Effect of postnatal development on calcium currents and slow charge movement in mammalian skeletal muscle

Single- (whole-cell patch) and two-electrode voltage-clamp techniques were used to measure transient (Ifast) and sustained (Islow) calcium currents, linear capacitance, and slow, voltage-dependent charge movements in freshly dissociated fibers of the flexor digitorum brevis (FDB) muscle of rats of various postnatal ages. Peak Ifast was largest in FDB fibers of neonatal (1-5 d) rats, having a magnitude in 10 mM external Ca of 1.4 +/- 0.9 pA/pF (mean +/- SD; current normalized by linear fiber capacitance). Peak Ifast was smaller in FDB fibers of older animals, and by approximately 3 wk postnatal, it was so small as to be unmeasurable. By contrast, the magnitudes of Islow and charge movement increased substantially during postnatal development. Peak Islow was 3.6 +/- 2.5 pA/pF in FDB fibers of 1-5-d rats and increased to 16.4 +/- 6.5 pA/pF in 45-50-d-old rats; for these same two age groups, Qmax, the total mobile charge measurable as charge movement, was 6.0 +/- 1.7 and 23.8 +/- 4.0 nC/microF, respectively. As both Islow and charge movement are thought to arise in the transverse-tubular system, linear capacitance normalized by the area of fiber surface was determined as an indirect measure of the membrane area of the t-system relative to that of the fiber surface. This parameter increased from 1.5 +/- 0.2 microF/cm2 in 2-d fibers to 2.9 +/- 0.4 microF/cm2 in 44-d fibers. The increases in peak Islow, Qmax, and normalized linear capacitance all had similar time courses. Although the function of Islow is unknown, the substantial postnatal increase in its magnitude suggests that it plays an important role in the physiology of skeletal muscle.     

Molecular evolution of voltage-sensitive ion channel genes: on the origins of electrical excitability

We have analyzed nucleic acid and amino acid sequence alignments of a variety of voltage-sensitive ion channels, using several methods for phylogenetic tree reconstruction. Ancient duplications within this family gave rise to three distantly related groups, one consisting of the Na+ and Ca++ channels, another the K+ channels, and a third including the cyclic nucleotide-binding channels. A series of gene duplications produced at least seven mammalian homologues of the Drosophila Shaker K+ channel; clones of only three of these genes are available from all three mammalian species examined (mouse, rat, and human), pointing to specific genes that have yet to be recovered in one or another of these species. The Shaw-related K+ channels and the Na+ channel family have also undergone considerable expansion in mammals, relative to flies. These expansions presumably reflect the needs of the high degree of physiological and neuronal complexity of mammals. Analysis of the separate domains of the four-domain channels (Ca++ and Na+) supports their having evolved by two sequential gene duplications and implies the historical existence of a functional two-domain channel.      

Microbiological quality of frozen cauliflower, corn, and peas obtained at retail markets.

The microbiological quality of blanched frozen cauliflower, cut corn, and peas at the retail level was determined. At 35 degrees C, mean aerobic plate count (APC) values for cauliflower, corn, and peas, respectively, were 30,000, 6,100, and 4,700 per g; at 30 degrees C, the mean APC values were 45,000, 8,500, and 6,800 per g, respectively. Geometric means for coliform, Escherichia coli, and Staphylococcus aureus counts for all three vegetables were less than 10 per g.     

Microbiological quality of some spices and herbs in retail markets.

The microbiological quality of 10 spices or herbs was determined by a national survey at the retail level. Aerobic plate count values for the 10 products ranged from less than 100 to 3.1 X 10(8) per g; mean values of the individual spices or herbs ranged from 1,400 to 820,000 per g. Coliform counts ranged from less than 3 to 1.1 X 10(6) per g; however, mean values were less than 20 per g for all products. Escherichia coli counts ranged from less than 3 to 2,300 per g. Except for celery seed, which had a mean value of 7 per g, all mean values were less than 3 per g. Yeast and mold counts were made for 5 of the 10 products. Mean values were generally low; the highest mean (290 per g) was obtained for cinnamon.     

Microbiological quality of frozen breaded fish and shellfish products.

A survey was made of the microbiological quality of seven frozen, breaded, precooked fish and shellfish products and of frozen, breaded, uncooked shrimp at the retail level. Geometric mean aerobic plate counts per gram (and number of units examined) were as follows: fish sticks, 8,300 (1,539); fish cakes, 5,600 (1,378); crab cakes, 4,900 (1,226); scallops, 1,700 (1,392); clams, 450 (1,384); haddock, 15,000 (1,306); fish in fish and chips dinner, 7,200 (1,485); and uncooked shrimp, 220,000 (1,462). Geometric mean coliform, Escherichia coli, and Staphylococcus aureus counts for all eight products ranged from 1 to 10/g.     

