Human Babesiosis, Bolivia, 2013

To investigate human babesiosis in the Bolivian Chaco, in 2013 we tested blood samples from 271 healthy persons living in 2 rural communities in this region. Microscopy and PCR indicated that 3.3% of persons were positive for Babesia microti parasites (US lineage); seroprevalence was 45.7%. Appropriate screening should mitigate the risk for transfusion-associated babesiosis

Rat Brain Cortex Mitochondria Release Group II Secretory Phospholipase A2 under Reduced Membrane Potential

Activation of brain mitochondrial phospholipase(s) A(2) (PLA(2)) might contribute to cell damage and be involved in neurodegeneration. Despite the potential importance of the phenomenon, the number, identities, and properties of these enzymes are still unknown. Here, we demonstrate that isolated mitochondria from rat brain cortex, incubated in the absence of respiratory substrates, release a Ca(2+)-dependent PLA(2) having biochemical properties characteristic to secreted PLA(2) (sPLA(2)) and immunoreacting with the antibody raised against recombinant type IIA sPLA(2) (sPLA(2)-IIA). Under identical conditions, no release of fumarase in the extramitochondrial medium was observed. The release of sPLA(2) from mitochondria decreases when mitochondria are incubated in the presence of respiratory substrates such as ADP, malate, and pyruvate, which causes an increase of transmembrane potential determined by cytofluorimetric analysis using DiOC(6)(3) as a probe. The treatment of mitochondria with the uncoupler carbonyl cyanide 3-chlorophenylhydrazone slightly enhances sPLA(2) release. The increase of sPLA(2) specific activity after removal of mitochondrial outer membrane indicates that the enzyme is associated with mitoplasts. The mitochondrial localization of the enzyme has been confirmed by electron microscopy in U-251 astrocytoma cells and by confocal laser microscopy in the same cells and in PC-12 cells, where the structurally similar isoform type V-sPLA(2) has mainly nuclear localization. In addition to sPLA(2), mitochondria contain another phospholipase A(2) that is Ca(2+)-independent and sensitive to bromoenol lactone, associated with the outer mitochondrial membrane. We hypothesize that, under reduced respiratory rate, brain mitochondria release sPLA(2)-IIA that might contribute to cell damage

Long-Standing International Cooperation in Parasitology Research: A Summary of 35 Years of Activities in the Bolivian Chaco

The Bolivian Chaco is a semiarid region with a low population density, situated in the southeast part of the Plurinational State of Bolivia. Here, despite the improvements of the last 15 years, poverty remains high in rural areas, where social vulnerability is widespread. The Guaraní ethnic group often lives in isolated communities with a low standard of hygiene and sanitation. This epidemiological scenario favors the spread of transmissible diseases, including several parasitic infections belonging to the neglected tropical diseases (NTDs) group. In this area, a long-standing research activity, built upon the synergism between local and foreign institutions, has been established since the late 1980s and helps to fill in the knowledge gap about the epidemiology dynamics of soil-transmitted helminths, vector-borne parasites, and other parasitic diseases. A 35-year history of cooperation programs in parasitology research has contributed to informing local health authorities of the NTD burden in the Bolivian Chaco and, ultimately, supports local healthcare providers in the management of parasitic diseases

A Calsequestrin-1 mutation associated with a skeletal muscle disease alters sarcoplasmic Ca2+ release

An autosomal dominant protein aggregate myopathy, characterized by high plasma creatine kinase and calsequestrin-1 (CASQ1) accumulation in skeletal muscle, has been recently associated with a missense mutation in CASQ1 gene. The mutation replaces an evolutionarily-conserved aspartic acid with glycine at position 244 (p.D244G) of CASQ1, the main sarcoplasmic reticulum (SR) Ca2+ binding and storage protein localized at the terminal cisternae of skeletal muscle cells. Here, immunocytochemical analysis of myotubes, differentiated from muscle-derived primary myoblasts, shows that sarcoplasmic vacuolar aggregations positive for CASQ1 are significantly larger in CASQ1-mutated cells than control cells. A strong co-immuno staining of both RyR1 and CASQ1 was also noted in the vacuoles of myotubes and muscle biopsies derived from patients. Electrophysiological recordings and sarcoplasmic Ca2+ measurements provide evidence for less Ca2+ release from the SR of mutated myotubes when compared to that of controls. These findings further clarify the pathogenic nature of the p.D244G variant and point out defects in sarcoplasmic Ca2+ homeostasis as a mechanism underlying this human disease, which could be distinctly classified as "CASQ1-couplonopathy".peer-reviewe

Evaluation of the gene expression and Delta 6-desaturase activity in different chicken strains.

