Reliable determination of training-induced alterations in muscle fiber composition in human skeletal muscle using quantitative polymerase chain reaction.

Determination of muscle fiber composition in human skeletal muscle biopsies is often performed using immunohistochemistry, a method that tends to be both time consuming, technically challenging, and complicated by limited availability of tissue. Here, we introduce quantitative reverse transcriptase polymerase chain reaction (qRT-PCR)-based Gene-family profiling (GeneFam) of myosin heavy chain (MyHC) mRNA expression as a high-throughput, sensitive, and reliable alternative. We show that GeneFam and immunohistochemistry result in similar disclosures of alterations in muscle fiber composition in biopsies from musculus vastus lateralis and musculus biceps brachii of previously untrained young women after 12 weeks of progressive strength training. The adaptations were evident as (a) consistent increases in MyHC2A abundance; (b) consistent decreases in MyHC2X abundance; and (c) consistently stable MyHC1 abundance, and were not found using traditional reference gene-based qRT-PCR analyses. Furthermore, muscle fiber composition found using each of the two approaches was correlated with each other (r = 0.50, 0.74, and 0.78 for MyHC1, A, and X, respectively), suggesting that GeneFam may be suitable for ranking of individual muscle phenotype, particularly for MyHC2 fibers. In summary, GeneFam of MyHC mRNA resulted in reliable assessment of alterations in muscle fiber composition in skeletal muscle of previously untrained women after 12 weeks of strength training.

Irisin and FNDC5: effects of 12-week strength training, and relations to muscle phenotype and body mass composition in untrained women.

PURPOSE: To investigate the effects of strength training on abundances of irisin-related biomarkers in skeletal muscle and blood of untrained young women, and their associations with body mass composition, muscle phenotype and levels of thyroid hormones. METHODS: Eighteen untrained women performed 12 weeks of progressive whole-body heavy strength training, with measurement of strength, body composition, expression of irisin-related genes (FNDC5 and PGC1α) in two different skeletal muscles, and levels of serum-irisin and -thyroid hormones, before and after the training intervention. RESULTS: The strength training intervention did not result in changes in serum-irisin or muscle FNDC5 expression, despite considerable effects on strength, lean body mass (LBM) and skeletal muscle phenotype. Our data indicate that training affects irisin biology in a LBM-dependent manner. However, no association was found between steady-state serum-irisin or training-associated changes in serum-irisin and alterations in body composition. FNDC5 expression was higher in m.Biceps brachii than in m.Vastus lateralis, with individual expression levels being closely correlated, suggesting a systemic mode of transcriptional regulation. In pre-biopsies, FNDC5 expression was correlated with proportions of aerobic muscle fibers, a relationship that disappeared in post-biopsies. No association was found between serum-thyroid hormones and FNDC5 expression or serum-irisin. CONCLUSION: No evidence was found for an effect of strength training on irisin biology in untrained women, though indications were found for a complex interrelationship between irisin, body mass composition and muscle phenotype. FNDC5 expression was closely associated with muscle fiber composition in untrained muscle.

Prevalence of iron deficiency and anemia among healthy women of reproductive age in Bhaktapur, Nepal.

OBJECTIVE: To determine the prevalence of anemia and iron status as assessed by biochemical markers and to explore the associations between markers of iron status and iron intake. STUDY AREA AND POPULATION: Five hundred healthy women of reproductive age from the Bhaktapur district of Nepal were included in the study. METHODS: A cluster sampling procedure was applied for this cross-sectional study. Women without any ongoing infection aged 13-35 years were selected randomly from the population. We measured the plasma concentration of hemoglobin (Hb), ferritin and transferrin receptors. Dietary information was obtained by a food frequency questionnaire and two 24-h dietary recalls. RESULTS: The prevalence of anemia (Hb concentration <12 g/dl) was 12% (n=58). The prevalence of depleted iron stores (plasma ferritin <15 microg/l) was 20% (n=98) whereas the prevalence of iron deficiency anemia (anemia, depleted iron stores with elevated transferrin receptor i.e. >1.54 mg/l) was 6% (n=30). Seven percent (n=35) of women were having iron-deficient erythropoiesis (depleted iron stores and elevated transferrin receptor but normal Hb). Out of the 58 anemic women, 41 (71%) and 31 (53%) were also having elevated plasma transferrin receptor and depleted iron stores, respectively. Fifty-four percent of the women ate less than the recommended average intake of iron. The main foods contributing to dietary iron were rice, wheat flour and green and dry vegetables. CONCLUSIONS: The prevalence of anemia in our study was substantially lower than the national figure for non-pregnant women. Only about half of the women with anemia were also having depleted iron stores, suggesting that other causes of anemia may be prevalent in this population. SPONSORSHIP: Norwegian Universities Committee for Development, Research and Education (NUFU).

