Frequent mutation of BAP1 in metastasizing uveal melanomas

Metastasis is a defining feature of malignant tumors and is the most common cause of cancer-related death, yet the genetics of metastasis are poorly understood. We used exome capture coupled with massively parallel sequencing to search for metastasis-related mutations in highly metastatic uveal melanomas of the eye. Inactivating somatic mutations were identified in the gene encoding BRCA1-associated protein 1 (BAP1) on chromosome 3p21.1 in 26 of 31 (84%) metastasizing tumors, including 15 mutations causing premature protein termination and 5 affecting its ubiquitin carboxyl-terminal hydrolase domain. One tumor harbored a frameshift mutation that was germline in origin, thus representing a susceptibility allele. These findings implicate loss of BAP1 in uveal melanoma metastasis and suggest that the BAP1 pathway may be a valuable therapeutic target.     

Phosphorylation by p38 MAPK as an alternative pathway for GSK3beta inactivation

Glycogen synthase kinase 3beta (GSK3beta) is involved in metabolism, neurodegeneration, and cancer. Inhibition of GSK3beta activity is the primary mechanism that regulates this widely expressed active kinase. Although the protein kinase Akt inhibits GSK3beta by phosphorylation at the N terminus, preventing Akt-mediated phosphorylation does not affect the cell-survival pathway activated through the GSK3beta substrate beta-catenin. Here, we show that p38 mitogen-activated protein kinase (MAPK) also inactivates GSK3beta by direct phosphorylation at its C terminus, and this inactivation can lead to an accumulation of beta-catenin. p38 MAPK-mediated phosphorylation of GSK3beta occurs primarily in the brain and thymocytes. Activation of beta-catenin-mediated signaling through GSK3beta inhibition provides a potential mechanism for p38 MAPK-mediated survival in specific tissues.     

Synchronizing rock clocks of Earth history

Calibration of the geological time scale is achieved by independent radioisotopic and astronomical dating, but these techniques yield discrepancies of approximately 1.0% or more, limiting our ability to reconstruct Earth history. To overcome this fundamental setback, we compared astronomical and 40Ar/39Ar ages of tephras in marine deposits in Morocco to calibrate the age of Fish Canyon sanidine, the most widely used standard in 40Ar/39Ar geochronology. This calibration results in a more precise older age of 28.201 +/- 0.046 million years ago (Ma) and reduces the 40Ar/39Ar method's absolute uncertainty from approximately 2.5 to 0.25%. In addition, this calibration provides tight constraints for the astronomical tuning of pre-Neogene successions, resulting in a mutually consistent age of approximately 65.95 Ma for the Cretaceous/Tertiary boundary.     

Insulin and IGF-I receptors and tyrosine kinase activity in carp ovaries: changes with reproductive cycle

Insulin and insulin-like growth factor I (IGF-I) receptors from carp ovaries were semipurified with wheat germ agglutinin at different moments of the reproductive cycle and their binding characteristics and tyrosine kinase activity were studied. Specific receptors for insulin and IGF-I were found. IGF-I receptors presented higher binding (23.8 ± 1.5%), number of receptors (965 ± 20fm/mg) and affinity (K_D 0.24 ± 0.03nM) than those shown for insulin receptors (4.1 ± 1%, 530 ± 85fm/mg and 0.85 ± 0.1nM, respectively). Insulin and IGF-I receptors have a tyrosine kinase activity which is not different from that found in muscle of the same species. Seasonal changes were found in binding, with maximum values for insulin and IGF-I reached at the end of pre-spawning period (June). However, while IGF-I binding was observed in all stages, insulin binding decreased in autumn and disappeared in winter, which suggests a different role for the two peptides in ovarian physiology.     

Development and characterization of a cell line TTCF from endangered mahseer Tor tor (Ham.)

Tor tor is an important game and food fish of India with a distribution throughout Asia from the trans-Himalayan region to the Mekong River basin to Malaysia, Pakistan, Bangladesh and Indonesia. A new cell line named TTCF was developed from the caudal fin of T. tor for the first time. The cell line was optimally maintained at 28°C in Leibovitz-15 (L-15) medium supplemented with 20% fetal bovine serum (FBS). The propagation of TTCF cells showed a high plating efficiency of 63.00%. The cytogenetic analysis revealed a diploid count of 100 chromosomes at passage 15, 30, 45 and 60 passages. The viability of the TTCF cell line was found to be 72% after 6 months of cryopreservation in liquid nitrogen (-196°C). The origin of the cell lines was confirmed by the amplification of 578- and 655-bp sequences of 16S rRNA and cytochrome oxidase subunit I (COI) genes of mitochondrial DNA (mtDNA) respectively. TTCF cells were successfully transfected with green fluorescent protein (GFP) reporter plasmids. Further, immunocytochemistry studies confirm its fibroblastic morphology of cells. Genotoxicity assessment of H?O? in TTCF cell line revealed the utility of TTCF cell line as in vitro model for aquatic toxicological studies.     

