Epitope Analysis and Efficacy Evaluation of Phosphatase 2C (PP2C) DNA Vaccine Against Toxoplasma gondii Infection.

Toxoplasma gondii infects almost all warm-blooded animals and negatively affects the health of a wide range of these animals, including humans. Protein phosphatase 2C (PP2C) is a T. gondii protein secreted by rhoptry organelles during host cell invasion. However, very little is known about whether this protein can induce protective immunity against T. gondii. In this study, bioinformatics analysis of PP2C revealed some useful information in the context of anti-toxoplasmosis treatments and vaccine research. In addition, the PP2C gene was amplified, and a eukaryotic expression vector (pEGFP-PP2C) was successfully constructed to express PP2C. Finally, the constructed pEGFP-PP2C was injected into mice to evaluate whether it could induce immunoprotection. Compared with the control groups, we found that immunizations with the pEGFP-PP2C plasmid could elicit specific IgG antibodies and cytokines against T. gondii infection. The survival of mice immunized with the pEGFP-PP2C plasmid was significantly prolonged compared with that of the control group mice. Based on the ability of pEGFP-PP2C to induce specific immune responses against T. gondii, we propose that PP2C merits consideration as a potential vaccine candidate against toxoplasmosis.

Prenatal lead exposure in relation to age at menarche: results from a longitudinal study in Mexico City.

Animal and cross-sectional epidemiological studies suggest that prenatal lead exposure is related to delayed menarche, but this has not been confirmed in longitudinal studies. We analyzed this association among 200 girls from Mexico City who were followed since the first trimester of gestation. Maternal blood lead levels were analyzed once during each trimester of pregnancy, and daughters were asked about their first menstrual cycle at a visit between the ages of 9.8 and 18.1 years. We estimated hazard ratios (HRs) and 95% confidence intervals (CI) for probability of menarche over the follow-up period using interval-censored Cox models, comparing those with prenatal blood lead level ⩾5 µg/dl to those with prenatal blood lead <5 µg/dl. We also estimated HRs and 95% CI with conventional Cox regression models, which utilized the self-reported age at menarche. In adjusted analyses, we accounted for maternal age, maternal parity, maternal education, and prenatal calcium treatment status. Across trimesters, 36-47% of mothers had blood lead levels ⩾5 µg/dl. Using interval-censored models, we found that during the second trimester only, girls with ⩾5 µg/dl prenatal blood lead had a later age at menarche compared with girls with prenatal blood lead levels <5 µg/dl (confounder-adjusted HR=0.59, 95% CI 0.28-0.90; P=0.05). Associations were in a similar direction, although not statistically significant, in the conventional Cox regression models, potentially indicating measurement error in the self-recalled age at menarche. In summary, higher prenatal lead exposure during the second trimester could be related to later onset of sexual maturation.

AIM2 contributes to the maintenance of intestinal integrity via Akt and protects against Salmonella mucosal infection.

The mechanism regulating the gastrointestinal epithelial barrier remains poorly understood. We herein demonstrate that Absent in melanoma-2 (AIM2) contributes to the maintenance of intestinal barrier integrity and defense against bacterial infection. AIM2-deficient mice displayed an increased susceptibility to mucosal but not systemic infection by Salmonella typhimurium, indicating a protective role for AIM2 in the gastrointestinal tract. In a Salmonella colitis model, compared with wild-type mice, AIM2(-/-) mice exhibited more severe body weight loss, intestinal damage, intestinal inflammation, and disruption of basal and activated epithelial cell turnover. In vivo and in vitro data showed that AIM2 restricted the early epithelial paracellular invasion of Salmonella and decreased epithelial permeability. The decreased epithelial barrier in AIM2(-/-) mice might be attributed to the altered expression of tight junction proteins that contribute to epithelial integrity. AIM2 promoted the expression of tight junction proteins through Akt activation. Together, these results suggest that AIM2 is required for maintaining the integrity of the epithelial barrier.

Effects of Graded Levels of Isomaltooligosaccharides on the Performance, Immune Function and Intestinal Status of Weaned Pigs.

