Integration of 60,000 exomes and ACMG guidelines question the role of Catecholaminergic Polymorphic Ventricular Tachycardia-associated variants.

Catecholaminergic Polymorphic Ventricular Tachycardia (CPVT) is a highly lethal cardiac arrhythmia disease occurring during exercise or psychological stress. CPVT has an estimated prevalence of 1:10,000 and has mainly been associated with variants in calcium-regulating genes. Identification of potential false-positive pathogenic variants was conducted by searching the Exome Aggregation Consortium (ExAC) database (n = 60,706) for variants reported to be associated with CPVT. The pathogenicity of the interrogated variants was assessed using guidelines from the American College of Medical Genetics and Genomics (ACMG) and in silico prediction tools. Of 246 variants 38 (15%) variants previously associated with CPVT were identified in the ExAC database. We predicted the CPVT prevalence to be 1:132. The ACMG standards classified 29% of ExAC variants as pathogenic or likely pathogenic. The in silico predictions showed a reduced probability of disease-causing effect for the variants identified in the exome database (p < 0.001). We have observed a large overrepresentation of previously CPVT-associated variants in a large exome database. Based on the frequency of CPVT in the general population, it is less likely that the previously proposed variants are associated with a highly penetrant monogenic form of the disease.

Genotypic and phenotypic characterization of garlic-fermenting lactic acid bacteria isolated from som-fak, a Thai low-salt fermented fish product.

AIMS: To evaluate the importance of garlic for fermentation of a Thai fish product, and to differentiate among garlic-/inulin-fermenting lactic acid bacteria (LAB) at strain level. METHODS AND RESULTS: Som-fak was prepared by fermentation of a mixture of fish, salt, rice, sucrose and garlic. pH decreased to 4.5 in 2 days, but omitting garlic resulted in a lack of acidification. LAB were predominant and approximately one third of 234 isolated strains fermented garlic and inulin (the carbohydrate reserve in garlic). These strains were identified as Lactobacillus pentosus and Lact. plantarum. Randomly Amplified Polymorphic DNA (RAPD) analysis revealed one major RAPD type (29 strains) isolated from all stages of fermentation. CONCLUSION: Garlic was essential for acidification of som-fak and garlic-fermenting strains constituted a significant, homogeneous part of the LAB flora. SIGNIFICANCE AND IMPACT OF THE STUDY: The present study indicates the role of fructans (garlic/inulin) as carbohydrate sources for LAB. Fructan fermenters may have several biotechnological applications, for example, as probiotics.

Characterization of lactic acid bacteria isolated from a Thai low-salt fermented fish product and the role of garlic as substrate for fermentation.

Lactic acid bacteria (LAB) isolated from raw materials (fish, rice, garlic and banana leaves) and processed som-fak (a Thai low-salt fermented fish product) were characterized by API 50-CH and other phenotypic criteria. Lactococcus lactis subsp. lactis and Leuconostoc citreum were specifically associated with fish fillet and minced fish, Lactobacillus paracasei subsp. paracasei with boiled rice and Weisella confusa with garlic mix and banana leaves. In addition, Lactobacillus plantarum, Lactobacillus pentosus and Pediococcus pentosaceus were isolated from raw materials. A succession of aciduric, homofermentative lactobacillus species, dominated by Lb. plantarum/pentosus, was found during fermentation. In total, 9% of the strains fermented starch and 19% fermented garlic, the two main carbohydrate components in som-fak. The ability to ferment garlic was paralleled by a capacity to ferment inulin. An increased percentage of garlic fermenting strains was found during fermentation of som-fak, from 8% at day 1 to 40% at day 5. No starch fermenting strains were isolated during fermentation. Three mixed LAB cultures, composed of either starch fermenting Lc. lactis subsp. lactis and Lb. paracasei subsp. paracasei, or garlic fermenting Lb. plantarum and Pd. pentosaceus, or a combination of these strains were inoculated into laboratory prepared som-fak with or without garlic. In som-fak without garlic, pH was above 4.8 after three days, irrespective of addition of mixed LAB cultures. The starch fermenting LAB were unable to ferment som-fak and sensory spoilage occurred after three days. Fermentation with the combined mix of starch and garlic fermenting strains led to production of 2.5% acid and a decrease in pH to 4.5 in two days. The fermentation was slightly slower with the garlic fermenting strains alone. This is the first report describing the role of garlic as carbohydrate source for LAB in fermented fish products.

