A regulatory SNP causes a human genetic disease by creating a new transcriptional promoter

We describe a pathogenetic mechanism underlying a variant form of the inherited blood disorder alpha thalassemia. Association studies of affected individuals from Melanesia localized the disease trait to the telomeric region of human chromosome 16, which includes the alpha-globin gene cluster, but no molecular defects were detected by conventional approaches. After resequencing and using a combination of chromatin immunoprecipitation and expression analysis on a tiled oligonucleotide array, we identified a gain-of-function regulatory single-nucleotide polymorphism (rSNP) in a nongenic region between the alpha-globin genes and their upstream regulatory elements. The rSNP creates a new promoterlike element that interferes with normal activation of all downstream alpha-like globin genes. Thus, our work illustrates a strategy for distinguishing between neutral and functionally important rSNPs, and it also identifies a pathogenetic mechanism that could potentially underlie other genetic diseases.     

The effects of drug detection training on behavioral reactivity and blood neurotransmitter levels in drug detection dogs: A preliminary study

The aim of the present study was to analyze the effects of drug detection training on behavior and blood neurotransmitter levels in drug detection dogs so as to investigate some variables influencing dog reactivity and responsiveness to training. In all, 20 dogs were sampled out of the Guardia di Finanza canine population. All the subjects were born, reared, housed, and trained in the same facility and followed the same training sessions. Dogs’ behavioral reactivity was scored according to a standardized working dogs test to evaluate natural dog attitudes. Plasma samples were analyzed by the high-performance liquid chromatography method to evaluate adrenaline, noradrenaline, L-3,4-dihydroxyphenylalanine, homovanillic acid (HVA), 3-methoxy-4-hydroxyphenylglycol acid (MHPG), 5-hydroxyindole acetic acid (5-HIAA), and 5-hydroxytryptamine (5-HT) levels. 5-HT and 5-HIAA were also analyzed from platelets. The analysis was carried out considering training, breed, and sex as independent variables. From a behavioral point of view, significant differences were recorded before and after training in “sociability,” “playfulness,” “predatory instinct,” and “aggressiveness” scores. Lower levels of platelet 5-HT and 5-HIAA were found after training. Plasma L-3,4-dihydroxyphenylalanine levels differed between sexes, with males showing higher concentrations. These results underline the importance of complete and objective evaluations protocols of the dogs before, during, and after drugs search training to determine effective and successful selection strategies and training procedures.     

A quantitative assessment of primary and secondary immune responses in cattle using a B cell ELISPOT assay

The aim of the study was to build a comprehensive picture of the appearance in the blood stream of Ag-specific plasma cells and memory B cells in the bovine model. For this purpose, we have developed a method allowing the detection and quantification of both cell types within individual calves immunised with ovalbumin. During the primary response, we detected a burst of ovalbumin-specific plasma cells at days 6 and 7 post-immunisation, which was followed by the production of specific Ab, whereas a gradual increase of memory B cells was only detected from day 15. As expected, a boost immunisation performed 7 weeks later induced a quicker and stronger secondary response. Indeed, a burst of plasma cells was detected in the blood at days 3 and 4, which was followed by a strong increase in Ab titres. Furthermore, a burst of memory B cells, and not a gradual increase, was detected at days 5 and 6 post-boost immunisation. Importantly, we showed a strong correlation between the anti-ovalbumin-specific IgG titres detected 5 months after secondary immunisation and the plasma cell numbers detected in the blood at the peak response after secondary immunisation. The detection and quantification of plasma cells following an mmunisation/vaccination strategy could constitute a very effective means for predicting the magnitude and longevity of an Ab response.     

Salmonella isolated from individual reptiles and environmental samples from terraria in private households in Sweden

Background

This study investigates Salmonella spp. isolated from privately kept reptiles and from environmental samples such as bedding materials or water from the floor of the enclosures (terraria). It also compares isolation of Salmonella using Modified Semisolid Rappaport-Vassiliadis (MSRV) medium or selective enrichment in Rappaport-Vassiliadis-Soya (RVS) pepton broth. Cloacal swabs or swabs from the cloacal area were collected from 63 individual reptiles belonging to 14 households. All reptiles were from different terraria and from 62 of these, environmental samples were also collected. Sampling were done by the reptile owners according to written instructions and sent by mail immediately after sampling. All but three samples were analyzed within 24 h after collection. Colonies suspected for Salmonella were tested for agglutination and serotyped using the White-Kauffmann-Le Minor scheme. The relative sensitivity (se) and specificity (sp) for MSRV compared with RVS, and the agreement coefficient kappa (κ) were calculated.

Results

Salmonella was isolated from 50/63 (80%) terraria, either from the reptiles (31/63; 49%) or from bedding material (39/62; 63%). The most common subspecies was Salmonella enterica subspecies enterica followed by S. enterica subspecies diarizonae. In reptiles, the most common S. enterica subspecies enterica serovars were Java (n = 4) and Fluntern (n = 4), compared with the serovars Tennessee (n = 10) and Fluntern (n = 10) in the environmental samples. The exact same set of Salmonella subspecies and serovars were not isolated from the individual reptiles and the environmental samples from any of the households. Isolation using MSRV yielded more Salmonella isolates 61/113 (54%) than enrichment in RVS 57/125 (46%). The se was 97.9% (95% Confidence Interval 93.9-100), the sp 78.5% (95% CI 68.5-88.5) and the κ 0.74, indicating substantial agreement between the tests.

Conclusions

Salmonella can be expected to be present in environments where reptiles are kept. This constitutes public health risks and should be considered during handling of the reptiles and during cleaning and disposal of bedding. A combination of different culturing techniques may be used to increase the isolation rate.     

A genomic regulatory network for development

Development of the body plan is controlled by large networks of regulatory genes. A gene regulatory network that controls the specification of endoderm and mesoderm in the sea urchin embryo is summarized here. The network was derived from large-scale perturbation analyses, in combination with computational methodologies, genomic data, cis-regulatory analysis, and molecular embryology. The network contains over 40 genes at present, and each node can be directly verified at the DNA sequence level by cis-regulatory analysis. Its architecture reveals specific and general aspects of development, such as how given cells generate their ordained fates in the embryo and why the process moves inexorably forward in developmental time.     

Processed sweet corn has higher antioxidant activity

Processed fruits and vegetables have been long considered to have lower nutritional value than the fresh produce due to the loss of vitamin C during processing. Vitamin C in apples has been found to contribute <0.4% of total antioxidant activity, indicating most of the activity comes from the natural combination of phytochemicals. This suggests that processed fruits and vegetables may retain their antioxidant activity despite the loss of vitamin C. Here it is shown that thermal processing at 115 degrees C for 25 min significantly elevated the total antioxidant activity of sweet corn by 44% and increased phytochemical content such as ferulic acid by 550% and total phenolics by 54%, although 25% vitamin C loss was observed. Processed sweet corn has increased antioxidant activity equivalent to 210 mg of vitamin C/100 g of corn compared to the remaining 3.2 mg of vitamin C in the sample that contributed only 1.5% of its total antioxidant activity. These findings do not support the notion that processed fruits and vegetables have lower nutritional value than fresh produce. This information may have a significant impact on consumers' food selection by increasing their consumption of fruits and vegetables to reduce the risk of chronic diseases.