The essential role of clathrin-mediated endocytosis in yellow head virus propagation in the black tiger shrimp Penaeus monodon

Yellow head virus (YHV) is one of the most widespread viruses seriously affecting black tiger shrimp (Penaeus monodon) cultivation. A previous microarray study demonstrated that clathrin coat assembly protein 17 (AP17) was significantly up-regulated after YHV infection (Pongsomboon et al., 2011). Clathrin coat AP17 is a part of the assembly protein σ2 (AP-2) complex which is involved in clathrin-mediated endocytosis. Quantitative RT-PCR (qRT-PCR) revealed that the clathrin coat AP17 gene was up-regulated 3-fold at 12 h post YHV infection. In addition, immunofluorescence microscopy showed that clathrin coat AP17 was highly expressed in the cytoplasm of the YHV-infected hemocytes. Knockdown of the clathrin coat AP17 gene dramatically reduced YHV replicativity by 32-fold. Interestingly, shrimp pre-treated with chlorpromazine, a commercial drug that inhibits clathrin-dependent endocytosis, exhibited significantly low levels of YHV infection. Taken together, these results suggest that clathrin-mediated endocytosis is involved in YHV propagation in P. monodon.     

Frontal EEG delta/alpha ratio and screening for post-stroke cognitive deficits: The power of four electrodes

This study analysed correlations between post-stroke, quantitative electroencephalographic (QEEG) indices, and cognition-specific, functional outcome measures. Results were compared between QEEG indices calculated from the standard 19 versus 4 frontal (or 4 posterior) electrodes to assess the feasibility and efficacy of employing a reduced electrode montage. Resting-state EEG was recorded at the bedside within 62–101 h after onset of symptoms of middle cerebral artery, ischaemic stroke (confirmed radiologically). Relative power for delta, theta, alpha and beta, delta/alpha ratio (DAR) and pairwise-derived brain symmetry index (pdBSI) were averaged; over all electrodes (global), over F3, F4, F7, F8 (frontal) and P3, P4, T5, T6 (posterior). The functional independence measure and functional assessment measure (FIM–FAM) was administered at mean 105 days post-stroke. Total (30 items) and cognition-specific (5 items) FIM–FAM scores were correlated with QEEG indices using Spearman's coefficient, with a Bonferroni correction. Twenty-five patients were recruited, 4 died within 3 months and 1 was lost to follow-up. Hence 20 cases (10 female; 9 left hemisphere; mean age 68 years, range 38–84) were analysed. Two QEEG indices demonstrated highly-significant correlations with cognitive outcomes: frontal DAR (ρ = − 0.664, p ≤ 0.001) and global, relative alpha power (ρ = 0.67, p ≤ 0.001). After correction there were no other significant correlations. Alpha activity – particularly frontally – may index post-stroke attentional capacity, which appears to be a key determinant of functional and cognitive outcomes. Likewise frontal delta pathophysiology influences such outcomes. Pending further studies, DAR from 4 frontal electrodes may inform early screening for post-MCA stroke cognitive deficits, and thereby, clinical decisions.     

表达H9N2禽流感病毒HA1蛋白的转基因和缓艾美耳球虫的构建

H9N2亚型禽流感病毒已经被证实可以传播到人类,对它存在的潜在危害已受到越来越多的关注。因此,研究安全而有效的H9N2禽流感病毒疫苗已迫在眉睫。为研究和缓艾美耳球虫作为活载体表达及运输禽流感病毒抗原的潜能,本研究构建了可稳定表达H9N2禽流感病毒HA1蛋白的转基因和缓艾美耳球虫。利用黄色荧光蛋白（YFP）及融合的乙胺嘧啶抗性基因（DHFR-TSm2m3）作为报告基因及筛选基因,我们将表达H9N2禽流感病毒HA1抗原的质粒载体核转球虫子孢子,接种鸡后在乙胺嘧啶药物的选择压力下进行筛选并连续传代获得转基因和缓艾美耳球虫系。利用染色体步移和免疫印迹证实了HA1基因的成功插入和表达;利用间接免疫荧光证实HA1蛋白定位于球虫子孢子细胞膜表面和头部。研究进一步发现,转基因球虫的繁殖力与野生球虫相当,感染后亦于第6d达到排卵囊高峰。该转基因和缓艾美耳球虫株具有作为疫苗活载体的潜能。     

Robust Noninferiority Tests for Potency of a Test Drug Against a Reference Drug

In comparing test formulations of a drug product to reference formulations it is necessary to check that the test formulation has as much biologic activity as the reference within some indifference zone. A modern method for testing this relative potency is to fit a nonlinear model of the dose–response curve, constrained such that the dose–response of the test is modeled with the same function as the reference except for a multiplicative parameter on the dose. This method works well as long as the nonlinear model fits the data. Although tests of this fit are mentioned in the literature, how relative potency should be measured when those tests fail has not been addressed. We study the use of the sandwich estimator of variance for this noninferiority potency problem when the model does not fit the data. We focus on the simple linear model. Using the sandwich variance, the inferences are asymptotically valid even if the model does not hold. We discuss interpretation of the results under the misspecified linear model, and show by simulation that the method performs well in some practical cases. Finally, we give advice for designing a study that uses these methods.     

