<Emphasis Type="Italic">Ixodes </Emphasis>(<Emphasis Type="Italic">Ixodes</Emphasis>) <Emphasis Type="Italic">pararicinus</Emphasis> Keirans &; Clifford, 1985 (Acari: Ixodidae): description of the immature stages,distribution, hosts and medical/veterinary importance

The immature stages of the Neotropical tick Ixodes (Ixodes) pararicinus Keirans & Clifford, 1985 are described from specimens in a laboratory colony initiated from engorged females collected on cattle and larvae fed on mice and chickens. The larva and nymph of I. pararicinus are described using SEM micrographs as well as drawings for some features of the larva. Additionally, immature stages of I. pararicinus collected on wild mice and birds in Uruguay and Argentina were compared with specimens from the laboratory colony. A taxonomic key to the nymphs of the species of the ricinus complex established in the Western Hemisphere is presented. The distribution of I. pararicinus comprises Argentina, Colombia and Uruguay, but it is also probably established in Bolivia, Brazil, Chile and Peru. Most adult ticks of this species have been found on introduced domestic artiodactyles, although Neotropical deer species must have been the ancestral host. Larvae and nymphs of I. pararicinus have also been found on sigmodontine rodents and passeriform birds. Although I. pararicinus is a member of the ricinus complex, which contains the main vectors of Borrelia burgdorferi (sensu lato), there are few studies concerning its potential for pathogen transmission.     

Decrease in protein solubility and cataract formation caused by the Pro23 to Thr mutation in human gamma D-crystallin

The P23T mutation in the human gammaD-crystallin gene has in recent years been associated with a number of well known cataract phenotypes. To understand the molecular mechanism of lens opacity caused by this mutation, we expressed human gammaD-crystallin (HGD), the P23T mutant, and other related mutant proteins in Escherichia coli and compared the structures and thermodynamic properties of these proteins in vitro. The results show that the cataract-causing mutation P23T does not exhibit any significant structural change relative to the native protein. However, in marked contrast to the native protein, the mutant shows a dramatically lowered solubility. The reduced solubility results from the association of the P23T mutant to form a new condensed phase that contains clusters of the mutant protein. The monomer-cluster equilibrium is represented by a solubility curve in the phase diagram. When the solubility limit is exceeded, the mutant protein forms the condensed phase after a nucleation time of 10-20 min. We found that the solubility of the P23T mutant exhibits an inverse dependence on temperature, i.e., the protein clusters are increasingly soluble as the temperature of the solution decreases. The solubility of P23T can be substantially altered by the introduction of specific mutations at or in the immediate vicinity of residue 23. We examined the mutants P23S, P23V, P23TInsP24, and P23TN24K and found that the latter two mutations can restore the solubility of the P23T mutant. These findings may help develop a strategy for the rational design of small molecule inhibitors of this type of condensed phase.     

Effect of Family History of Type 2 Diabetes on White Blood Cell Count in Adult Women

Objective: To evaluate the effect of a first‐degree family history of type 2 diabetes on white blood cell (WBC) count, a risk factor for atherosclerotic vascular disease, in glucose‐tolerant adult women Research Methods and Procedures: WBC count was measured in 174 normal weight, overweight, and obese female offspring of type 2 diabetic patients (FH⁺) and 174 age‐ and BMI‐matched female controls with no family history of type 2 diabetes (FH^?). Other measurements included fat mass (FM), measured by body impedance analysis; central fat accumulation, evaluated by waist circumference; insulin resistance, estimated by homeostatic model assessment for insulin resistance (HOMA_IR); systolic and diastolic blood pressure; and fasting concentrations of glucose, insulin, and lipids. Results: WBC count, waist circumference, systolic blood pressure, and fasting levels of glucose, insulin, and triglycerides were significantly higher in FH⁺ than in FH^? subjects. In FH⁺ individuals, WBC count was positively associated with BMI, FM, waist circumference, HOMA_IR, and triglyceride and insulin concentrations, and negatively correlated with age and high‐density lipoprotein‐cholesterol. In FH^? subjects, WBC count was directly associated with BMI, FM, waist circumference, and triglyceride and insulin concentrations, and inversely correlated with age and high‐density lipoprotein‐cholesterol. After multivariate analyses, WBC count maintained a significant association with age, systolic blood pressure, and HOMA_IR in FH⁺ subjects and with age, BMI, FM, and triglycerides in FH^? individuals. Discussion: This study indicates that WBC count is increased in adult women with genetic predisposition to type 2 diabetes, and its main correlates are insulin resistance in FH⁺ and adiposity in FH^? individuals.     

