Evidence for two transferrin loci in the Salmo trutta genome.

To determine the organization of transferrin (TF) locus in the Salmo trutta genome, partial DNA and cDNA sequencing, fluorescent in situ hybridization (FISH) and Salmo salar BAC analysis were performed. TF expression levels and copy number prediction were assessed using real-time PCR. In addition to two previously reported DNA TF variant sequences of S. trutta and Salmo marmoratus (TF1), two novel variant sequences (TF2) were revealed in both species. Variant-specific sequence tags, characterizing two variants for each TF type (TF1 and TF2), were identified in genomic clones from each of the F1 hybrids between S. trutta and S. marmoratus. These clearly documented double heterozygote status at the TF loci. The real-time PCR data showed that each of the two TF types (TF1 and TF2) existed in one copy only and that the transcription of TF2 was considerably lower compared with TF1. Using FISH, hybridization signals were observed on two medium-sized acrocentric chromosomes of S. trutta karyotype. A TF type-specific PCR followed by a restriction analysis revealed the presence of two TF loci in the majority of analysed BAC clones. It was concluded that the TF gene is duplicated in the genome of S. trutta, and that the two TF loci are located adjacent to one another on the same chromosome. The differing transcription levels of TF1 and TF2 appear to depend on the corresponding promoter activity, which at least for TF2 seems to vary between different Salmo congeners.

In vitro comparison of antifungal effects of a coal tar gel and a ketoconazole gel on Malassezia furfur.

Malassezia furfur seems to be a major pathogenetic factor in seborrhoeic dermatitis, a frequent human skin disease. To estimate the antifungal properties of a coal tar gel (5 mg ml-1 coal tar) which is used in the treatment of seborrhoeic dermatitis of the scalp, we compared its effects on the in vitro growth of M. furfur with those of a ketoconazole gel (20 mg ml-1 ketoconazole). None of the gels was fungicidal within incubation times up to 20 min. During a single application, both gels remain on the skin for only 5 min. Fungicidal effects are consequently unlikely to play a substantial therapeutic role. Fungistatic effects were observed with both gels. In cultures inoculated with 1 x 10(3) cells ml-1, a 1:49 152 dilution of the ketoconazole gel and a 1:768 dilution of the coal tar gel still showed inhibitory effects. At inoculum densities of 1 x 10(5) ml-1, both gels were fungistatic only in dilutions of a maximum of 1:40. Our results suggest that under clinical treatment conditions the fungistatic activities of both preparations should be comparable.

[Inhibition of testosterone metabolism by 17-alpha-estradiol in rat liver slices]. / Hemmung des Testosteron-Stoffwechsels durch 17 alpha-Estradiol in Rattenleberschnitten.

Inhibition of the Testosterone Metabolism in Rat Liver Slices by 17 alpha-Estradiol. The influence of 17 alpha-estradiol (CAS 57-91-0), a hormonally almost inactive isomer of physiological 17 beta-estradiol, on the metabolism of [14C]-labeled testosterone in rat liver slices was investigated. The analysis of extracts from incubates (3.0 ml medium, 100 mg liver slices, 416 nmol [14C]-testosterone, 0.1-30 micrograms 17 alpha-estradiol, 37 degrees C, 30 min) by thin layer chromatography showed, that 30 micrograms of 17 alpha-estradiol inhibited the testosterone turnover in liver slices of female animals. The failure of a significant inhibitory effect in liver slices of male animals is attributed to the known, much smaller total turnover of testosterone in male liver cells. The amount of unchanged 4-en-3-oxo-steroid (testosterone and 4-androstene-3,17-dione) was increased by a factor of 2.65 and 2.25, respectively. With high probability, the inhibition was the result of a decreased hydrogenation of testosterone to dihydrotestosterone (DHT, 17 beta-hydroxy-5 alpha-androstan-3-one), catalyzed by 5 alpha-reductase, since the production rates of DHT and the DHT-transformation metabolites (5 alpha-androstane-3 alpha,17 beta-diol and 5 alpha-androstane-3,17-dione) were significantly lowered (factors: 0.16, 0.61, 0.61, respectively). In further experiments 17 beta-estradiol and 17 alpha-ethinylestradiol could be shown to inhibit the testosterone turnover in liver slices of female rats, too, but to a lower extent that 17 alpha-estradiol (relative inhibitory effects: 17 alpha-estradiol:17 beta-estradiol:17 alpha-ethinylestradiol = 100 : 73 : 58).(ABSTRACT TRUNCATED AT 250 WORDS)

Toxicity of 8-methoxypsoralen in cynomolgous monkeys (Macaca fascicularis).

