Continuous-flow thermal gradient PCR

Continuous-flow thermal gradient PCR is a new DNA amplification technique that is characterized by periodic temperature ramping with no cyclic hold times. The device reported in this article represents the first demonstration of hold-less thermocycling within continuous-flow PCR microfluidics. This is also the first design in which continuous-flow PCR is performed within a single steady-state temperature zone. This allows for straightforward miniaturization of the channel footprint, shown in this device which has a cycle length of just 2.1 cm. With a linear thermal gradient established across the glass device, the heating and cooling ramp rates are dictated by the fluid velocity relative to the temperature gradient. Local channel orientation and cross-sectional area regulate this velocity. Thus, rapid thermocycling occurs while the PCR chip is maintained at steady state temperatures and flow rates. Glass PCR chips (25 x 75 x 2 mm) of both 30 and 40 serpentine cycles have been fabricated, and were used to amplify a variety of targets, including a 181-bp segment of a viral phage DNA (PhiX174) and a 108-bp segment of the Y-chromosome, amplified from human genomic DNA. With this unique combination of hold-less cycling and gradient temperature ramping, a 40-cycle PCR requires less than 9 min, with the resulting amplicon having high yield and specificity.     

Assessment of risks and dose thresholds for some effects of acute exposure

Findings from the analyses of the dose-response relationship are reviewed with regard to different effects of acute radiation exposure. The analyses have been performed based on the dosimetry and clinical data for the nuclear workers acutely exposed to gamma rays or gamma rays and neutrons as a result of radiation accidents at the Mayak Production Association (Russia). The statistically significant risk curves for morbidity and mortality from acute radiation syndrome (ARS), as well as risks of the onset of vomiting at the prodromal phase and agranulocytosis, have been obtained. The Weibull model appropriately describes the corresponding risk curves. Estimates of the dose thresholds have been obtained for ARS morbidity (～0.7 Gy) and mortality (～6-7 Gy), vomiting at the prodromal phase (～1.5 Gy), and agranulocytosis (～3.5 Gy). The statistically significant power dependence between the onset of vomiting at the prodromal phase and the onset of agranulocytosis, as well as the dose dependence for the onset of agranulocytosis, has been revealed.     

Estimating the effective reproduction number for pandemic influenza from notification data made publicly available in real time: A multi-country analysis for influenza A/H1N1v 2009

In the early phase of an emerging pandemic such as A/H1N1v 2009, it is essential to have a good understanding of its transmissibility, which is often summarized by the reproductive number. Before a country is affected, its government may want to make their own assessment of what is going on in areas of the world that have previously been affected by the disease. However, having access to detailed data is problematic. The only publicly available international dataset with information for a large number of countries was the WHO cumulated case counts per country. In this paper, we show how and in which situations the recorded history of cumulated case counts provides valuable information to estimate the effective reproductive number in an early phase and for a large number of countries.     

School site and the potential to walk to school: the impact of street connectivity and traffic exposure in school neighborhoods

The impact of neighborhood walkability (based on street connectivity and traffic exposure) within 2 km of public primary schools on children regularly walking to school was examined. The most (n=13) and least walkable (n=12) schools were selected using a school-specific 'walkability' index and a cross sectional study undertaken of Year 5, 6 and 7 children (n=1480) and consenting parents (n=1332). After adjustment, regularly walking to school was higher in children attending schools in high walkable neighborhoods (i.e, high street connectivity and low traffic volume) (Odds ratio (OR) 3.63; 95% Confidence Interval (CI) 2.01-6.56), and less likely in neighborhoods with high connectivity but high traffic volume (OR 0.32; 95% CI 0.22-0.47). Connected street networks provide direct routes to school but when designed for heavy traffic, the potential for children to walk to school is reduced. This highlights the importance of carefully considering school siting and, particularly, street design in school neighborhoods.     

