Cystine storage in cultured myotubes from patients with nephropathic cystinosis.

Sorted muscle cells, cultured from a patient with nephropathic cystinosis, stored 100 times normal amounts of cystine. Subcellular fractionation and density-gradient centrifugation confirmed that the cystine was located in a lysosomal compartment. 2. Myoblasts from cystinotic patients in culture underwent fusion to myotubes in a normal fashion. 3. The free thiol cysteamine effectively depleted cystinotic-muscle cells of cystine. 4. Cultured myoblast and myotubes offered a unique system for investigating the effects of lysosomal storage on differentiated cell functions.     

The Salmonella wien virulence plasmid pZM3 carries Tn1935, a multiresistance transposon containing a composite IS1936-kanamycin resistance element

Tn1935, a 23.5-kb transposon mediating resistance to ampicillin, kanamycin, mercury, spectinomycin, and sulfonamide was isolated from pZM3, an IncFIme virulence plasmid from Salmonella wien. Tn1935 possesses the entire sequence of Tn21 and contains two additional DNA segments of 0.95 and 2.7 kb carrying the ampicillin and kanamycin resistance genes, respectively. The latter is part of a composite element since it is flanked by two IS15-like insertion sequences (IS1936) in direct orientation. IS1936 is about 800 bp long and is closely related to IS15 delta, IS26, IS46, IS140, and IS176. Functional analysis of IS1936-mediated cointegrates shows that both insertion sequences are active and able to form cointegrates at the same frequency. Resolution of the cointegrates requires the presence of the host Rec system. The presence of the composite IS1936-element within Tn1935 supports the hypothesis that multidrug resistance transposons evolved by insertion of antibiotic determinants which are themselves transposable.     

Xenopus spermatozoon: Is there any correlation between motility and oxygen consumption?

The motility status of Xenopus laevis spermatozoa does not affect their respiration rate. Oxygen consumption for 10⁹ spermatozoa is approximately 0.4 μmol/minute. Oxygen consumption is not increased by gramicidin D, an uncoupler, and it is not blocked by KCN or NaN₃. The adenosine triphosphate (ATP) content of spermatozoa that have been activated is definitely less than that in the spermatozoa that remained immotile. Incubation in KCN, NaN₃, and gramicidin decreases the ATP content and impairs motility. The conclusions of the present study are that in Xenopus spermatozoa motility and oxygen consumption are not correlated, and the composition of the respiratory chain of these spermatozoa presents noteworthy peculiarities.     

Rat liver lowMr phosphotyrosine protein phosphatase isoenzymes: Purification and amino acid sequences

Two lowM _r phosphotyrosine protein phosphatases have been isolated from rat liver. The enzymes were previously known as lowM _r acid phosphatases, but several recent studies have demonstrated that this family of enzymes possesses specific phosphotyrosine protein phosphatase activity. We determined the complete amino acid sequences of the two isoenzymes and named them AcP1 and AcP2. Both consist of 157 amino acid residues, are acetylated at the NH₂-terminus, and have His as the COOH-terminus. The molecular weights calculated from the sequences are 18,062 for AcP1 and 17,848 for AcP2. They are homologous except in the 40–73 zone, where about 50% of residues are different. This fact suggests that the two isoenzymes are produced by an alternative splicing mechanism. There is no homology between these two isoenzymes and the receptor-like phosphotyrosine protein phosphatases LAR, CD45, human placenta PTPase 1B, and rat brain PTPase-1. AcP1 and AcP2 are also distinct from rat liver PTPase-1 and PTPase-2, since these last enzymes have higher molecular weights. AcP1 differs from AcP2 with respect to (1) substrate affinity and (2) its sensitivity to activators and inhibitors, thus suggesting a their different physiological function.     

