Associated changes in Ca2+ and Sr2+ activation properties and fiber proteins in cross-reinnervated rabbit soleus

Changes in the Ca²⁺ and Sr²⁺ activation properties of functionally skinned slow-twitch soleus fibers were measured and compared with those of normal fast-twitch extensor digitorum longus (EDL) following cross-reinnervation of soleus with the nerve to EDL. Most of the fibers showed either complete transformation of activation properties (66%) or remained unchanged (34%). The change in sensitivity to divalent cations was correlated with changes in the proteins present in fibers pooled on the basis of their activation properties. The banding patterns of the 35,000- and 37,500-dalton proteins (tropomyosin and troponin T) in cross-reinnervated soleus were correspondingly transformed to those of normal EDL. Slow and fast myosin light chains were present in the pooled cross-reinnervated fibers. Fiber distributions based on activation properties were confirmed by histochemical features. For the first time it has been demonstrated that cross-reinnervation produced changes in the activation properties of soleus fibers and associated changes in the regulatory proteins measured.     

Quantitative trait loci (QTL) for circadian rhythms of locomotor activity in mice

The loomotor activity of male mice (Mus musculus) was monitored by infrared photoelectric beams under three lighting regimens: LD (12 h of light and 12 h of dark), DD (constant dark), and LL (constant broad-spectrum light, 10 lux). Circadian period of locomotor activioty (τ) was compared among 3 inbred strains of mice, C57BL/6J (B6), BALB/c (C), and DBA/2J (D2), and 26 recombinant inbred strains B×D (B6×D2). the τ under both continuous low-intensity light and continuous darkenss varied significantly among strains. Under DD the mean τ was 23.8 h for B6, 23.7 h for D2, and 23.6 h for C. Under LL the mean τ was 25.1 for B6, 23.9 h for D2, and 25.5 h for C. Frequency histograms of the mean τ of 26B×D RI mouse strains (three to seven animals per strain) in either DD or LL and the difference between them, Δτ, had distributions which appeared unimodal, suggesting polygenic inheritances. The narrow-sense heritability determined using 26 strains of B×D RI mice was about 55% for τ and about 38% for both τ in LL and Δτ. An estimated four loci contribute to the variance of τ in constant darkness and five to the variance of τ in constant low-intensity light among the strains studied. Quantitative trait locus (QTL) analysis identified several potential genetic loci associated with τ in constant darkness, τ in constant low-intensity light, and Δτ. The associations of highest probability for each of these traits were theD1Nds4 locus (p<0.001) on mouse chromosome 1, theD5Ncvs52 locus (p<.05) on mouse chromosome 5, and thePmv12 locus (p<.01) at 70 cM on mouse chromosome 5, respectively. A QTL identified for τ was associated (p<.05) with theD2NDS1 marker at 45 cM on chromsome 2 near the Ea 6 marker at 46 cM associated (p<.05) with that reported for the period of wheel running activity in seven C×B RI strains (Schwartz, W. J., and Zimmerman, P.,J. Neurosci. 10:3685 1990).     

A calmodulin-binding peptide relaxes skinned muscle from guinea-pig taenia coli

During smooth muscle activation the calcium calmodulin complex interacts with myosin light chain kinase (MLCK) whereby activating it. A synthetic peptide analogue (RS20) corresponding to the calmodulin recognition sequence of MLCK has been synthesized and previously found to inhibit the calmodulin stimulated light chain kinase activity. Here we studied the effect of this peptide on skinned fibers from guinea pig taenia coli. Maximal contractions induced by 30 M Ca²⁺ at 0.1 M calmodulin could be completely relaxed by the peptide at 1 M. The inhibitory effect was accompanied by partial dephosphorylation only of the regulatory myosin light chain. Relaxation could be reversed by addition of calmodulin which also increased the extent of light chain phosphorylation.The calmodulin concentration required for reversing the inhibition depended on the concentration of the inhibitory peptide suggesting that the peptide competed with MLCK for the calmodulin binding site. As the calcium-calmodulin-peptide mixture constitutes a calmodulin buffer, our results suggest, that the peptide is a calmodulin antagonist unique in terms of its potency and that less than nanomolar concentrations of free calmodulin may be required for inducing smooth muscle contractions.     

