Genetic variants in SCNN1B and AHCYL1 are associated with eggshell quality in Chinese domestic laying ducks (Anas platyrhynchos).

1. The objective of this study was to investigate the evolution of SCNN1B and AHCYL1 proteins among 10 domestic avian and mammalian animal species, to uncover the expression patterns of SCNN1B and AHCYL1 genes in ducks, identify the genetic variants of the SCNN1B and AHCYL1 genes and analyse their effects on eggshell quality.2. Expression profiles of the SCNN1B and AHCYL1 genes in Sansui female ducks were determined using real-time fluorescence quantitative PCR to identify SNPs. The duck SCNN1B and AHCYL1 genes were amplified to identify SNPs. A total of 502 Sansui female ducks were genotyped by sequencing, and the associations between the mRNA expression/SNP genotypes and 6 eggshell quality indices were analysed using PASW Statistics 18.0.3. The results showed that the SCNN1B and AHCYL1 proteins are highly conserved in different mammalian or domestic animals, especially the AHCYL1 protein. The SCNN1B and AHCYL1 genes were widely expressed in different tissues of male and female ducks, and expression level in the uterus was greater than in other tissues. The expression of SCNN1B and AHCYL1 during the oviposition cycle indicated that expression levels were related to the eggshell mineralisation stage.4. The mRNA expression levels of the uterine SCNN1B and AHCYL1 genes were positively correlated with eggshell strength (ESS), percentage (ESP) and weight (ESW) (P < 0.05), respectively. Ten novel SNPs in SCNN1B and AHCYL1 genes from Chinese domestic laying ducks were identified through PCR amplicon sequencing.5. Genetic association analysis indicated g.797509 C > T, g.797573 C > T and g.797834 C > T in SCNN1B gene and g.169244 T > A, g.169265 T > C and g.175311 T > C in AHCYL1 gene had a significant effect on eggshell quality. Correlation analysis between the SNP genotype and SCNN1B and AHCYL1 genes expression in the uterus showed that the genotypes of g.797509 C > T, g.797573 C > T, g.797834 C > T, g.169244 T > A and g.175311 T > C sites affected the expression of SCNN1B and AHCYL1 genes in utero (P < 0.05).6. The study indicated SCNN1B and AHCYL1 as candidate genes to improve eggshell traits in ducks.

Association of PRKCA expression and polymorphisms with layer duck eggshell quality.

1. Eggshell quality is important for the poultry industry. Calcium is deposited during eggshell formation, and protein kinase C alpha (PRKCA) is involved in transmembrane transport of calcium ions in cells. However, the biological function of PRKCA in poultry is still not understood. Therefore, the aim of this study was to explore the association of mRNA expression and single nucleotide polymorphisms (SNPs) of the PRKCA gene with eggshell quality in laying ducks. 2. The mRNA expression and SNPs of the PRKCA gene were detected by real-time fluorescence quantitative PCR (qRT-PCR) and sequencing of PCR products in 45-week-old female Sansui ducks, which is a high production layer duck breed in China. The association of mRNA expression and SNPs in the PRKCA gene with layer duck eggshell traits was analysed using SPSS (v18.0) software. 3. The results demonstrated that PRKCA mRNA was widely expressed in all examined tissues, and expression was highest in kidney and lowest in the gizzard. Furthermore, the PRKCA mRNA level in uterus was significantly positively correlated with eggshell strength and eggshell weight (P < 0.05). Three novel SNPs, the synonymous mutations of g.9571770 T > C in exon 5, g.9583222 C > T and g.9583227 G > A in exon 7, were found in the PRKCA gene, giving four haplotypes and 10 diplotypes, which affected the mRNA secondary structure and free energy. The g.9583222 C > T and g.9583227 G > A mutations were significantly associated with eggshell strength (P < 0.05). Diplotype H1H1 was advantageous for increasing the strength and thickness of an eggshell. 4. In conclusion, the study showed that the mRNA transcription and genetic variation in the PRKCA gene could significantly affect the strength of duck eggshell and that the PRKCA gene is an important candidate gene for improving eggshell quality in poultry.

Enhancing expression of the pseudorabies virus glycoprotein E in yeast and its application in an indirect sandwich ELISA.

