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The intracellular transport and distribution of cysteamine phosphate derivatives.

Radioautography and extractive techniques were used to analyze the transport of cysteamine phosphate and its derivatives in salamander oocytes. The quantitative relations among the processes involved - membrane permeation, enzymatic dephosphorylation, binding through mixed disulfide formation, and cytoplasmic diffusion - were elucidated. Within the detection limits, all of the intracellular material is present as dephosphorylated derivatives. Cytoplasmic diffusion is effectively slowed by binding (the "chromatographic" effect) and makes an appreciable contribution to cellular flux rates. As a consequence, one can observe by radioautography a cortical diffusion ring which spreads inward as a function of influx time, while also increasing in peak density because of the finite membrane permeability. Good agreement was found between the transport parameters determined by radioautography and those from influx data for the whole oocyte. The ratio of nuclear to cytoplasmic concentrations of the cysteamine phosphate derivatives at equilibrium is about 0.4. The nuclear membrane is, however, a negligible barrier to transport, and the asymmetry appears to arise primarily from the quantity and sulfhydryl content of the binding proteins in the two compartments.

Animals↗

Effect of cysteamine hydrochloride on secretion of growth hormone in male swine.

The objective of this study was to determine the effect of cysteamine hydrochloride (CSH) on growth hormone (GH) secretion in male swine. Twelve Poland China x Yorkshire boars, weighing 103.4 +/- 3.0 kg and fitted with indwelling jugular vein catheters, were individually penned in an environmentally controlled room. Boars received i.v. injections of either 0, 25, 50, or 75 mg CSH/kg body weight (BW) at h 0 (n = 3/treatment). Blood samples were collected every 15 min from h 0 to h 4. Serum concentrations of GH were determined by radioimmunoassay. There was an effect of treatment (P < .05) on mean GH concentrations. Mean GH concentrations (ng/ml) were 1.97 +/- .46, 2.24 +/- .59, .91 +/- .06, and .62 +/- .08 for boars receiving 0, 25, 50, and 75 mg CSH/kg BW, respectively. The dose of CSH-mean GH response had a linear (P < .01) component. Cysteamine hydrochloride at the 75 mg/kg BW dose decreased mean GH concentrations (P < .05) compared to the 0 and 25 mg/kg BW groups. The frequency and amplitude of GH pulses were similar (P > .1) among treatments. Overall, GH pulse amplitude was 2.35 +/- .58 ng/ml and GH pulse frequency was .75 +/- .07 pulses/h. Results from this experiment indicate that CSH suppresses circulating GH concentrations in a dose dependent fashion in boars.

Animals↗

(S)-(2-(2'-Pyridyl)ethyl)cysteamine and (S)-(2-(2'-pyridyl)ethyl)-d,l-homocysteine as ligands for the "fac-[M(CO)(3)](+)" (M = Re, (99m)Tc) core.

The reaction of fac-[NEt(4)](2)[Re(CO)(3)Br(3)] with (S)-(2-(2'-pyridyl)ethyl)cysteamine, L(1), in methanol leads to the formation of the cationic fac-[Re(CO)(3)(NSN)][Br] complex, 1, with coordination of the nitrogen of the pyridine, the sulfur of the thioether, and the nitrogen of the primary amine. When fac-[NEt(4)](2)[Re(CO)(3)Br(3)] reacts with the homocysteine derivative (S)-(2-(2'-pyridyl)ethyl)-d,l-homocysteine, L(2), the neutral fac-Re(CO)(3)(NSO) complex, 2, is produced with coordination of the nitrogen of the primary amine, the sulfur of the thioether, and the oxygen of the carboxylate group, while the pyridine ring remains uncoordinated. The analogous technetium-99m complexes, 1' and 2', were also prepared quantitatively by the reaction of L(1) and L(2) with the fac-[(99m)Tc(CO)(3)(H(2)O)(3)](+) precursor at 70 degrees C in water. Given that both (S)-(2-(2'-pyridyl)ethyl)cysteamine and homocysteine can be easily N- or S-derivatized by a bioactive molecule of interest, both the NSN or NSO ligand systems could be used to develop target-specific radiopharmaceuticals for diagnosis and therapy.

Crystallography, X-Ray↗

A study of cysteamine ionization in solution by Raman spectroscopy and theoretical modeling.

Different cysteamine (H2N-CH2-CH2-SH) ionization forms have been studied by polarized Raman spectroscopy in solutions prepared with H2O and D2O and by DFT calculations at the B3LYP/6-31++G(d,p) level. To account for solvation effects, we employed the integral equation formalism polarizable continuum model (IEFPCM) option and explicit water molecules. Calculated relative energies and Raman spectra revealed that gauche rotamers around the C-C bond are the most stable conformers in solution. The experimental pKa values and Raman spectra of various ionization forms were best predicted by using a model with three explicit water molecules and the IEFPCM option. In general, the use of IEFPCM tends to lower the calculated frequencies for a few bands, but in some cases (S-H stretching mode) this effect is expressed very strongly. Potential energy distribution (PED) analysis of gauche conformers of various cysteamine ionization forms provided the possibility of discriminating spectroscopically methylene groups adjacent to sulfur, (CH2)S, and nitrogen, (CH2)N, sites. In general, stretching and scissoring modes as well as wagging and twisting vibrations of the (CH2)N group were found to be at higher frequencies. The influence of ionization of SH and NH2 groups on the vibrational spectrum is discussed, and Raman markers for further amine group ionization studies are suggested.

