Nano-indentation and laser-induced damage testing in optical multilayer-dielectric gratings.

We demonstrate how a nanomechanical test identifies areas of mechanical field concentration as being comparable to areas where optical fields are known to be concentrated, in the special context of laser-induced damage testing (LIDT) of diffractive gratings of silica deposited on optical multilayers. The nano-indentation response of the diffraction gratings is measured in a new mode that allows for the extraction of a measurable metric characterizing the brittleness of the gratings, as well as their ductility. We show that lower LIDTs are positively correlated with an increased grating brittleness, and therefore identify a nanomechanical approach to describe LIDTs. We present extensive numerical simulations of nano-indentation tests and identify different deformation modes including stretching, shear concentration, and bending as precursors to mechanical failure in the nano-indentation test. The effects of geometrical inhomogeneities on enhanced stress generation in these gratings are specifically examined and addressed, and we show the agreement between nanomechanical testing and analytical interpretation of these inhomogeneities.

Surveillance of nosocomial transmission of Candida albicans in an intensive care unit by DNA fingerprinting.

The frequency of nosocomial transmission of the opportunistic fungal pathogen Candida albicans in an intensive care unit was tested by DNA fingerprinting of 91 isolates from 32 hospitalized patients with the mid-repetitive Ca3 DNA probe. This showed that serial isolates of C. albicans from individual patients belonged to genetically distinct strains. In comparison with nosocomial bacterial pathogens, the transmission of C. albicans in an intensive care unit occurred at a much lower frequency. In conclusion, the threat of C. albicans infection does not lie within the hospital, but in commensal isolates. These findings are relevant for infection control practices.

Signaling through adenylyl cyclase is essential for hyphal growth and virulence in the pathogenic fungus Candida albicans.

The human fungal pathogen Candida albicans switches from a budding yeast form to a polarized hyphal form in response to various external signals. This morphogenetic switching has been implicated in the development of pathogenicity. We have cloned the CaCDC35 gene encoding C. albicans adenylyl cyclase by functional complementation of the conditional growth defect of Saccharomyces cerevisiae cells with mutations in Ras1p and Ras2p. It has previously been shown that these Ras homologues regulate adenylyl cyclase in yeast. The C. albicans adenylyl cyclase is highly homologous to other fungal adenylyl cyclases but has less sequence similarity with the mammalian enzymes. C. albicans cells deleted for both alleles of CaCDC35 had no detectable cAMP levels, suggesting that this gene encodes the only adenylyl cyclase in C. albicans. The homozygous mutant cells were viable but grew more slowly than wild-type cells and were unable to switch from the yeast to the hyphal form under all environmental conditions that we analyzed in vitro. Moreover, this morphogenetic switch was completely blocked in mutant cells undergoing phagocytosis by macrophages. However, morphogenetic switching was restored by exogenous cAMP. On the basis of epistasis experiments, we propose that CaCdc35p acts downstream of the Ras homologue CaRas1p. These epistasis experiments also suggest that the putative transcription factor Efg1p and components of the hyphal-inducing MAP kinase pathway depend on the function of CaCdc35p in their ability to induce morphogenetic switching. Homozygous cacdc35 Delta cells were unable to establish vaginal infection in a mucosal membrane mouse model and were avirulent in a mouse model for systemic infections. These findings suggest that fungal adenylyl cyclases and other regulators of the cAMP signaling pathway may be useful targets for antifungal drugs.

The TEA/ATTS transcription factor CaTec1p regulates hyphal development and virulence in Candida albicans.

The temporal and spatial expression of stage-specific genes during morphological development of fungi and higher eukaryotes is controlled by transcription factors. In this study, we report the cloning and functional analysis of the Candida albicans TEC1 (CaTEC1) gene, a new member of the TEA/ATTS family of transcription factors that regulates C. albicans virulence. The promoters of the type 4, 5 and 6 proteinase isogenes (SAP4-6) contain repetitive TEA/ATTS consensus sequence motifs. This finding suggests a possible role for a homologue of Saccharomyces cerevisiae TEC1 during the activation of proteinase gene expression in C. albicans. CaTEC1 is predominantly expressed in the hyphal form of C. albicans. In vitro, serum-induced hyphal formation as well as evasion from MPhi after phagocytosis is suppressed in catec1/catec1 mutant cells. Furthermore, expression of the proteinase isogenes SAP4-6 is no longer inducible in these mutant cells. The deletion of the CaTEC1 gene attenuates virulence of C. albicans in a systemic model of murine candidiasis, although both mutant and revertant cells that were prepared from infected tissues or the vaginal mucosa grew in a hyphal morphology in vivo. CaTEC1 complements the pseudohyphal and invasive growth defect of haploid and diploid S. cerevisiae tec1/tec1 mutant cells and strongly activates the promoter of FLO11, a gene required for pseudohyphal growth. This study provides the first evidence pointing to an essential role for a member of the TEA/ATTS transcription factor family that had so far only been ascribed to function during development as a virulence regulator in microbial pathogenesis.

