MicroRNA-206 is differentially expressed in Brca1-deficient mice and regulates epithelial and stromal cell compartments of the mouse mammary gland.

Depletion of Brca1 leads to defects in mouse mammary gland development and mammary tumors in humans and mice. To explore the role of microRNAs (miRNAs) in this process, we examined the mammary glands of MMTV-Cre Brca1(Co/Co) mice for differential miRNA expression using a candidate approach. Several miRNAs were differentially expressed in mammary tissue at day 1 of lactation and in mammary epithelial cell lines in which Brca1 messenger RNA (mRNA) levels have been reduced. Functional studies revealed that several of these miRNAs regulate mammary epithelial cell function in vitro, including miR-206. Creation and analysis of MMTV-miR-206 transgenic mice showed no effect on lactational mammary development and no tumors, but indicates a role in mammary tissue remodeling in mature mice, potentially involving Igf-1 and Sfrp1. These results indicate the potential of miRNAs to mediate the consequences of Brca1 loss and suggest a novel function for miR-206.

Humanization of the mouse mammary gland.

Although mouse models have provided invaluable information on the mechanisms of mammary gland development, anatomical and developmental differences between human and mice limit full understanding of this fundamental process. Humanization of the mouse mammary gland by injecting immortalized human breast stromal cells into the cleared murine mammary fat pad enables the growth and development of human mammary epithelial cells or tissue. This facilitates the characterization of human mammary gland development or tumorigenesis by utilizing the mouse mammary fat pad. Here we describe the process of isolating human mammary stromal and epithelial cells as well as their introduction into the mammary fat pads of immunocompromised mice.

Analysis of Brca1-deficient mouse mammary glands reveals reciprocal regulation of Brca1 and c-kit.

Disruption of the breast cancer susceptibility gene Brca1 results in defective lobular-alveolar development in the mammary gland and a predisposition to breast tumourigenesis in humans and in mice. Recent evidence suggests that BRCA1 loss in humans is associated with an expansion of the luminal progenitor cell compartment in the normal breast and tumours with a luminal progenitor-like expression profile. To further investigate the role of BRCA1 in the mammary gland, we examined the consequences of Brca1 loss in mouse mammary epithelial cells in vitro and in vivo. Here, we show that Brca1 loss is associated with defective morphogenesis of SCp2 and HC11 mouse mammary epithelial cell lines and that in the MMTV-Cre Brca1(Co/Co) mouse model of Brca1 loss, there is an accumulation of luminal progenitor (CD61(+)CD29(lo)CD24(+)) cells during pregnancy. By day 1 of lactation, there are marked differences in the expression of 1379 genes, with most significantly altered pathways and networks, including lactation, the immune response and cancer. One of the most differentially expressed genes was the luminal progenitor marker, c-kit. Immunohistochemical analysis revealed that the increase in c-kit levels is associated with an increase in c-kit positivity. Interestingly, an inverse association between Brca1 and c-kit expression was also observed during mammary epithelial differentiation, and small interfering RNA-mediated knockdown of Brca1 resulted in a significant increase in c-kit mRNA levels. We found no evidence that c-kit plays a direct role in regulating differentiation of HC11 cells, suggesting that Brca1-mediated induction of c-kit probably contributes to Brca1-associated tumourigenesis via another cellular process, and that c-kit is likely to be a marker rather than a mediator of defective lobular-alveolar development resulting from Brca1 loss.

Aspects of abstinence after morphine ingestion.

Sprague-Dawley male rats were intoxicated with morphine, using an ingestion method where exposed and control rats received equivalent amounts of calories and nutrients. The degree of physical dependence on morphine was demonstrated by studying and quantifying abstinence symptoms after withdrawal or after administration of opiate antagonists. The aims of the study were (1) to further enlighten the specificity and validity of the intoxication method concerning physical dependence, and (2) to determine whether some of the abstinence signs might be of value to facilitate quantitation of the degree of physical dependence on morphine, with diet and fluid intake being maintained under control. Withdrawn rats showed a decreased fluid diet intake and a body weight loss, the latter partly due to anorexia. Other mild abstinence signs were irritation, tremor and some motor excitation. The body weight loss during the first day of morphine withdrawal was proportional to the accumulated drug dose (between 25 and 300 mg morphine PO/kg b.wt.). However, prolonged morphine treatment on one dose (340 mg/kg b.wt.) did not reinforce the body weight changes caused by morphine withdrawal. The succeeding weight gain some days after morphine withdrawal was not entirely dependent on the amount of fluid diet intake. Methadone was shown to partially block the decrease in diet intake and the weight loss seen during morphine withdrawal. The naloxone-precipitated withdrawal symptoms were motor excitation, cholinergic signs, body weight loss, diarrhoea and decreased diet intake. The weight loss 2 hr after naloxone administration to long-term intoxicated rats was proportional to the naloxone dose.(ABSTRACT TRUNCATED AT 250 WORDS)

Ciprofloxacin in acute male gonorrhea.

