The congenital cataract-causing mutations P20R and A171T are associated with important changes in the amyloidogenic feature,structure and chaperone-like activity of human αB-crystallin

Cataract is the major reason for human blindness worldwide. α-Crystallin, as a key chaperone of eye lenses, keeps the lenticular tissues in its transparent state over time. In this study, cataract-causing familial mutations, P20R and A171T, were introduced in CRYАB gene. After successful expression in Escherichia coli and subsequent purification, the recombinant proteins were subjected to extensive structural and functional analyses using various spectroscopic techniques, gel electrophoresis, and electron microscopy. The results of fluorescence and Raman assessments suggest important but discreet conformational changes in human αB-Cry upon these cataractogenic mutations. Furthermore, the mutant proteins exhibited significant secondary structural alteration as revealed by FTIR and Raman spectroscopy. An increase in conformational stability was seen in the human αB-Cry bearing these congenital cataractogenic mutations. The oligomeric size distribution and chaperone-like activity of human αB-Cry were significantly altered by these mutations. The P20R mutant protein was observed to loose most of the chaperone-like activity. Finally, these cataractogenic mutant proteins exhibited an increased propensity to form the amyloid fibrils when incubated under environmental stress. Overall, the structural and functional changes in mutated human αB-Cry proteins can shed light on the pathogenic development of congenital cataracts.     

Synthesis and anti-inflammatory activity of celecoxib like compounds

Nine novel 4-[3-(4-Dimethylamino-phenyl)-5-aryl-4,5-dihydro-pyrazol-1-yl]-benzenesulfonamides (2a-i) were synthesized and evaluated for their anti-inflammatory and antiproliferative activities. These compounds (2a-i) showed moderate to strong anti-inflammatory activity in carrageenan rat paw oedema test. Compounds 2b, 2d and 2g showing comparable anti-inflammatory activity to that of reference drug celecoxib were evaluated for their ulcerogenic and analgesic activities. The effect of 2b, 2d and 2g on the content of NO, TNF-α and PGE₂ in exudates from rat paw stimulated by carrageenan was also evaluated. The compound 2c showed considerable antitumor activities against all 60 human tumor cell lines with effective GI₅₀ (MG-MID) value of 3.63 µM. It exhibited maximum activity against melanoma (LOX IMVI and SK-MEL-5) cancer cell lines with GI₅₀ value less than 2 μM.     

Uric acid modulates vascular endothelial function through the down regulation of nitric oxide production

Abstract

Endothelial dysfunction characterized by decreased nitric oxide (NO) bioavailability is the first stage of coronary artery disease. It is known that one of the factors associated with an increased risk of coronary artery disease is a high plasma level of uric acid. However, causative associations between hyperuricaemia and cardiovascular risk have not been definitely proved. In this work, we tested the effect of uric acid on endothelial NO bioavailability. Electrochemical measurement of NO production in acetylcholine-stimulated human umbilical endothelial cells (HUVECs) revealed that uric acid markedly decreases NO release. This finding was confirmed by organ bath experiments on mouse aortic segments. Uric acid dose-dependently reduced endothelium-dependent vasorelaxation. To reveal the mechanism of decreasing NO bioavailability we tested the effect of uric acid on reactive oxygen species production by HUVECs, on arginase activity, and on acetylcholine-induced endothelial NO synthase phosphorylation. It was found that uric acid increases arginase activity and reduces endothelial NO synthase phosphorylation. Interestingly, uric acid significantly increased intracellular superoxide formation. In conclusion, uric acid decreases NO bioavailability by means of multiple mechanisms. This finding supports the idea of a causal association between hyperuricaemia and cardiovascular risk.     

Fruiting-inducing activity of cerebrosides observed with Schizophyllum commune

Isolation and identification of substances having an activity to stimulate the fruiting body formation of Schizophyllum commune were attempted. The active principles in its mycelia were divided into four fractions by sequential purification with silica gel column and reverse-phase HPLC column chromatography. By infrared spectra and thin-layer chromatography, the active substances in these four fractions were revealed as cerebrosides. About 0.1 μg of the cerebroside fractions showed a discriminative stimulating activity on S. commune when tested by the method these authors adopted. The active substance in the fraction II was N-2′-hydroxypalmitoyl-1-O-glucosyl-nonadecasphingadienine. The cerebrosides from pea seeds and Fusicoccum amygdali showed the similar activity on S. commune, but some commercial synthetic cerebrosides and cerebrosides from bovine and porcine brains exhibited no stimulating activity. Only definite cerebrosides with special structures seem to be able to induce the fruiting of S. commune.     