In chickens, body fatty acids (FA) are derived from dietary uptake, de novo synthesis and/or bioconversion. Among the different fatty acid classes, n-3 long-chain PUFA (n-3 LC-PUFA) are of particular interest due to their positive role on human health. Domestic animals are unable to synthesize long chain-PUFA, but they can convert dietary linoleic and \u3b1-linolenic acid through a pathway catalyzed by elongation and desaturation enzymes. Meat from animal species is characterized by different FA composition; within the same species the FA profile reflects the endogenous biosynthesis as well as the composition of the diet. The relatively low efficacy of the desaturation enzymes in poultry allows for further consideration on the selection of genotypes able to synthesize a higher PUFA amount. In this study, the liver messenger RNA copies and enzyme activity of the \u3946-desaturase were evaluated in three chicken strains (slow-SG, fast-growing- FG and SG x FG crossing-SFG). Three groups of laying hens for each genotype were fed with a standard diet. Chickens were slaughtered at hatch, and liver was taken for analysis. A relationship between genotype and desaturation ability was evidenced. The FG strain showed a lower mRNA copies of FADS2 gene than the SG one (P>0.01). However, RNA expression was two or three-hundred times higher in SFG than SG and FG strains, respectively (P<0.01). Even the enzyme activity confirmed what shown by mRNA. The \u3946-desaturase activity was significantly higher in SG than FG (172.0 vs. 63.56 pmol in 30 min/mg prot), whereas in SFG was at intermediate level (136.66 pmol in 30min/mg prot). However, the enzyme activity of SFG was not significantly different from SG (P>0.01). SG chicken in comparison to FG strain showed a higher desaturase activity (P<0.05), whereas there were no significantly differences between \u3946- desaturase activity in SG and SFG. Even mRNA copy number was widely affected by genotype. Indeed, SFG showed a much higher mRNA abundance, whereas SG does not show significant differences. Data herein reported showed that the \u3946-destaurase activity is strongly affected by genotype. However, several other factors involved in the process of expression/translation, can be assessed. Gene expression may follow certain design principles for optimal evolutionary fitness as demonstrated by the high mRNA copies of the SFG

Botulinum-A toxin injections into the detrusor muscle decrease nerve growth factor bladder tissue levels in patients with neurogenic detrusor overactivity

Purpose: We investigated the effects of BTX-A on visceral afferent nerve transmission by measuring bladder tissue NGF levels in patients with neurogenic detrusor overactivity before and after intravesical treatment with BTX-A. We also compared the bladder tissue NGF content with clinical and urodynamic data. Materials and Methods: A total of 23 patients underwent clinical evaluation and urodynamics with detection of the UDC threshold, maximum pressure and maximum cystometric capacity before, and at the 1 and 3-month followups. Endoscopic bladder wall biopsies were also obtained at the same time points. NGF levels were measured in tissue homogenate by enzyme-linked immunosorbent assay (Promega, Madison, Wisconsin). Results: At 1 and 3 months mean catheterization and incontinent episodes were significantly decreased (p ��0.05 and ��0.001, respectively). On urodynamics we detected a significant increase in the UDC threshold and maximum cystometric capacity, and a significant decrease in UDC maximum pressure at the 1 and 3-month followups compared to baseline (each p ��0.001). At the same time points we detected a significant decrease in NGF bladder tissue content (each p ��0.02). Conclusions: BTX-A intravesical treatment induces a state of NGF deprivation in bladder tissue that persists at least up to 3 months. As caused by BTX-A, the decrease in acetylcholine release at the presynaptic level may induce a decrease in detrusor contractility and in NGF production by the detrusor muscle. Alternatively BTX-A can decrease the bladder level of neurotransmitters that normally modulate NGF production and release

