Improved method for production of antibodies against T-2 toxin and diacetoxyscirpenol in rabbits.

A new, improved approach for the production of antibodies against T-2 toxin and diacetoxyscirpenol (DAS) was developed. The method involves the use of immunogens which were prepared by conjugating O-carboxymethoxyl oxime (CMO) derivatives of both toxins to bovine serum albumin (BSA). Isomers a and b of CMO-T-2 toxin and isomer b of CMO-DAS were tested. Antibodies against both toxins were demonstrated as early as 4 weeks after immunization. a-CMO-T-2-BSA conjugate was a better immunogen than the b isomer, and the highest titers (6,000) were reached 14 weeks after immunization and one booster injection. Antibody titers for rabbits immunized with the b isomer of CMO-T-2 never reached more than 2,000. The specificity of antibodies obtained from rabbits after immunization with CMO-T-2-BSA was similar to that of hemisuccinate-T-2-BSA. Anti-b-T-2 antibodies had slightly higher cross-reactivity with H-T-2 toxin than did the antibody obtained from rabbits immunized with the conjugate of the a isomer. The relative cross-reactivities of anti-a-CMO-T-2 antibody with T-2, acetyl-T-2, H-T-2, T-2-triol, 3'-OH-T-2, and T-2 tetraol were 1, 4.5, 5.7, 250, 500, and 3,000, respectively. The relative cross-reactivities of anti-b-T-2 antibody with T-2, acetyl-T-2, H-T-2, and T-2 triol were 1, 2, 3, and 488, respectively. Antibodies against b-CMO-DAS showed a high degree of cross-reactivity with monoacetoxyscirpenols (MAS). The relative cross-reactivities of anti-B-DAS antibody with DAS, 4-MAS, 15-MAS, acetyl-deoxynivalenol, T-2-toxin, acetyl-T-2, and neosolaniol were 1, 4, 5, 76, 107, 147, and 266, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Increased 5-Hydroxytryptamine and Norepinephrine Release from Rat Brain Slices by the Red Sea Flatfish

The effect of the Red Sea flatfish toxin pardaxin was examined on K+-evoked and on basal release of either [3H]norepinephrine or [3H]5-hydroxytryptamine from preloaded rat cortical slices. The K+-induced release of the neurotransmitters was stimulated in a dose-related manner at concentrations ranging from 0.5 to 4 micrograms/ml. Basal release of the two transmitters was elevated to a lesser extent. Although the stimulation of evoked release was approximately equivalent for the two neurotransmitters, the response to 5-hydroxytryptamine was reversible whereas that of norepinephrine was not washed by 20 min of superfusion. The mechanisms involved in producing these actions of pardaxin are not known; however, they may be mediated by changes in electrolyte fluxes across the neuronal membranes.     

伸展素在黄瓜胞壁中的转化及其与羟脯氨酸和异联酪氨酸的关系

用~(14)C-Pro和~3H_2-Tyr离体暗培养黄瓜子叶,发现细胞质、SECW和RCW中的Hyp/Pro和Idt/Tyr都随时间呈线性增加。这两种比值后两者高于前者,而两种比值增加速度之比在胞质部分最大、RCW中最小。表明在胞质和胞壁中都有Pro羟化和Tyr异联化过程,但羟化作用主要发生在胞质中,异联化主要在RCW中。 理化处理(高渗溶液和CHM)和放射性示踪证明胞质中存在HRGP库;它被分泌到胞壁后,先以离子键与壁结合,后转变为共价键与壁结合的伸展素。     

Direct measurement of carbon substrate oxidation and incorporation patterns in RuMP-type methylotrophs: Chemostatic cultures of Methylomonas L3

A technique using C-14 isotope tracers to probe the branching of carbon flow in methylotrophic bacteria has been devised and applied to continuous steady-state cultures. Methylomonas L3, a strain which utilizes the KDPG/TA variant of the ribulose monophosphate cycle for carbon fixation, was employed in the experimental studies. The actual in vivo rates of substrate-carbon incorporation into biomass, both direct and via CO(2), and of the two carbon oxidation schemes were determined in three different steady-state cultures. The results show that the carbon substrate is oxidized predominantly via formate (the linear oxidation scheme), and that the cyclic scheme of oxidation is minimally, if at all, utilized. The carbon incorporation and oxidation patterns appear to vary considerably with the dilution rate and the inoculum history. The experimental accuracy of the new technique is discussed in detail.     

