A practical and reliable method for determination of urinary 3-methylhistidine

A practical and reliable semiautomated method for analysis of urinary 3-methylhistidine (3-MH) was designed combining the isolation of 3-MH by ion-exchange chromatography with the color reaction given by ninhydrin-orthopthalaldehyde (ninhydrin-OPT) reagent after alkalinization. 2 ml of urine were passed through disposable columns packed with an ion-exchange resin (Dowex 50-X8, 200–400 mesh) and the acidic and neutral amino acids were eluted with 10 ml of 0.2 M pyridine solution. Then, the 3-MH was quantitatively eluted and separated from histidine with a volume of 9 ml of a 1.5 M pyridine solution. Standard Autoanalyzer equipment was used for the automation of spectrophotometry. The method permits the analysis of 40 samples in duplicate per day. The 3-MH color reaction was linear for concentrations from 0.015 to 0.24 μ mol/ml. The mean recoveries of 3-MH from standards and urine were 98.6 ± 1.3 and 99.0 ± 1.3%, respectively. Duplicate determinations of urine samples showed a variation coefficient of 1.88%. An excellent agreement was obtained between urine samples analyzed by the present method and by an amino acid analyzer. The need for the elimanation of the interfering amino acids was clearly demonstrated.     

High-Purity Preparation of HSV-2 Vaccine Candidate ACAM529 Is Immunogenic and Efficacious In Vivo

Genital herpes is a sexually transmitted infection (STI) caused by herpes simplex virus 2 (HSV-2) and to a lesser extent herpes simplex virus 1 (HSV-1). Infection by HSV-2 is life-long and is associated with significant cost to healthcare systems and social stigma despite the highly prevalent nature of the disease. For instance, the proportion of HSV-2 seropositive to seronegative adults is approximately 1 in 5 in the US and greater than 4 in 5 in some areas of sub-Saharan Africa. The replication-defective vaccine strain virus dl5-29 was re-derived using cells appropriate for GMP manufacturing and renamed ACAM529. Immunization with dl5-29 was previously reported to be protective both in mice and in guinea pigs, however these studies were performed with vaccine that was purified using methods that cannot be scaled for manufacturing of clinical material. Here we describe methods which serve as a major step towards preparation of ACAM529 which may be suitable for testing in humans. ACAM529 can be harvested from infected cell culture of the trans-complementing cell line AV529 clone 19 (AV529-19) without mechanical cell disruption. ACAM529 may then be purified with respect to host cell DNA and proteins by a novel purification scheme, which includes a combination of endonuclease treatment, depth filtration, anion-exchange chromatography and ultrafiltration/diafiltration (UF/DF). The resultant virus retains infectivity and is ∼ 200-fold more pure with respect to host cell DNA and proteins than is ACAM529 purified by ultracentrifugation. Additionally, we describe a side-by-side comparison of chromatography-purified ACAM529 with sucrose cushion-purified ACAM529, which shows that both preparations are equally immunogenic and protective when tested in vivo.     

Glucagon-like immunoreactivity in hypothalamic neurons of the rat

Summary Antisera specific for three different regions of pancreatic proglucagon were used to examine the distribution of such immunoreactivity in rat hypothalamus. Neurons in the supraoptic and paraventricular nuclei were immunoreactive with an antiserum against glucagon, but not with antisera directed towards the aminoterminal region of proglucagon (glicentin) or the glucagon-like peptide I sequence in the carboxyl-terminal region of proglucagon. These findings confirm a previous report of glucagon-like immunoreactivity in the supraoptic and paraventricular nuclei, but indicate that, while this material is immunochemically related to glucagon, it is not derived from a proglucagon-like precursor.     

Mini-review: toxicity of mercury as a consequence of enzyme alteration

Mercury, in both its elemental and bonded states, is noted for its negative effects on biological organisms. Recent anthropogenic and environmental disasters have spurred numerous comparative studies. These studies attempted to detail the biochemical implications of mercury ingestion, in low, persistent concentrations as well as elevated acute dosages. The studies propose models for mercuric action on healthy cells; which is centered on the element’s disruption of key enzymatic processes at deposition sites. Mercury’s high affinity for the sulfhydryl moieties of enzyme catalytic sites is a common motif for enzyme inactivation. These permanent covalent modifications inactivate the enzyme, thereby inducing devastating effects on an organism’s metabolic functions. Mercury has been shown to be highly nonspecific in its binding to sulfhydryl moieties, and highly varied in terms of how it is encountered by living organisms. This review focuses on mercury’s effects on a wide swath of enzymes, with emphasis on how these alterations deleteriously affect several metabolic pathways.     