AIMS: The purpose of this study was to produce a recombinant pseudorabies virus (PRV) glycoprotein E (gE) protein with the correct antigenicity for use as a low-cost diagnostic antigen. METHODS AND RESULTS: The gene fragment encoding the amino-terminal immunodominant region of PRV gE (codons 31-270) (gEN31-270) was codon optimized and expressed constitutively and secreted using a Pichia pastoris expression system. Yeast-expressed gEN31-270 (ygEN31-270) was harvested from the culture supernatant, and ygEN31-270 was shown to exhibit N-linked glycosylation. An indirect sandwich enzyme-linked immunosorbent assay (ELISA) was developed using ygEN31-270 as a coating antigen, and the results showed that the assay had high sensitivity and specificity, as well as almost perfect concordance with a commercial gE ELISA kit. CONCLUSIONS: The immunodominant region (amino acids 31-270) of gE was expressed successfully in P. pastoris using a codon optimization strategy. ygEN31-270 was secreted and N-glycosylated. The ygEN31-270-based indirect sandwich ELISA showed high sensitivity and specificity to detect gE-specific antibodies in swine serum samples. SIGNIFICANCE AND IMPACT OF THE STUDY: The ygEN31-270-based indirect sandwich ELISA may provide an alternative method for developing a diagnostic kit with easy manipulation and low cost.

Assessing coughing-induced influenza droplet transmission and implications for infection risk control.

Indoor transmission of respiratory droplets bearing influenza within humans poses high risks to respiratory function deterioration and death. Therefore, we aimed to develop a framework for quantifying the influenza infection risk based on the relationships between inhaled/exhaled respiratory droplets and airborne transmission dynamics in a ventilated airspace. An experiment was conducted to measure the size distribution of influenza-containing droplets produced by coughing for a better understanding of potential influenza spread. Here we integrated influenza population transmission dynamics, a human respiratory tract model, and a control measure approach to examine the indoor environment-virus-host interactions. A probabilistic risk model was implemented to assess size-specific infection risk for potentially transmissible influenza droplets indoors. Our results found that there was a 50% probability of the basic reproduction number (R0) exceeding 1 for small-size influenza droplets of 0·3-0·4 µm, implicating a potentially high indoor infection risk to humans. However, a combination of public health interventions with enhanced ventilation could substantially contain indoor influenza infection. Moreover, the present dynamic simulation and control measure assessment provide insights into why indoor transmissible influenza droplet-induced infection is occurring not only in upper lung regions but also in the lower respiratory tract, not normally considered at infection risk.

Genetic analysis of maize kernel thickness by quantitative trait locus identification.

Kernel thickness is one of the most important traits in kernel structure, and is related to yield. To ascertain its genetic information more clearly, an immortal recombinant inbred line segregation population was used to map the quantitative trait loci (QTLs) for kernel thickness. As a result, two QTLs were identified on chromosome 9; both of them had negative additive effects, and could decrease kernel thickness to some extent. The QTLs explained 25.8% of the total phenotypic variation. These results advance our understanding of the genetic basis of kernel thickness in maize-breeding programs.

Behavioural response in educated young adults towards influenza A(H1N1)pdm09.

The purpose of this paper was to determine how contact behaviour change influences the indoor transmission of influenza A(H1N1)pdm09 among school children. We incorporated transmission rate matrices constructed from questionnaire responses into an epidemiological model to simulate contact behaviour change during an influenza epidemic. We constructed a dose-response model describing the relationships between contact rate, viral load, and respiratory symptom scores using published experimental human infection data for A(H1N1)pdm09. Findings showed that that mean numbers of contacts were 5.66 ± 6.23 and 1.96 ± 2.76 d-1 in the 13-19 and 40-59 years age groups, respectively. We found that the basic reproduction number (R 0) was <1 during weekends in pandemic periods, implying that school closures or class suspensions are probably an effective social distancing policy to control pandemic influenza transmission. We conclude that human contact behaviour change is a potentially influential factor on influenza infection rates. For substantiation of this effect, we recommend a future study with more comprehensive control measures.

Assessing dengue infection risk in the southern region of Taiwan: implications for control.

Dengue, one of the most important mosquito-borne diseases, is a major international public health concern. This study aimed to assess potential dengue infection risk from Aedes aegypti in Kaohsiung and the implications for vector control. Here we investigated the impact of dengue transmission on human infection risk using a well-established dengue-mosquito-human transmission dynamics model. A basic reproduction number (R 0)-based probabilistic risk model was also developed to estimate dengue infection risk. Our findings confirm that the effect of biting rate plays a crucial role in shaping R 0 estimates. We demonstrated that there was 50% risk probability for increased dengue incidence rates exceeding 0.5-0.8 wk-1 for temperatures ranging from 26°C to 32°C. We further demonstrated that the weekly increased dengue incidence rate can be decreased to zero if vector control efficiencies reach 30-80% at temperatures of 19-32°C. We conclude that our analysis on dengue infection risk and control implications in Kaohsiung provide crucial information for policy-making on disease control.

