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Bovine CD18 identified as a species specific receptor for Pasteurella haemolytica leukotoxin.

Ligand blotting of Pasteurella haemolytica leukotoxin (LKT) susceptible BL3 bovine lymphoma cell membranes with LKT detected two putative receptors with Mr of 95 and 100 kDa, whereas no LKT binding to membrane proteins was detected for LKT non-susceptible human leukemic cells. Anti-bovine CD18 and CD11a/CD18 mAb recognized 95 and 100kDa bands from BL3 cell membranes. CD18 isolated from BL3 cell membranes bound LKT. Pre-incubation of BL3 cells with anti-bovine CD18 or CD11a/CD18 mAb caused partial inhibition of LKT-induced leukolysis. Therefore, we propose that bovine CD18 acts as a species-specific leukocyte receptor for P. haemolytica LKT.

Animals↗

Fimbriae and determination of host species specificity of Bordetella bronchiseptica.

A monoclonal antibody, designated CF8 and prepared against fimbrial protein enrichments of Bordetella bronchiseptica 110H, was determined by immunogold electron microscopy to bind to some but not all fimbrial filaments on intact bacterial cells. Comparison of the reactivity of this antibody with that of monoclonal antibody BPF2, which is specific for Bordetella pertussis serotype 2 fimbriae, indicated that CF8 recognizes an epitope similar to that recognized by BPF2. By Western blot (immunoblot), it was determined that monoclonal antibody CF8 does not react with proteins denatured by treatment with sodium dodecyl sulfate and beta-mercaptoethanol and by boiling for 5 min but that it does recognize fimbrial proteins in their native, nondenatured state. This antibody was used to compare fimbriae between strains of B. bronchiseptica isolated from different species. Strains from pigs, dogs, guinea pigs, and four other species were compared by an enzyme immunoassay. Strains isolated from pigs were found to express significantly more CF8-reactive and B. pertussis serotype 2 cross-reactive fimbriae than strains isolated from guinea pigs. Strains from dogs were more variable in reactivity than those from pigs or guinea pigs. The reactivity with antifimbrial monoclonal antibody CF8 did not correlate with enzyme electromorphotype but did correlate with the host species, suggesting a role for fimbriae in the determination of host species specificity of B. bronchiseptica.

Animals↗

Mutational analysis of centromeric DNA elements of Kluyveromyces lactis and their role in determining the species specificity of the highly homologous centromeres from K. lactis and Saccharomyces cerevisiae.

The centromere of Kluyveromyces lactis was delimited to a region of approximately 280 bp, encompassing KlCDEI, II, and III. Removal of 6 bp from the right side of KlCDEIII plus flanking sequences abolished centromere function, and removal of 5 bp of KlCDEI and flanking sequences resulted in strongly reduced centromere function. Deletions of 20-80 bp from KlCDEII resulted in a decrease in plasmid stability, indicating that KlCDEII must have a certain length for proper centromere function. Centromeres of K. lactis do not function in Saccharomyces cerevisiae and vice versa. Adapting the length of KlCDEII to that of ScCDEII did not improve KlCEN function in S. cerevisiae, while doubling the ScCDEII length did not improve ScCEN function in K. lactis. Thus the difference in CDEII length is not in itself responsible for the species specificity of the centromeres from each of the two species of budding yeast. A chimeric K. lactis centromere with ScCDEIII instead of KlCDEIII was no longer functional in K. lactis, but did improve plasmid stability in S. cerevisiae, although to a much lower level than a wild-type ScCEN. This indicates that the exact CDEIII sequence is important, and suggests that the flanking AT-rich CDEII has to conform to specific sequence requirements.

Base Sequence↗

[Baculovirus p74 gene is a species-specific gene].

The p74 gene of Autographa californica multicasid nucleopolyhedrovirus (AcMNPV) bacmid was knockouted and substituted by the p74 gene of Spodoptera litura multicapsid nucleopolyhedrovirus (SpltMNPV), using RecA-mediated homologous recombination in the E. coli. No selection marker, which might influence the expression and function of p74 gene, was left in the modified p74 locus. The promoter of AcMNPV p74 gene directly controlled the expression of SpltMNPV p74 gene in the recombinant AcMNPV bacmid-polhSL74. RT-PCR showed that the substituted p74 gene was transcribed. Bioassay showed that the recombinant virus AcMNPV bacmid-polhSL74 could not infect the Argyrogramma agnata larvae per os, and thus showing the p74 gene is species-specific.

