Epitope mapping of antibodies to alpha-synuclein in LRRK2 mutation carriers, idiopathic Parkinson disease patients, and healthy controls

Alpha-synuclein (Snca) plays a major role in Parkinson disease (PD). Circulating anti-Snca antibodies has been described in PD patients and healthy controls, but they have been poorly characterized. This study was designed to assess the prevalence of anti-Snca reactivity in human subjects carrying the LRRK2 mutation, idiopathic PD (iPD) patients, and healthy controls and to map the epitopes of the anti-Snca antibodies. Antibodies to Snca were detected by ELISA and immunoblotting using purified recombinant Snca in plasma from individuals carrying LRRK2 mutations (104), iPD patients (59), and healthy controls (83). Epitopes of antibodies were mapped using recombinant protein constructs comprising different regions of Snca. Clear positive anti-Snca reactivity showed no correlation with age, sex, years of evolution, or the disability scores for PD patients and anti-Snca reactivity was not prevalent in human patients with other neurological or autoimmune diseases. Thirteen of the positive individuals were carriers of LRRK2 mutations either non-manifesting (8 out 49 screened) or manifesting (5 positive out 55), three positive (out of 59) were iPD patients, and five positive (out of 83) were healthy controls. Epitope mapping showed that antibodies against the N-terminal (a.a. 1-60) or C-terminal (a.a. 109-140) regions of Snca predominate in LRRK2 mutation carriers and iPD patients, being N122 a critical amino acid for recognition by the anti-C-terminal directed antibodies. Anti-Snca circulating antibodies seem to cluster within families carrying the LRRK2 mutation indicating possible genetic or common environmental factors in the generation of anti-Snca antibodies. These results suggest that case-controls' studies are insufficient and further studies in family cohorts of patients and healthy controls should be undertaken, to progress in the understanding of the possible relationship of anti-Snca antibodies and PD patholog

National Fisheries Laboratory Category 2 parasite guide

This report provides a guide into Category 2 parasites affecting freshwater fish and salmonids. First a brief summary is given of distinctions between parasites of Category 1 and 2. The Guide then provides a list of category 2 parasites, highlighting damage they can cause, species of fish affected, if it can be treated, how widespread the parasite is and how it is transferred.

Lipid Nanoparticles as Carriers for RNAi against Viral Infections: Current Status and Future Perspectives

The efforts made to develop RNAi-based therapies have led to productive research in the field of infections in humans, such as hepatitis C virus (HCV), hepatitis B virus (HBV), human immunodeficiency virus (HIV), human cytomegalovirus (HCMV), herpetic keratitis, human papillomavirus, or influenza virus. Naked RNAi molecules are rapidly digested by nucleases in the serum, and due to their negative surface charge, entry into the cell cytoplasm is also hampered, which makes necessary the use of delivery systems to exploit the full potential of RNAi therapeutics. Lipid nanoparticles (LNP) represent one of the most widely used delivery systems for in vivo application of RNAi due to their relative safety and simplicity of production, joint with the enhanced payload and protection of encapsulated RNAs. Moreover, LNP may be functionalized to reach target cells, and they may be used to combine RNAi molecules with conventional drug substances to reduce resistance or improve efficiency. This review features the current application of LNP in RNAi mediated therapy against viral infections and aims to explore possible future lines of action in this field.

LDLR and PCSK9 Are Associated with the Presence of Antiphospholipid Antibodies and the Development of Thrombosis in aPLA Carriers

