Analyzing B-vitamins in Human Milk: Methodological Approaches

According to the World Health Organization (WHO), infants should be exclusively breastfed for the first six months of life. However, there is insufficient information about the concentration of nutrients in human milk. For some nutrients, including B-vitamins, maternal intake affects their concentration in human milk but the extent to which inadequate maternal diets affect milk B-vitamin content is poorly documented. Little is known about infant requirements for B-vitamins; recommendations are generally set as Adequate Intakes (AI) calculated on the basis of the mean volume of milk (0.78 L/day) consumed by infants exclusively fed with human milk from well-nourished mothers during the first six months, and the concentration of each vitamin in milk based on reported values. Methods used for analyzing B-vitamins, commonly microbiological, radioisotope dilution or more recently chromatographic, coupled with UV, fluorometric and MS detection, have rarely been validated for the complex human milk matrix. Thus the validity, accuracy, and sensitivity of analytical methods is important for understanding infant requirements for these nutrients, the maternal intakes needed to support adequate concentrations in breast milk. This review summarizes current knowledge on methods used for analyzing the B-vitamins thiamin, riboflavin, niacin, vitamin B-6 and pantothenic acid, vitamin B-12, folate, biotin, and choline in human milk, their chemical and physical properties, the different forms and changes in concentration during lactation, and the effects of deficiency on the infant.     

Combining land use information and small stream sampling with PCR-based methods for better characterization of diffuse sources of human fecal pollution

Diffuse sources of human fecal pollution allow for the direct discharge of waste into receiving waters with minimal or no treatment. Traditional culture-based methods are commonly used to characterize fecal pollution in ambient waters, however these methods do not discern between human and other animal sources of fecal pollution making it difficult to identify diffuse pollution sources. Human-associated quantitative real-time PCR (qPCR) methods in combination with low-order headwatershed sampling, precipitation information, and high-resolution geographic information system land use data can be useful for identifying diffuse source of human fecal pollution in receiving waters. To test this assertion, this study monitored nine headwatersheds over a two-year period potentially impacted by faulty septic systems and leaky sanitary sewer lines. Human fecal pollution was measured using three different human-associated qPCR methods and a positive significant correlation was seen between abundance of human-associated genetic markers and septic systems following wet weather events. In contrast, a negative correlation was observed with sanitary sewer line densities suggesting septic systems are the predominant diffuse source of human fecal pollution in the study area. These results demonstrate the advantages of combining water sampling, climate information, land-use computer-based modeling, and molecular biology disciplines to better characterize diffuse sources of human fecal pollution in environmental waters.     

Interlaboratory comparison of real-time PCR protocols for quantification of general fecal indicator bacteria

The application of quantitative real-time PCR (qPCR) technologies for the rapid identification of fecal bacteria in environmental waters is being considered for use as a national water quality metric in the United States. The transition from research tool to a standardized protocol requires information on the reproducibility and sources of variation associated with qPCR methodology across laboratories. This study examines interlaboratory variability in the measurement of enterococci and Bacteroidales concentrations from standardized, spiked, and environmental sources of DNA using the Entero1a and GenBac3 qPCR methods, respectively. Comparisons are based on data generated from eight different research facilities. Special attention was placed on the influence of the DNA isolation step and effect of simplex and multiplex amplification approaches on interlaboratory variability. Results suggest that a crude lysate is sufficient for DNA isolation unless environmental samples contain substances that can inhibit qPCR amplification. No appreciable difference was observed between simplex and multiplex amplification approaches. Overall, interlaboratory variability levels remained low (<10% coefficient of variation) regardless of qPCR protocol.     

