A competitive enzyme linked immunosorbent assay for recombinant Bm86 produced in Pichia pastoris

A competitive enzyme linked immonosorbent assay (ELISA) was developed for quantification of recombinant Bm86 protein (rBm86), antigen of the vaccine GavacTM against the cattle tick Boophilus microplus. Monoclonal antibody (Mab) SSBm1 showed identical recognition to folded and denatured antigen and was used in the competition assay. This ELISA was sensitive enough to detect 60 ng/ml of rBm86. Within-assay coefficient of variation was 5.7 to 6.2 % and between-assay variation was 7.7 to 11.6 %. The level of expression of recombinant Bm86 in Pichia pastoris reached about 2.7 g per liter of culture after 80 hour of fermentation. © Rapid Science Ltd. 1998     

A new weather factor predicting airborne pollen concentration: Péczely’s macrosynoptic weather types

In the present study determinative factors for ragweed (Ambrosia elatior) pollination were studied in Budapest between 1991 and 1996. The aim was to create a model to predict the day-to-day pollen count variation. The pollen concentration is determined mostly by the potential concentration and the mean concentration of the three previous days. These two variables can explain 56% of the total variance. Daily mean and maximum temperature, daily temperature fluctuation and the number of hours of sunshine in the previous day have a significant positive effect on the pollen count. The amount of precipitation on the previous day, relative humidity on the actual and on the previous day influence it negatively. When a cyclone prevails, pollen concentration is usually lower than the seasonal average. Some anticyclonal situations have a similar effect, while other anticyclonal types promote pollination. A predictive model was then created by multiple regression using the potential concentration, the mean concentration of the three previous days, the daily temperature fluctuation and the synoptic type as independent variables. This model can explain more than 68% of the total variance, and its accuracy is >71%. The model seems to predict accurately the trends during the pollen seasons, thus it will probably be a good tool in the practical prediction in Budapest, and the methodology will hopefully be applicable to other sites of the Carpathian Basin too.     

Biosynthesis and ultrasonic degradation of bacterial poly(gamma-glutamic acid)

A study of the production of poly(gamma-glutamic acid) (PGGA) by Bacillus licheniformis NCIMB 11709 grown on medium E in shake flasks at 30 degrees C is reported. The enantiomeric composition of PGGA was found to be highly sensitive to the concentration of Mn++, especially when the ion is present in small amounts (相似文献   

Biochemical and genetic characteristics of TEM-29B, a novel extended spectrum beta-lactamase

A clinical strain of Escherichia coli (strain Ec 41553) that was resistant to ceftazidime produced a TEM-type beta-lactamase with a pI of 5.4. Clavulanic acid restored the ceftazidime activity, thus suggesting an extended spectrum beta-lactamase (ESBL). The gene encoding ESBL was located in a plasmid of 57 kb. After cloning and sequencing, the ESBL (TEM-29B) showed one amino acid replacement with respect to the TEM-1 sequence, Arg-164 to His. This change increased mainly the rate of hydrolysis of ceftazidime but not of cefotaxime and aztreonam. The relevance of this substitution in the increase of ceftazidime MIC is thus stressed.     

Molecular analysis of a ring chromosome X in a family with fragile X syndrome

The phenotypically normal sister of a patient affected by fragile X syndrome was referred for genetic counselling and was found to carry a mosaic karyotype 46,X,r(X)/45,X. Because the probability of the simultaneous chance occurrence of fragile X syndrome and a ring chromosome X in the same family is very low, we postulated that the breakpoint of the ring chromosome X originated in the cytogenetic break in Xq27.3 responsible for fragile X syndrome. In order to determine the relative positions of the breakpoint on the ring chromosome X and the (CGG)n unstable sequence responsible for the fragile X mutation, we used molecular markers to analyse the telomeric regions of chromosome X in this family. The results showed that the ring chromosome X was the maternal fragile X chromosome and that the telomeric deletion on the long arm encompassed the (CGG)n sequence. This suggests that the cytogenetic break in Xq27.3 is distinct from the unstable (CGG)n sequence, or that the break followed by the end-to-end fusion creating the ring chromosome was not completely conservative. Analysis of DNA markers on the short arm of chromosome X evidenced a deletion of a large part of the pseudoautosomal region, allowing us to position the genes involved in stature and in some syndromes associated with telomeric deletions of Xp on the proximal side of the pseudoautosomal region.     