Trypanosoma cruzi Infection Is a Potent Risk Factor for Non-alcoholic Steatohepatitis Enhancing Local and Systemic Inflammation Associated with Strong Oxidative Stress and Metabolic Disorders

Background

The immune mechanisms underlying experimental non-alcoholic steatohepatitis (NASH), and more interestingly, the effect of T. cruzi chronic infection on the pathogenesis of this metabolic disorder are not completely understood.

Methodology/Principal Findings

We evaluated immunological parameters in male C57BL/6 wild type and TLR4 deficient mice fed with a standard, low fat diet, LFD (3% fat) as control group, or a medium fat diet, MFD (14% fat) in order to induce NASH, or mice infected intraperitoneally with 100 blood-derived trypomastigotes of Tulahuen strain and also fed with LFD (I+LFD) or MFD (I+MFD) for 24 weeks. We demonstrated that MFD by itself was able to induce NASH in WT mice and that parasitic infection induced marked metabolic changes with reduction of body weight and steatosis revealed by histological studies. The I+MFD group also improved insulin resistance, demonstrated by homeostasis model assessment of insulin resistance (HOMA-IR) analysis; although parasitic infection increased the triglycerides and cholesterol plasma levels. In addition, hepatic M1 inflammatory macrophages and cytotoxic T cells showed intracellular inflammatory cytokines which were associated with high levels of IL6, IFNγ and IL17 plasmatic cytokines and CCL2 chemokine. These findings correlated with an increase in hepatic parasite load in I+MFD group demonstrated by qPCR assays. The recruitment of hepatic B lymphocytes, NK and dendritic cells was enhanced by MFD, and it was intensified by parasitic infection. These results were TLR4 signaling dependent. Flow cytometry and confocal microscopy analysis demonstrated that the reactive oxygen species and peroxinitrites produced by liver inflammatory leukocytes of MFD group were also exacerbated by parasitic infection in our NASH model.

Conclusions

We highlight that a medium fat diet by itself is able to induce steatohepatitis. Our results also suggest a synergic effect between damage associated with molecular patterns generated during NASH and parasitic infection, revealing an intense cross-talk between metabolically active tissues, such as the liver, and the immune system. Thus, T. cruzi infection must be considered as an additional risk factor since exacerbates the inflammation and accelerates the development of hepatic injury.     

Isolation of protoplasts from in vitro-grown quince BA29 leaves

Summary The aim of this research was to isolate protoplasts from in vitro-grown quince BA 29 leaves. Effects of various concentrations of cellulase and pectinase and different incubation periods were evaluated. The most effective treatment included (per ml) 50 units cellulase and 10 units pectinase, in the incubation medium and incubation for 21 h at 50 rpm in the dark at 27°C. This treatment produced a protoplast yield of 8.2×10⁷ gfw⁻¹ of which 90% were viable. Protoplast viability was influenced by length of the incubation period and ranged from about 40 to 100% among the various experiments.     

A 45,X male with molecular evidence of a translocation of Y euchromatin onto chromosome 1

Summary A 45,X complement was found in lymphocyte and fibroblast cultures of a male infant with severe growth and mental retardation and mild dysmorphism. Lymphocyte DNA from this patient was found to contain Yp chromosome sequences. In situ hybridization (ISH) with the 50f2 probe led to a clear assignment of euchromatic material on the short arm of chromosome 1. This observation and others from the literature argue in favour of the conclusion that all 45.X males are probably either the result of undetected mosaicism or are carriers of Y translocated material.     

Sandwich enzyme-linked immunosorbent assay for the quantification of human serum albumin fragment 408–423 in bodily fluids

Urinary levels of human serum albumin (hSA) fragment 408–423 have been proposed to represent an early marker for graft-versus-host disease (GvHD) and chronic kidney diseases. Here, we developed an enzyme-linked immunosorbent assay (ELISA) for the quantification of hSA(408–423). The sandwich ELISA has a detection limit of 0.5 ng/ml and is highly specific for hSA(408–423) because it does not cross-react with other albumin fragments or the full-length precursor. This ELISA allows rapid and convenient quantification of hSA(408–423) in bodily fluids, further clarifying the prognostic and diagnostic value of this peptide in GvHD, kidney disease, and other disorders.     

Occurrence of Ornithodoros brasiliensis Aragão (Acari: Argasidae) in São Francisco de Paula, RS, Southern Brazil

There have been no reports of the endemic Ornithodoros brasiliensis (Arag?o) in Rio Grande do Sul, southern Brazil, since the 1950s. In January 2007, 21 O. brasiliensis ticks were collected in a rural area named "Cruzinha" in the municipality of S?o Francisco de Paula, RS, and another population was sampled later that year (October) in Vargem do Cedro, another rural area of S?o Francisco de Paula, following reports of human parasitism by ticks. The reappearance of this tick is a reason for concern in terms of public health.