Animals↗

Identification of novel species-specific antigens of Mycoplasma hyopneumoniae by preparative SDS-PAGE ELISA profiling.

Mycoplasma hyopneumoniae, M. hyorhinis and M. flocculare are commonly isolated from the respiratory tract of pigs and are phylogenetically related. The identification and characterization of antigens specific for M. hyopneumoniae is crucial for the development of serological reagents and for understanding the mechanisms of pathogenicity of this pathogen. Protein and antigen profiles of six strains of M. hyopneumoniae, four strains of M. hyorhinis and a type strain of M. flocculare were compared using SDS-PAGE and immunoblotting. Five strains of M. hyopneumoniae originally isolated from diverse geographical regions produced similar protein and antigen profiles. One strain, C1735/2, produced a unique protein profile and was poorly immunoreactive, suggesting that some strains of M. hyopneumoniae may possess a structurally modified repertoire of antigens. Major M. hyopneumoniae antigens with molecular masses of approximately 36, 43, 48, 52, 76, 78, 80, 82, 94, 106, 114 and 200 kDa were identified by immunoblotting using hyperimmune pig sera raised against both high and low passage strains of M. hyopneumoniae. Porcine hyperimmune sera raised against the GDL type strain of M. hyorhinis reacted strongly with all M. hyorhinis strains although the profiles displayed considerable variation. Major antigens of molecular mass 42, 49, 52, 78, 80 and 82 kDa were identified in type strains GDL and BTS-7 and field strain 2; however, field strain 1 produced a unique profile. A preparative SDS-PAGE profiling (PPP) technique was developed which enabled quantification of the immunoreactivity of denatured antigens with porcine serum by ELISA. PPP facilitated the rapid identification of species-specific and cross-reactive antigens among the three mycoplasma species. PPP studies revealed several strongly immunoreactive M. hyopneumoniae-specific antigens of 43, 76, 94, 114 and 200 kDa as well as antigens of molecular mass between 52 and 62 kDa which were not apparent in immunoblotting studies. Rabbit monospecific anti-42 kDa serum reacted specifically with a 43 kDa antigen in whole cell lysates of geographically diverse strains of M. hyopneumoniae and failed to cross-react with M. flocculare or M. hyorhinis whole cell lysates. This study has identified a number of M. hyopneumoniae-specific antigens which warrant further investigation to determine their potential as diagnostic reagents and the role they play, if any, in pathogenicity.

Animals↗

Trypanosoma brucei: growth differences in different mammalian sera are not due to the species-specificity of transferrin.

The heterodimeric transferrin receptors of Trypanosoma brucei (Tf-Rs) are encoded by two genes termed ESAG7 and ESAG6. These genes belong to polycistronic transcription units contained in the multiple expression sites for the variant surface glycoprotein (VSG ESs), only one of which is active at a time. Each VSG ES carries a different copy of these genes, leading to alternative expression of Tf-Rs with quite distinct binding affinities for transferrins from various mammals. T. brucei clones exhibit marked growth differences depending on the species-specificity of the serum. Since transferrin is a vital growth factor for the parasite, we investigated whether it could be responsible for these observations. We analyzed the cases of Tf-Rs from two ESs preferentially selected for expression in man and mouse, respectively. We show that serum-dependent growth variations of trypanosomes expressing these receptors are independent of transferrin, and that both high- and low-affinity Tf-Rs allow efficient trypanosome growth in various sera, either in vivo or in vitro.

Animals↗

Species specificity in the blood cholesterol-lowering effect of YM-16638.