100 male adults suffering from acute gonococcal urethritis were hospitalised and treated with decreasing doses from 2000 mg to 100 mg of ciprofloxacin (1-cyclopropyl-6-fluoro-1,4-dihydro-4-oxo-7-(1-piperazinyl)-3-quinoline carboxylic acid, Bay o 9867; designated tradename: Ciprobay). Clinical signs (burning, exudate, edema, inflammation) and bacteriology (native and culture) were assessed hourly during the first 9 h and at 12, 24 and 72 h after start of therapy. Most patients were controlled during 10 days. 99 patients showed clinical and bacteriological cure. One patient with subacute gonococcal urethritis (protocol violation) did not react on 250 mg. No postgonococcal urethritis was seen. No side effects were reported. For the acute male gonococcal urethritis a 100 mg single dose treatment with ciprofloxacin can be recommended.

[Mixed infections of the urogenital tract with Trichomonas vaginalis and yeast-like fungi in humans with reference to treatment]. / Mischinfektionen des Urogenitaltraktes mit Trichomonas vaginalis und hefeähnlichen Pilzen beim Menschen unter Berücksichtigung der Behandlung.

Comparative studies on the incidence of mixed infections with T. vaginalis and yeast-like fungi in man carried out within the years 1969-1980 and 1981-1984 showed a progressive increase of this kind of disease. Clinical treatment with Clotrimazole was performed in 35 patients. Local therapy with Clotrimazole was effective, if 3-5 days later the patients receive Tinidazole at one dosage. In some patients, the therapy has to be continued with Metronidazole.

Long lasting changes in gerbil brain after chronic ethanol exposure: a quantitative study of the glial cell marker S-100 and DNA.

Glial S-100 protein, soluble protein, and DNA were quantitatively studied in brains of gerbils chronically exposed to ethanol in a nutritionally complete fluid diet. Eight different brain areas were studied. After exposure to ethanol for 3 months followed by a 4-month post-treatment ethanol-free period, increased amounts of S-100 protein per wet weight were found in the frontal cerebral cortex, the sensory-motor cerebral cortex, the posterior cerebellar vermis, and the brainstem. The increase of S-100 in the posterior cerebellar vermis was paralleled by an increase in DNA per wet weight, which was also increased in the anterior cerebellar vermis. However, a decreased content of DNA was observed in the frontal cerebral cortex, despite the increase of S-100 protein, suggesting a cell loss affecting cells other than astroglial in this area. In the cerebellar vermis, elevated concentrations of soluble proteins per wet weight were found, whereas a decreased amount was found in the anterior cerebellar hemispheres. It is suggested that the S-100 protein acts as a marker for astroglial cell volume and that a concomitant increase of S-100 protein and DNA might indicate an increase in the number of astroglial cells. Thus, our results obtained after ethanol exposure and subsequent ethanol abstinence are compatible with changes consisting of astroglial hypertrophy in the cortex areas and brainstem, as well as astroglial hypertrophy and/or proliferation in the posterior cerebellar vermis, a clear sign of the preceding noxae.

[Trichomoniasis of the urogenital tract in patients hospitalized in 1976-1983]. / Trichomoniase des Urogenitaltraktes bei hospitalisierten Kranken in den Jahren 1976-1983.

We report on investigations carried out on T. vaginalis and mixed infection (T. vaginalis and yeast-like fungi). Special attention was paid to metronidazol resistance on T. vaginalis strains isolated from patients who had been repeatedly treated with this medicament. Therefore we point to the necessity of defining the patients' susceptibility to this remedy.

Decreased morphine intake by opiate addicted rats administered zimelidine, a 5-HT uptake inhibitor.

Zimelidine, a specific 5-HT uptake inhibitor, reduced peroral morphine consumption by morphine-addicted adult male and female Sprague-Dawley rats and old male rats in choice tests. The effect was dose dependent in male rats. Thus, the availability of central 5-HT appears to be important for the regulation of morphine preference in rat. The results are discussed in relation to recent literature where ethanol preference has been found to be attenuated by zimelidine. The results may provide insights into the complex cellular mechanisms underlying opiate addiction.