The prevalence of anemia and iron deficiency is more common in breastfed infants than their mothers in Bhaktapur, Nepal.

BACKGROUND/OBJECTIVES: Iron deficiency anemia is a widespread public health problem, particularly in low- and middle-income countries. Maternal iron status around and during pregnancy may influence infant iron status. We examined multiple biomarkers to determine the prevalence of iron deficiency and anemia among breastfed infants and explored its relationship with maternal and infant characteristics in Bhaktapur, Nepal. SUBJECTS/METHODS: In a cross-sectional survey, we randomly selected 500 mother-infant pairs from Bhaktapur municipality. Blood was analyzed for hemoglobin, ferritin, total iron-binding capacity, transferrin receptors and C-reactive protein. RESULTS: The altitude-adjusted prevalence of anemia was 49% among infants 2-6-month-old (hemaglobin (Hb) <10.8 g/dl) and 72% among infants 7-12-month-old (Hb <11.3 g/dl). Iron deficiency anemia, defined as anemia and serum ferritin <20 or <12 µg/l, affected 9 and 26% of infants of these same age groups. Twenty percent of mothers had anemia (Hb <12.3 g/dl), but only one-fifth was explained by depletion of iron stores. Significant predictors of infant iron status and anemia were infant age, sex and duration of exclusive breastfeeding and maternal ferritin concentrations. CONCLUSIONS: Our findings suggest that iron supplementation in pregnancy is likely to have resulted in a low prevalence of postpartum anemia. The higher prevalence of anemia and iron deficiency among breastfed infants compared with their mothers suggests calls for intervention targeting newborns and infants.

The immune response to pneumococcal polysaccharide vaccine in zinc-depleted mice.

Zinc depletion affects several facets of the immune system and the resistance to infections. We assessed the effect of zinc deprivation on the immune response to the pneumococcal polysaccharide antigens in the commercially available Pneumovax pneumococcal vaccine. Young female BALB/c mice were fed diets with 2.7, 5.8 or 25 micro g of elemental zinc per mg diet. After six weeks of pair feeding, there were significant differences in the mean body weights between the feeding groups and we demonstrated a dose response of the zinc level in the diet on growth. The induced zinc deficiency had no discernible effect on the antipneumococcal polysaccharide immunoglobulin M (IgM) response following immunization with the pneumococcal vaccine. Although zinc depletion has a detrimental effect on the immune system, the murine T-cell-independent response to antigens such as those in the pneumococcal polysaccharide capsule does not seem to be affected.

Pneumococcal pulmonary infection, septicaemia and survival in young zinc-depleted mice.

The aim of the present study was to explore whether mice fed a diet low in Zn (2.0 mg Zn/kg diet) for a relatively short period of time were more prone to severe Streptococcus pneumoniae infection than mice fed a normal diet (25 mg elemental Zn/kg). The Zn-deficient mice were compared with mice in two Zn-adequate control groups; one pair-fed and another with free access to the diet. After 2 weeks feeding, the mice were infected intranasally under anaesthesia with a suspension containing about 10(7) pneumococci. Clinical status was observed every day and blood samples were examined for S. pneumoniae every second day for a week. All infected mice examined carried the infecting strain intranasally. The survival time and time before positive blood culture were significantly shorter in the Zn-depleted group than in the pair-fed Zn-adequate group (hazard ratios 15.6 and 3.2, and respectively). At the end of the observation period, ten of the twelve mice in the Zn-deficient group were dead while one of twelve and two of twelve were dead in the two Zn-adequate control groups. This study shows that even acutely-induced Zn deficiency dramatically increases the risk of serious pneumococcal infection in mice.