Experimental modulation of capsule size in Cryptococcus neoformans

Experimental modulation of capsule size is an important technique for the study of the virulence of the encapsulated pathogen Cryptococcus neoformans. In this paper, we summarize the techniques available for experimental modulation of capsule size in this yeast and describe improved methods to induce capsule size changes. The response of the yeast to the various stimuli is highly dependent on the cryptococcal strain. A high CO(2) atmosphere and a low iron concentration have been used classically to increase capsule size. Unfortunately, these stimuli are not reliable for inducing capsular enlargement in all strains. Recently we have identified new and simpler conditions for inducing capsule enlargement that consistently elicited this effect. Specifically, we noted that mammalian serum or diluted Sabouraud broth in MOPS buffer pH 7.3 efficiently induced capsule growth. Media that slowed the growth rate of the yeast correlated with an increase in capsule size. Finally, we summarize the most commonly used media that induce capsule growth in C. neoformans

Synthesis of MgB4O7:Dy3+and Thermoluminescent Characteristics at Low Doses of Beta Radiation

The synthesis and thermoluminescent characteristics of dysprosium-doped MgB4O7 are analyzed. The phosphor at different concentrations (0, 0.1, 0.5, 1, 2 and 4 mol%) of the dopant was prepared by the solution-assisted method. The magnesium borate compound was confirmed by X-ray diffraction. The annealing and dopant concentrations effects on the crystalline matrix were investigated. The highest thermoluminescent sensitivity was found with 450°C of annealing temperature and at high Dy3+ concentration too. The un-doped MgB4O7 phosphor shows a broad glow curve which peaked at 199°C and about 306 °C. Introducing Dy3+ dopant in the matrix that behavior was strongly changed. The wide glow curve shows three glow peaks; two small shoulders at 124 and 195 °C, and a highest peak between 323 and 336 °C temperature range. A large linear dose-response (5 – 2000 mGy) beta dose was obtained. The complex glow curves were deconvolved and the kinetics parameters were determined considering the general order kinetics model

Phospholipids Trigger Cryptococcus neoformans Capsular Enlargement during Interactions with Amoebae and Macrophages

A remarkable aspect of the interaction of Cryptococcus neoformans with mammalian hosts is a consistent increase in capsule volume. Given that many aspects of the interaction of C. neoformans with macrophages are also observed with amoebae, we hypothesized that the capsule enlargement phenomenon also had a protozoan parallel. Incubation of C. neoformans with Acanthamoeba castellanii resulted in C. neoformans capsular enlargement. The phenomenon required contact between fungal and protozoan cells but did not require amoeba viability. Analysis of amoebae extracts showed that the likely stimuli for capsule enlargement were protozoan polar lipids. Extracts from macrophages and mammalian serum also triggered cryptococcal capsular enlargement. C. neoformans capsule enlargement required expression of fungal phospholipase B, but not phospholipase C. Purified phospholipids, in particular, phosphatidylcholine, and derived molecules triggered capsular enlargement with the subsequent formation of giant cells. These results implicate phospholipids as a trigger for both C. neoformans capsule enlargement in vivo and exopolysaccharide production. The observation that the incubation of C. neoformans with phospholipids led to the formation of giant cells provides the means to generate these enigmatic cells in vitro. Protozoan- or mammalian-derived polar lipids could represent a danger signal for C. neoformans that triggers capsular enlargement as a non-specific defense mechanism against potential predatory cells. Hence, phospholipids are the first host-derived molecules identified to trigger capsular enlargement. The parallels apparent in the capsular response of C. neoformans to both amoebae and macrophages provide additional support for the notion that certain aspects of cryptococcal virulence emerged as a consequence of environmental interactions with other microorganisms such as protists

Molecular Mechanism and Potential Targets for Blocking HPV-Induced Lesion Development

Persistent infection with high-risk HPV is the etiologic agent associated with the development of cervical cancer (CC) development. However, environmental, social, epidemiological, genetic, and host factors may have a joint influence on the risk of disease progression. Cervical lesions caused by HPV infection can be removed naturally by the host immune response and only a small percentage may progress to cancer; thus, the immune response is essential for the control of precursor lesions and CC. We present a review of recent research on the molecular mechanisms that allow HPV-infected cells to evade immune surveillance and potential targets of molecular therapy to inhibit tumor immune escape