Network information analysis reveals risk perception transmission in a behaviour-influenza dynamics system.

Influenza poses a significant public health burden worldwide. Understanding how and to what extent people would change their behaviour in response to influenza outbreaks is critical for formulating public health policies. We incorporated the information-theoretic framework into a behaviour-influenza (BI) transmission dynamics system in order to understand the effects of individual behavioural change on influenza epidemics. We showed that information transmission of risk perception played a crucial role in the spread of health-seeking behaviour throughout influenza epidemics. Here a network BI model provides a new approach for understanding the risk perception spread and human behavioural change during disease outbreaks. Our study allows simultaneous consideration of epidemiological, psychological, and social factors as predictors of individual perception rates in behaviour-disease transmission systems. We suggest that a monitoring system with precise information on risk perception should be constructed to effectively promote health behaviours in preparation for emerging disease outbreaks.

Inpatient expenditures on alcohol-attributed diseases and alcohol tax policy: a nationwide analysis in Taiwan from 1996 to 2010.

OBJECTIVES: To assess the two opposing effects of alcohol tax policy interventions (tax rate increase in 2002 and decrease in 2009) on hospitalization in monetary terms of alcohol-attributed diseases (AADs) in Taiwan. STUDY DESIGN: An interrupted time-series analysis. METHODS: Admissions data from 1996 to 2010 were retrieved from the National Health Insurance Research Database claims file and analysed in this study. Data for 430,388 males and 34,874 females aged 15 or above who were admitted due to an AAD were collected. An interrupted time-series analysis examining the effects of the implementation of alcohol tax policy on quarterly adjusted hospital inpatient charges (HICs) for AADs was employed. RESULTS: The study showed significant (p < 0.001) changes in the adjusted HICs for AADs in 2002. Quarterly HICs showed an abrupt 14.8% decline (i.e., a 1.3 million US dollar reduction) after the first tax policy was implemented. No change in quarterly HICs for AADs was found after the alcohol tax increase. The total cost of treating these AAD inpatients over the course of the 15-year period was 640.9 million US dollars. Each inpatient with an AAD costs an average of $900-$2000 depending on the patient's sex and age with the cost increasing gradually after the two tax interventions. More than 80% of the HICs were attributed to alcoholic liver diseases. Psychoses accounted for 6%-18% of the total HICs. Alcohol abuse and alcohol poisoning accounted for less than 2% of the total HICs. CONCLUSIONS: This study provides evidence that alcohol taxation has resulted in an immediate reduction of medical expenditures related to AADs. The policy of increasing alcohol tax rates may have favourable influences on health care resources related to treating AADs.

Cloning and sequence analysis of an actin gene in aloe.

Aloe (Aloe spp), containing abundant polysaccharides and numerous bioactive ingredients, has remarkable medical, ornamental, calleidic, and edible values. In the present study, the total RNA was extracted from aloe leaf tissue. The isolated high-quality RNA was further used to clone actin gene by using reverse transcription-polymerase chain reaction (RT-PCR). The result of sequence analysis for the amplified fragment revealed that the cloned actin gene was 1012 bp in length (GenBank accession No. KC751541.1) and contained a 924-bp coding region and encoded a protein consisting of 307 amino acids. Homologous alignment showed that it shared over 80 and 96% identity with the nucleotide and amino acid sequences of actin from other plants, respectively. In addition, the cloned gene was used for phylogenetic analyses based on the deduced amino acid sequences, and the results suggested that the actin gene is highly conserved in evolution. The findings of this study will be useful for investigating the expression patterns of other genes in Aloe.

Optimization of the procedure for extracting nucleic acids from aloe.

Aloe, an important folk herbal drug, includes abundant polysaccharides and secondary metabolites, which make it difficult to isolate high-quality DNA or RNA. In this paper, one and two improved methods were used to isolate the genomic DNA and RNA from the leaf of aloe, respectively. The obtained samples presented good quality and integrity; thus, they could be further used for many downstream molecular experiments. These reported protocols for DNA and RNA extraction offered a valuable reference for other related studies.

An effective method for extracting total RNA from Dioscorea opposita Thunb.

