Growth and production of a tropical predatory shrimp, Macrobrachium hainanense (Palaemonidae), in two Hong Kong streams

1. Macrobrachium hainanense is a predatory palaemonid shrimp (total length >7 cm) that can be abundant [density 3–5 m^?2; biomass 484–606 mg ash‐free dry mass (AFDM) m^?2] in forest streams in Hong Kong, China. This study investigated the growth and production of M. hainanense during 2001 and 2002 in pools of two forested streams (one third‐ and one fourth‐order). 2. The growth of tagged individuals was recorded in situ and compared with that of tagged and untagged shrimps in laboratory tanks. Field and laboratory estimates yielded similar growth rates of 0.7 mm carapace length (CL) per month, and instantaneous growth rate was 0.004 g AFDM g^?1 day^?1. Tagging did not affect growth in the laboratory. Cohort analysis of field populations produced similar estimates of growth to that of tagged individuals, and the growth of M. hainanense was generally slower than has been reported for other Macrobrachium species. Mass‐specific growth rate of M. hainanense in the field varied with size and was two to five times higher in small individuals (<10 mm CL). In addition, growth rate varied with season and was 40% lower in the dry season when temperature was at the annual minimum. 3. Males grew bigger than females (36 versus 25 mm CL). The minimum lifespan of M. hainanense in the field, calculated from size‐specific growth rates, ranged from 29.3 months (females) to 47.6 months (males). Male lifespan derived from cohort analysis was estimated as 48 and 46 months in the two streams. Females reached maturity in 17–18 months (at 15–17 mm CL) while males matured at 24–26 months (at 18–22 mm CL). Females bred twice (at 2 and 3 years of age) while males probably bred three times (at 2, 3 and 4 years) in both streams. 4. Macrobrachium hainanense production in the fourth‐order stream, calculated by the size‐frequency method, was 900 and 1096 mg AFDM m^?2 year^?1 (for 2001 and 2002, respectively) with a production/biomass (P/B) of 2.1–2.3 year^?1. In the third‐order stream, production was 987 and 1304 mg AFDM m^?2 year^?1 (for 2001 and 2002, respectively) with a P/B of 1.7–2.1 year^?1. Production estimates based on the instantaneous growth method were half of those obtained by the size‐frequency method. 5. Although M. hainanense production at the third‐order stream exceeded that in the fourth‐order, growth rates showed the opposite pattern and were 0.31–0.43 mm CL month^?1 and 0.56–0.65 mm CL month^?1 in the third‐ and fourth‐order streams, respectively. Greater mortality in the latter may account for low production at a site where growth rate was high. 6. Production of M. hainanense in both streams was lower during 2001 when rainfall was higher. This may reflect the influence of spates associated with monsoonal rains, which could have reduced M. hainanense production through spate‐induced mortality or by reducing the abundance of prey. This study provides the first in situ estimate of secondary production by a non‐commercial Macrobrachium species in Asia or elsewhere. It involved a whole‐pool approach to sampling that allowed the estimation of production and population parameters on a realistic scale.     

Burden of Invasive Pneumococcal Disease in Children Aged 1 Month to 12 Years Living in South Asia: A Systematic Review

Objective

The primary objective was to estimate the burden of invasive pneumococcal disease (IPD) in children aged 1 month to 12 years in South Asian countries.

Methods

We searched three electronic databases (PubMed, Embase and the Cochrane Library) using a comprehensive search strategy, we manually searched published databases (Index Medicus and Current Contents) and we also searched the bibliographies of the included studies and retrieved reviews. The searches were current through June 2013. Eligible studies (community-based and hospital-based) were pooled and a separate analysis for India was also completed. A meta-regression analysis and heterogeneity analysis were performed. The protocol was registered with PROSPERO registration number CRD42013004483.

Results

A total of 22 studies surveying 36,714 children were included in the systematic review. Hospital-based prospective studies from South Asia showed that 3.57% of children had IPD, and 15% of all bacterial pneumonia cases were due to Streptococcus pneumoniae. Indian studies showed that the incidence of IPD was 10.58% in children admitted to hospitals with suspected invasive bacterial diseases, and 24% of all bacterial pneumonia cases were due to S. pneumonia. Population-based studies from South Asian countries showed that 12.8% of confirmed invasive bacterial diseases were caused by S. pneumonia whereas retrospective hospital-based studies showed that 28% of invasive bacterial diseases were due to S. pneumoniae. Meta-regression showed that there was a significant influence of the antigen testing method for diagnosing IPD on IPD prevalence.