Effects of evening bright light exposure on melatonin, body temperature and sleep

SUMMARY  Five male subjects were exposed to a single 2-h period of bright (2500 lux) or dim (<100 lux) light prior to sleep on two consecutive nights. The two conditions were repeated the following week in opposite order. Bright light significantly suppressed salivary melatonin and raised rectal temperature 0.3°C (which remained elevated during the first 1.5 h of sleep), without affecting tympanic temperature. Bright light also increased REM latency, NREM period length, EEG spectral power in low frequency, 0.75-8 Hz and sigma, 12–14 Hz (sleep spindle) bandwidths during the first hour of sleep, and power of all frequency bands (0.5–32 Hz) within the first NREMP. Potentiation of EEG slow wave activity (0.5-4.0 Hz) by bright light persisted through the end of the second NREMP. The enhanced low-frequency power and delayed REM sleep after bright light exposure could represent a circadian phase-shift and/or the effect of an elevated rectal temperature, possibly mediated by the suppression of melatonin.     

Polymorphic metabolizing genes and susceptibility to atherosclerosis among cigarette smokers

Atherosclerosis (AR) is the leading cause of morbidity and mortality in the US and cigarette smoking is a major contributing factor to the disease. Like cigarette smoking in lung cancer, genetic susceptibility may be an important factor in determining who is more likely to develop AR. However, the current emphasis has been on susceptibility based on altered cardiovascular homeostasis. In this investigation, we studied 120 AR patients and 90 matched controls to elucidate the association between polymorphisms in some metabolizing genes (GSTM1, GSTT1, CYP2E1, mEH, PON1, and MPO) and susceptibility to AR. We found that the GSTT1 null allele and the fast allele of mEH(*) (exon 4) are associated with risk for AR. Furthermore, the combined genotypes GSTM1 null/ CYP2E1(*)5B, GSTM1 null/mEH YY, and GSTT1 null/mEH YY are significantly associated with susceptibility to AR (OR = 15.42, 95% CI = 1.33-77.93, P = 0.021; OR = 3.48, 95% CI = 1.63-8.04, P = 0.0008; OR = 3.4; 95% CI = 0.99-17.38, P = 0.05; respectively). We have also conducted cytogenetic analysis to elucidate if induction of chromosome aberrations (CAs) is a biomarker of AR susceptibility. We found that among cigarette smokers (AR patients and smoker controls), individuals having the GSTM1 null allele had a significantly higher frequency of CAs compared to those with the normal allele (P < 0.05). This association was not found among nonsmokers. In addition, individuals who had inherited the CYP2E1(*)5B allele exhibited a significantly higher CA frequency (8.0 +/- 0.82) compared to those with the CYP2E1 wild-type genotype (4.31 +/- 0.35). Since the analysis of genetic susceptibility factors is still in its infancy, our study may stimulate additional investigations to understand the roles of genetic susceptibility and cigarette smoking in AR.     

Neutral Polymeric Surfactants Derived from Dextran: A Study of Their Aqueous Solution Behavior

Summary: Amphiphilic polysaccharides are obtained by hydrophobic modification of a neutral bacterial polysaccharide, dextran. By reacting the polysaccharide with aliphatic epoxides (epoxyoctane and epoxydodecane) in dimethyl sulfoxide, a series of amphiphilic polymers is obtained which covers a large range of structural parameters (length of the polysaccharide, number and nature of hydrocarbon moieties). The solution behavior of dextran derivatives is first characterized by viscometric measurements in dilute and semi‐dilute domains. The effects of molecular parameters on polymer viscosity behavior are evidenced and discussed. Information on the state of aggregation of polymers is obtained by the use of static and dynamic light scattering. The presence of aggregates in the dilute domain is clearly evidenced and their structural characteristics are estimated (size, molecular weight and number of aggregation). The aggregates are shown to account for the viscometric results in the examined concentration range, relating their chemical parameters (hydrodynamic radius and molecular weight) to the macroscopic behavior of the solutions.