Eye factors and lens-forming transformations of outer cornea in Xenopus laevis larvae

Outer cornea of lensectomized Xenopus laevis tadpoles at state 50 (according to Nieuwkoop, P.D. and Faber, J., ('56) Normal Table of Xenopus laevis, Daudin, North-Holland, Amsterdam, pp. 1-243) was removed 3, 7 and 10 days after lensectomy and implanted between the outer and the inner cornea of larvae of the same species at stage 51-52. In these conditions, the implanted outer cornea remained isolated from the retinal factor of the vitreous chamber, although it received the nutritional factors normally reaching the outer cornea. Results show that lens-forming transformation process of the outer cornea is arrested, and lens-forming structures undergo regression at speed which increases with increasing precocity of the stage of lens-forming transformation undergone by the implanted cornea. These data suggest that the process of lens-forming transformation is not a single-step process, but a sequence of interactions extending over a long period of time requiring the continuous presence of the retinal factor in the vitreous chamber until complete differentiation of the lens is achieved.     

Oxidative destruction of DNA by the adriamycin-iron complex

The 2:1 adriamycin-Fe(III) complex is able to bind to DNA and to catalyze its oxidative destruction. The binding of the drug-metal complex to DNA is indicated by characteristic spectral changes which are different from those seen with adriamycin intercalation and by the propensity of the drug-metal complex to precipitate DNA. Furthermore, intercalated adriamycin appears not to be available for iron binding. The resulting ternary complex is quite stable: it is not disrupted by incubation in the presence of EDTA and can be isolated by using Sephadex G-50 column chromatography. Disruption of the ternary complex requires vigorous conditions (extraction with phenol at 60 degrees C). The adriamycin-iron complex in free solution has the capacity to catalyze the reduction of oxygen by thiols. The DNA-bound drug-metal complex preserves this capacity over a wide range of complex/DNA ratios. As a consequence of this thiol-dependent oxygen reduction, DNA is cleaved. This thiol-dependent DNA cleavage has been shown to require hydrogen peroxide as an intermediate product. These results have led us to propose that the thiol-dependent DNA cleavage reaction has two stages involving (1) reduction of oxygen leading to hydrogen peroxide and then (2) peroxide-dependent DNA cleavage. An unusual property of this reaction is that the cleavage is not random but gives rise to a defined 2300 base pair fragment.     

Analysis of lumped and distributed elements models of cut muscle fibers in vaseline or sucrose gap preparations

A general method of finding the time course and the steady state distribution of potential in Vaseline or sucrose gap preparations is given by making use of the linear cable equation. The general solution has been found analytically in terms of its Laplace transform and then numerically inverted. Two particular experimental situations, namely the single gap and the double gap preparations, have been analyzed. The results have been compared with the solutions of the commonly used lumped elements models. While for the double gap no large errors are introduced by the lumped model, for the single gap there are significant differences. The dependence of the voltage distribution on various electrical and geometrical parameters has been examined. It is suggested that the proposed mathematical treatment might be used by experimenters as a reference to assess the validity of simplified lumped models.     

Neuroendocrine modulation of haloperidol-induced catalepsy

Evidence has been accumulated implicating sex hormones as possible modulators of extrapyramidal motor function. In the present study we have investigated the effects of estrogens, progesterone, testosterone, prolactin and calcitonin on behavioral parameters related to nigro-striatal dopaminergic system, such as haloperidol-induced catalepsy in male rats. It was found that 7-days estradiol benzoate treatment (5 micrograms/rat/day) significantly increases haloperidol-induced catalepsy, suggesting a possible antidopaminergic activity of estrogens. On the other hand, prolactin facilitates nigro-striatal dopaminergic transmission. Interestingly, 7 day treatment with medroxy-acetate progesterone (MAP, 5 mg/Kg, i.p.) brings about a trend to a decrease in haloperidol-induced catalepsy, while no significantly effect was observed following acute MAP administration at the same dose. So, it is tempting to speculate that chronic progestinic treatment may result in an increase in dopaminergic tonus. Testosterone, acutely administered (5mg/kg.s.c.) induces changes similar to those observed following progesterone administration. Finally, also calcitonin is able to influence haloperidol-induced catalepsy by markedly increasing it.