The expression of ceruloplasmin, an angiogenic glycoprotein, by mouse embryonic fibroblasts

Balb/c 3T3, Swiss 3T3 and Rous sarcoma virus transformed Balb/c 3T3 mouse embryonic fibroblasts produced ceruloplasmin in vitro, whereas primary cultures prepared from the Balb/c mouse embryos did not produce ceruloplasmin. The amount of ceruloplasmin synthesis by the Balb/c 3T3 cell line is enhanced by Rous sarcoma virus-transformation (1.5-3 fold) and by treatment with dexamethasone (about 2.4 fold). The protein was identified as ceruloplasmin by immunoprecipitation with ceruloplasmin-specific polyclonal antibody, and by similarity of peptide maps, and subunit molecular weight (135,000 dalton) to that of authentic ceruloplasmin from primary cultures of mouse hepatocytes.     

Cloning and DNA sequence of a plasmid-determined citrate utilization system in Escherichia coli.

The citrate utilization determinant from a large 200-kilobase (kb) naturally occurring plasmid was previously cloned into the PstI site of plasmid vector pBR325 creating the Cit+ tetracycline resistance plasmid pWR61 (15 kb). Tn5 insertion mutagenesis analysis of plasmid pWR61 limited the segment responsible for citrate utilization to a 4.8-kb region bordered by EcoRI and PstI restriction nuclease sites. The 4.8-kb fragment was cloned into phage M13, and the DNA sequence was determined by the dideoxyribonucleotide method. Within this sequence was a 1,296-base-pair open reading frame with a preceding ribosomal binding site. The 431-amino-acid polypeptide that could be translated from this open reading frame would be highly hydrophobic. A second long open reading frame with the potential of encoding a 379-amino-acid polypeptide preceded the larger open reading frame. Portions of the 4.8-kb fragment were further subcloned with restriction endonucleases BglII and BamHI, reducing the minimum size needed for a citrate-positive phenotype to a 1.9-kb BamHI-BglII fragment (which includes the coding region for the 431-amino-acid polypeptide, but only the distal 2/3 of the reading frame for the 379-amino-acid polypeptide). Citrate utilization results from a citrate transport activity encoded by the plasmid. With the 4.8-kb fragment (as with larger fragments) the citrate transport activity was inducible by growth on citrate. On transfer from glucose, succinate, malate, or glycerol medium to citrate medium, the Cit+ Escherichia coli strains showed a delay of 36 to 48 h before growth.     

Sister chromatid differential staining and NOR activity of BrdU-resistant sublines

Summary Two 30 g/ml BrdU-resistant sublines and two 60 g/ml BrdU-resistant sublines are induced from a Chinese hamster cell line Wg3h (HGPRT^–) by one-step and two-step selections, respectively. By inoculating the cells into BrdU-free medium or by adding more BrdU into the culture medium for 26–27 h, it was found that the two BrdU-resistant sublines analysed have very clear sister chromatid differential (SCD) staining patterns. This indicates that some of the nuclear DNA of the BrdU-resistant cells incorporate with BrdU to reach a kinetic balance. Frequencies of sister chromatid exchange (SCE) of the resistant cells are twice to four times as high as those of the Wg3h cells, depending on which BrdU-resistant subline is analysed. The SCE frequencies of the resistant cells also increase with the BrdU concentration in the medium. Analysis of silver-stained nucleolar organizer regions (NORs) indicates that the NOR activity of three out of the four BrdU-resistant sublines is significantly suppressed, i.e., averages of the Ag-NOR number and number of the chromosomes bearing Ag-NORs per cell decrease significantly. The degree of suppression for different BrdU-resistant sublines may be quite different. The suppressed NOR activity of the resistant cells can gradually be restored when the cells are inoculated into BrdU-free medium, but the recovery speed is far lower than that of the Wg3h cells. The suppression of the NOR activity of the BrdU-resistant sublines should be due to BrdU toxicity.     

猕猴(Macaca mulatta)主动脉弓的分支类型

随着心血管系统研究的进展和猿猴在医学中的广泛应用,猴主动脉弓分支的情况引起人们的重视。虽然张胜泉等（1974）对猕猴的心脏冠状动脉的解剖学作过调查,但有关主动脉弓分支的资料很少。为此,我们进行了本题研究,以供参考。     

Carbon oxidation and incorporation patterns in batch cultures ofMethylomonas L3

Summary A recently developed methodology of directly measuring the oxidation and incorporation patterns of carbon substrate in continuous cultures of RuMP-type methylotrophs is extended to batch cultures of the obligate methylotrophMethylomonas L3. The ratio of cyclic to total substrate oxidation varies with the initial methanol concentration from 0 to 68%. Formaldehyde, as a methanol cosubstrate, enhances the net substrate oxidation. The substrate oxidation and incorporation pattern is also affected by the state of the culture inoculum.