The objective of this study was to investigate the effects of graded levels of isomaltooligosaccharides (IMO) on the performance, immune function and intestinal microflora and intestinal mucosal morphology of weaned pigs. In a 28-day experiment, one hundred eighty, twenty eight-day-old, crossbred (Duroc×Large White×Landrace), weaned pigs, with an initial body weight of 8.19±1.45 kg, were fed either an unsupplemented corn-soybean meal based diet or similar diets supplemented with 0.2%, 0.4%, 0.6%, or 0.8% IMO added at the expense of corn. Each treatment was replicated six times with six pigs (three barrows and three gilts) per pen. From day 0 to 14, weight gain was linearly increased (p<0.05), while gain:feed (p<0.05) was linearly improved and diarrhea rate (p = 0.05) linearly declined as the IMO level increased. On d 14, the level of the immunoglobulins IgA, IgM, and IgG in the serum of pigs were linearly increased (p<0.05) with increasing IMO supplementation. Interleukin-6 (IL-6) was linearly (p<0.05) and quadratically (p<0.05) decreased as IMO intake increased. From day 15 to 28, there was a trend for weight gain to be linearly increased, and IL-2 was linearly (p<0.05) increased as IMO supplementation increased on d 28. Over the entire experiment, weight gain was linearly increased (p<0.05), while gain:feed (p<0.05) was linearly improved and diarrhea rate (p<0.05) was linearly decreased as the IMO level increased. Supplementation with IMO had no effect on the intestinal microflora of pigs in the ileum and cecum of pigs, as well as the villus height and crypt depth in the ileum and jejunum (p>0.05). These results indicate that dietary inclusion of IMO increases weight gain, gain:feed and enhanced the immune status of pigs, and could be a valuable feed additive for use in weaned pigs, particularly during the period immediately after weaning.

Structural and antigenic analysis and cloning of surface antigen protein 5 in Toxoplasma gondii.

Toxoplasma gondii is identified as an obligate intracellular apicomplexan parasite that infects warm blooded animals and humans worldwide. SAG5 protein includes SAG5A, -5B, -5C, -5D, and -5E five subtypes. SAG5A, -5B, -5C, and -5D are expressed on the surface of Toxoplasma gondii. In this study, we used online T-Coffee tool to analyze SAG5 proteins sequence alignment. SMART software was used to predict secondary structures of SAG5A, -5B, -5C, and -5D. The 3D models of SAG5 proteins were constructed and analyzed with SWISS-MODEL server and VMD software. Results indicated that SAG5A, -5B, -5C, and -5D are highly homologous proteins. Furthermore, liner-B cell epitopes and Th-cell epitopes of the four proteins were predicted using DNAMAN software and Epitope Database online service. The bioinformatics analysis of SAG5A, -5B, -5C, and -5D proteins could provide valuable information on prevention and treatment of toxoplasmosis. In addition, the four genes were obtained by PCR and inserted into an eukaryotic expression vector pEGFP-C1 respectively. Identified by restriction enzyme digestion, the four recombinant plasmids were transfected into HEK 293-T cells and tested by RT-PCR. Results showed that the constructed plasmids were all transfected to HEK 293-T cells successfully.

Planning for Uncertainty and Fallbacks Can Increase the Number of Transplants in a Kidney-Paired Donation Program.

A kidney-paired donation (KPD) pool consists of transplant candidates and their incompatible donors, along with nondirected donors (NDDs). In a match run, exchanges are arranged among pairs in the pool via cycles, as well as chains created from NDDs. A problem of importance is how to arrange cycles and chains to optimize the number of transplants. We outline and examine, through example and by simulation, four schemes for selecting potential matches in a realistic model of a KPD system; proposed schemes take account of probabilities that chosen transplants may not be completed as well as allowing for contingency plans when the optimal solution fails. Using data on candidate/donor pairs and NDDs from the Alliance for Paired Donation, the simulations extend over 8 match runs, with 30 pairs and 1 NDD added between each run. Schemes that incorporate uncertainties and fallbacks into the selection process yield substantially more transplants on average, increasing the number of transplants by as much as 40% compared to a standard selection scheme. The gain depends on the degree of uncertainty in the system. The proposed approaches can be easily implemented and provide substantial advantages over current KPD matching algorithms.

Statistical analysis of repeated measurements with informative censoring times.

Incomplete repeated measurement data often arise in medical studies. A problem that has recently drawn much attention in the literature in this situation is that the incompleteness or missingness is informative or related to the underlying variable of interest. In this paper we propose a non-parametric global test for treatment comparison in the presence of informative incompleteness. A semi-parametric regression model is also presented for assessing conditional treatment effects given the drop-out patterns, adopting the idea similar to that behind the pattern-mixture modelling approach and discussed in Shih and Quan. The proposed methods can be easily implemented and are conceptually simple and similar too, but can be applied to more general cases than those given in Yao et al. They are evaluated by numerical studies and applied to data from a clinical trial of adult schizophrenics.

Marginal models for longitudinal continuous proportional data.

Continuous proportional data arise when the response of interest is a percentage between zero and one, e.g., the percentage of decrease in renal function at different follow-up times from the baseline. In this paper, we propose methods to directly model the marginal means of the longitudinal proportional responses using the simplex distribution of Barndorff-Nielsen and Jørgensen that takes into account the fact that such responses are percentages restricted between zero and one and may as well have large dispersion. Parameters in such a marginal model are estimated using an extended version of the generalized estimating equations where the score vector is a nonlinear function of the observed response. The method is illustrated with an ophthalmology study on the use of intraocular gas in retinal repair surgeries.