Surface-Associated Plasminogen Binding of Cryptococcus neoformans Promotes Extracellular Matrix Invasion

BACKGROUND:The fungal pathogen Cryptococcus neoformans is a leading cause of illness and death in persons with predisposing factors, including: malignancies, solid organ transplants, and corticosteroid use. C. neoformans is ubiquitous in the environment and enters into the lungs via inhalation, where it can disseminate through the bloodstream and penetrate the central nervous system (CNS), resulting in a difficult to treat and often-fatal infection of the brain, called meningoencephalitis. Plasminogen is a highly abundant protein found in the plasma component of blood and is necessary for the degradation of fibrin, collagen, and other structural components of tissues. This fibrinolytic system is utilized by cancer cells during metastasis and several pathogenic species of bacteria have been found to manipulate the host plasminogen system to facilitate invasion of tissues during infection by modifying the activation of this process through the binding of plasminogen at their surface. METHODOLOGY:The invasion of the brain and the central nervous system by penetration of the protective blood-brain barrier is a prerequisite to the establishment of meningoencephalitis by the opportunistic fungal pathogen C. neoformans. In this study, we examined the ability of C. neoformans to subvert the host plasminogen system to facilitate tissue barrier invasion. Through a combination of biochemical, cell biology, and proteomic approaches, we have shown that C. neoformans utilizes the host plasminogen system to cross tissue barriers, providing support for the hypothesis that plasminogen-binding may contribute to the invasion of the blood-brain barrier by penetration of the brain endothelial cells and underlying matrix. In addition, we have identified the cell wall-associated proteins that serve as plasminogen receptors and characterized both the plasminogen-binding and plasmin-activation potential for this significant human pathogen. CONCLUSIONS:The results of this study provide evidence for the cooperative role of multiple virulence determinants in C. neoformans pathogenesis and suggest new avenues for the development of anti-infective agents in the prevention of fungal tissue invasion

How insects survive the cold: molecular mechanisms - a review

Insects vary considerably in their ability to survive low temperatures. The tractability of these organisms to experimentation has lead to considerable physiology-based work investigating both the variability between species and the actual mechanisms themselves. This has highlighted a range of strategies including freeze tolerance, freeze avoidance, protective dehydration and rapid cold hardening, which are often associated with the production of specific chemicals such as antifreezes and polyol cryoprotectants. But we are still far from identifying the critical elements behind over-wintering success and how some species can regularly survive temperatures below -20°C. Molecular biology is the most recent tool to be added to the insect physiologist’s armoury. With the public availability of the genome sequence of model insects such as Drosophila and the production of custom-made molecular resources, such as EST libraries and microarrays, we are now in a position to start dissecting the molecular mechanisms behind some of these well-characterised physiological responses. This review aims to provide a state of the art snapshot of the molecular work currently being conducted into insect cold tolerance and the very interesting preliminary results from such studies, which provide great promise for the future

Antagonistic control of fluid secretion by the Malpighian tubules of<i>Tenebrio molitor</i>: effects of diuretic and antidiuretic peptides and their second messengers

SUMMARYFluid secretion by insect Malpighian tubules is controlled by haemolymph-borne factors. The mealworm Tenebrio molitor provides the first known example of antagonistic interactions between endogenous neuropeptides acting on Malpighian tubules. The two corticotropin-releasing-factor (CRF)-related diuretic peptides previously isolated from Tenebrio molitor, Tenmo-DH37 and Tenmo-DH47, were found to stimulate Tenebrio molitor tubules in vitro in a dose-dependent manner with EC50 values of 0.12 nmol l–1 and 26 nmol l–1 respectively. However, no synergistic or additive effect was observed when these two peptides were tested simultaneously. We then investigated antagonism between second messengers: dose–response curves were constructed for stimulation of Tenebrio molitor tubules by cyclic AMP and their inhibition by cyclic GMP. When both cyclic nucleotides were included in the bathing Ringer, the stimulatory effect of cyclic AMP was neutralised by cyclic GMP. Similarly, the stimulatory effect of Tenmo-DH37 was reversed on addition of an antidiuretic peptide (Tenmo-ADF), which was recently isolated from Tenebrio molitor and acts via cyclic GMP. The cardioacceleratory peptide CAP2b, originally isolated from Manduca sexta, also increases intracellular cyclic GMP levels and inhibited fluid secretion by Tenebrio molitor tubules, with an EC50 value of 85 nmol l–1. This inhibitory effect was reversed by Tenmo-DH37. Endogenous diuretic and antidiuretic peptides, effective at low concentrations and acting via antagonistic second messengers, have the potential for fine control of secretion rates in the Malpighian tubules of Tenebrio molitor.</jats:p

