Expression of novel genes linked to the androgen-induced, proliferative shutoff in prostate cancer cells

Androgens control cell numbers in the prostate through three separate pathways: (a) inhibition of cell death, (b) induction of cell proliferation (Step-1) and (c) inhibition of cell proliferation (Step-2, proliferative shutoff). The mechanisms underlying these phenomena are incompletely understood. The human prostate carcinoma LNCaP variants express these pathways as follows: LNCaP-FGC express both steps, LNCaP-LNO expresses Step-2, LNCaP-TAC expresses Step-1, and LNCaP-TJA cells express neither step. These cells facilitated the search for mediators of the androgen-induced proliferative shutoff pathway. Androgen exposure for 24 h or longer induced an irreversible proliferative shutoff in LNCaP-FGC cells. The Wang and Brown approach for identifying differentially expressed mRNAs was used to search for mediators of Step-2. Ten unique inserts were identified and from those ten, three genes were further studied. The basal expression of these genes in shutoff-negative variants was not affected by androgen exposure. They were induced by androgens in shutoff-positive LNCaP variants and the androgen receptor-transfected, shutoff-positive, MCF7-AR1 cells. These genes were induced only in the range of androgen concentrations that elicited the shutoff response. Time course analysis showed that their induction precedes the commitment point by 12–18 h. In addition, they were expressed in the normal prostate during proliferative shutoff. These features suggest that the candidate genes have a role in the regulation cascade for proliferative shutoff.     

Hydrological cycle and interannual variability of the aquatic community in a temporary saline lake (Fuente de Piedra,Southern Spain)

Fuente de Piedra is a shallow, temporary saline lake whoseseasonal behavior is strongly dependent on the annual hydrologicalbudget. In this study, we outline the characteristics of Fuente dePiedra Lake for two years that had different hydrological budgets.The high precipitations in 1989–90 caused the lake not to dry asusual, and decreased both salinity and the amplitude of changes.There were also differences in nutrient dynamics, with generallylower concentrations of soluble reactive phosphorus and ammonium,whereas in the more humid year nitrate showed a distinct maximum inwinter. Winter bloom chlorophyll a concentrations were alsomuch higher in 1989–90 (>600 g l-1) but there wasalso a winter productive phase that was presumably poorly coupledwith consumption processes that predominate in spring. Planktonicassemblages were different between years. Highly halotolerantphytoplankton species (Dunaliella salina and D. viridis) became scarcer, and especially two previouslyunrecorded diatoms (Cyclotella sp. and Chaetoceros sp.)became dominant in the bloom time in the wet year. The speciesrichness of the zooplankton increased in the wet year, with newspecies appearing that were not collected during 1987–88(Branchinectella media, Daphnia mediterranea, Macrothrix sp.,Arctodiaptomus salinus, Cyclops sp., Sigara sp...).There was also a much higher development of macrophytes (Ruppiadrepanensis, Althenia orientalis, Lamprothamnium papulosum)and bird populations, especially flamingoes (Phoenicopterusruber).Important interannual variations in this sort of system pointto the need for long term studies to observe the whole range ofstates that define the lake as an entity.     

RTN TRH causes prolonged respiratory stimulation

Cream, Carlos L., Aihua Li, and Eugene E. Nattie. RTNTRH causes prolonged respiratory stimulation. J. Appl.Physiol. 83(3): 792-799, 1997.We injectedthyrotropin-releasing hormone (TRH; 10 nl; 0.25, 0.5, 1.0, or 10 mM),its inactive free acid form (TRHOH; 1 mM), or a metabolite with lowTRH-receptor binding affinity, histidine-proline diketopiperazine (cHP;1 mM), into the retrotrapezoid nucleus of anesthetized rats. Injectionlocation was verified by anatomic analysis. Lower doses (0.25-0.5mM) significantly increased both the product of integrated phrenicamplitude and frequency(Phr · f) and f for 20-30min compared with artificial cerebrospinal fluid control injections. Higher doses (1.0-10 mM) produced greater and long-lastingstimulation of Phr · f,Phr, and f and of blood pressure. Thisstimulation reached values 150% of baseline and durations of 270 minafter a single injection. TRHOH (1 mM ) or cHP (1 mM) had no effect onPhr but increased f, as did 1 mM TRH. We concludethat TRH has a very powerful stimulatory effect in the retrotrapezoidnucleus region on Phr · f, withthe Phr response seemingly specific for TRHreceptors. Similar responses of f to TRHOH and cHP suggest it may benonspecific.

     

XY gonadal dysgenesis and gonadoblastoma: a study in two sisters with a cryptic deletion of the Y chromosome involving the SRY gene

This paper reports a case of XY gonadal dysgenesis in two sisters. Both patients presented an eunochoid female phenotype with normal external genitalia. At laparotomy, the elder sister was found to have bilateral gonadoblastoma. Cytogenetic studies, which included G and C banding and in situ hybridization, showed that the patients had an apparently normal 46, XY karyotype. PCR analyses revealed absence of the conserved portion (HMG box) of the SRY gene and of the Y chromosome pseudoautosomal boundary region sequence in both patients. The presence of the ZFY sequence was detected by Southern hybridization in the two affected sisters. The patients' father (46, XY, no mosaicism detected in peripheral blood lymphocytes) was positive for SRY and ZFY sequences. The occurrence of gonadoblastoma is discussed in terms of the genetic factors that may lead to tumor development.     