Overview: Japanese encephalitis

Japanese encephalitis (JE) is one of the most important endemic encephalitis in the world especially in Eastern and Southeastern Asia. JE affects over 50,000 patients and results in 15,000 deaths annually. JE virus is a single stranded positive sense RNA virus belonging to family flaviviridae. JE virus is transmitted through a zoonotic cycle between mosquitoes, pigs and water birds. Humans are accidentally infected and are a dead end host because of low level and transient viremia. In the northern region, large epidemics occur during summers whereas in the southern region JE tends to be endemic: cases occur throughout the year with a peak in the rainy season. Occurrence of JE is more closely related to temperature than to humidity. JE is regarded as a disease of children in the endemic areas but in the newly invaded areas, it affects both the adults and children because of the absence of protective antibodies. For every patient of JE, there are large numbers of subclinical cases (25–1000). Symptomatic JEV infection manifests with nonspecific febrile illness, aseptic meningitis or encephalitis. Encephalitis manifests with altered sensorium, seizures and focal neurological deficit. Acute flaccid paralysis may occur due to anterior horn cell involvement. A wide variety of movement disorders especially transient Parkinsonian features and dystonia (limb, axial, orofacial) are reported in 20–60% patients. JE mainly affects thalamus, corpus striatum, brainstem and spinal cord as revealed by MRI and on autopsy studies. Coinfection of JE and cysticercosis occurs because of the important role of pigs in the life cycle of both JEV and cysticercosis.     

Suitability of five cladoceran species from Mexico for in situ experimentation

Although they are highly suited for site-specific risk assessment, little attention has been given to in situ ecotoxicological experimentation in tropical regions. This study intended to assess the suitability of five freshwater cladocerans from Mexico for in situ experiments. Juveniles of Ceriodaphnia cornuta, Macrothrix triserialis group, Simocephalus vetulus, Diaphanosoma birgei, and Pseudosida cf. ramosa, from two populations, were used to perform seven day laboratory and in situ experiments. Pseudosida cf. ramosa was very sensitive to handling and was discarded from further evaluations. Ceriodaphnia cornuta reproduced significantly more neonates, with smaller coefficients of variation, than M. triserialis and S. vetulus. Also due to its worldwide distribution in the tropics, C. cornuta is suggested to be highly suitable for in situ experiments.     

Cygnet River Virus,a Novel Orthomyxovirus from Ducks,Australia

A novel virus, designated Cygnet River virus (CyRV), was isolated in embryonated eggs from Muscovy ducks in South Australia. CyRV morphologically resembles arenaviruses; however, sequencing identified CyRV as an orthomyxovirus. The high mortality rate among ducks co-infected with salmonellae suggests that CyRV may be pathogenic, either alone or in concert with other infections.     

Phenotypic and genetic characterization of a next generation live-attenuated yellow fever vaccine candidate

We assessed the genetic and phenotypic characteristics of a yellow fever vaccine candidate, which was cloned from a YF-VAX substrain selected for growth in Vero cells (vYF-247), during the manufacturing process from the master seed lot (MSL) and working seed lot (WSL) through to the drug substance (DS) stage. There were nine minor nucleotide variants observed from the MSL to the DS stage, of which five led to amino acid changes. The variant positions were, however, not known risks for any virulence modification. vYF-247 exhibits a homogenous plaque size profile (as expected for a cloned vaccine candidate) composed of small plaques (<1 mm) that remained consistent throughout the manufacturing process. In addition, there was no change in the viral replication rate. Of note, the DS sequences across the two manufacturing campaigns (2018 and 2019) were very similar suggesting a high batch-to-batch consistency. All MSL, WSL and DS batches exhibited similar neurovirulence profiles in mice and had a more attenuated neurovirulence phenotype than the YF-VAX (egg-based vaccine) comparator. Overall, the neurovirulence phenotype of vYF-247 does not change from MSL, WSL to DS. These data collectively support the safety and genetic stability of vYF-247 during the production process.     

Effects of physical positions on sleep architectures and post-nap functions among habitual nappers

The post-lunch sleepiness is considered to be part of biological rhythm. Many people take nap as a countermeasure to this afternoon circadian nadir. The aim of the study is to investigate the effect of a short-term midday nap in different physical positions after a full night sleep on subjective and physiological measures. Thirty-six young healthy habitual nappers (18 men, 18 women) aged 18-23 participated in the study. They were randomly assigned to three experimental conditions: to nap in a seat (NS), to nap in a bed (NB) and no-nap (NN). Measures of subjective sleepiness, mood, fatigue, and P300 component were taken before and after a 20 min nap. Results showed that sleepiness, fatigue, and mood for both NS and NB were improved after napping. Moreover, objective alertness was enhanced in NB relative to NS and NN, which showed the larger P300 amplitude after nap. The delta power during sleep stage 2 of NS was inferior to that of NB, which induced a lower alertness after the sitting nap. Consequently, the result demonstrated the function of delta wave during stage 2 on arousal level following the brief midday nap. In general, taking a nap in a seat is a way to subjective restoration but taking a nap in a bed benefits the habitual nappers subjectively and physiologically.