Evaluation of the role of Carnobacterium piscicola in spoilage of vacuum- and modified-atmosphere-packed cold-smoked salmon stored at 5 degrees C.

The microflora on spoiled cold-smoked salmon often consists of a mixture of lactic acid bacteria (LAB) and Gram-negative bacteria. To elucidate the role of the different groups, a storage trial was carried out in which nisin and CO2 were used for the selective inhibition of the two bacterial groups. The shelf-life of vacuum-packed cold-smoked salmon, recorded by sensory evaluation, was four weeks at 5 degrees C and the microflora was composed of LAB (10(6)-10(7) cfu/g) with an associate Gram-negative flora in varying levels (10(5)-10(7) cfu/g). The addition of nisin and/or a CO2-atmosphere increased the shelf-life to five or six weeks and limited the level of LAB to about 10(4)-10(6), 10(3)-10(6) and 10(2)-10(4) cfu/g, respectively. CO2-atmosphere +/- nisin inhibited the growth of Gram-negative bacteria, whereas nisin had no effect on these in vacuum packages. The Gram-negative flora on vacuum-packed salmon was dominated by a Vibrio sp., resembling V. marinus, Enterobacteriaceae (Enterobacter agglomerans, Serratia liquefaciens and Rahnella aquatilis) and occasionally Aeromonas hydrophila. Irrespective of the addition of nisin and/or CO2-atmosphere, the LAB microflora was dominated by Carnobacterium piscicola, which was found to account for 87% of the 255 LAB isolates characterized. Whole-cell-protein patterns analysed by SDS-PAGE confirmed the Carnobacterium species identification. The spoilage potential of C. piscicola isolates was further studied by inoculation of approx. 10(6) cfu/g in cold-smoked salmon stored at 5 degrees C. The salmon did not spoil within 4 weeks of storage in vacuum- or CO2-atmosphere, and it is concluded that despite high levels (> 10(7) cfu/g) of C. piscicola, sensory rejection was caused by autolytic changes. This was supported by the development of soft texture and sour, rancid and bitter off-flavours at the point of spoilage, irrespective of the length of shelf-life and low or high total counts of LAB and Gram-negative bacteria.

Analysis of starvation conditions that allow for prolonged culturability of Vibrio vulnificus at low temperature.

The response of the estuarine human pathogen Vibrio vulnificus to starvation for carbon, nitrogen or phosphorus, or all three nutrients simultaneously (multiple-nutrient), was examined with respect to the maintenance of culturability during incubation at low temperature. V. vulnificus showed similar survival patterns during starvation for the individual nutrients when kept at 24 degrees C. On the other hand, cultures prestarved at 24 degrees C and then shifted to 5 degrees C maintained culturability at low temperature in a starvation-condition-dependent manner. Carbon and multiple-nutrient starvation were indistinguishable in their ability to mediate maintenance of culturability in the cold. Prolonged starvation for phosphorus had a similar effect, but nitrogen starvation did not allow for maintenance of culturability. Extracellular factors produced during starvation were not observed to have an effect on the culturability of cells incubated at low temperature. Protein synthesis during starvation for individual nutrients was analysed by two-dimensional PAGE of pulse-labelled proteins. Carbon and multiple-nutrient starvation gave nearly identical protein induction patterns involving at least 34 proteins, indicating that carbon starvation determines both responses. Nitrogen starvation for 1 h induced 24 proteins, while phosphorus starvation induced a set of 10 proteins after 1 h and about 40 proteins after 18 h. It is suggested that starvation for carbon or phosphorus induces maintenance of culturability of V. vulnificus incubated at low temperature via the synthesis of distinct sets of starvation-specific proteins.