Neurochemical effects ofl-pyroglutamic acid

The effect ofl-pyroglutamic acid, a metabolite that accumulates in pyroglutamic aciduria, on different neurochemical parameters was investigated in adult male Wistar rats. Glutamate binding, adenylate cyclase activity and G protein coupling to adenylate cyclase were assayed in the presence of the acid.l-pyroglutamic acid decreased Na⁺-dependent and Na⁺-independent glutamate binding Basal and GMP-PNP stimulated adenylate cyclase activity were not affected by the acid. Furthermore, rats received unilateral intrastriatal injections of 10–300 nmol of bufferedl-pyroglutamic acid. Vehicle (0.25 M Tris-Cl, pH 7.35–7.4) was injected into the contralateral striatum. Neurotoxic damage was assessed seven days after the injection by histological examination and by weighing both cerebral hemispheres. No difference in histology or weight could be identified between hemispheres. These results suggest that, although capable of interfering with glutamate binding, pyroglutamate did not cause a major lesion in the present model of neurotoxicity.     

Antibiotic treatment induces an increase of the specific antibody levels in Brucella melitensis infected mice

Abstract The effects of doxycycline (DOX) and streptomycin (SM) treatment of Brucella melitensis infected mice on humoral immune response were examined. In female BALB/c mice, DOX was administered at a dose of 50 mg/kg/12 h, for 21 or 45 consecutive days, alone or combined with SM (10 mg/kg/12 h) for 14 days. All treatments induced a significant increase in specific IgG levels (ELISA and CIEP) against LPS and cytosolic antigens of Brucella during treatment. This was not related with therapeutic failure or relapse since all treatments induced a significant reduction in the degree of infection.     

Batch culture of Candida utilis in a medium deprived of a phosphorous source

Summary Candida utilis var. major NRRL-Y-1084 was grown in a defined medium without a phosphorous (P) source. During the exponential phase, cells divided according to a specific growth rate of 0.32 h^-1, which is lower than the usual rate for a balanced medium (0.4–0.6 h^-1). The relative P content of the biomass decreased from 2.70% to 0.75% over a period of 6 h, including 2 h of cell division arrest. At the end of this period there was another interruption of cell division. After that, multiplication restarted at a considerably lower rate and it deviated slightly from the exponential pattern. The stationary phase began when biomass P content reached 0.4%–0.5%, slowly decreasing afterwards to 0.25–0.20%. Biomass synthesis was less affected than cell division by the relative decrease of endogenous P, the two processes differing partially in their kinetics. Cell lysis started shortly before the stationary phase and affected about 20% of the population by the end of the assay. RNA and P content of the resulting biomass were 2.4% and 0.25% respecitvely, P being mainly incorporated to RNA.The relationship of biomass production to glucose uptake was very low, probably because the marked P deficiency called for an increase in energy consumption for growth and specially for maintenance. Compared with yeasts grown in a balanced medium, 40% increase in glycogen was observed, whereas no mean changes in the content of cell wall carbohydrates (glucan and mannan) and that of true protein were found.Member of the Scientific Researcher's Career of the Consejo Nacional de Investigaciones Cientificas y Técnicas (CONICET). Agrentina     

Pseudomonas aeruginosa acid phosphatase and cholinesterase induced by choline and its metabolic derivatives may contain a similar anionic peripheral site

Summary Different compounds derived from choline, and obtained by demethylation or by oxidation of the primary alcohol group with subsequent N-demethylation, were tested as inducer agents of acid phosphatase and cholinesterase in Ps. aeruginosa. It was found that betaine and dimethylglycine were the most effective inducers of both enzyme activities. These metabolites including choline itself, were not inducers of acid phosphatase and cholinesterase in other Gram-negative bacteria such as: Escherichia coli, Salmonella typhimurium, Shigella flexneri, Enterobacter liquefacciens and Proteus mirabilis. The acid phosphatase activities found in these bacteria were not inhibited in vitro by choline, betaine and phosphorylcholine. From these results it may be concluded that the acid phosphatase activity from Ps. aeruginosa is different from the same activity observed in the other bacteria. In addition, it is also shown that Ps. aeruginosa acid phosphatase and cholinesterase were inhibited by a number of compounds containing a positively charged amino group, with methyl or ethyl groups bound to it. These results seem to confirm that Ps. aeruginosa acid phosphatase and cholinesterase may contain a similar anionic site.     

Five new filamentous,glucose-fermenting Candida isolated from decayed wood in the evergreen rainy Valdivian forest of southern Chile

Five novel representatives of the yeast genus Candida isolated from advanced stages of wood degradation from fallen trunks of Eucryphia cordifolia Cav., Laurelia sempervirens Weim., Laurelia philippiana Weim., Nothofagus dombeyii (Mirb.) Blume, and Nothofagus obliqua (Mirb.) Blume, in the evergreen rainy Valdivian forest of southern Chile, are described and illustrated. They clearly differ from all accepted Candida species to warrant their establishment as five new species of yeasts: Candida coipomensis sp. nov., Candida ralunensis sp. nov., Candida laureliae sp. nov., Candida osornensis sp. nov., and Candida llanquihuensis sp. nov.     

Four new species of Apiotrichum isolated from decayed wood in the evergreen rainy Valdivian forest of southern Chile

Four unusual Apiotrichum, isolated from decayed wood of Eucryphia cordifolia Cav., Nothofagus obliqua (Mirb.) Blume, and Laurelia sempervirens Wein., one of which was also isolated from the intestinal tract of Scaptomyza multispinosa Malloch (Diptera), are described and illustrated. These species differ from all the accepted Apiotrichum species (1–3) to warrant their establishment as four new species: Apiotrichum eucryphiae, Apiotrichum osvaldii, Apiotrichum futronensis and Apiotrichum nothofagi.