Analysis of microsatellite markers in a Cuban water buffalo breed

The aim of this Regional Research Communication was to validate a panel of 30 microsatellite markers recommended by FAO/ISAG for studies of biodiversity in cattle to improve the characterisation of Cuban buffalo populations. The water buffalo (Bubalus bubalis) is an economically important livestock species. Therefore, research focused on the study of the genetic relationships among water buffalo populations is useful to support conservation decisions and to design breeding schemes. Twenty-eight of the 30 tested regions were amplified, one of which (ETH10) turned out to be monomorphic. A total of 143 alleles were observed in the Cuban water buffalo population. The average number of alleles per locus was 5·04. The number of alleles per polymorphic locus ranged from two (INRA 63 and MM12) to nine (ETH185). The observed and expected heterozygosity ranged from 0·108 (HAUT24) to 0·851 (CSSM66) and 0·104 (MM12) to 0·829(INRA32), respectively. The polymorphic information content (PIC) ranged from 0·097 (MM12) to 0·806 (INRA32), and the overall value for these markers was 0·482. Within the population, inbreeding estimates (F IS) was positive in 14 of the 30 loci analysed. This study thus highlights the usefulness of heterologous bovine microsatellite markers to assess the genetic variability in Cuban water buffalo breeds. Furthermore, the results can be utilised for future breeding strategies and conservation

Ecological niche modeling to calculate ideal sites to introduce a natural enemy: the case of apanteles opuntiarum (hymenoptera: Braconidae) to control cactoblastis cactorum (lepidoptera: Pyralidae) in North America

The cactus moth, Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae), is an invasive species in North America where it threatens Opuntia native populations. The insect is expanding its distribution along the United States Gulf Coast. In the search for alternative strategies to reduce its impact, the introduction of a natural enemy, Apanteles opuntiarum Martínez and Berta (Hymenoptera: Braconidae), is being pursued as a biological control option. To identify promising areas to intentionally introduce A. opuntiarum for the control of C. cactorum, we estimated the overlap of fundamental ecological niches of the two species to predict their common geographic distributions using the BAM diagram. Models were based on native distributional data for both species, 19 bioclimatic variables, and the Maxent algorithm to calculate the environmental suitability of both species in North America. The environmental suitability of C. cactorum in North America was projected from Florida to Texas (United States) along the Gulf coastal areas, reaching Mexico in northern regions. Apanteles opuntiarum environmental suitability showed a substantial similarity with the calculations for C. cactorum in the United States. Intentional introductions of A. opuntiarum in the actual distribution areas of the cactus moth are predicted to be successful; A. opuntiarum will find its host in an environment conducive to its survival and dispersal.Fil: Pérez De la O, Nidia Bélgica. Universidad Autónoma de Chiapas. Facultad de Ciencias Agrícolas; MéxicoFil: Espinosa Zaragoza, Saúl. Universidad Autónoma de Chiapas. Facultad de Ciencias Agrícolas; MéxicoFil: López Martínez, Víctor. Universidad Autónoma del Estado de Morelos; MéxicoFil: Hight, Stephen D.. US Department of Agriculture. Agricultural Research Service; Estados UnidosFil: Varone, Laura. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fundación para el Estudio de Especies Invasivas; Argentin

Cryptococcus neoformans Capsular Enlargement and Cellular Gigantism during Galleria mellonella Infection

We have studied infection of Cryptococcus neoformans in the non-vertebrate host Galleria mellonella with particular interest in the morphological response of the yeast. Inoculation of C. neoformans in caterpillars induced a capsule-independent increase in haemocyte density 2 h after infection. C. neoformans manifested a significant increase in capsule size after inoculation into the caterpillar. The magnitude of capsule increase depended on the temperature, being more pronounced at 37°C than at 30°C, which correlated with an increased virulence of the fungus and reduced phagocytosis at 37°C. Capsule enlargement impaired phagocytosis by haemocytes. Incubation of the yeast in G. mellonella extracts also resulted in capsule enlargement, with the polar lipidic fraction having a prominent role in this effect. During infection, the capsule decreased in permeability. A low proportion of the cells (<5%) recovered from caterpillars measured more than 30 µm and were considered giant cells. Giant cells recovered from mice were able to kill the caterpillars in a manner similar to regular cells obtained from in vivo or grown in vitro, establishing their capacity to cause disease. Our results indicate that the morphological transitions exhibited by C. neoformans in mammals also occur in a non-vertebrate host system. The similarities in morphological transitions observed in different animal hosts and in their triggers are consistent with the hypothesis that the cell body and capsular responses represent an adaptation of environmental survival strategies to pathogenesis