Physical and chemical characters,phytoplankton and primary production of Ezequiel Ramos Mexía Reservoir (Argentina)

We describe the distribution in space and time of a series of physical and chemical variables, phyto-plankton, and primary production in Ezequiel Ramos Mexía Reservoir (Argentina). Its waters are soft, poor in nutrients and with a low transparency that greatly depresses primary production. Phytoplankton data indicate the presence of 79 taxa with Bacillariophyceae, Cyanophyta and Chlorophyta alternatively dominant. Chlorophyll a was always low and never exceeded 3 mg m⁻³. Based on these results, the trophic status of this ecosystem is discussed.     

Genetics of life span in mice

Thymic involution that occurs earlier in some individuals than others may be the result of complex interactions between genetic factors and the environment. Such interactions may produce defects of thymus-dependent immune regulation associated with susceptibility to developing autoimmune diseases, malignancy, and an increased number of infections associated with aging.The major histocompatibility complex may be important in determining profiles of cause of death and length of life in mice. Genetic influences on life span involve interactions between loci and allelic interactions during life which may change following viral infections or exposure to other environmental factors. We have used different experimental protocols to study the influence of H-2 on life span and found that interactions between genetic regions, are inconsistent, particularly when comparing mice infected or not infected with Sendai virus.Genes important for life span need to be studied against many genetic backgrounds and under differing environmental conditions because of the complexity of the genetics of life span. Several genetic models were used to demonstrate that the MHC is a marker of life span in backcross and intercross male mice of the H-2^d and H-2^b genotypes in B10 congenic mice. Females lived longer than males in backcross and intercross mice, while males lived longer than females in B10 congenics. H-2^d was at a disadvantage for life span in backcross mice of the dilute brown and brown males exposed to Sendai infection, but intercross mice not exposed to Sendai virus of the same genotype were not at a disadvantage. H-2^d mice were not disadvantaged when compared to H-2^b in B10 congenics that had not been exposed to Sendai virus infection but the reverse was true when they were exposed. Overall, all our studies suggest that genetic influences in life span may involve interactions between loci and many allelic interactions in growing animals or humans. These genetic influences on life span may vary after they are exposed to infections or other environmental conditions. This paper emphasizes the need to use several genetic models, especially animals that have been monitored for infections, to study the genetics of life span.     

Effects of specific versus cross-training on running performance

The cross-training (XT) hypothesis suggests that despite the principle of specificity of training, athletes may improve performance in one mode of exercise by training using another mode. To test this hypothesis we studied 30 well-trained individuals (10 men, 20 women) in a randomized longitudinal trail. Subjects were evaluated before and after 8 weeks of enhanced training (+10%/week), accomplished by adding either running (R) or swimming (XT) to baseline running, versus continued baseline running (C). Both R ( – 26.4s) and XT (– 13.2s) improved time trial (3.2 km) performance, whereas C did not (– 5.4s). There were no significant changes during treadmill running in maximum oxygen uptake (O_2peak; – 0.2, – 6.0, and + 2.7%), steady state submaximal O₂ at 2.68 m · s^–1 ( – 1.2, – 3.3 and + 0.2 ml · kg^–1 · min^–1), velocity at O_2peak (+0.05, +0.25 and +0.09 m · s^–1) or accumulated O₂ deficit (+ 11.2, – 6.1 and + 9.4%) in the R, XT or C groups, respectively. There was a significant increase in velocity associated with a blood lactate concentration of 4 mmol · l^–1 in R but not in XT or C ( + 0.32, + 0.07 and + 0.08 m · s^–1). There were significant changes in arm crank O_2peak ( + 5%) and arm crank O₂ at 4 mmol · l^–1 ( + 6.4%) in XT. There was no significant changes in arm crank O_2peak ( + 1.3 and – 7.7%) or arm crank O₂ at 4 mmol · l^–1 ( + 0.8 and + 0.4%) in R or C, respectively. The data suggest that muscularly non-similar XT may contribute to improved running performance but not to the same degree as increased specific tranining.