Conclusion

S. pneumoniae is responsible for a substantial bacterial disease burden in children of South Asian countries including India despite the presence of high heterogeneity in this meta-analysis. Treatment guidelines must be formulated, and preventive measures like vaccines must also be considered.     

Production,HPLC analysis,and in situ apoptotic activities of swainsonine toward lepidopteran,Sf‐21 cell line

Swainsonine, a secondary metabolite from Metarhizium anisopliae has been extensively studied in the complementary areas of therapeutics and toxicology. This work aims to develop a simple UV‐HPLC method of analyses for swainsonine in Metarhizium fermentation broth and to explore its in situ entomotoxic activities. The partially purified broth was quantitatively analyzed using middle UV (205 nm)‐reverse phase HPLC method with different mobile phases and gradient programmes. Swainsonine was eluted as single peak at (t_e) 6.0–6.9 min with average concentration of 4.04 ± 0.52 μg/mL using optimal mobile phase (0.1% trifluoroacetic acid in water and acetonitrile). The mass spectrometry analysis further indicated the characteristic MS1 species for swainsonine, [M+H]⁺ 174.30 in corresponding HPLC peaks. The antiproliferative effects of swainsonine on lepidopteran, Sf‐21 cells were determined through 3‐(4, 5‐dimethylthia‐zol‐2‐yl)?2, 5‐diphenyl tetrazolium bromide (IC_{50 standard} = 3.90 μM and IC_{50 purified} = 5.27 μM) and trypan blue dye exclusion (IC_{50 standard} = 6.91 μM and IC_{50 purified} = 8.67 μM) assays. The fluorescence activated cell sorting evaluation of Sf‐21 cells showed nearly 35% and 42% of population in various apoptotic stages at 36 h, when treated with standard and purified swainsonine, respectively. The morphodimensional field emission scanning electron and atomic force microscopic analyses further confirmed the characteristic apoptotic features like membrane blebbings, ruptures and volume shrinkage in the lepidopteran cells after 24–36 h of post‐treatment incubation. The study describes the potential entomotoxic activities of swainsonine and its role in the virulence of Metarhizium spp. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:1196–1205, 2014     

Fitting Membrane Resistance along with Action Potential Shape in Cardiac Myocytes Improves Convergence: Application of a Multi-Objective Parallel Genetic Algorithm

Fitting parameter sets of non-linear equations in cardiac single cell ionic models to reproduce experimental behavior is a time consuming process. The standard procedure is to adjust maximum channel conductances in ionic models to reproduce action potentials (APs) recorded in isolated cells. However, vastly different sets of parameters can produce similar APs. Furthermore, even with an excellent AP match in case of single cell, tissue behaviour may be very different. We hypothesize that this uncertainty can be reduced by additionally fitting membrane resistance (R_m). To investigate the importance of R_m, we developed a genetic algorithm approach which incorporated R_m data calculated at a few points in the cycle, in addition to AP morphology. Performance was compared to a genetic algorithm using only AP morphology data. The optimal parameter sets and goodness of fit as computed by the different methods were compared. First, we fit an ionic model to itself, starting from a random parameter set. Next, we fit the AP of one ionic model to that of another. Finally, we fit an ionic model to experimentally recorded rabbit action potentials. Adding the extra objective (R_m, at a few voltages) to the AP fit, lead to much better convergence. Typically, a smaller MSE (mean square error, defined as the average of the squared error between the target AP and AP that is to be fitted) was achieved in one fifth of the number of generations compared to using only AP data. Importantly, the variability in fit parameters was also greatly reduced, with many parameters showing an order of magnitude decrease in variability. Adding R_m to the objective function improves the robustness of fitting, better preserving tissue level behavior, and should be incorporated.     