Male and female cynomolgous monkeys were administered 0, 2, 6 or 18 mg/kg 8-methoxypsoralen (8-MOP) 3 times a week orally for 26 consecutive weeks. Dose-dependent emesis was the most sensitive indicator of 8-MOP toxicity. The lowest dose to elicit emesis was 3 x 6 mg/kg/week of 8-MOP. Among the histological findings proliferation of Kupffer cells was the only recurring observation. However, these finding as well as some hematological and serum electrolyte changes lacked a dose-response relationship. In the highest dosage group one female monkey was found in moribund condition on the 39th day of the study and was killed. Histopathological evidence indicated beginning shock as the cause of the rapidly deteriorating health of the monkey. Similar to effects in man and rats, 8-MOP displayed nonlinear pharmacokinetics in the cynomolgous monkey, saturation occurring between 3 x 2 and 3 x 6 mg/kg/week. Increased clearance of 8-MOP in the lowest dosage group after 26 test weeks was attributed to a combination of enzyme induction and saturable first pass effect. Since the plasma profile of 8-MOP at the lowest dose (3 x 2 mg/kg/week) in cynomolgous monkeys closely resembles that in humans after therapeutic doses (0.4-0.6 mg/kg) and because of other similarities (vomiting as earliest sign of toxicity, saturable first pass effect), it is reasonable to assume that chronic toxicity of 8-MOP as defined in this study is quite predictive for man.

Corticosterone modulates acute toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in male Sprague-Dawley rats.

Bilateral adrenalectomy or adrenal demedullation was performed on male Sprague-Dawley rats by established surgical techniques. Subsequently, the dose-response (mortality and mean time to death) to TCDD was determined in adrenalectomized (10, 20, 40 micrograms/kg TCDD ip in 95:5 corn oil:acetone) or demedullated (15, 30, 60 micrograms/kg TCDD) rats. Adrenalectomy drastically increased mortality and greatly shortened mean time to death after dosing with TCDD. More importantly, adrenalectomized TCDD-treated rats died of hypoglycemic shock without losing much body weight. Conversely, adrenal demedullation had no effect on mortality or mean time to death caused by TCDD when compared to nondemedullated TCDD-treated controls. Thus, it was concluded that the factor(s) modulating the acute toxicity of TCDD resides in the adrenal cortex and not in the medulla. Administration of corticosterone (25 micrograms/ml in drinking water) to adrenalectomized rats returned the toxicity of TCDD to levels seen in nonadrenalectomized rats suggesting that this hormone is another key factor (in addition to the thyroid hormones) in the modulation of the acute toxicity of TCDD. Corticosterone supplementation (25, 50, or 100 micrograms/ml) to nonadrenalectomized rats, or to thyroidectomized-adrenalectomized rats (25 micrograms/ml), resulted in no additional beneficial effect indicating that a factor(s) other than thyroid hormones and corticosterone is also involved in the acute toxicity of TCDD.

[Topical treatment of glucocorticoid-sensitive dermatoses with diflorasone diacetate]. / Ausserliche Behandlung von Glukokortikoid-empfindlichen Dermatosen mit Diflorason-diacetat.

Treatment with diflorasone diacetate ointment and cream (0.05%) was strikingly successful in a multicenter clinical trial involving 4,651 patients in total suffering from various corticosteroid-sensitive dermatoses. According to the clinical evaluation, 93% of the patients were cured or much improved. Approximately 7% of the patients questioned rated the local tolerance of the two forms of administration as good to very good. Only 4% of the patients rated the tolerance of the ointment or cream as moderate or bad. None of the patients showed any systemic side-effects. The molecular structure of diflorasone-17,21-diacetate and the specific pharmaceutical properties of the ointment and cream evidently guarantee a good bioavailability of the glucocorticosteroid in the skin. Clinical and experimental results confirm that diflorasone diacetate belongs to the group of highly active corticosteroids.

Effect of partial jejunectomy and colectomy on the disposition of hexachlorobenzene in rats treated or not treated with hexadecane.

The disposition of hexachlorobenzene (HCB) was studied in partially jejunectomized (middle section) or colectomized (excision of cecum and proximal colon) rats after iv or ip dosage (1.5 to 2.0 mg/kg). Excision of about 50% of the jejunum had no effect on body weight, feed intake, volume of urine, weight of feces, or urinary and fecal excretion of HCB as demonstrated by a comparison of sham-operated and jejunectomized animals. Similarly colectomy did not affect body weight, feed intake, volume of urine, or urinary excretion of HCB. However, the wet weight of feces was significantly higher and the amount of HCB in feces significantly lower in colectomized than in sham-operated rats. Hexadecane increased fecal excretion of HCB about two- to threefold without affecting its urinary excretion. The effect of jejunectomy and colectomy was similar in hexadecane-treated animals to that seen in untreated rats. Concentration of HCB in adipose tissue was significantly higher in colectomized rats than in sham-operated controls. Data represent in vivo evidence that the major site of nonbiliary, intestinal excretion of HCB is the large intestine.