In vivo virulence of Candida albicans isolates causing mucosal infections in people infected with the human immunodeficiency virus.

Mucosal candidiasis is common in human immunodeficiency virus (HIV) infection. Susceptibility to such infections may be attributed to reduced host defense mechanisms and/or virulence of the organism. In the present study, we compared the virulence of mucosal Candida albicans isolates from HIV-infected people, with and without fluconazole-refractory infection, in established murine models of systemic and vaginal candidiasis. Compared with the mortality rate ( approximately 70%) after intravenous challenge with 2 virulent reference isolates, challenge with most clinical isolates (66%-77%) resulted in prolonged survival. In contrast, fungal burden induced by intravaginal challenge of nearly all (97%) isolates was similar to that of the virulent controls. There were no differences in in vitro growth rates for any of the isolates, and there was no association between reduced mortality and clinical failure to fluconazole, in vitro antifungal susceptibility, site of infection, or other host factors. These results suggest that virulence of C. albicans is tissue specific and is not a factor in the development of fluconazole-refractory infections in advanced HIV disease.

Local Th1/Th2 cytokine production during experimental vaginal candidiasis: potential importance of transforming growth factor-beta.

Host defense mechanisms against vaginal Candida albicans infections are poorly understood. Despite the protective role of T helper (Th)1-type cell-mediated immunity (CMI) against mucosal C. albicans infections, studies using an estrogen-dependent murine model of vaginal candidiasis have shown a lack of effect of systemic Th1-type CMI against a vaginal C. albicans infection, and a lack of changes in local T cells during infection. In the present study, the local Thl- (interleukin [IL]-2, interferon [IFN]-gamma and IL-12) and Th2- (IL-4, IL-10 and transforming growth factor [TGF]-beta1) type cytokines were evaluated in vaginal tissue during an experimental C. albicans infection. Results showed constitutive expression of TGF-beta1 in vaginal tissue of naive mice that was two-fold higher than the levels of the other cytokines examined. These high levels of TGF-beta1 were further increased as a result of pseudoestrus and/or infection, and were corroborated at the messenger RNA level. Furthermore, the levels of TGF-beta in naive or infected mice were significantly higher in the vagina compared to other areas of the genital tract. Finally, TGF-beta1 predominated as well in the draining, but not non-draining, lymph nodes during infection. These results suggest that TGF-beta1, a potent immunoregulatory cytokine, may be important in the lack of demonstrable CMI at the vaginal mucosa against C. albicans.

Special Report on Electrical Standards: New Internationally Adopted Reference Standards of Voltage and Resistance.

This report provides the background for and summarizes the main results of the 18th meeting of the Consultative Committee on Electricity (CCE) of the International Committee of Weights and Measures (CIPM) held in September 1988. Also included are the most important implications of these results. The principal recommendations originating from the meeting, which were subsequently adopted by the CIPM, establish new international reference standards of voltage and resistance based on the Josephson effect and the quantum Hall effect, respectively. The new standards, which are to come into effect starting January 1, 1990, will result in improved uniformity of electrical measurements worldwide and their consistency with the International System of Units or SI. To implement the CIPM recommendations in the U.S. requires that, on January 1, 1990, the value of the U.S. representation of the volt be increased by about 9.26 parts per million (ppm) and the value of the U.S. representation of the ohm be increased by about 1.69 ppm. The resulting increases in the U.S. representations of the ampere and watt will be about 7.57 ppm and 16.84 ppm, respectively. The CCE also recommended a particular method, affirmed by the CIPM, of reporting calibration results obtained with the new reference standards that is to be used by all national standards laboratories.

Possible Changes in the U.S. Legal Units Of Voltage and Resistance.

The Consultative Committee on Electricity of the International Committee on Weights and Measures is considering adopting sometime in the future 1) a new value for the Josephson frequency-voltage ratio 2e/h (e is the elementary charge and h is the Planck constant) and 2) a value for the quantized Hall resistance R H≡h/e 2. Both values are to be chosen as consistent with their International System of Units (SI) values as possible and would be used by every national standards laboratory which employs the Josephson and quantum Hall effects to define and maintain their national or legal units of voltage and resistance. Based on current knowledge, this would lead to an increase in the U.S. Legal Volt of about nine parts-per-million (ppm) and an increase in the U.S. Legal Ohm of about 1.5 ppm. Comparable changes would be required in the voltage and resistance units of most other national, governmental, and industrial standards laboratories throughout the world. Many high-precision instruments would also have to be readjusted to make them consistent with the new units. It is the purpose of this paper to review in some detail the basis for these proposed and potentially significant changes.

New Results From Previously Reported NBS Fundamental Constant Determinations.

A new treatment of previously reported results of three electric-unit-dependent fundamental constant experiments carried out at NBS over the last decade or so yields accurate, indirect values in SI units for a number of important quantities. These include the fine-structure constant α, the Avogadro constant N A, the Josephson frequency-voltage ratio 2e/h, and the quantized Hall resistance R H≡h/e 2.