[Diagnostic and epidemiologic significance of multinuclear and round forms of Trichomonas vaginalis]. / Diagnostik und epidemiologische Bedeutung der multinuklearen und Rundformen von Trichomonas vaginalis.

Trichomonas are analyzed in the material taken directly from the urogenital tract of male and female patients cultivated on Roiron medium and using different methods for staining the preparations was carried out. It is observed that some of the preparations examined show round nuclear forms of T. vaginalis impossible the detect by laboratory methods used up to now. The techniques used in this study decrease the risk of false diagnosis of these forms of trichomoniasis by less experienced examiners thus facilitating correct diagnosis and therapy. It should be also said, that they are simple and easy to carry out in the laboratory.

[Effect of different cytostatic agents and antibiotics on the biology of Trichomonas in vitro]. / Einfluss verschiedener Zytostatika und Antibiotika auf die Biologie der Trichomonaden in vitro.

The purpose of these examinations was an attempt to elucidate the influence of some cytostatic preparations (Fluorouracil, Triaziquone (Treminon), Mechlorethaminee Oxide Hydrochloride (Mitomen), Methotroxate, Mitomycins, Cyclophosphamide (Endoxan), Mechlorethamin (Nitrogranulogen) and antibiotics (Penicillin, Streptomycin, Aureomycin, Nystatin) on formation of the multinuclear forms of Trichomonas under the conditions of culture on artificial medium and to investigate the cidal and static effects of chosen cytostatics and antibiotics of different species of Trichomonas in vitro. It was found that Mechlorethamine (Nitrogranulogen) and Mitomycins have distinct influence on formation of the multinuclear forms of Trichomonas in culture. Similar effects were observed in the antibiotic group with aureomycin only if applied pure or combined with other antibiotic drugs. In addition, it was shown that Triaziquone (Treminon) exerts the strongest effects on the examined species of Trichomonas.

Clinical studies on the effectiveness of Wratizolin in the treatment of some dermatoses.

Clinical studies on therapeutic effectiveness of ITCL applied as 2-3% ointment, 2% cream and 2-3% lotion were carried out. The study material included 96 patients at the age of 9-80. Of this group, 19 patients were with herpes zoster, 16 with herpes simplex, 29 with eczema complicated by secondary bacterial infection, 30 with palm and foot pustulosis, 1 patient with actinic cheilitis and 1 with varicella. The results obtained have revealed that the preparation is particularly effective in the treatment of dermatoses induced by herpes zoster and herpes simplex virus.

Identification and characterization of sulfhydryl-containing proteolytic fragments involved in the Ca2+-induced conformational change of beef brain S-100.

The Ca2+-dependent conformational alteration of the brain-specific S-100 protein was studied by reacting the protein with N-ethyl[2,3-14C]maleimide in the absence and presence of Ca2+ and under denaturing conditions. Peptic hydrolysates of the 14C-labeled protein were analyzed and fractionated by high-performance liquid chromatography. Labeled peptide fractions were characterized by high-voltage electrophoresis and TLC. A clear distinction could be made between two classes of sulfhydryl-containing fragments: (a) neutral, hydrophobic, and (b) acidic. Ca2+ markedly favored 14C incorporation into the former components, whereas the latter were readily available only under denaturing conditions.

A new ingestion method for long-term morphine intoxication in rat.

A new method for long-term morphine intoxication in rat was developed. It was designed to deal with the nutritional imbalance and body weight loss that generally occurs using conventional techniques for morphine treatment. The morphine is administered in a nutritionally complete diet. Also pair-feeding is used to deal with intoxicated rats that do not eat the same amount of food as controls. The technique was validated during the study of different intoxication conditions, using different initial doses, dose increments and final doses. An initial dose of 25 mg morphine/kg b.w., raised exponentially up to 340 mg/kg b.w. in 8 days, made the rats dependent, as tested by withdrawal signs, precipitated by excluding morphine from the diet, or by administration of antagonists. A final dose of up to 715 mg morphine/kg b.w. was reached in 13 days without decreased food intake. However, initial doses of 340 or 715 mg/kg led to impaired weight gain and diet consumption. Plasma morphine levels of 3 micrograms/ml serum were reached on a dose of 340 mg/kg b.w. Also, preference for morphine diet over control diet was evaluated by choice tests. The technique is simple, time-saving and inexpensive, allowing the treatment of numerous animals for long periods under standardized intoxication conditions. No animals get ill or die.