Introduction The identification of the genetic risk factors that could discriminate non-thrombotic from thrombotic antiphospholipid antibodies (aPLA) carriers will improve prognosis of these patients. Several human studies have shown the presence of aPLAs associated with atherosclerotic plaque, which is a known risk factor for thrombosis. Hence, in order to determine the implication of atherosclerosis in the risk of developing thrombosis in aPLA positive patients, we performed a genetic association study with 3 candidate genes, APOH, LDLR and PCSK9. Material & Methods For genetic association study we analyzed 190 aPLA carriers -100 with non-thrombotic events and 90 with thrombotic events-and 557 healthy controls. Analyses were performed by chi(2) test and were corrected by false discovery rate. To evaluate the functional implication of the newly established susceptibility loci, we performed expression analyses in 86 aPLA carrier individuals (43 with thrombotic manifestations and 43 without it) and in 45 healthy controls. Results Our results revealed significant associations after correction in SNPs located in LDLR gene with aPLA carriers and thrombotic aPLA carriers, when compared with healthy controls. The most significant association in LDLR gene was found between SNP rs129083082 and aPLA carriers in recessive model (adjusted P-value = 2.55 x 10(-3); OR = 2.18; 95% CI = 1.49-3.21). Furthermore, our work detected significant allelic association after correction between thrombotic aPLA carriers and healthy controls in SNP rs562556 located in PCSK9 gene (adjusted P-value = 1.03 x 10(-2); OR = 1.60; 95% CI = 1.24-2.06). Expression level study showed significantly decreased expression level of LDLR gene in aPLA carriers (P-value < 0.0001; 95% CI 0.16-2.10; SE 0.38-1.27) in comparison to the control group. Discussion Our work has identified LDLR gene as a new susceptibility gene associated with the development of thrombosis in aPLA carriers, describing for the first time the deregulation of LDLR expression in individuals with aPLAs. Besides, thrombotic aPLA carriers also showed significant association with PCSK9 gene, a regulator of LDLR plasma levels. These results highlight the importance of atherosclerotic processes in the development of thrombosis in patients with aPLA.

Estudo do metabolismo mitocondrial e da resposta anti-apoptótica de células endoteliais humanas durante a evolução da infecção por taquizoítos de Toxoplasma gondii

A toxoplasmose é uma zoonose amplamente distribuída que afeta mais de um terço da população mundial e de grande importância na saúde pública. A maioria das infecções em humanos por Toxoplasma gondii é assintomática. A toxoplasmose é amplamente investigada visto que se apresenta como uma doença grave em pessoas imunodeprimidas (portadores da síndrome da imunodeficiência adquirida (SIDA), não tratados, indivíduos transplantados, paciente em tratamento quimioterápico ou em uso de drogas supressoras e gestantes). A toxoplasmose congênita frequentemente pode levar ao aborto espontâneo ou até mesmo resultar na formação de crianças com algum grau de atraso no desenvolvimento mental e/ou físicos, deste modo, a transmissão congênita pode ser muito mais importante do que se pensava, pois os parasitos encontrados na circulação sanguinea são capazes de infectar as células endoteliais dos vasos e os tecidos circunjacentes, podendo resultar no encistamento do T. gondii. Atualmente a toxoplasmose vem sendo investigada devido a sua associação a inúmeras outras doenças, assim, estudos sobre a evolução da infecção por T. gondii em diferentes tipos de células hospedeiras se fazem necessários para uma abordagem terapêutica adequada. Ao invadir a célula hospedeira o parasito possui a capacidade de recrutar as mitocôndrias promovendo mudanças na organização mitocondrial ao longo da progressão da infecção, garantindo um ambiente favorável a sua multiplicação. Diante disso, investigamos se o parasito possui a capacidade de interferir no metabolismo mitocondrial e na resposta apoptótica da célula endotelial. O presente trabalho teve como objetivo analisar o metabolismo mitocondrial através da respirometria de alta-resolução e da resposta apoptótica através do western blotting das células endoteliais da veia umbilical humana (HUVEC) infectadas por 2, 6 e 20 horas por taquizoítos de T. gondii. A respirometria de alta-resolução revelou que o parasito interfere no metabolismo energético da célula hospedeira. A análise do conteúdo de proteínas da família Bcl-2 por western blotting revelou maior estímulo apoptótico no tempo inicial de infecção, quando comparado aos demais tempos. Os resultados dos conteúdos de caspase 3, proteína efetora da apoptose, não demonstrou diferença nos tempos iniciais de infecção Entretanto, em tempos mais tardios, o conteúdo de caspase 3 mostrou-se significativamente aumentado quando comparado às HUVEC não infectadas. A dinâmica de replicação do parasito foi observada através do monitoramento pelo sistema Time-Lapse Nikon BioStation IMQ em tempo real das células infectadas por T.gondii. Portanto, nossos resultados sugerem que o protozoário ao recrutar as mitocôndrias da célula hospedeira interfere no metabolismo mitocondrial e na modulação da apoptose para garantir um ambiente favorável a sua multiplicação.