     

Swelling,Elasticity and Spatial Inhomogeneity of Poly(N,N‐dimethylacrylamide) Hydrogels Formed at Various Polymer Concentrations

Summary: The equilibrium swelling degree, modulus of elasticity and the spatial inhomogeneity of poly(N,N‐dimethylacrylamide) (PDMAAm) hydrogels were investigated over the entire range of the initial monomer concentration. The degree of dilution of the networks after their preparation was denoted by ν, the volume fraction of crosslinked polymer after the gel preparation. The linear swelling ratio of the gels increased linearly with increasing ν. Depending on the value of ν, three different gel regimes were observed: (1) For ν < 0.3, increasing ν decreases the extent of cyclization during crosslinking so that the effective crosslink density of gels increases with rising ν. (2) For 0.3 < ν < 0.7, increasing ν reduces the accessibility of the pendant vinyl groups during crosslinking due to steric hindrance at high polymer concentrations. As a result, the effective crosslink density of gels decreases with increasing ν. (3) For ν > 0.7, the modulus of elasticity increases sharply with increasing ν due to the increasing extent of chain entanglements in this high concentration regime. Static light scattering measurements on the gels show that the degree of spatial gel inhomogeneity in PDMAAm gels attains a maximum value at ν = 0.06. The appearance of a maximum as well as the ν‐dependence of scattered light intensities from gels was successfully reproduced by the theory proposed by Panyukov and Rabin.

Effective crosslink density ν_e of the hydrogels shown as a function of ν.     

Fibre-type specific expression of fast and slow essential myosin light chain mRNAs in trained human skeletal muscles

The fibre-type specific expression patterns of fast and slow isoforms of essential (alkali) myosin light chains (ELC) was analysed in trained, untrained and pathological human muscles. Biopsies from m. vastus lateralis of moderately trained and untrained persons, as well as highly trained endurance and strength athletes were analysed, by in situ hybridization, for the expression of the `fast' ELC 1f/3f and the `slow' ELC 1sb. We wanted to investigate if changes in the fibre-type specific ELC mRNA pattern could be used as markers for training adaptation, especially, if the mRNA of the slow ELC 1sb isoform would appear in type IIA fibres as a result of endurance training (Baumann et al. 1987). We found the fast/slow ELC expression patterns in the fibre types to be remarkably stable. Physiological stress, even high training loads, did not affect it. No IIA fibres expressing ELC 1sb mRNA were found. They could be detected, however, in pathological muscle samples, where fast/slow ELC patterns not found in normal muscles were frequent. Our data suggest that in healthy muscles, only a subset of the theoretically possible combinations of myosin heavy and light chain isoforms are expressed at the level of their mRNAs.     

Preferential VH/V lambda pairings occur in the available B cell repertoire of adult BALB/c mice.

To gain insights into the composition of the B cell repertoire, we have investigated VH gene family expression associated with individual light chains. For this purpose, we have examined the use of 12 VH gene families in a large collection of hybridomas expressing one of the four lambda light chains [lambda 1 (V1J1), lambda 2 (V2J2 and V x J2) and lambda 3 (V1J3)]. Our results show that the distribution of the VH families is very different from one lambda subtype to another. This suggests that a few substitutions between VL regions are sufficient to generate very different associated repertoires by strong selection mechanisms. Moreover, we assume that the global VH expression pattern is not random but rather composed of many preferential VH/VL associations.     

Effect of the phenol antioxidant katavidan on autooxidation of microsomes exposed to visible light

A study is performed of the effect of the phenol antioxidant katavidan on autooxidation of microsomes from rat liver exposed to visible light. It is shown that katavidan in a concentration of 10⁻³ M inhibits but in concentrations of 10⁻⁵–10⁻⁷ M stimulates autooxidation of microsomes. No stimulation is observed under conditions of dark incubation. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 118, № 10, pp. 393–394, October, 1994 Presented by I. P. Ashmarin, Member of the Russian Academy of Medical Sciences