Protective effects of green tea extracts (polyphenon E and EGCG) on human cervical lesions

We investigated clinical efficacy of green tea extracts (polyphenon E; poly E and (-)-epigallocatechin-3-gallate [EGCG]) delivered in a form of ointment or capsule in patients with human papilloma virus (HPV) infected cervical lesions. Fifty-one patients with cervical lesions (chronic cervicitis, mild dysplasia, moderate dysplasia and severe dysplasia) were divided into four groups, as compared with 39 untreated patients as a control. Poly E ointment was applied locally to 27 patients twice a week. For oral delivery, a 200 mg of poly E or EGCG capsule was taken orally every day for eight to 12 weeks. In the study, 20 out of 27 patients (74%) under poly E ointment therapy showed a response. Six out of eight patients under poly E ointment plus poly E capsule therapy (75%) showed a response, and three out of six patients (50%) under poly E capsule therapy showed a response. Six out of 10 patients (60%) under EGCG capsule therapy showed a response. Overall, a 69% response rate (35/51) was noted for treatment with green tea extracts, as compared with a 10% response rate (4/39) in untreated controls (P<0.05). Thus, the data collected here demonstrated that green tea extracts in a form of ointment and capsule are effective for treating cervical lesions, suggesting that green tea extracts can be a potential therapy regimen for patients with HPV infected cervical lesions.     

NMR structure and mutagenesis of the FADD (Mort1) death-effector domain

When activated, membrane-bound receptors for Fas and tumour-necrosis factor initiate programmed cell death by recruiting the death domain of the adaptor protein FADD to the membrane. FADD then activates caspase 8 (also known as FLICE or MACH) through an interaction between the death-effector domains of FADD and caspase 8. This ultimately leads to the apoptotic response. Death-effector domains and homologous protein modules known as caspase-recruitment domains have been found in several proteins and are important regulators of caspase (FLICE) activity and of apoptosis. Here we describe the solution structure of a soluble, biologically active mutant of the FADD death-effector domain. The structure consists of six antiparallel, amphipathic alpha-helices and resembles the overall fold of the death domains of Fas and p75. Despite this structural similarity, mutations that inhibit protein-protein interactions involving the Fas death domain have no effect when introduced into the FADD death-effector domain. Instead, a hydrophobic region of the FADD death-effector domain that is not present in the death domains is vital for binding to FLICE and for apoptotic activity.     

Mechanism of simultaneous solutes separation and concentration by size exclusion cyclic separation

The semicontinuous separation/concentration of two solutes with different molecular size by the size exclusion cyclic separation method is based on the opposite swelling responses of two gels to a temperature change. Experimental results of separation and concentration of solutes are verified qualitatively by the theoretical models based on the local equilibrium assumption. Separation of two solutes is shown by the breakthrough curves in coupled gel columns. In closed coupled columns, the volumetric space for the large molecules which are totally excluded from the gels becomes smaller, creating a concentrating effect as the gels swell when temperature changes. A mechanistic model is suggested to predict the large molecule concentration to increase to its solubility limit as cycle repeats.     

Role of conserved Asn-Tyr-Asp-Tyr sequence in bacterial copper/2,4, 5-trihydroxyphenylalanyl quinone-containing histamine oxidase

Copper amine oxidase contains a covalently bound quinonoid cofactor, 2,4,5-trihydroxyphenylalanyl quinone (TPQ), which is synthesized by post-translational modification of a specific tyrosyl residue occurring in the highly conserved sequence, Asn-Tyr-(Asp/Glu)-Tyr. To elucidate the role(s) of the conserved sequence in the biogenesis of TPQ, each of the corresponding residues at positions 401-404 in the recombinant histamine oxidase from Arthrobacter globiformis has been replaced with other amino acids by site-directed mutagenesis. When Asn-401 was changed to Asp or Gln, the rate of TPQ formation by copper-dependent self-processing was 10(3)- to 10(4)-fold slower than in the wild-type enzyme. When Tyr-402 was replaced by Phe, TPQ was not formed at all, showing that Tyr-402 is essential as the precursor to TPQ. In contrast, Asp-403 could be replaced by Glu without changes in the rate of TPQ formation, whereas its replacement by Asn led to a marked decrease. Furthermore, when Tyr-404 was changed to Phe, TPQ was formed swiftly on incubation with copper ions, but the TPQ enzyme exhibited very low activity with altered substrate specificity. These results collectively indicate that a very rigorous structural motif is required for efficient formation of TPQ and for the catalytic activity in the active site of copper amine oxidases.     

Phylogenetic relationship within Fusarium sambucinum Fuckel sensu lato, determined from ribosomal RNA sequences

The importance of abnormalities of function and epididymis structure in the etiology of male infertility is still not well understood. We studied 52 individuals distributed in five age groups: fetuses, children, adolescents, adults and the elderly. The region of the body of epididymis was obtained by autopsy and immediately fixed by immersion in a solution of 10% buffered formaldehyde, embedded in paraffin and histologically prepared. The samples were observed under an optic microscope. Test-points were counted in 12 random microscopic fields with the M42 test-system. The following stereological parameters were determined: ductal area, volumetric densities (Vv) of the duct, smooth muscle, connective tissue, epithelial duct and blood vessels. The main results distinguished by those whose averages were statistically significant (p < 0.05), showed that the ductal area is 9.7 times greater in the adolescent/adult/elderly group than the children's group. The Vv of the lumen of the epididymis duct occupies 11.7% of the epididymis body in the fetal period, 5.3% in the child and in individuals after puberty this figures reaches more than 15%. The Vv of smooth muscle occupies 28.3% of the body of the epididymis in the fetus and 35.9% in children, but after puberty this figures stays around 22%. The Vv of the connective tissue occupies 26% in prenatal life, 37% in children, and after puberty these figures range from 21 to 27.5%. Comparing the results of the adult group with that of the elderly group there is an increase in the volumetric density of the connective tissue by 18.1%. In conclusion, the epididymal duct area and the Vv of the ductal lumen, smooth muscle and connective tissue were significant comparing the different groups. However, the quantitative relative differences of the duct's epithelium and the blood vessels were not significant comparing these groups. The study of quantitative aspects of the normal human epididymis can increase our knowledge about male fertility.     

Activity of quinolones in the Ames Salmonella TA102 mutagenicity test and other bacterial genotoxicity assays

Eight quinolones were examined for their bacterial mutagenicity in the Ames Salmonella TA102 assay and for their effects in other bacterial genotoxicity assays. In the quantitative Ames plate incorporation assay, all eight quinolones induced His+ deletion reversion in Salmonella tester strain TA102, with maximum reversion observed at about two to eight times the MIC. The quinolones also induced the SOS response. At quinolone concentrations close to the MIC, SOS cell filamentation gene sulA was induced in sulA::lacZ fusion strain Escherichia coli PQ37. RecA-mediated cleavage of lambda repressor in lambda::lacZ fusion strain E. coli BR513 was measurable at about 10 times the MIC, though no induction occurred at 20 micrograms of nalidixic or oxolinic acid per ml. Genotoxicity of quinolones also was observed in the Bacillus subtilis DNA repair assay, in which the mutant strain M45 (recA) was more susceptible to quinolones than its parent strain, H17 (rec+). The results from these analyses indicate that quinolones induce SOS functions and are mutagenic in bacteria; these properties correspond to their antimicrobial activities.