Altered methylation at microRNA-associated CpG islands in hereditary and sporadic carcinomas: a methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA)-based approach

MicroRNAs (miRNAs) are small noncoding RNAs that contribute to tumorigenesis by acting as oncogenes or tumor suppressor genes and may be important in the diagnosis, prognosis and treatment of cancer. Many miRNA genes have associated CpG islands, suggesting epigenetic regulation of their expression. Compared with sporadic cancers, the role of miRNAs in hereditary or familial cancer is poorly understood. We investigated 96 colorectal carcinomas, 58 gastric carcinomas and 41 endometrial carcinomas, occurring as part of inherited DNA mismatch repair (MMR) deficiency (Lynch syndrome), familial colorectal carcinoma without MMR gene mutations or sporadically. Methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) assays were developed for 11 miRNA loci that were chosen because all could be epigenetically regulated through the associated CpG islands and some could additionally modulate the epigenome by putatively targeting the DNA methyltransferases or their antagonist retinoblastoma-like 2 (RBL2). Compared with the respective normal tissues, the predominant alteration in tumor tissues was increased methylation for the miRNAs 1-1, 124a-1, 124a-2, 124a-3, 148a, 152 and 18b; decreased methylation for 200a and 208a; and no major change for 373 and let-7a-3. The frequencies with which the individual miRNA loci were affected in tumors showed statistically significant differences relative to the tissue of origin (colorectal versus gastric versus endometrial), MMR proficiency versus deficiency and sporadic versus hereditary disease. In particular, hypermethylation at miR-148a and miR-152 was associated with microsatellite-unstable (as opposed to stable) tumors and hypermethylation at miR-18b with sporadic disease (as opposed to Lynch syndrome). Hypermethylation at miRNA loci correlated with hypermethylation at classic tumor suppressor promoters in the same tumors. Our results highlight the importance of epigenetic events in hereditary and sporadic cancers and suggest that MS-MLPA is an excellent choice for quantitative analysis of methylation in archival formalin-fixed, paraffin-embedded samples, which pose challenges to many other techniques commonly used for methylation studies.     

Transgenic maize plants expressing the Totivirus antifungal protein, KP4, are highly resistant to corn smut

The corn smut fungus, Ustilago maydis, is a global pathogen responsible for extensive agricultural losses. Control of corn smut using traditional breeding has met with limited success because natural resistance to U. maydis is organ specific and involves numerous maize genes. Here, we present a transgenic approach by constitutively expressing the Totivirus antifungal protein KP4, in maize. Transgenic maize plants expressed high levels of KP4 with no apparent negative impact on plant development and displayed robust resistance to U. maydis challenges to both the stem and ear tissues in the greenhouse. More broadly, these results demonstrate that a high level of organ independent fungal resistance can be afforded by transgenic expression of this family of antifungal proteins.     

Association of human TLR1 and TLR6 deficiency with altered immune responses to BCG vaccination in South African infants

The development of effective immunoprophylaxis against tuberculosis (TB) remains a global priority, but is hampered by a partially protective Bacillus Calmette-Guérin (BCG) vaccine and an incomplete understanding of the mechanisms of immunity to Mycobacterium tuberculosis. Although host genetic factors may be a primary reason for BCG''s variable and inadequate efficacy, this possibility has not been intensively examined. We hypothesized that Toll-like receptor (TLR) variation is associated with altered in vivo immune responses to BCG. We examined whether functionally defined TLR pathway polymorphisms were associated with T cell cytokine responses in whole blood stimulated ex vivo with BCG 10 weeks after newborn BCG vaccination of South African infants. In the primary analysis, polymorphism TLR6_C745T (P249S) was associated with increased BCG-induced IFN-γ in both discovery (n = 240) and validation (n = 240) cohorts. In secondary analyses of the combined cohort, TLR1_T1805G (I602S) and TLR6_G1083C (synonymous) were associated with increased IFN-γ, TLR6_G1083C and TLR6_C745T were associated with increased IL-2, and TLR1_A1188T was associated with increased IFN-γ and IL-2. For each of these polymorphisms, the hypo-responsive allele, as defined by innate immunity signaling assays, was associated with increased production of TH1-type T cell cytokines (IFN-γ or IL-2). After stimulation with TLR1/6 lipopeptide ligands, PBMCs from TLR1/6-deficient individuals (stratified by TLR1_T1805G and TLR6_C745T hyporesponsive genotypes) secreted lower amounts of IL-6 and IL-10 compared to those with responsive TLR1/6 genotypes. In contrast, no IL-12p70 was secreted by PBMCs or monocytes. These data support a mechanism where TLR1/6 polymorphisms modulate TH1 T-cell polarization through genetic regulation of monocyte IL-10 secretion in the absence of IL-12. These studies provide evidence that functionally defined innate immune gene variants are associated with the development of adaptive immune responses after in vivo vaccination against a bacterial pathogen in humans. These findings could potentially guide novel adjuvant vaccine strategies as well as have implications for IFN-γ-based diagnostic testing for TB.