The temporal variation of intertidal nematodes in the Westerschelde I. The importance of an estuarine gradient

Biannual meiobenthic sampling (Spring and Autumn) was carried out in 1983–1989 at two fine sandy intertidal stations in the Westerschelde estuary. Both stations are exposed daily for more than one hour and are situated in the polyhaline and the mesohaline zone of the estuary. Average density data of non-selective deposit-feeders > predators > epigrowth-feeders > selective deposit-feeders for both stations are presented in spite of different nematode species composition. No difference between Spring and Autumn nor trend over 7 years could be detected. Higher total nematode densities are found at the polyhaline station (average 3200 ind. 10 cm^?2) in comparison with the mesohaline station (average 2300 ind. 10 cm^?2), a difference mainly due to higher non-selective deposit-feeders and predators densities in the polyhaline station. Each year, heterogeneous variance is found for all feeding types at the mesohaline station, but only for epigrowth-feeders and predators at the polyhaline station. The higher nematode density at the polyhaline station is probably caused by the more stable nematode structure. An unstable nematode temporal pattern at the mesohaline station is suggested to be combined with the detritus food chain system in the mesohaline zone. The unstable estuarine habitats are mainly caused by their upstream effects: the River Schelde, which clearly influences the stability of the nematode communities.     

Mechanism of Free Fatty Acid Effects on Hepatocyte Insulin Receptor Binding and Processing

We determined whether the paimitate effects on hepatocyte insulin receptor binding and post-receptor trafficking were mediated by accelerated mitochondrial (β-oxidation or accumulation of intracellular fatty acyl-CoA derivatives and possibly protein acyiation. Preincubation of hepatocytes with moderate concentrations of paimitate (0.5 mM) resulted in a 23% decline in cell-surface binding and proportional decreases in receptor-mediated insulin internalization and degradation. Brief pretreatment of hepatocytes with the carnitine palmityltransferase-I inhibitor, methyl paimoxirate (MP), prevented 70% of the paimitate effects. At higher paimitate concentrations (2.0 mM), cell-surface binding was reduced by 34%, whereas internalization of the receptor complex was reduced by 78%. These effects were only partially prevented by MP pretreatment. Receptor-mediated insulin degradation increased by 34% and was uninfluenced by MP pretreatment. Octanoate, which is rapidly shunted into mitochondrial oxidation, produced a dose-dependent reduction in insulin binding, with proportional decreases in internalization and degradation. Similarly preincubation with 2.0 mM oleate, which, unlike palmitate, is not known to produce protein acylation, resulted in proportional decreases in insulin receptor binding and receptor-mediated internalization and degradation. High concentrations of octanoate or oleate (2.0 mM) did not reproduce the additive postreceptor effects of palmitate. We conclude that the receptor and post-receptor effects of moderate palmitate concentrations are closely linked to accelerated fatty acid oxidation. The post-receptor effects observed at higher concentrations involve other mechanisms, possibly relating to intracellular levels of palmityl-CoA derivatives.     

Body wall muscles in the haplotaxidsHaplotaxis gordioides andPelodrilus leruthi (Annelida,Oligochaeta)

Summary The muscle organization of the body wall in two species of Haplotaxida is described.Haplotaxis gordioides andPelodrilus leruthi show significant differences in the longitudinal muscle layer. The observations suggest that inH. gordioides, the muscle fibres could foreshadow the flat circomyarian organization of microdrile muscles, while theP. leruthi organization recalls the pinnate arrangement of megadrile body wall.     

Molecular studies of a translocated (X;22) DiGeorge patient using somatic cell hybridization.

In order to better characterize the chromosomic rearrangement of an unbalanced 45XX t(X;22) (q28;q11) DiGeorge patient, a somatic hybrid clone segregating the translocated chromosome was constructed and investigated using X and 22 linked markers. Our study demonstrated that this de novo translocation was from paternal origin. The breakpoint was assigned between DXS296 and IDS loci at Xq28 and between D22S9 and BCRL2 at 22q11. This observation and published data allow to locate a "critical region" for DiGeorge syndrome between these two last loci on 22q11. Our hybrid clone may be a useful tool for mapping new probes arising in this region.     

Morphogenesis of body wall muscle fibers in Enchytraeus minutus

The helical muscle fibers of Enchytraeus minutus (Annelida, Enchytraeidae) have been investigated with respect to their development. We have shown that longitudinal fibers, in the cocoon, begin to develop as platymyarian fibers, then develop into shallow coelomyarian ones and finally, just before hatching, reach a flat circomyarian stage. After hatching, the fibers grow only in size due to an increase in contractile material in the helical sarcomere system. Indications are given as to the ways this process can take place.