Urethral reconstruction after total penectomy for urethral cancer.

A 59-year-old white man developed a ventral ulcer with irregular limits in the middle portion of the penis. The result of the pathologic analysis was compatible with invasive squamous cell urethral carcinoma. A total penectomy was performed. In these cases, the usually recommended urinary diversion is perineal urethrostomy. However, due to the specifications of the case, perineal urethrostomy could not be performed. The literature did not offer any other alternative for patients with this same condition. Therefore, a urethral reconstruction using a groin skin flap had to be performed.

Effects of melatonin on paraquat or ultraviolet light exposure-induced DNA damage.

The effect of paraquat or ultraviolet (UV) light exposure on calf thymus DNA was investigated in vitro. When paraquat (0.1, 0.25, 0.5, or 1.0 mM) was incubated with brain or lung homogenates prepared from mice in the presence of calf thymus DNA at 37 degrees C for 90 min, paraquat inflicted damage to DNA in a concentration-dependent manner. However, DNA damage was completely abolished by co-treatment with melatonin (1.5 mM), a hydroxyl radical (.OH) scavenger. In addition, when paraquat (1.0 or 2.5 mM) was incubated with liver microsomes in the presence of calf thymus DNA and Fe3+ (3.0 microM) at 37 degrees C for 90 min, DNA damage also occurred and was prevented by the co-treatment of melatonin (1.5 mM). DNA was also damaged by UV light exposure or when the Fenton reaction was induced; the Fenton reaction generates .OH; again, the damage was blocked by the co-treatment of melatonin. These results suggest that .OH induced by paraquat or UV light probably account for the DNA damage. In short, DNA damage induced by paraquat and UV radiation were completely prevented by co-treatment with the .OH scavenger, melatonin.

Preventive effect of melatonin against DNA damage induced by cyanide, kainate, glutathione/Fe(3+)/O(2), or H(2)O(2)/Fe(2+).

We have investigated hydroxyl free radical (.OH)-mediated damage to calf thymus DNA produced by potassium cyanide, kainate, H(2)O(2)/Fe(2+), or glutathione/Fe(3+) by in vitro method. When calf thymus DNA was exposed to potassium cyanide (0.5, 0.75, 1.0, 1.5, or 2.0 mM) or kainate (0.25, 0.5, or 1.0 mM) for 90 min at 37 degrees C in homogenate or the 9000g supernatant of mice brain, the quantity of DNA damage was observed to be concentration-dependent. Similarly, glutathione (1.0, 2.0, 4.0, 5.0, or 6.0 mM) inflicted damage on calf thymus DNA in the presence of Fe(3+) (3.0 microM). In addition, hydrogen peroxide (0.15, 0.30, 0.75, 1.50, or 3.0 mM) also caused damage to calf thymus DNA in the presence of Fe(2+) (3.0 microM) in the same manner. Furthermore, it was observed that the DNA damage induced by potassium cyanide (2.0 mM), kainate (0.5 mM), glutathione (4.0 mM)/Fe(3+), and H(2)O(2) (1.5 mM)/Fe(2+) was prevented by the treatment with melatonin (1.0 or 1.5 mM), a potent .OH scavenger. These results suggest that cyanide, kainate, glutathione/Fe(3+), and H(2)O(2)/Fe(2+)-mediated .OH may play a cardinal role for DNA damage induced by these chemicals. Hence the conclusion of the present study is that melatonin protects against DNA damage induced by the .OH produced by these chemicals.

A possible mechanism for the increase in brain tyrosine levels induced by cyanide in mice.

Subcutaneous administration of cyanide significantly increased blood tyrosine levels of mice in a dose dependent manner. Tyrosine aminotransferase activity in liver of mice was significantly decreased in the presence of cyanide (8, 10, 20, 40, 50, 65, 80 and 100 microM), also in a concentration-dependent manner, with a positive correlation between the percentage increase of blood tyrosine levels and the percentage decrease of hepatic tyrosine aminotransferase activity. These results suggest that the increased tyrosine levels induced in blood by cyanide may be related to its inhibition of tyrosine aminotransferase activity in the liver. Cyanide decreased hepatic ATP content and increased blood ammonia levels and brain tyrosine in a dose-dependent manner. As it is known that hyperammonaemia increases the uptake of neutral amino acids such as tyrosine into the brain from blood, the mechanism by which tyrosine levels increase in the brain may be based on increases of both tyrosine and ammonia levels in blood.

Nitroprusside intoxication: protection of alpha-ketoglutarate and thiosulphate.

Protection against the lethal effects of sodium nitroprusside (SNP) was observed in mice after treatment with alpha-ketoglutarate (AKG), either alone or in combination with sodium thiosulphate (STS). The LD50 of SNP was 12.0 (11.0-13.0) mg/kg in mice. Ip injection of AFG (500 mg/kg twice in 20 min) increased the LD50 1.7-fold in mice. STS (1 g/kg, ip) alone increased the LD50 5.5-fold. Furthermore, combined administration of AKG and STS increased the LD50 6.9-fold. SNP elicited increased cyanide levels in blood of mice in a dose-dependent manner. SNP (10 mg/kg, sc) administration gave rise to blood cyanide levels of 73.2 +/- 3.0 microM, 30 min after treatment. Ip injection of AKG significantly decreased blood cyanide levels by 30% in mice 30 min after treatment with 10 mg SNP/kg. A single injection of STS (1 g/kg) or a combination of AKG and STS reduced in blood cyanide levels by 88 or 98%, respectively, in mice after treatment with 10 mg SNP/kg. In addition, the increase in blood cyanide levels induced by injection of 50 mg SNP/kg was markedly inhibited by a combination of AKG and STS or (to a lesser extent) by STS alone. These results suggest that the combined administration of AKG and STS, by preventing the increase in blood cyanide levels induced by SNP, may afford protection against the toxic effects of SNP.