1. The compound YM-16638, [[5-[[3-(4-acetyl-3-hydroxy-2-propylphenoxy)propyl] thio]-1,3,4-thiadiazol-2-yl]thio] acetic acid was developed in a series of in vitro and in vivo studies as a leukotriene D4 receptor antagonist. 2. In a clinical trial as a leukotriene antagonist drug, this compound was found to have a potent serum cholesterol lowering effect in normolipidaemic healthy male volunteers. 3. In the present study, we investigated the serum cholesterol lower effect of this compound in various species of experimental animals. 4. Administration of YM-16638 did not cause a significant decrease in serum total cholesterol (TC) in mice (up to 200 mg kg-1, body weight per day for 28 days), rats (200 mg kg-1 for 15 days) or rabbits (90 mg kg-1 for 18 days). In hamsters, administration of YM-16638 orally or by peritoneal injection at 50 mg kg-1 or more daily for 7 days caused a significant decrease in serum TC and the rate of body weight gain. In monkeys, serum TC did not change in YM-16638-administered squirrel monkeys (50 mg kg-1 daily for 3 weeks), but a significant decrease in serum TC was observed in cynomolgus monkeys (33% decrease at 30 mg kg-1 for 4 weeks) and rhesus monkeys (27% decrease at 30 mg kg-1 for 3 weeks) without any serious decrease in body weight. These results were consistent with those in a phase I study with human subjects. In contrast, serum alanine aminotransferase (ALT) level decreased in all animals after YM-16638 treatment. 5. From these results, we conclude that YM-16638 has a potent hypocholesterolaemic effect, but that this effect if species-specific and is only recognized clearly in human subjects and old-world monkeys.

Alanine Transaminase↗

Killing of Proteus mirabilis by polymorphonuclear leukocyte granule proteins: evidence for species specificity by antimicrobial proteins.

Low-molecular-weight (Mr, ca. 3,800) polypeptides containing human defensins HNP-1 and HNP-2 (T. Ganz, M. S. Selsted, D. Szlarek, S. L. Harwig, K. Daher, D. F. Bainton, and R. I. Lehrer, J. Clin. Invest. 76:1427-1434, 1985) prepared in our laboratory from acid extracts of human polymorphonuclear granulocyte granules and purified human defensins were found to exert potent bactericidal action against Proteus mirabilis. The antimicrobial action of the extracts of human polymorphonuclear leukocytes granules against P. mirabilis appears to be due to the presence of the defensins. Because P. mirabilis resists the antimicrobial action of other granule proteins, we interpret the present results to mean that the various antimicrobial proteins display species specificity in their microbicidal action.

Blood Bactericidal Activity↗

Carbohydrate-mediated sperm-egg interaction and species specificity: a clue from the Unio elongatulus model.

As a first step fertilization requires that sperm bind to the extracellular coat of the egg. It is generally accepted that binding is mediated by complex carbohydrates on the egg coat, recognized by carbohydrate-binding proteins on the sperm surface. In the mollusc bivalve Unio elongatulus, the main functional epitope of the carbohydrate ligand has been determined, and it shares several characteristics with other sugar structures involved in the fertilization process in different animal models. A polyclonal antibody against the Unio epitope reacts with the human ZP3 glycoprotein and the Unio ligand binds human spermatozoa in an in vitro assay. These findings are discussed in the light of the species specificity of sperm-egg binding at fertilization.

Animals↗

Monoclonal antibodies directed against human tissue-type plasminogen activator: a characterization of their species specificity, affinity and heavy-chain binding.

Six monoclonal antibodies (mIgG) and a polyclonal antibody (pIgG) directed against human tissue-type plasminogen activator (t-PA) were tested for their species specificity towards human or murine t-PA. Whereas pIgG as well as several mIgGs discriminated poorly between these two t-PA species, one mIgG (clone E3) was highly specific for human t-PA. Inhibition and binding studies of human t-PA by mIgGs revealed high affinity-high inhibitory (E3) as well as high affinity-poor inhibitory (B1) mIgGs. The relative affinity of two mIgGs for human t-PA was found to be equal or even superior to that of pIgG. Immunoblotting of reduced two-chain t-PA and of an isolated heavy chain of t-PA prepared by recombinant DNA technology, showed that the E3 antibody was directed against the heavy chain of t-PA.

Animals↗

Isolation of a Pseudomonas solanacearum-specific DNA probe by subtraction hybridization and construction of species-specific oligonucleotide primers for sensitive detection by the polymerase chain reaction.

A subtraction hybridization technique was employed to make a library enriched for Pseudomonas solanacearum-specific sequences. One cloned fragment, PS2096, hybridized under stringent conditions to DNA of 82 P. solanacearum strains representing all subgroups of the species. Other plant-associated bacteria, including closely related species such as Pseudomonas capacia, Pseudomonas picketti, or Pseudomonas syzygii, did not hybridize to PS2096. A minimum number of between 4 x 10(5) and 4 x 10(6) P. solanacearum cells could routinely be detected with PS2096 labelled either with [32P]dCTP or with digoxigenin-11-dUTP. To improve the sensitivity of detection, PS2096 was sequenced to allow the construction of specific oligonucleotide primers to be used for polymerase chain reaction (PCR) amplification. After 50 cycles of amplification, 5 to 116 cells, depending on the strain, could reproducibly be detected by visualization of a 148-bp PCR product on an agarose gel. A preliminary field trial in Burundi with the probe and PCR primers has confirmed that they are sensitive tools for specifically detecting low-level infections of P. solanacearum in potato tubers.