Thyroid hormones modulate the toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD).

These experiments examine the role of thyroxine (T4) and triiodothyronine (T3) on the toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The first experiment is continuation of a study reported previously (Rozman et al., 1984). In this experiment, 60 male Sprague-Dawley rats were divided into 6 equal groups. Four groups of rats were thyroidectomized by 3 mCi Na131 l/kg rat. Five weeks later 2 of the thyroidectomized and 1 of the nonthyroidectomized groups of rats received ip 100 micrograms TCDD/kg body weight in corn oil/acetone, whereas 3 corresponding groups of rats served as vehicle controls. Two days after dosing and every 7 d thereafter, 1 thyroidectomized control group and 1 thyroidectomized TCDD-dosed group were given ip 105 micrograms T4/kg body weight. Mortality and body weight were monitored. The course of TCDD toxicity was similar in nonthyroidectomized and thyroidectomized T4-treated rats but was different in thyroidectomized animals without T4 replacement therapy. At d 90 after TCDD dosage, mortality was still lower and the mean time to death was increased (p less than 0.01) in this group of rats compared to nonthyroidectomized or thyroidectomized T4-treated rats. However, administration of T4 starting at d 91 after dosing with TCDD resulted within 2 wk in the same final mortality in thyroidectomized rats as in nonthyroidectomized or thyroidectomized T4-treated animals, indicating that thyroid hormones modulate the time course of the wasting syndrome but do not affect the ultimate mortality figure. Body weight loss was much slower in thyroidectomized (approximately 1 g/d) than in nonthyroidectomized or thyroidectomized T4-treated rats (approximately 8 g/d). In the second experiment the three vehicle control groups of the first experiment were used. Nonthyroidectomized vehicle controls and thyroidectomized T4-treated controls were maintained as before, whereas thyroidectomized controls received T3 at 5 micrograms/kg daily. One month later each rat was dosed with TCDD at 100 micrograms/kg in corn oil/acetone. Toxicity of TCDD was similar in nonthyroidectomized, thyroidectomized T4-treated, and thyroidectomized T3-treated rats as judged by mortality, body weight, and food intake, indicating no difference between T3 and T4 in the modulation of TCDD toxicity.

[Double-blind group comparison of topical meclocycline, erythromycin and placebo in the treatment of papulo-pustulosa acne]. / Doppelblinder Gruppenvergleich von topischem Meclocyclin, Erythromycin und Placebo in der Behandlung der Akne papulo-pustulosa.

Ten institutions participated in a controlled clinical trial in order to evaluate the efficacy of topical meclocycline and erythromycin in comparison to placebo with regard to papulopustular acne. Both drugs had been incorporated in the same galenic formulation that served as placebo. The vehicle employed in this study guaranteed equally favorable drug relies for both preparations. At the end of the trial, 419 patients could be evaluated for efficacy. As impartial criterion for evaluation, the number of inflammatory lesions on the right side of the face was counted before and after three months of treatment. In addition, we recorded the patients' and physicians' overall judgment at the end of the study. As compared with placebo, meclocycline as well as erythromycin brought about statistically significant improvement already after two months of treatment. After three months, the results were statistically very highly significant (p less than 0.001). At any time of the study, there could not be demonstrated any difference between the two groups treated with meclocycline and erythromycin.

Effect of thyroidectomy and thyroxine on 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induced toxicity.

Chemical thyroidectomy effectively protected athyroid rats from mortality during 45 days after dosing with 100 micrograms 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)/kg, whereas 70 to 80% of nonthyroidectomized-euthyroid and thyroidectomized-T4 (thyroxine)-maintained-euthyroid rats died within the same period of time. There was a significant decrease in body weight of all TCDD-treated groups compared to vehicle controls. However, body weight loss was much slower in thyroidectomized-athyroid (congruent to 1 g/day) than in nonthyroidectomized-euthyroid or in thyroidectomized-T4-euthyroid (congruent to 8 g/day) rats. TCDD significantly reduced feed intake in nonthyroidectomized-euthyroid and thyroidectomized-T4-euthyroid rats, but no altered feed consumption was observable in thyroidectomized-athyroid animals. These data indicate that thyroid hormone(s) play(s) an important role in mediating the toxicity of TCDD.

Stimulation of nonbiliary, intestinal excretion of hexachlorobenzene in rhesus monkeys by mineral oil.