Dioscorea opposita Thunb., included in the genus Dioscorea of the family Dioscoreaceae, is an important herb with great edible and medicinal value. In this study, the total RNA from leaves of Lichuan Dioscorea opposita Thunb. was isolated by an improved Trizol method. The results showed that the RNA extracted by the improved Trizol method had good integrity, and the RNA could be used for down-stream molecular biology operations including reverse transcription-polymerase chain reaction.

Seasonal dynamics of tuberculosis epidemics and implications for multidrug-resistant infection risk assessment.

Understanding how seasonality shapes the dynamics of tuberculosis (TB) is essential in determining risks of transmission and drug resistance in (sub)tropical regions. We developed a relative fitness-based multidrug-resistant (MDR) TB model incorporated with seasonality and a probabilistic assessment model to assess infection risk in Taiwan regions. The model accurately captures the seasonal transmission and population dynamics of TB incidence during 2006-2008 and MDR TB in high TB burden areas during 2006-2010 in Taiwan. There is ~3% probability of having exceeded 50% of the population infected attributed to MDR TB. Our model not only provides insight into the understanding of the interactions between seasonal dynamics of TB and environmental factors but is also capable of predicting the seasonal patterns of TB incidence associated with MDR TB infection risk. A better understanding of the mechanisms of TB seasonality will be critical in predicting the impact of public control programmes.

Cost-effectiveness of influenza control measures: a dynamic transmission model-based analysis.

We investigated the cost-effectiveness of different influenza control strategies in a school setting in Taiwan. A susceptible-exposure-infected-recovery (SEIR) model was used to simulate influenza transmission and we used a basic reproduction number (R 0)-asymptomatic proportion (θ) control scheme to develop a cost-effectiveness model. Based on our dynamic transmission model and economic evaluation, this study indicated that the optimal cost-effective strategy for all modelling scenarios was a combination of natural ventilation and respiratory masking. The estimated costs were US$10/year per person in winter for one kindergarten student. The cost for hand washing was estimated to be US$32/year per person, which was much lower than that of isolation (US$55/year per person) and vaccination (US$86/year per person) in containing seasonal influenza. Transmission model-based, cost-effectiveness analysis can be a useful tool for providing insight into the impacts of economic factors and health benefits on certain strategies for controlling seasonal influenza.

Using experimental human influenza infections to validate a viral dynamic model and the implications for prediction.

The aim of this work was to use experimental infection data of human influenza to assess a simple viral dynamics model in epithelial cells and better understand the underlying complex factors governing the infection process. The developed study model expands on previous reports of a target cell-limited model with delayed virus production. Data from 10 published experimental infection studies of human influenza was used to validate the model. Our results elucidate, mechanistically, the associations between epithelial cells, human immune responses, and viral titres and were supported by the experimental infection data. We report that the maximum total number of free virions following infection is 10(3)-fold higher than the initial introduced titre. Our results indicated that the infection rates of unprotected epithelial cells probably play an important role in affecting viral dynamics. By simulating an advanced model of viral dynamics and applying it to experimental infection data of human influenza, we obtained important estimates of the infection rate. This work provides epidemiologically meaningful results, meriting further efforts to understand the causes and consequences of influenza A infection.

A simple allometric diffusion-based biokinetic model to predict Cu(II) uptake across gills of freshwater clam Corbicula fluminea.

The purpose of this study was to link Fick's type mass transfer and biokinetics together with Michaelis-Menten kinetics to arrive at a simple predictive framework for quantifying biouptake mechanisms in gills of freshwater clam Corbicula fluminea exposed to Cu(II). A diffusion-based Cu(II) influx and permeability can be calculated using physiological and allometric-related parameters. Simulations indicate that Cu(II) bioconcentration factor of gills was 42. Estimated steady-state Cu(II) gill uptake influx and permeability were 0.097 nmol cm(-2) s(-1) and 0.48 cm s(-1), respectively. The proposed simple allometric diffusion-based biokinetic model meets the need for describing nonequilibrium aspects of biouptake mechanisms in bivalve gills.

Understanding influenza virus-specific epidemiological properties by analysis of experimental human infections.

This study aimed to estimate the natural history and transmission parameters based on experimental viral shedding and symptom dynamics in order to understand the key epidemiological factors that characterize influenza (sub)type epidemics. A simple statistical algorithm was developed by combining a well-defined mathematical scheme of epidemiological determinants and experimental human influenza infection. Here we showed that (i) the observed viral shedding dynamics mapped successfully the estimated time-profile of infectiousness and (ii) the profile of asymptomatic probability was obtained based on observed temporal variation of symptom scores. Our derived estimates permitted evaluation of relationships between various model-derived and data-based estimations, allowing evaluation of trends proposed previously but not tested fully. As well as providing insights into the dynamics of viral shedding and symptom scores, a more profound understanding of influenza epidemiological parameters and determinants could enhance the viral kinetic studies of influenza during infection in the respiratory tracts of experimentally infected individuals.