Base Sequence↗

[Polymorphisms and species specificity of D2S2944 and D1S2134 loci in Chinese Han population in Chengdu].

OBJECTIVE: To obtain the data of polymorphism distribution of the two STR loci D2S2944, D1S2134 in Chinese Han population in Chengdu and evaluate their usefulness in the field of forensic science. METHODS: PCR, polyacrylamide gel electrophoresis (PAGE) and silver staining were used to analyze 120 unrelated individuals of Chinese Han ethnic group in Chengdu. Fourteen different animals: monkey, pig, dog, bull, goat, chicken, duck, fish, cat, rabbit, Guinea pig, mouse, eel and frog were selected as controls in this study for evaluating the species specificity of the two STR loci. RESULTS: Eight alleles and twenty-two genotypes were found in D2S2944. The observed heterozygosity (h), discrimination power (DP), polymorphism information content (PIC), chance of paternity exclusion power (EP) were 0.824, 0.925, 0.78, 0.644 respectively. Ten alleles and twenty-six genotypes were observed in D1S2134. The h, DP, PIC and EP were 0.769, 0.920, 0.79, 0.543 respectively. The genotype distributions of the two loci were analyzed by some related software and no deviation from the Hardy-Weinberg equilibrium was observed. Evaluated by way of using different animals as controls, the two STR loci of human beings were found to have good specificity. CONCLUSION: These data indicate that D1S2134 and D2S2944 are of good polymorphism, high EP and DP, and can be applied as the candidate genetic marks to forensic parentage testing and identification. The methods to analyze them are simple, dependable and repeatable.

Alleles↗

Analysis of fish ZP1/ZPB homologous genes--evidence for both genome duplication and species-specific amplification models of evolution.

The vertebrate egg envelope is composed of a family of related proteins, the zona pellucida (ZP) proteins, which are characterized by the presence of a conserved zona pellucida domain. Analysis of teleost fish ZP gene sequences has demonstrated that there are no direct orthologues of the mammalian ZPB and ZP1 genes, but that teleost fish contain multiple copies of two classes of genes (ZPXa and ZPXb) that are equally related to ZPB and ZP1. The two classes of genes are further distinguished by expression in liver or ovary, respectively, indicating there was probably an initial duplication event, followed by a switch to hepatic expression of one of the paralogues. This switch was followed in some species by additional amplification of one of the paralogues with the subsequent loss of the other. It is proposed that the expansion of the number of ZPXa and ZPXb genes and the acquisition of dual sites of synthesis are the result of an ancient polyploidization event, followed by additional species-specific gene amplifications.

Animals↗

Species specificity of pharmacological characteristics of CCK-B receptors.

Novel CCK-B receptor antagonists, tetronothiodin and L-156,586, showed different affinities for CCK-B receptors in brain membranes from human, rat, guinea pig and mouse. [125I]CCK-8 bound to these membranes with a similar affinity. However, tetronothiodin was most potent in rat (IC50 = 3.6 nM), followed by guinea pig (96 nM), human (210 nM) and mouse (280 nM). L-156,586 bound with highest affinity to membranes from guinea pig (11 nM), and with lowest affinity to membranes from mouse (220 nM). These results suggest the existence of species specificity of CCK-B receptors, and that these two compounds are useful tools for discrimination between these receptors.

Aged↗

Species-specific inhibition by glycophorins of complement activation via the alternative pathway.

Glycophorin, one of the major glycoproteins of erythrocytes (E), was extracted from human E (glycophorin-Hu) and guinea pig E (glycophorin-GP) and adsorbed to rabbit-E. The adsorption of glycophorin-Hu and glycophorin-GP to rabbit-E made the E resistant to hemolysis by human serum and guinea pig serum, respectively, via the alternative complement pathway (ACP). However, it did not make the rabbit-E resistant to hemolysis by serum heterologous to the glycophorin adsorbed. This species-specific inhibition by glycophorin of ACP activation should play a role in restricting ACP activation on self cell membranes. By recognizing the self-cell surface as the place where the complement reaction must be prevented, ACP will be able to accomplish the discrimination of non-self constituents without diversity of recognition sites for a variety of foreign substances.