Hepatic ATP content and hyperammonemia induced by CCl4 in rats.

An investigation of the mechanism of development of hyperammonemia observed in CCl4-induced hepatic encephalopathy was performed in rats. CCl4 (1.0 ml/kg 3 times per week for over 10 weeks) caused a severe hyperammonemia and depletion of hepatic ATP contents in only those rats with hepatic encephalopathy. However, CCl4 (1.0 ml/kg 3 times per week for 7 weeks) did not cause hepatic encephalopathy and did not change in blood ammonia levels. Administration of 2,4-dinitrophenol (2,4-DNP) in these CCl4-treated rats caused hepatic encephalopathy within 30 min after injection and then the increase of 140 micrograms/dl in blood ammonia levels and the decrease of 80% in hepatic ATP contents were observed. However, the administration of 2,4-DNP in CCl4-untreated rats did not cause hepatic encephalopathy within 30 min after injection although the increase of 70 micrograms/dl in blood ammonia levels and the decrease of 80% in hepatic ATP contents were observed. Hepatic activities of carbamylphosphate synthetase (CPS) and argininosuccinate synthetase (ASS), important enzymes of the urea cycle, were significantly inhibited by 85% and 60% respectively, in rats treated with CCl4 plus 2,4-DNP. However, in rats treated with 2,4-DNP and without CCl4, the hepatic activities of CPS and ASS were inhibited only 25% and 0%, respectively. These findings suggest that the severe hyperammonemia, which may be produced by the decrease of hepatic ATP content and the inhibition of CPS and ASS, may play an important role in induction of hepatic encephalopathy.

Effects of ethanol and barbiturates on Ca2+-ATPase activity of erythrocyte and brain membranes.

Exposure to ethanol or pentobarbital in vitro stimulated the ATP-dependent efflux of calcium from human red blood cells (RBC) and the Ca2+-ATPase activity of RBC and rat brain synaptic plasma membranes (SPM). These effects were obtained with concentrations of ethanol (50 mM) and pentobarbital (60 microM) associated with intoxication in vivo. The enhancement of SPM Ca2+-ATPase by ethanol was due to an increase in the apparent affinity of the enzyme for calcium with no change in the maximum velocity. SPM Ca2+-ATPase was also stimulated by an unsaturated fatty acid, cis-vaccenic acid methyl ester (cis-VAME). The membrane-disordering effects of ethanol, four barbiturates and cis-VAME were evaluated in SPM using the fluorescent probe molecule 1,6-diphenyl-1,3,5-hexatriene (DPH). All the compounds decreased the fluorescence polarization of DPH, and these decreases were proportional to the increase in Ca2+-ATPase produced by these drugs. These findings suggest that the increase in Ca2+-ATPase and calcium efflux produced by ethanol and pentobarbital results from the membrane-disordering effects of these drugs.

Calcium-dependent 86 Rb efflux and ethanol intoxication: studies of human red blood cells and rodent brain synaptosomes.

Effects of ethanol on calcium-dependent potassium efflux were investigated in red blood cells (RBC) from humans and brain synaptosomes from rats and mice. 86 Rb was used as a tracer for potassium. Synaptosomes and RBC were lysed and resealed with 86 Rb and calcium-EGTA buffers to regulate intracellular levels of ionized calcium. In vitro addition of ethanol (100 mM) stimulated the calcium-dependent 86 Rb efflux of synaptosomes. This stimulation was blocked by apamin, an inhibitor of the calcium-dependent potassium current of nerve cells. In addition, intracerebroventricular injection of apamin inhibited ethanol-induced narcosis in mice, providing behavioral evidence for the importance of calcium-stimulated potassium efflux in alcohol intoxication. In vitro addition of ethanol, propanol or butanol increased calcium-dependent 86 Rb efflux of human RBC at low concentrations of free calcium, but did not change the calcium-independent efflux of 86 Rb. These results suggest that the calcium-dependent 86 Rb efflux of nerve endings may have an important role in the pharmacological and toxicological effects of ethanol.

Effects of amino acids, especially taurine and gamma-aminobutyric acid (GABA), on analgesia and calcium depletion induced by morphine in mice.

The analgesic effect of morphine was antagonized in mice by intracerebroventricular pretreatment with taurine, gamma-aminobutyric acid (GABA) or glycine and was potentiated by ethylene glycol tetra-acetic acid (EGTA) but not altered by L-glutamate or L-aspartate. The potentiation of morphine analgesia by EGTA was reversed by a concentration of taurine that did not alter the tail-flick response. The selective depletion of 45Ca2+ from synaptic vesicles observed with morphine administration was significantly inhibited by taurine injection (1.2 mumol/brain, i.vt.) but was not altered by the same dose of GABA. Inhibition of ATP-dependent 45Ca2+ uptake in synaptosomes by morphine was also completely reversed by taurine (10(-2)M which by itself did not alter 45Ca2+ uptake. These results suggest that antagonism of morphine analgesia by taurine may be caused by blockade of the morphine-induced inhibition of both ATP-dependent synaptosomal 45Ca2+ uptake and changes in synaptic vesicular 45Ca2+ localization, while the antagonism by GABA was not associated with synaptosomal Ca2+.