Seminalplasmin: A protein with many biological properties

Proteins present in the seminal plasma of mammals are known to influence functions associated with ejaculated spermatozoa such as motility, capacitation, acrosome reaction and fertilising ability. The proteins isolated and characterised so far influence only one of the above functions of spermatozoa. Seminalplasmin, a protein isolated from the seminal plasma of bull is exceptional in that it influences many of the above spermatozoal functions. It is also a potent antimicrobial protein and capable of lysing microbial and mammalian cells. The physiological function of seminalplasmin as nature's own antifertility agent is discussed.     

Performance of Two Fruit Fly (Diptera: Tephritidae) Pupal Parasitoids (Coptera haywardi [Hymenoptera: Diapriidae] and Pachycrepoideus vindemiae [Hymenoptera: Pteromalidae]) under Different Environmental Soil Conditions

We evaluated the performance of Coptera haywardi (Ogloblin) (Diapriidae) and Pachycrepoideus vindemiae (Rondani) (Pteromalidae), both hymenopteran pupal parasitoids of Anastrepha spp. (Diptera: Tephritidae). Performance was studied by manipulating the following environmental conditions in the laboratory: (1) soil type, (2) soil moisture content, (3) soil compaction, and (4) depth at which pupae were buried in the soil. There were two experiments: in the first, exposure time of pupae was held constant and in the second, it varied. In the first experiment, C. haywardi was significantly more effective than P. vindemiae in parasitizing fly pupae. With exposure time held constant (36 h), only soil type and pupal burial depth were significantly related to parasitism rates. While P. vindemiae only parasitized pupae located on the soil surface, C. haywardi attacked pupae that were buried up to 5 cm deep, performing better in clayey than in loamy soil. In the second experiment, exposure time (24, 36, 48, and 72 h) had no significant effect on parasitism rates, but soil type did. P. vindemiae again only attacked pupae on the soil surface while C. haywardi was also able to parasitize pupae that were buried up to 5 cm deep. We conclude that C. haywardi represents a viable candidate to replace the environmentally unfriendly P. vindemiae in augmentative biological control programs against fruit flies.     

Influence of activity in a heterogeneous environment on the dynamics of fish growth: an individual-based model of roacl

A spatio-temporal individual-based model (IBM), including bioenergetic principles, is used to investigate how energy costs coupled to activity represent a variable fraction in a heterogeneous environment and how this is related to another energetic criterion, the potential individual growth, under the conditions of a structured environment. With this approach, it is possible to connect simulated spatio-temporal activity patterns with the energetic needs required for these activities. By using simple foraging rules and a modified random walk model it is possible to reproduce spatial distributions and length frequency distributions. The simulated spatial distribution of roach Rutilus rutilus in Lake Belau, Germany, results in a mean weekly activity multiplier [(standard metabolic rate + activity costs) (standard metabolic rate)⁻¹] of 1–9 with deviations >100% during a simulated year. These deviations are of key importance to differences in the growth rate of individual simulated roach.     

The haemagglutination activity in the different developmental stages and changes of this activity caused by analogues of insect hormones in two members of the Culex pipiens complex of mosquitoes

Abstract. Haemagglutination activity was studied in last larval instars, pupae and adults of both sexes of the Culex pipiens mosquito complex. In females of Culex pipiens quinquefasciatus Say, an anautogenous member of C. pipiens complex, activity was detected in the homogenate of the head‐thorax complex, gut and remaining abdomen (after removing the gut), before and after sucking water, glucose solution or blood. Similar investigations were made in females of autogenous Culex pipiens molestus Forskal, a member of the same mosquito complex. The results were compared with data obtained for males of these subspecies, plus the larvae and pupae. The results show that haemagglutination activity in adult females depends not on the sucking of blood, but on the age of the females. In newly emerged females, the level of activity was very low, but after 24 h or 48 h of activity was highly enhanced, even in females that had no possibility to suck water or other liquids including blood. Furthermore, in C. p. molestus, the haemagglutination activity does not change in hungry or engorged females, and a high level of activity was found in the gut of last larval instars, a developmental stage that never takes blood. Treatment with methoprene reduces haemagglutination activity in both sexes, but 20‐hydroxycdysone application decreased activity in the gut of females. It is suggested that haemagglutination activity may be regulated indirectly by the endocrine system. The possibility that haemagglutination activity may depend on some aspect of the digestive system that has no direct connection to blood uptake is discussed. In addition, the activity may be important not only for the elimination of infections, but also for the processing of food and the utilization and transport of nutrients.