Cysteamine↗

A randomised placebo-controlled trial of topical cysteamine therapy in patients with nephropathic cystinosis.

Five patients with nephropathic cystinosis were evaluated to assess the ability of topical cysteamine to clear corneal cystine crystals. All patients were randomised to receive topical cysteamine 0.2% six times a day in one eye with normal saline in the other eye as a control. All five patients showed some improvement in visual symptoms (photophobia, blepharospasm and visual acuity) together with an improvement in corneal crystal density. Three of these also had an improvement in Snellen visual acuity and contrast sensitivity.

Adolescent↗

Effect of cysteamine administration on growth and efficiency of food utilisation in chicks.

1. Cysteamine hydrochloride, which is a potent inhibitor of somatostatin secretion, was fed to male broiler chicks at dietary inclusion rates of 0, 25, 125, 250, 600, 1200, 1800 and 2400 mg/kg during three separate experiments. 2. Weight gain, food intake and efficiency of food utilisation were measured weekly over a two (experiment 1 and 2) or three-week period (experiment 3) during which chicks were 3 to 10 d old (week 1) to 17 to 24 d old (week 3). 3. Cysteamine hydrochloride included in the food at 1200 and 1800 mg/kg resulted in a significant increase in efficiency of food utilisation, attributable to a significant decrease in food intake. 4. There was no difference in weight gains between chicks in the control group and those receiving 1200 mg/kg. However a significant decrease in weight gain was observed in chicks receiving 1800 mg/kg.

Animal Nutritional Physiological Phenomena↗

Effect of salt solutions on the radiosensitivity of mammalian cells. IV. Treatment with NaCl solutions containing ouabain, NEM, PNAP, cysteamine or DMSO.

The effects of a wide concentration range of NaCl solutions containing either ouabain, ethanol, para-nitroacetophenone (PNAP), N-ethylmaleimide (NEM), cysteamine or dimethyl sulphoxide (DMSO) on cellular radiosensitivity have been examined. Ouabain and NEM treatment increased the radiosensitivity of V79 Chinese hamster cells, but the action of these chemicals did not depend on the concentration of NaCl. PNAP increased cellular radiosensitivity with increasing NaCl concentration reaching a maximum effect at 0.6 to 0.7 M NaCl. The radioprotective properties of cysteamine, DMSO and ethanol were all strongly dependent on the NaCl concentration in a complex but qualitatively similar manner. DMSO (2.0 M) increased radiation survival of cells after a 1380 rad dose by a factor of about 10(4) when present in 0.075 M NaCl and by a factor of 8.7 when present in 1.2 M NaCl.

Acetophenones↗

Suppression of induced beta-galactosidase synthesis by cysteamine and its reversion by gamma-irradiation in the presence of ascorbate.

The induced synthesis of beta-galactosidase in E. coli was found to be inhibited by cysteamine. This inhibitory effect of the SH compound was antagonized by the addition of ascorbate followed by gamma-irradiation with relatively low doses. The cAMP level which, it has been suggested, plays a role in the radioprotective action of cysteamine, is stabilized by ascorbate against changes induced by irradiation.

Ascorbic Acid↗

A radioprotector: cysteamine, inhibits oxygen transport in lipidic membranes.

Spin labeling techniques make possible the observation of oxygen diffusion or concentrations in phospholipid membranes. In such a system, cysteamine, depending upon the molecular cysteamine/DPPC ratio and the pH conditions, inhibits oxygen transport, and this result provides an original explanation for cellular hypoxia after cysteamine administration.

Biological Transport↗

Range of toxicity of topical cysteamine in rabbit eyes.

Cysteamine eye drops in concentrations from 0.1% to 10% were tested in albino rabbits in both short and long-term experiments. Toxicity, consisting of an inflammatory response, was proportional to the cysteamine concentration and absent at concentrations at or below 0.5%.

Animals↗

The penetration of topical cysteamine into the human eye.

The penetration of cysteamine into human aqueous was measured to assess its suitability as a topical agent for the prevention of radiation-induced cataract. Cysteamine drops (0.5% and 1.0%) were administered preoperatively to cataract patients every half hour, for two, four, six or eight hours. Aqueous humor was collected at the start of surgery and assayed for total free thiol. There was no significant difference in the aqueous thiol level of any of the treatment groups compared with the controls. No side effects were observed.

Administration, Topical↗

Effect of dimethylsulphoxide on cysteamine-induced duodenal ulceration in rats.