Intensified Tuberculosis Case Finding among Malnourished Children in Nutritional Rehabilitation Centres of Karnataka,India: Missed Opportunities

Background

Severe acute malnutrition (SAM) is the most serious form of malnutrition affecting children under-five and is associated with many infectious diseases including Tuberculosis (TB). In India, nutritional rehabilitation centres (NRCs) have been recently established for the management of SAM including TB. The National TB Programme (NTP) in India has introduced a revised algorithm for diagnosing paediatric TB. We aimed to examine whether NRCs adhered to these guidelines in diagnosing TB among SAM children.

Methods

A cross-sectional study involving review of records of all SAM children identified by health workers during 2012 in six tehsils (sub-districts) with NRCs (population: 1.8 million) of Karnataka, India.

Results

Of 1927 identified SAM children, 1632 (85%) reached NRCs. Of them, 1173 (72%) were evaluated for TB and 19(2%) were diagnosed as TB. Of 1173, diagnostic algorithm was followed in 460 (37%). Among remaining 763 not evaluated as per algorithm, tuberculin skin test alone was conducted in 307 (41%), chest radiography alone in 99 (13%) and no investigations in 337 (45%). The yield of TB was higher among children evaluated as per algorithm (4%) as compared to those who were not (0.3%) (OR: 15.3 [95%CI: 3.5-66.3]). Several operational challenges including non-availability of a full-time paediatrician, non-functioning X-ray machine due to frequent power cuts, use of tuberculin with suboptimal strength and difficulties in adhering to a complex diagnostic algorithm were observed.

Conclusion

This study showed that TB screening in NRCs was sub-optimal in Karnataka. Some children did not reach the NRC, while many of those who did were either not or sub-optimally evaluated for TB. This study pointed to a number of operational issues that need to be addressed if this collaborative strategy is to identify more TB cases amongst malnourished children in India.     

Microbiological quality of frozen shrimp and lobster tail in the retail market.

The microbiological quality of three frozen shrimp products and frozen lobster tail at the retail level was determined. The number of retail units of the four products examined and the geometric means for aerobic plate counts at 30 and 35 degrees C, respectively, were: 1,464 units of cooked, peeled shrimp--13,000 and 7,200 per g; 1,468 units of raw, peeled shrimp--860,000 and 300,000 per g; 1,300 units of raw, in-shell shrimp--800,000 and 300,000 per g; 1,315 units of lobster tail--140,000 and 42,000 per g. Geometric means for coliform, Escherichia coli, and Staphylococcus aureus counts for all products were < 10 per g.     

Inhibition of hepatocyte proteolysis and lactate gluconeogenesis by chloroquine

Chloroquine (50 μm) is rapidly taken up by isolated hepatocytes in a temperature-dependent manner. It inhibits glucose synthesis from lactate, but not from pyruvate or dihydroxyacetone. The inhibition is reversed by lysine or ammonia but not by oleate or carnitine. Ammonia inhibits chloroquine uptake by the hepatocytes but lysine does not. Chloroquine also inhibits urea synthesis, the release of ninhydrin-reacting substances, the accumulation of amino acids, and the lactate-dependent accumulation of glutamate. Ethanol oxidation in the presence of lactate is also inhibited, and this too is reversed by lysine. Chloroquine increases the redox state of the cytosolic compartment, as evidenced by lactate-to-pyruvate ratios, of hepatocytes prepared from both 48-h fasted and meal-fed rats. The above findings are consistent with chloroquine entering the lysosomes of the hepatocytes and inhibiting proteolysis by raising the lysosomal pH. Isolated hepatocytes are deficient in amino acids and, chloroquine inhibition of proteolysis prevents replenishment of the amino acid pools. Thus, chloroquine prevents reconstitution of the malate-aspartate shuttle required for the movement of reducing equivalents into the mitochondrion during lactate gluconeogenesis, ethanol oxidation, and glycolysis. The metabolic competency of freshly isolated hepatocytes, therefore, depends on the replenishment of amino acid pools by lysosomal breakdown of endogenous protein. Furthermore, chloroquine uptake may be an index of lysosomal function with isolated hepatocytes.     

Microbiological quality of five potato products obtained at retail markets.

The microbiological quality of frozen hash brown potatoes, dried hash brown potatoes with onions, frozen french fried potatoes, dried instant mashed potatoes, and potato salad was determined by a national sampling at the retail level. A wide range of results was obtained, with most sampling units of each products having excellent microbiological quality. Geometric mean aerobic plate counts were as follows: dried hash brown potatoes, 270/g; frozen hash brown potatoes with onions, 580/g; frozen french fried potatoes 78/g; dried instant mashed potatoes, 1.1 x 10(3)/g; and potato salad, 3.6 x 10(3)/g. Mean values of coliforms, Escherichia coli, and Staphylococcus aureus were less than 10/g.