A record of Probopyrus bithynis (Richardson, 1904) in Macrobrachium rosenbergii (De Man) from coastal Andhra Pradesh, India, with special reference to host-parasite relationship

An incidence of bopyrid isopod infestation was observed in giant freshwater prawn, Macrobrachium rosenbergii (de Man) juveniles (40-60 mm/0.9-1.5 g) in a scampi culture farm in East Godavari district of Andhra Pradesh. The presence of parasite was observed by conspicuous boil like swelling of the branchial chamber where the parasite was found lodged on the gills. The infested gill was highly compressed and necrosed. Only one branchial chamber was infested by the parasite while the other gill was normal. The infested prawns were thin and emaciated and showed retarded growth. The parasite was identified as Probopyrus bithynis (Richardson, 1904) which caused inhibition of ventilation due to its permanent lodging in the branchial chamber and impaired the gaseous exchange by gills. It was also observed that this parasite caused parasitic castration in the infested prawns.

Dynamics of parasite population and its histopathological and histophysiological effects in the stomach of a freshwater fish

The caryophyllaeid cestode Lytocestoides fossilis infects the freshwater catfish Heteropneustes fossilis. The study was conducted for two consecutive years (2004-06) to record the bio-statistical data of the parasite. The incidence, intensity, density and index of infection of the parasite have been recorded. The infection was more during June to September, moderate during February to May and low during October to January. The parasite brought about severe histopathological changes in the stomach of infected fish. The changes observed in the stomach of fish included structural damage of the villi, inflammation, and fibrosis associated with hyperplasia and metaplasia. The hypertrophy of mucous layer led to vacuolation and necrosis. Histochemical changes were noticed with enhanced carbohydrate, protein and lipid contents. The enhanced substrate content in the infected organ might be due to the disfunctioning of the digestive tract, which results in the accumulation of various metabolites. Mucus secretion was triggered as a protective interaction against parasitic invasion. The parasitic infection affects the general metabolic state of the host and as the result, the fish becomes sluggish and moribund.

A suctorean parasite of Penaeus monodon larvae

A pathogenic suctorean, identified as Ephelota gemmipara was observed in P. monodon larvae spawned and reared in tanks. Commonly found to inhabit hydroid colonies, E. gemmipara has a stalked body with two types of tentacles, the sucking and piercing types, and was observed to reproduce by multiple exogenous budding.

Lamproglena notopterai sp. nov.- a copepod parasite from a freshwater fish of Sindh, Pakistan

The study was conducted at the Central Experimental Station, Philippine Rice Research Institute, Maligaya, Science City of Munoz, Nueva Ecija, Philippines during the wet season to determine the suitable stocking density(s) for better growth and yield of fish under rice-fish production systems. Recovery rate of GIFT tilapia in different stocking densities ranged from 75.74 to 83.47%. Among different treatments, rice +5,000 fingerlings/ha and rice +10,000 fingerlings/ha resulted in the highest recovery rate of 83.33% and 83.47%, respectively. The lowest recovery rate of 75.75% was obtained from rice +20,000 fingerlings/ha, but similar to that was obtained (78.56%) from rice +15,000 fingerlings/ha. Significantly higher rate of gain in body weight and that of specific growth rate were recorded in the treatment from rice +5,000 fingerlings/ha, while other treatments resulted in similar absolute and specific growth rate. Fish yield increased significantly with relatively higher stocking densities, but higher densities produced maximum number of smaller fishes and also lower recovery rate.