1. Rats were pretreated with dimethylsulphoxide (DMSO) at concentrations of 2, 4 and 6% v/v in the drinking water for 7 days, then injected with a single subcutaneous dose of cysteamine (28 mg/100 g bodyweight) to induce duodenal ulceration. 2. DMSO reduced, in a dose-dependent manner, the incidence and extent of the ulceration. 3. Mucus duodenal content was not significantly affected by either DMSO or cysteamine treatments.

Animals↗

Cysteamine: a potent and specific depletor of pituitary prolactin.

Cysteamine rapidly reduces the concentration of prolactin in pituitary tissue in vivo and in vitro. The effect is dose-dependent, reversible, and cannot be accounted for by prolactin release. Cysteamine does not appear to exert its effect through dopamine receptors and does not alter lactotrope morphology, as determined by electron microscopy.

Animals↗

Influence of cysteamine on differential staining of BUdR-substituted human chromosomes.

In 5-bromodeoxyuridine (BUdR)-substituted human chromsomes stained with 4'-6-diamidino-2-phenylindole (DAPI) differential staining is suppressed totally by the H+-donor cysteamine (concentration 0.08 M). We propose that differential staining appears because the double BUdR-substituted chromatid will be disintegrated via a photosensitive dye-visible light system. It is suggested that cysteamine prevents the production of strand breaks in DNA and, consequently, differential staining in BUdR-substituted chromosomes. Furthermore it is shown that differential staining with DAPI causes irreversible changes in the double BUdR-substituted chromatid. This finding can be explained with the above mentioned mechanism.

Bromodeoxyuridine↗

Reactivation by cysteamine of vascular alpha adrenoceptors blocked by the tetramine disulfide benextramine and interaction of benextramine with phenoxybenzamine.

The tetramine disulfide benextramine irreversibly blocked the noradrenaline-induced contraction of the isolated rabbit aorta in a manner similar to that observed with phenoxybenzamine. Cysteamine abolished the benextramine blockade but not the phenoxybenzamine blockade. Incubation of aortic tissue with benextramine prior to phenoxybenzamine exposure and followed by cysteamine treatment led to a recovery of the noradrenaline response that depended on the dose of benextramine. Thus benextramine protected the alpha adrenoceptors against phenoxybenzamine blockade.

Animals↗

Adverse reactions to oral cysteamine use in nephropathic cystinosis.

Cysteamine (2-aminoethanethiol) has been given orally to 19 patients with nephropathic cystinosis for periods of 8-24 months in doses ranging from 50 to 70 mg base/kg/day. Adverse reactions were noted in 3 patients early in the study when a rapidly increasing dosage schedule was followed. The reactions included hyperthermia, lethargy and rash. These reactions were not seen when patients were started on a very low dosage which was increased gradually at 3-week intervals to a level which depleted leukocytes of about 90% of their free cystine. All three reactions resolved within 24 h or cessation of therapy and in these cases successful readministration of drug was achieved. Chronic cysteamine administration to pediatric patients with cystinosis is feasible. The efficacy of this therapy is still being evaluated.

Child↗

Effects of cysteamine and of irradiation on microsomal membrane-bound enzymes.

Changes in lipid peroxidation, drug-metabolizing enzymes and kinetic constants of ethylmorphine demethylation and of ethylmorphine interaction with cytochrome P-450 were examined after gamma-irradiation. The radiation-induced lipid peroxidation was increased with increasing radiation dose, while cytochrome P-450 content and ethylmorphine demethylase activity decreased. Of the enzyme kinetics parameters, Vmax was modified. Cysteamine protected against the effects of irradiation. This compound interacts with cytochrome P-450 and inhibits the drug-metabolizing enzyme system. Mechanisms of radiation damage, protective and toxic properties of cysteamine are discussed.

Animals↗

Depletion of bovine pituitary prolactin by cysteamine involves a thiol:disulfide mechanism.

Cysteamine [2-aminoethanethiol (CySH)] reduces measurable PRL concentrations in vivo and in vitro. Since secretion is also inhibited, CySH may block conversion from a poorly assayable hormone storage form(s) to readily assayable, releasable PRL. This would represent a previously unrecognized mechanism for secretory regulation. We undertook the present study to identify the sites involved in the loss of measurable PRL (depletion) induced by cysteamine. The disulfide cystamine was ineffective on secretory granules unless combined with reduced glutathione, indicating the generation of the active CySH-thiol form. Pretreatment of granules with thiol-blocking agents resulted in dose-dependent enhancement of CySH inhibition, achieving nearly complete inhibition with 5 mM iodoacetamide. In contrast, pretreatment with reduced glutathione or dithiothreitol, respectively, impaired or abolished the CySH effect. Similar results were obtained when tissues were exposed to thiols and/or alkaline conditions after CySH treatment; for example, exposure to 50 mM glutathione reduced the lowering of measured pituitary homogenate PRL from 76% to 37%. These data suggest that the mechanism by which CySH causes PRL depletion is mediated by granule disulfides and the -SH of CySH. The regulation of thiol:disulfide equilibria appears to be an important determinant of the detectability of PRL storage forms and of their secretion.

Animals↗