Four rhesus monkeys were administered various doses of hexachlorobenzene (HCB) po, to achieve widely varying adipose tissue levels. One month later, each animal was provided with a bile duct bypass allowing for interruption of the enterohepatic circulation (EHC). Effects of mineral oil-supplemented diet and/or interruption of the EHC on urinary, biliary, and fecal excretion of HCB and its metabolites were quantified. Urinary excretion of HCB was not affected by mineral oil but was reduced 20 to 60% by interruption of the EHC. Similarly, biliary excretion of HCB was also reduced 25 to 60% by interruption of the EHC and was not altered by mineral oil. Fecal excretion was increased about fivefold by mineral oil, whereas interruption of the EHC had no effect on the amount of HCB in feces. Results demonstrate that interruption of the EHC reduced urinary and biliary excretion of HCB metabolites, whereas mineral oil specifically stimulated intestinal excretion of the parent compound.

Relationship of body weight to disposition of hexachlorobenzene in rats.

Male rats of various body weights were dosed twice with [14C]hexachlorobenzene (50 mg/kg, p.o.) in olive oil by gavage on 2 consecutive days. During 2 weeks after dosage, cumulative excretion into urine was about 1% of the dose and unrelated to body weight. Cumulative excretion into feces was 30 +/- 10% of the dose and decreased with increasing body weight. In contrast, the concentration of hexachlorobenzene in urine, feces, kidney, liver and adipose tissue 14 days after dosing were higher in larger than in smaller animals. However, the relative concentration, i.e. the concentration of hexachlorobenzene in urine, feces or tissue divided by the concentration of hexachlorobenzene in adipose tissue, eliminated most of the variability among individual animals. This allows direct comparison of dispositional data of animals with greatly differing body burdens.

Intraluminal hexadecane enhances large intestinal excretion of tissue hexachlorobenzene in rats.

In young adult female rats with a cecal stoma, administration of hexadecane into the stomach (750 or 500 mg) and into the ligated colon (250 mg) enhanced 2- to 3-fold the intestinal excretion of [14C]hexachlorobenzene. Apparent sites of increased transferral from the blood into luminal contents were (caudal) ileum, cecum and colon. Presence of nonabsorbed hexadecane in luminal contents seems to be a prerequisite for the effect. Results support previous reports that enhancement of intestinal excretion of lipophilic chemicals by liquid paraffins takes place in the large intestine.

[Comparative study of diflorasone diacetate and clobetasol-17-propionate in PUVA-resistant psoriasis]. / Vergleichsstudie mit Diflorasondiacetat und Clobetasol-17-Propionat bei PUVA-resistenter Psoriasis.

In a double-blind controlled trial, we have compared the therapeutic effectiveness of Diflorason-Diacetate and Clobetasol-17-propionate with 50 PUVA-resistent psoriatics. Both externals resulted in a highly significant decline of psoriatic symptoms. The difference in therapeutic achievement between the two respective groups was statistically not significant. According to the physician's overall judgement, however, Clobetasol was favored significantly over Diflorasone-Diacetate, while the patients only showed a slight preference for Clobetasol. Based on these findings and previously published results, Diflorasone-Diacetate may be classified as one of the most effective skin corticosteroids, ranging just behind Clobetasol.

Enhanced intestinal excretion of hexachlorobenzene in rats by intraluminal injection of hexadecane.

The effect of hexadecane on the intestinal excretion of hexachlorobenzene was studied in female Sprague-Dawley rats dosed twice with 14C-hexachlorobenzene at 50 mg kg-1 per os. Injection of 75 mg n-hexadecane into ligated and unligated segments of the intestine increased concentrations of hexachlorobenzene in intestinal contents by about two- or three-fold in jejunal and ileal segments, and about two-fold in the cecal-colon segment. The jejunum appeared to be the site of greatest excretion of hexachlorobenzene followed by the ileum, the cecum and the colon. This order is opposite to our previous data from animals with an undisturbed intestinal passage. The apparently greater excretion of hexachlorobenzene into the small intestine is probably due to its much larger surface area than that of the large intestine. However, the residency time of luminal contents in the large intestine normally exceeds that in the small intestine by about 20--40-fold, which apparently more than compensates for the difference in relative surface area between small and large intestine. Thus, residency time appears to be a more important factor than surface area in determining the intestinal elimination of hexachlorobenzene. These results with hexachlorobenzene are probably typical of physiological disposition of lipophilic halogenated hydrocarbons generally.

Mineral oil in the diet enhances fecal excretion of DDT in the rhesus monkey.

Seven days after dosing of two rhesus monkeys with 14C-DDT (50 mg/kg) per os, one monkey was put on a diet containing 5% mineral oil for 35 days, whereas the other animal served as control. During 5 weeks both urinary and fecal excretion of radioactivity was more than doubled in the treated as compared to the untreated animal. At the end of the treatment, concentration of DDT in adipose tissue of the mineral oil treated animal was about half of that found in the control. The data represent further support of previous reports that mineral oil reduces body burdens of refractory lipophilic chemicals.