An investigation into the bacterial aetiology of 'black patch necrosis' in Dover sole, Solea solea L.

Abstract. A recurring disease known as 'black patch necrosis' (BPN) has been the cause of severe mortalities in 0-group and 1-group Dover sole during the period 1974-1978 at the Sea Fish Industry Authority's Marine Cultivation Unit at Hunter-ston, Ayrshire, Scotland. Samples of diseased and healthy Dover sole tissue were examined microbiologically and a long thin filamentous Gram-negative organism, strongly resembling Flexibacter columnar's , was repeatedly isolated from diseased tissue but was not found in healthy tissue. This organism was pathogenic to Dover sole, producing 100% mortality in 96 h at 17.5°C (±2°C) and the organism was reisolated from all the infected fish. The pathogenic isolate and reisolate were examined and compared with reference strain NCMB 1038 of Flexibacter columnarts , formerly known as Chondrococcus columnaris or Cytophaga columnaris. The isolate, reisolate and type culture were identical with respect to cell morphology, oxidase test, catalase production, production of H₂S, casein hydrolysis, gelatin liquefaction, hydrolysis of Tween 20, degradation of tyrosine and starch hydrolysis. All bacterial types showed similar patterns of sensitivity to a range of antibiotics. It is proposed that a Flexibacter columnari's-like organism is associated with the condition in farmed Dover sole known as 'black patch necrosis'.     

Soybean meal alters autochthonous microbial populations, microvilli morphology and compromises intestinal enterocyte integrity of rainbow trout, Oncorhynchus mykiss (Walbaum)

Rainbow trout were fed either a diet containing fishmeal (FM) as the crude protein source or a diet containing 50% replacement with soybean meal (SBM) for 16 weeks. An enteritis-like effect was observed in the SBM group; villi, enterocytes and microvilli were noticeably damaged compared with the FM group. The posterior intestine microvilli of SBM-fed fish were significantly shorter and the anterior intestine microvilli significantly less dense than the FM-fed fish. Electron microscopy confirmed the presence of autochthonous bacterial populations associated with microvilli of both fish groups. Reduced density of microvilli consequently led to increased exposure of enterocyte tight junctions, which combined with necrotic enterocytes is likely to diminish the protective barrier of the intestinal epithelium. No significant differences in total viable counts of culturable microbial populations were found between the groups in any of the intestinal regions. A total of 1500 isolates were tentatively placed into groups or genera, according to standard methods. Subsequent partial 16S rRNA sequencing revealed species that have not been identified from the rainbow trout intestine previously. Compared with the FM group levels of Psychrobacter spp. and yeast were considerably higher in the SBM group; a reduction of Aeromonas spp. was also observed.     

Method for Determining Apparent Digestibility of Carbohydrate and Protein Sources for Artificial Diets for Juvenile Sea Cucumber,Australostichopus mollis

Effective nursery diets can greatly reduce the cost of producing juvenile sea cucumbers for release to aquaculture or restocking programs. However, methods for systematically testing the bioavailability of artificial diet ingredients for sea cucumbers are poorly developed, and consequently, there has been little research in this field. The current study presents methods developed to test the suitability of common carbohydrate and protein sources for inclusion in artificial diets for juvenile Australostichopus mollis. Apparent digestibility of carbohydrates was moderate and did not exceed 50% for any carbohydrates assayed. Wheat starch and carrageenans showed the highest digestibility amongst carbohydrates. Differences in apparent digestibility were not reflected in growth performance of juvenile A. mollis fed carbohydrate diets; overall growth performance was poor for all diets. Artificial protein sources consistently exhibited higher apparent digestibility, ranging from 75.1% for fish meal to 98.1% for casein. Low‐cost protein sources, like meat meal, show promise for future use in artificial diet formulation. However, delivering high protein content may reduce ingestion rates and thereby lower overall digestive efficiency in juveniles. Results show that artificial carbohydrate sources have some potential as diet constituents for juvenile sea cucumbers but are unsuitable as primary energy sources. Future testing of artificial carbohydrate sources for A. mollis may require predigestion to improve digestibility.     

Growth of Juvenile Crayfish Procambarus llamasi (Villalobos 1955) Fed Different Farm and Aquaculture Commercial Foods

In this study, a range of commercially available animal and fish feeds were tested in the production of juveniles of the crayfish Procambarus llamasi, an endemic species of the Yucatan Peninsula, Mexico. A randomized block design was implemented to observe the effect of commercial shrimp (Crude Protein, CP = 38%), trout (CP = 43.2%), tilapia (CP = 31.8%), rabbit (CP = 16.1%), turkey (CP = 18%), and pig (CP = 15.2%) diets in a recirculating aquaculture system. Shrimp food showed the best results in terms of growth performance, while lowest growth values (P<0.05) were obtained with farm animal diets. Trout and tilapia diets were intermediate. Pig feed was the most cost-effective at US$ 0.44 per kg of crayfish growth. Where no feeds specifically for crayfish are available, terrestrial animal feeds have the advantage of low price, ready availability in the market, and acceptable performance.