Mitigation Effects of Bentonite and Yeast Cell Wall Binders on AFB1, DON,and OTA Induced Changes in Laying Hen Performance,Egg Quality,and Health

The objective of this study was to evaluate the efficacy of mycotoxin binders in reducing the adverse effects of co-occurring dietary aflatoxin B₁ (AFB₁), deoxynivalenol (DON) and ochratoxin A (OTA) on laying hens. Three hundred and sixty 26-week-old Roman laying hens were randomly allocated into four experimental groups with 10 replicates of nine birds each. The four groups received either a basal diet (BD; Control), a BD supplemented with 0.15 mg/kg AFB₁ + 1.5 mg/kg DON + 0.12 mg/kg OTA (Toxins), a BD + Toxins with Toxo-HP binder (Toxins + HP), or a BD + Toxins with TOXO XL binder (Toxins + XL) for 12 weeks. Compared to the control, dietary supplementation of mycotoxins decreased (P < 0.10) total feed intake, total egg weight, and egg-laying rate, but increased feed/egg ratio by 2.5–6.1% and mortality during various experimental periods. These alterations induced by mycotoxins were alleviated by supplementation with both TOXO HP and XL binders (P < 0.10). Furthermore, dietary mycotoxins reduced (P < 0.05) eggshell strength by 12.3% and caused an accumulation of 249 μg/kg of DON in eggs at week 12, while dietary supplementation with TOXO HP or XL mitigated DON-induced changes on eggshell strength and prevented accumulation of DON in eggs (P < 0.05). Moreover, dietary mycotoxins increased relative liver weight, but decreased spleen and proventriculus relative weights by 11.6–22.4% (P < 0.05). Mycotoxin exposure also increased alanine aminotransferase activity and reduced immunoglobulin (Ig) A, IgM, and IgG concentrations in serum by 9.2–26.1% (P < 0.05). Additionally, mycotoxin exposure induced histopathological damage and reduced villus height, villus height/crypt depth, and crypt depth in duodenum, jejunum and (or) ileum (P < 0.05). Notably, most of these histological changes were mitigated by supplementation with both TOXO HP and XL (P < 0.05). In conclusion, the present study demonstrated that the mycotoxin binders TOXO HP and XL can help to mitigate the combined effects of AFB₁, DON, and OTA on laying hen performance, egg quality, and health.     

Vagal control of lower oesophageal sphincter motility in the cat

1. The effects of vagal efferent fibre stimulation on the smooth muscle of the lower oesophageal sphincter have been studied on the anaesthetized animal and on the isolated and perfused organ.2. In both muscle layers (longitudinal and circular) vagal stimulation elicits two types of electromyographic (e.m.g.) potentials: (a) excitatory junction potentials (e.j.p.s) where there is a depolarization of the smooth muscle fibres. E.j.p.s can give rise to spike potentials inducing a contraction of the sphincter; (b) inhibitory junction potentials (i.j.p.s) where there is hyperpolarization of the smooth muscle fibres, often followed by a transient depolarization which may initiate spikes (post-inhibitory rebound).3. Pure i.j.p.s are observed after atropine treatment which suppresses e.j.p.s. Under these conditions, a long lasting vagal stimulation induces a long duration hyperpolarization concomitant with an opening of the lower oesophageal sphincter followed after the cessation of stimulation by a powerful rebound leading to a strong contraction which closes the sphincter.4. Several arguments, pharmacological (action of acetylcholine (ACh), atropine and hexamethonium) and physiological (threshold and latency of responses) lead to the following conclusions.Preganglionic vagal fibres are cholinergic and they activate (a) intramural excitatory cholinergic neurones; (b) intramural non-adrenergic inhibitory neurones (purinergic neurones). Preganglionic fibres leading to inhibition have a higher threshold than those leading to excitation.Both excitatory and inhibitory pathways are interconnected inside the intramural network. In particular, activation of intramural inhibitory neurones, by relaxing the oesophagus orally to the lower oesophageal sphincter, inhibits intramural excitatory neurones and subsequently blocks vagal excitatory responses.5. Two functions may be attributed to the vagal extrinsic innervation: (a) closure of the lower oesophageal sphincter by maintaining the basal tone of the sphincter; this would imply that at rest the inhibitory control is supplanted by the excitatory one; (b) sphincter opening during swallowing by suppressing the excitatory stimulus and reinforcing the inhibitory one (it may be recalled that after bilateral vagotomy, swallowing is no longer followed by a relaxation of the sphincter).