DNA Promoter Methylation-dependent Transcription of the Double C2-like Domain β (DOC2B) Gene Regulates Tumor Growth in Human Cervical Cancer

Double C2-like domain β (DOC2B) gene encodes for a calcium-binding protein, which is involved in neurotransmitter release, sorting, and exocytosis. We have identified the promoter region of the DOC2B gene as hypermethylated in pre-malignant, malignant cervical tissues, and cervical cancer cell lines by methylation-sensitive dimethyl sulfoxide-polymerase chain reaction and bisulfite genome sequencing; whereas, it was unmethylated in normal cervical tissues (p < 0.05). The promoter hypermethylation was inversely associated with mRNA expression in SiHa, CaSki, and HeLa cells and treatment with demethylating agent 5-aza-2-deoxycytidine restored DOC2B expression. The region −630 to +25 bp of the DOC2B gene showed robust promoter activity by a luciferase reporter assay and was inhibited by in vitro artificial methylation with Sss1 methylase prior to transient transfections. Overexpression of the DOC2B gene in SiHa cells when compared with controls showed significantly reduced colony formation, cell proliferation, induced cell cycle arrest, and repressed cell migration and invasion (p < 0.05). Ectopic expression of DOC2B resulted in anoikis-mediated cell death and repressed tumor growth in a nude mice xenograft model (p < 0.05). DOC2B expressing cells showed a significant increase in intracellular calcium level (p < 0.05), impaired AKT1 and ERK1/2 signaling, and induced actin cytoskeleton remodeling. Our results show that promoter hypermethylation and silencing of the DOC2B gene is an early and frequent event during cervical carcinogenesis and whose reduced expression due to DNA promoter methylation may lead to selective cervical tumor growth.     

Aminoparthenolides as novel anti-leukemic agents: Discovery of the NF-κB inhibitor,DMAPT (LC-1)

A series of aminoparthenolide analogs (6–37) were synthesized and evaluated for their anti-leukemic activity. Eight compounds exhibited good anti-leukemic activity with LD₅₀’s in the low μM range (1.5–3.0 μM). Compounds 16, 24 and 30 were the most potent compounds in the series, causing greater than 90% cell death at 10 μM concentration against primary AML cells in culture, with LD₅₀ values of 1.7, 1.8 and 1.6 μM.     

General relationships of mammalian orders and evolutionary development of primates inferred from best-fit α-globin phylogenies

A best-fit α-globin phylogeny was identified by means of a global minimization approach from among the topologies generated by a parsimony strategy. Zip parsimony method was used to derive a set of near-parsimonious trees. For each of these topologies, a difference matrix was computed; and the topology with the best goodness of fit with the original matrix was retained as the best tree. Based on this phylogenetic scheme inter-relationships among eutherian orders and the evolutionary development of primates has been discussed.     

Transient up-regulation of myotonic dystrophy protein kinase-binding protein, MKBP, and HSP27 in the neonatal myocardium.

Myotonic dystrophy protein kinase (DMPK)-binding protein, MKBP, has high homology with a small heat shock protein, HSP27. Western blotting analyses showed that MKBP level in rat heart rapidly increased, with a sharp peak at one week after birth (3-fold the level at the fetus), but that it rapidly decreased (1/10 of peak value at 13 weeks). Human myocardium also showed similar age-dependency. Similar but small increase of HSP27 was observed in the neonatal rat myocardium, but not in constitutive and inducible forms of HSP70. Immunofluorescence analysis localized MKBP at the Z lines and intercalated discs in the rat myocardium. MKBP may protect actin cytoskeleton or other proteins of heart muscle against oxidative stress in the neonate.     

Ultraviolet irradiation apparatus for disinfecting liquids of high ultraviolet absorptivities

A nozzle was used to generate freely supported totally enclosed thin liquid films, known as 'liquid bells', from suspensions of Escherichia coli in water and humic acid solutions. Disinfection was achieved by continuously recirculating the liquids to the nozzle and irradiating the bells generated with ultraviolet light. The absorptivities at 254 nm of the liquids treated ranged from 0.18 to 4.0 and after 30 min treatment E. coli viability, expressed in terms of surviving fractions, was reduced to 1.88 times 10-5 and 1.84 times 10-4 respectively.     

Fitness consequences of body melanization in Drosophila immigrans from montane habitats

We investigated eight populations of Drosophila immigrans from low to high montane localities (600–2202 m) for altitudinal variations in abdominal melanization and fitness-related traits (desiccation resistance, copulation duration, and fecundity). On the basis of common garden experiments, persistence between-population differences at 21°C suggests that observed variations in fitness-related traits have a genetic basis. Parent–offspring regression analyses showed higher heritability (h²= 0.77) for melanization patterns on all the abdominal tergites. All the traits showed significantly higher repeatability across generations. Under colder and drier environments in dispersed montane localities, abdominal melanization and desiccation resistance significantly increased (1.5–1.9 fold) along altitude. Thus, there are correlated effects of abdominal melanization on desiccation resistance. Genetic correlations, based on family means, were significantly high between abdominal melanization and other fitness traits. Furthermore, darker flies along increasing altitude resulted in a 35–40% increase in copulation duration as well as fecundity. There are significantly positive correlations of abdominal melanization with copulation duration as well as fecundity on the basis of within- as well as between-population variations. Such observations are in agreement with the thermal budget hypothesis. Present data suggest that changes in body melanization impact fitness-related traits in montane populations of Drosophila immigrans .     

Listeria monocytogenes adheres to many materials found in food-processing environments

AIMS: To investigate the adhesion of Listeria monocytogenes 10403S to 17 different, food-use approved materials representing metals, rubbers and polymers. METHODS AND RESULTS: Adhesion assays were conducted by placing 'coupons' of the materials in planktonic cultures at 30 degrees C, and then immediately withdrawing them ('short contact') or leaving them submerged in the cultures for 2 h. Adherent cells were recovered by sonication. In the short contact experiments, the logarithm of the mean viable counts ranged from 3.67 +/- 0.43 to 4.78 +/- 0.38. After 2 h contact time, the numbers of adherent cells had increased significantly for all materials with the exception of polypropylene. The highest count (6.33 +/- 0.31) recorded was for stainless steel 405. CONCLUSION: Adhesion to a wide range of materials was time-dependent and characterized by reversible and irreversible stages. SIGNIFICANCE AND IMPACT OF THE STUDY: Adhesion test protocols must account for cell carry-over and cells which are only weakly bound. Material selection may only have a limited role in reducing food contamination by listeria.