness for assessing patients with low back pain. Thus, it has been proposed that objective measures to complement the clinician's examination would be beneficial in the formulation of dependable diagnoses. METHODS: Randomly designated subjects, who in describing their conditions were objective or role playing, were assessed by clinicians and a machine for diagnosis of low back pain assessment versus normal backs. Each subject's pain assessment was compared with a gold standard that was established by experts in low back pain. Components of the clinical examination were analyzed to assess which were the most informative in making a reliable diagnosis. The information content of the machine assessment was also analyzed and a strategy to complement the clinical diagnosis with the machine diagnosis determined. RESULTS: Discordance among the various components of the clinical examination was a strong indicator of when the efficacy of the clinical examination dropped below a random level of decision making. When there was discordance, incorporating the functional evaluation by machine into the clinical diagnosis improved the performance of the clinician. Notably, in nonobjective subjects, the accuracy of diagnosis was enhanced by as much as 69%. CONCLUSIONS: It is possible to improve the accuracy of clinical diagnosis by incorporating a functional evaluation by machine when there is discordance between physical examination findings and reported pain.

Acute Disease↗

Zinc, copper, and iron metabolism during porcine fetal development.

Zinc, copper, and iron levels in maternal and fetal pig tissues and fluids were measured starting on d 30 of gestation and continuing to term (d 114) at 10-d intervals. Fetal hematocrit increased from a low of 19% on d 30 to 32% by d 50, after which it remained above 30% to term. Amniotic fluid zinc, copper, and iron all reached maximal levels by d 60 of gestation. Maternal serum zinc levels fluctuated little during gestation, but fetal serum zinc concentration was significantly elevated above maternal levels during the second trimester. Fetal serum copper levels were significantly lower than maternal values throughout gestation and this was also the case for ceruloplasmin oxidase activity. Maternal serum iron reached its lowest level by d 80 of gestation when rate of transfer of iron to the developing fetuses was high. Fetal serum iron declined throughout gestation, reaching its lowest level on d 100. In general, fetal liver concentrations of zinc, copper, and iron were higher than the corresponding maternal values throughout gestation. Distinct increases were noted for fetal hepatic zinc and copper concentrations during the second trimester of pregnancy and these were accompanied by increases in cytosolic and metallothionein-bound zinc and copper levels. Maternal hepatic iron declined during the second trimester, reaching its lowest point on d 80, indicative of the shunting of maternal iron reserves to fetal tissues. Fetal kidney metal levels did not demonstrate any distinctive developmental patterns with respect to zinc, copper, or iron concentrations, but a general accumulation of each metal was observed as gestation progressed. The results of this study highlight some of the distinct changes occurring in the metabolism of zinc, copper, and iron in both maternal and fetal tissues and fluids during gestation in the pig.

Animals↗