Viral kinetics and exhaled droplet size affect indoor transmission dynamics of influenza infection.

The purpose of this paper was to investigate the effects of viral kinetics and exhaled droplet size on indoor transmission dynamics of influenza infection. The target cell-limited model with delayed virus production was adopted to strengthen the inner mechanisms of virus infection on human epithelial cell. The particle number and volume involved in the viral kinetics were linked with Wells-Riley mathematical equation to quantify the infection risk. We investigated population dynamics in a specific elementary school by using the seasonal susceptible - exposed - infected - recovery (SEIR) model. We found that exhaled pulmonary bioaerosol of sneeze (particle diameter <10 microm) have 10(2)-fold estimate higher than that of cough. Sneeze and cough caused risk probabilities range from 0.075 to 0.30 and 0.076, respectively; whereas basic reproduction numbers (R(0)) estimates range from 4 to 17 for sneeze and nearly 4 for cough, indicating sneeze-posed higher infection risk. The viral kinetics and exhaled droplet size for sneeze affect indoor transmission dynamics of influenza infection since date post-infection 1-7. This study provides direct mechanistic support that indoor influenza virus transmission can be characterized by viral kinetics in human upper respiratory tracts that are modulated by exhaled droplet size. Practical Implications This paper provides a predictive model that can integrate the influenza viral kinetics (target cell-limited model), indoor aerosol transmission potential (Wells-Riley mathematical equation), and population dynamic model [susceptible - exposed - infected - recovery (SEIR) model] in a proposed susceptible population. Viral kinetics expresses the competed results of human immunity ability with influenza virus generation. By linking the viral kinetics and different exposure parameters and environmental factors in a proposed school setting with five age groups, the influenza infection risk can be estimated. On the other hand, we implicated a new simple means of inhaling to mitigate exhaled bioaerosols through an inhaled non-toxic aerosol. The proposed predictive model may serve as a tool for further investigation of specific control measure such as the personal protection masks to alter the particle size and number concentration characteristics and minimize the exhaled bioaerosol droplet to decrease the infection risk in indoor environment settings.

A human PBPK/PD model to assess arsenic exposure risk through farmed tilapia consumption.

The purpose of this study was to develop a biologically based risk assessment model for human health through consumption of arsenic (As) contaminated farmed tilapia (Oreochromis mossambicus) from blackfoot disease (BFD)-endemic area in Taiwan for estimating the consumption advice. We linked a physiologically based pharmacokinetic (PBPK) and a pharmacodynamic (PD) model to account for the exposure and dose-response profiles of As in human. Risk analysis indicates that consumption of farmed tilapia poses no significant threat from As-induced lung and bladder cancers. The predicted risk-based median consumption advice was no more than 5-17 meals month(-1) (or 2-6 g day(-1)).

Online detection of waterborne bioavailable copper by valve daily rhythms in freshwater clam Corbicula fluminea.

Freshwater clam Corbicula fluminea, a surrogate species in metal toxicity testing, is a promising bioindicator of impairment in aquatic ecosystems. Little is known, however, about the relationship between clam valve daily rhythmic response and metal bioavailability related to a metal biological early warning system (BEWS) design. The purpose of this study was to link biotic ligand model (BLM)-based bioavailability and valve daily rhythm in C. fluminea to design a biomonitoring system for online in situ detection of waterborne copper (Cu). We integrated the Hill-based dose-time-response function and the fitted daily rhythm function of valve closure into a constructed programmatic mechanism. The functional presentation of the present dynamic system was completely demonstrated by employing a LabVIEW graphic control program in a personal computer. We used site-specific effect concentration causing 10% of total valve closure response (EC10) as the detection threshold to implement the proposed C. fluminea-based Cu BEWS. Here our results show that the proposed C. fluminea-based BEWS could be deliberately synthesized to online in situ transmit rapidly the information on waterborne bioavailable Cu levels under various aquatic environmental conditions through monitoring the valve daily rhythmic changes. We suggested that the developed C. fluminea-based dynamic biomonitoring system could assist in developing technically defensible site-specific water quality criteria to promote more efficient uses in water resources for protection of species health in aquatic environments.