Growth, yield and seed composition of native Australian legumes with potential as grain crops

BACKGROUND: Many Australian native legumes grow in arid and nutrient‐poor environments. Yet few Australian herbaceous legumes have been investigated for domestication potential. This study compared growth and reproductive traits, grain yield and seed composition of 17 native Australian legumes with three commercial grain legumes. RESULTS: Seed yields of seven native legumes were > 40% of Cicer arietnum, with highest seed yields and harvest indices in Glycine sp. (14.4 g per plant, 0.54 g g^?1) and Lotus cruentus (10.2 g per plant, 0.65 g g^?1). Five native species flowered earlier than field pea (Pisum sativa) (109 days), though many were slower to flower and set seed. Largest seeds were found in Glycine canescens (17 mg), with seed of other native species 14 times smaller than commercial cultivars. Seed composition of many native legumes was similar to commercial cultivars (200–330 g protein kg^?1 dry weight (DW), 130–430 g dietary fibre kg^?1 DW). Two Cullen species had high fat content (>110 g kg^?1 DW) and Trigonella sauvissima had the highest crude protein content (370 g kg^?1 DW). CONCLUSION: The seed composition and reproductive traits of some wild native Australian legumes suggest they could offer potential as grain crops for soils and environments where the current grain legumes are uneconomic. Further evaluation of genetic diversity, especially for seed size, overall productivity, and reproductive development is needed. Copyright © 2011 Society of Chemical Industry     

Development and preliminary evaluation of a pediatric measure of community integration: The Pediatric Community Participation Questionnaire (PCPQ).

Objective: To develop and evaluate a new pediatric measure of community integration: the Pediatric Community Participation Questionnaire (PCPQ). Participants: 105 youths with physical disabilities. Measures: In addition to the PCPQ, demographic and disability-related questions and the Functional Disability Inventory (FDI) were administered. Results: The PCPQ demonstrated convergent validity through its high association with functional ability, discriminant validity through its ability to discriminate between ambulatory and nonambulatory participants, and excellent internal consistency. Conclusions: Community participation, an important component of positive overall development, may be particularly important for youths with physical disabilities. The findings from this study provide preliminary support for the validity and reliability of the PCPQ, supporting its use as a measure of community participation with this population. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Neural parameters contributing to temperature compensation in the flight CPG of the locust, Locusta migratoria

Elevated thoracic temperature increases the wingbeat frequency of flying locusts. We investigated the extent to which temperature-induced changes in resting membrane potential and postsynaptic potential amplitude contribute to the effects of increased temperature on the frequency of the central flight rhythm. Flight neurons were hyperpolarized by changing the K+ concentration of the superfusing saline from 10 mM to 2 mM. 5 min of low-K+ superfusion hyperpolarized flight motoneurons from -42.8 mV to -50.1 mV with a concomitant decrease of the frequency of the central flight rhythm from 11.6 Hz to 10.5 Hz. The amplitude of postsynaptic potentials was halved after 10 min of zero Ca2+/high Mg2+ superfusion, but the frequency of the central rhythm did not change significantly. GABAergic inhibitory connections were reduced in amplitude using picrotoxin. This treatment increased the frequency of the central rhythm from 11.6 Hz to 12.9 Hz, and increased the thermosensitivity of the rhythm frequency. We conclude that the excitatory effect of increased temperature on rhythm frequency is not mediated by temperature effects on membrane potential and/or synaptic potential amplitude. We propose that the inhibitory effect of temperature-induced hyperpolarization of the membrane potential compensates for the excitatory effect of temperature on rhythm frequency (e.g. via increased conduction velocity). We further suggest that some measure of temperature compensation is afforded by equal effects on the amplitudes of excitatory and inhibitory postsynaptic potentials, such that the net effect on the level of excitation is zero.     

Transient central hypothyroidism as a cause of failure to thrive in newborns and infants

The course of two neonates and one 4-month-old infant with laboratory and clinical evidence of central hypothyroidism is described. All three presented with failure to thrive and improved after L-T4 therapy. Early recognition and treatment of newborns and infants with central hypothyroidism is important to maximize the potential for growth and development. Two of the three infants have been documented to have transient central hypothyroidism of hypothalamic origin, not previously reported.     

Memorizing and recalling spatial-temporal patterns in an oscillator model of the hippocampus

We describe the model of the hippocampus consisting of interactive oscillators with input from the entorhinal cortex (modulating the main information flow by a theta rhythm) and the septum (a theta rhythm generator). When interconnections between oscillators are allowed to strengthen in an adaptive way, the network can be trained using a series of lessons. This results in a connection matrix that memorizes the temporal sequence of inputs. Presenting one of the lessons to the trained network results in reproduction of the remainder of the sequence. In this paper, we create such a connection matrix, derive from it an appropriate Markov chain and simulate the chain to illustrate its dynamics.