Nonsex Genes in the Mating Type Locus of Candida albicans Play Roles in a/α Biofilm Formation,Including Impermeability and Fluconazole Resistance

The mating type locus (MTL) of Candida albicans contains the mating type genes and has, therefore, been assumed to play an exclusive role in the mating process. In mating-incompetent a/α cells, two of the mating type genes, MTL a1 and MTLα2, encode components of the a1-α2 corepressor that suppresses mating and switching. But the MTL locus of C. albicans also contains three apparently unrelated “nonsex” genes (NSGs), PIK, PAP and OBP, the first two essential for growth. Since it had been previously demonstrated that deleting either the a/α copy of the entire MTL locus, or either MTLa1 or MTLα2, affected virulence, we hypothesized that the NSGs in the MTL locus may also play a role in pathogenesis. Here by mutational analysis, it is demonstrated that both the mating type and nonsex genes in the MTL locus play roles in a/α biofilm formation, and that OBP is essential for impermeability and fluconazole resistance.     

Identification of Genes Upregulated by the Transcription Factor Bcr1 That Are Involved in Impermeability,Impenetrability, and Drug Resistance of Candida albicans a/α Biofilms

Candida albicans forms two types of biofilm, depending upon the configuration of the mating type locus. Although architecturally similar, a/α biofilms are impermeable, impenetrable, and drug resistant, whereas a/a and α/α biofilms lack these traits. The difference appears to be the result of an alternative matrix. Overexpression in a/a cells of BCR1, a master regulator of the a/α matrix, conferred impermeability, impenetrability, and drug resistance to a/a biofilms. Deletion of BCR1 in a/α cells resulted in the loss of these a/α-specific biofilm traits. Using BCR1 overexpression in a/a cells, we screened 107 genes of interest and identified 8 that were upregulated by Bcr1. When each was overexpressed in a/a biofilms, the three a/α traits were partially conferred, and when each was deleted in a/α cells, the traits were partially lost. Five of the eight genes have been implicated in iron homeostasis, and six encode proteins that are either in the wall or plasma membrane or secreted. All six possess sites for O-linked and N-linked glycosylation that, like glycosylphosphatidylinositol (GPI) anchors, can cross-link to the wall and matrix, suggesting that they may exert a structural role in conferring impermeability, impenetrability, and drug resistance, in addition to their physiological functions. The fact that in a screen of 107 genes, all 8 of the Bcr1-upregulated genes identified play a role in impermeability, impenetrability, and drug resistance suggests that the formation of the a/α matrix is highly complex and involves a larger number of genes than the initial ones identified here.     

Latency, threshold and frequency selectivity of the crossed medial cochlear efferent pathways

The efferent innervation of guinea pig cochleas was sectioned medially, at the level of the floor of the fourth ventricle, to study the effects of the crossed part of the medial efferent pathway on the compound action potential (CAP) masking phenomenon. Sectioning reduced CAP masking for a masker level varying with the frequency of the masker and the time elapsed between the masker onset and the probe onset. Functional properties of the crossed part of the medial efferent tracts: latency, thresholds and frequency selectivity, could be deduced from these data. This intensification of the masking phenomenon permitting the improvement of the signal to noise ratio, may thus be attributed to the crossed part of the medial efferent bundle which innervates the outer hair cells.     

Microdialysis Monitoring of 3,4-Dihydroxyphenylalanine Accumulation After Decarboxylase Inhibition: A Means to Estimate In Vivo Changes in Tyrosine Hydroxylase Activity of the Rat Locus Ceruleus

Abstract: An on-line microdialysis approach was developed to estimate changes in tyrosine hydroxylase activity in the locus ceruleus noradrenergic neurons of anesthetized rats by measuring the 3,4-dihydroxyphenylalanine (DOPA) acumulation in the extracellular fluid during perfusion of an aromatic amino acid decarboxylase inhibitor through a dialysis probe. The aromatic amino acid decarboxylase inhibitor used was difluoromethyl-DOPA, which was shown to be more stable than NSD 1015 or Ro 4-4602 in the perfusion fluid. A 1-h perfusion of a 10⁻⁴ mol/L of difluoromethyl-DOPA solution induced a linear increase in DOPA concentration in the locus ceruleus dialysates that achieved a steady state within 1 h. The identity of DOPA accumulated in dialysates during aromatic amino acid decarboxylase inhibition was confirmed by the disappearance of the chromatographic peak when DOPA formation was blocked by the administration of α-methyl- p -tyrosine. Systemic administration of the α₂-antagonist piperoxane before difluoromethyl-DOPA perfusion markedly increased the DOPA concentration during both the accumulation and the steady-state periods, showing that the present technique is a suitable in vivo approach to monitor changes in tyrosine hydroxylase activity occurring in the locus ceruleus neurons.     

Los suplementos de calcio y el posible aumento del riesgo cardiovascular

The primary goal of osteoporosis treatment is to prevent the occurrence of fragility fractures, and thereby reduce morbidity and mortality. Among the various approaches to the treatment of this disease include ensuring proper calcium intake and to obtain adequate levels of vitamin D. Virtually all clinical trials with drugs used to treat osteoporosis systematically include calcium and vitamin D supplements. In light of the recent publication of clinical trials and meta-analyses, a possible increase in cardiovascular risk, particularly in the form of a myocrdial infarction, is hypothesised in patients taking calcium supplements. However, data published to date are inconclusive. Until the development of new scientific evidence, it seems reasonable to recommend, whenever practicable and individualized for each patient, increasing calcium intake with food and reserve supplements for patients with very low calcium intake in the diet. It would also be advisable for the administration of total daily dose to be fractionated throughout the day and with meals, and to obtain appropriate levels of vitamin D (25-hydroxycholecalciferol or calcidiol), along with the basic treatment for osteoporosis that is decided to be prescribed to patients.