FIRST DETECTION OF BABESIA MICROTI IN PEOPLE LIVING IN THE PLURINATIONAL STATE OF BOLIVIA

INTRODUCTION: Tick-transmitted hemoparasites of the genus Babesia (phylum Apicomplexa) show a worldwide distribution and have a wide spectrum of vertebrate hosts that includes, occasionally, humans (Gray et al, Ticks Tick Borne Dis, 2010, 1:3–10). Indeed, the cattle species B. divergens (and B. venatorum) in Europe and the rodent species B. microti (and B. duncani) in North America have been shown as responsible for most human infections. Babesiosis is a malaria-like illness that ranges from asymptomatic infection to an even fatal disease, associated to hemolytic anemia and severe systemic complications (Vannier et al., N Engl J Med, 2012, 366:2397–407). B. microti infection is generally mild, self-limiting or asymptomatic, but a fatal outcome may be occasionally observed. Deer, heavily infested by vector ticks that -at larval stage- feed on rodents, are important in the epidemiology of the infection (Telford et al., J Med Entomol, 1993, 30:223-7). In Bolivia, only B bovis and B. bigemina have been investigated in cattle and seem to be stably endemic in the Eastern part of the country (Carrique et al., Veterinary J, 2000, 160:162–4). No data are available about the presence of B. microti. MATERIALS AND METHODS: Research aims to investigate on human babesiosis in two rural communities of Southeastern Bolivia (Ivamirapinta and Bartolo), as part of a screening on hematic parasites carried out in 2013. In the study area, economy is based on agriculture and animal farming, and people live in close contact with pets and wild animals (deer included). Blood samples were taken from 273 healthy subjects (1-83 years-old) and used to arrange thick and thin smears, to extract DNAs, and to obtain sera. Smears were Giemsa stained and examined by microscopy. DNAs were amplified by PCR using a pair of generic apicomplexan 18S rRNA-specific primers. Positive amplicons were confirmed by sequencing. Several sera (from infected people and from older components of some families) were submitted to an IFI test available to detect B. microti infection in humans (B. microti-IFA IgG Antibody kit, Fuller Laboratories, Fullerton, CA). RESULTS: A total of 10 (3.7%) thin smears were positive to B. microti. Morphological diagnosis was confirmed by sequencing. As expected, preliminary serological results evidenced reactivity to B. microti antigens not only in all positive subjects (at >1:128 dilutions) but also in further 4/43 subjects negative to microscopy and PCR (at dilutions 1:64 or 1:128). CONCLUSIONS: This is the first record of human babesiosis in the Plurinational State of Bolivia, in areas where people is exposed to many other parasites that can induce severe anemia. More extensive studies are needed to improve knowledge about all zoonotic babesial species, and people should be alerted about sanitary risks originated by ticks. Finally, physicians should be aware of the presence of B. microti, and well trained on clinical features of babesiosis in order to intervene, when needed, with appropriate treatment

Crosstalk between Long-Term Sublethal Oxidative Stress and Detrimental Inflammation as Potential Drivers for Age-Related Retinal Degeneration

Age-related retinal degenerations, including age-related macular degeneration (AMD), are caused by the loss of retinal pigmented epithelial (RPE) cells and photoreceptors. The pathogenesis of AMD, deeply linked to the aging process, also involves oxidative stress and inflammatory responses. However, the molecular mechanisms contributing to the shift from healthy aging to AMD are still poorly understood. Since RPE cells in the retina are chronically exposed to a pro-oxidant microenvironment throughout life, we simulated in vivo conditions by growing ARPE-19 cells in the presence of 10 &mu;M H2O2 for several passages. This long-term oxidative insult induced senescence in ARPE-19 cells without affecting cell proliferation. Global proteomic analysis revealed a dysregulated expression in proteins involved in antioxidant response, mitochondrial homeostasis, and extracellular matrix organization. The analyses of mitochondrial functionality showed increased mitochondrial biogenesis and ATP generation and improved response to oxidative stress. The latter, however, was linked to nuclear factor-&kappa;B (NF-&kappa;B) rather than nuclear factor erythroid 2&ndash;related factor 2 (Nrf2) activation. NF-&kappa;B hyperactivation also resulted in increased pro-inflammatory cytokines expression and inflammasome activation. Moreover, in response to additional pro-inflammatory insults, senescent ARPE-19 cells underwent an exaggerated inflammatory reaction. Our results indicate senescence as an important link between chronic oxidative insult and detrimental chronic inflammation, with possible future repercussions for therapeutic interventions