Estudo comparativo de éguas repetidoras ou não de cio através da avaliação histológica do endométrio e das concentrações plasmáticas de progesterona

O trabalho teve como objetivo geral fazer um estudo comparativo da histologia endometrial e das concentrações plasmáticas de progesterona (P4) em éguas, repetidoras ou não de cio. A hipótese do presente estudo é que existe correlação entre o histórico de infertilidade, os achados histopatológicos e a concentração de P4 destes animais. Para tanto, foram utilizadas 36 éguas, em idade reprodutiva (3-23 anos), das raças Mangalarga Marchador e Campolina. Utilizaram-se éguas não repetidoras (n=11) como grupo controle, sendo 4 éguas doadoras e 7 receptoras; e repetidoras de cio (n=25), 15 doadoras e 10 receptoras. Tal classificação foi realizada de acordo com o histórico reprodutivo destes animais. Amostras de endométrio foram coletadas para a realização de avaliação histopatológica e amostras de sangue para a mensuração das concentrações plasmáticas de progesterona. Os fragmentos de tecido endometrial obtidos pela biópsia foram fixados no Fixador de Bouin. Posteriormente os fragmentos foram processados e incluídos em parafina. Os cortes foram corados por Hematoxilina-Eosina (HE) para exame histopatológico. As concentrações plasmáticas de P4 foram mensuradas pelo método de enzima-imunoensaio (ELISA). Não foi observada correlação entre as concentrações de P4 e a subfertilidade, sendo estas variáveis tratadas como independentes. Houve uma correlação positiva entre a idade e a subfertilidade (p<0.05), assim, quanto mais velha a égua, maior a subfertilidade. Foi observada uma correlação entre a categoria da biópsia e a subfertilidade (p<0.05), ou seja, quanto mais alterações histológicas, maiores as chances da égua ser subfértil. Contudo, nem todas as éguas classificadas na Categoria I e II levaram a gestação a termo e nem todas da Categoria III tiveram falhas na reprodução, pois, deve-se considerar que outros fatores podem influenciar na manutenção da gestação. A biópsia endometrial demonstrou ser uma técnica fácil, segura, barata e com um desconforto mínimo para o animal. Sugere-se que esta técnica, juntamente com outros dados, constitui uma importante ferramenta para a avaliação da fertilidade da égua

Zygomycotic Lymphadenitis in Slaughtered Feedlot Cattle

During the 12 months of 2006, zygomycotic lymphadenitis was diagnosed in 194 of 198 feedlot steers (0.04% of cattle slaughtered during that period) in a California slaughterhouse as part of bovine tuberculosis surveillance. Mesenteric lymph nodes were involved in 190 cases. Affected lymph nodes were enlarged (2 to 42 cm in greatest dimension), firm, and mottled gray-white to yellow with multiple granular or caseocalcareous foci. Histologically, nodal architecture was effaced by necrosis, granulomatous inflammation, and fibrosis. In approximately 20% of the cases, granulomas were mainly restricted to subcapsular sinuses and afferent lymphatic vessels, causing granulomatous lymphangitis. Nonseptate, irregularly branching hyphae with nonparallel walls and bulbous enlargements were common in necrotic areas and within the cytoplasm of multinucleated giant cells. Fungal cultures were performed on 124 affected lymph nodes using 7 media, but no zygomycetes were cultured. Fungal DNA was amplified from 20 lymph nodes. Amplicons from 16 nodes had nearly 100% homology with sequences for Rhizomucor pusillus; 4 amplicons had (&gt; 98%) homology with Absidia corymbifera sequences. Zygomycosis should be considered in the differential diagnosis for granulomatous lymphadenitis in feedlot steers. </jats:p