Animals↗

Enzymatic properties of the Caenorhabditis elegans Dna2 endonuclease/helicase and a species-specific interaction between RPA and Dna2.

In both budding and fission yeasts, a null mutation of the DNA2 gene is lethal. In contrast, a null mutation of Caenorhabditis elegans dna2+ causes a delayed lethality, allowing survival of some mutant C.elegans adults to F2 generation. In order to understand reasons for this difference in requirement of Dna2 between these organisms, we examined the enzymatic properties of the recombinant C.elegans Dna2 (CeDna2) and its interaction with replication-protein A (RPA) from various sources. Like budding yeast Dna2, CeDna2 possesses DNA-dependent ATPase, helicase and endonuclease activities. The specific activities of both ATPase and endonuclease activities of the CeDna2 were considerably higher than the yeast Dna2 (approximately 10- and 20-fold, respectively). CeDna2 endonuclease efficiently degraded a short 5' single-stranded DNA tail (<10 nt) that was hardly cleaved by ScDna2. Both endonuclease and helicase activities of CeDna2 were stimulated by CeRPA, but not by human or yeast RPA, demonstrating a species-specific interaction between Dna2 and RPA. These and other enzymatic properties of CeDna2 described in this paper may shed light on the observation that C.elegans is less stringently dependent on Dna2 for its viability than Saccharomyces cerevisiae. We propose that flaps generated by DNA polymerase delta-mediated displacement DNA synthesis are mostly short in C.elegans eukaryotes, and hence less dependent on Dna2 for viability.

Adenosine Triphosphatases↗

Expression of human TFIIA subunits in Saccharomyces cerevisiae identifies regions with conserved and species-specific functions.

The transcription factor TFIIA stabilizes the interaction between the TATA-binding protein (TBP) and promoter DNA and facilitates activator function. In yeast, TFIIA is composed of large (TOA1) and small (TOA2) subunits that interact to form a beta-barrel domain and a helix bundle domain. Here we report plasmid shuffle experiments showing that the human subunits (TFIIAalpha/beta, ALF, and TFIIAgamma) are not able to support growth in yeast and that the failure is associated with morphological abnormalities related to cell division. To determine the regions responsible for species specificity, we examined a series of chimeric yeast-human subunits. The results showed that yeast-human hybrids that contained the N-termini of TFIIAgamma or TFIIAalpha/beta were viable, presumably because they could form a functional interspecies alpha-helical bundle. Likewise, a TOA1 hybrid that contained the nonconserved internal region from TFIIAalpha/beta also had no effect on TFIIA function. However, hybrids that contained the acidic region III or C-terminal region IV from TFIIAalpha/beta grew more slowly than the wild-type TOA1 subunit, and if both regions were exchanged, this effect was far more severe. Although these hybrids exchanged sequences which are involved in beta-barrel formation and interactions with TBP, they were all active in a TBP-dependent mobility shift assay. The results suggest that the growth phenotypes of these hybrids might be due to a failure to interact with components of the yeast transcription machinery other than TBP. Finally, we show that sequences from region III of TFIIA large subunits fall into classes that are either highly acidic or that are divergent and nonacidic, and provide the first evidence to suggest that, at least in yeast, this region is important for TFIIA function.

Conserved Sequence↗

Trophic action of nerve extract on denervated skeletal muscle in vivo: dose dependency, species specificity, and timing of treatment.

Atrophy in a denervated muscle results from the disuse caused by paralysis of the muscle, and from the loss of special neurotrophic substances. Proteins extracted from rats' sciatic nerves have been shown to prevent the nondisuse atrophy of denervated rats' extensor digitorum longus (EDL) muscles when administered by intramuscular injection. The present investigation elicited more information about this neurotrophic phenomenon. Extract of liver failed to affect the rate of atrophy of denervated EDL muscles. Injection of various doses of extract of rats' or sheep's sciatic nerves indicated that the action of the extract was dose-dependent but not species-specific. Muscles denervated for 14 days and injected with extract on all 14 days or only on the first or last 7 days all exhibited significantly less atrophy than those that were not injected.

Animals↗