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PubMed1999

Species-specific production of microbial volatile organic compounds (MVOC) by airborne fungi from a compost facility.

Thirteen airborne fungal species frequently isolated in composting plants were screened for microbial volatile organic compounds (MVOC), i.e., Aspergillus candidus, A. fumigatus, A. versicolor, Emericella nidulans, Paecilomyces variotii, Penicillium brevicompactum, Penicillium clavigerum, Penicillium crustosum, Penicillium cyclopium, Penicillium expansum, Penicillium glabrum, Penicillium verruculosum, and Tritirachium oryzae. Air samples from pure cultures were sorbed on Tenax GR and analyzed by thermal desorption in combination with GC/MS. Various hydrocarbons of different chemical groups and a large number of terpenes were identified. Some compounds such as 3-methyl-1-butanol and 1-octen-3-ol were produced by a number of species, whereas some volatiles were specific for single species. An inventory of microbial metabolites will allow identification of potential health hazards due to an exposure to fungal propagules and metabolites in the workplace. Moreover, species-specific volatiles may serve as marker compounds for the selective detection of fungal species in indoor domestic and working environments.

Air Microbiology↗
PubMed1990

Monoclonal antibody recognizing a species-specific protein from Streptococcus pneumoniae.

Monoclonal antibodies (MAbs) against a nonencapsulated strain (R36A) of Streptococcus pneumoniae were produced to aid in a search for antigens common to this species. By Western immunoblot analysis, a species-specific 37-kilodalton (kDa) protein was found in lysates of 24 different encapsulated strains of S. pneumoniae. Monoclonal antibodies against the 37-kDa antigen did not react with 55 heterologous strains representing 19 genera and 36 species of bacteria that can also cause acute lower respiratory tract disease. Immunogold staining suggests that the antigen is synthesized inside the pneumococcal cell. However, MAbs to the 37-kDa antigen bound whole cells in the enzyme-linked immunosorbent assay and the indirect immunofluorescence assay. Antibody-binding epitopes of the antigen are probably exposed on the outer surface of the pneumococcus cell wall. The effectiveness of the 37-kDa antigen as a useful diagnostic marker is under study.

Antibodies, Bacterial↗
PubMed2005

Species-specific differences and similarities in the behavior of hand-raised dog and wolf pups in social situations with humans.

In order to reveal early species-specific differences, we observed the behavior of dog puppies (n = 11) and wolf pups (n = 13) hand raised and intensively socialized in an identical way. The pups were studied in two object-preference tests at age 3, 4, and 5 weeks. After a short isolation, we observed the subjects' behavior in the presence of a pair of objects, one was always the subject's human foster parent (caregiver) and the other was varied; nursing bottle (3 weeks), unfamiliar adult dog (3 and 5 weeks), unfamiliar experimenter (4 and 5 weeks), and familiar conspecific age mate (4 weeks). Dogs and wolves did not differ in their general activity level during the tests. Wolf pups showed preference for the proximity of the caregiver in two of the tests; Bottle-Caregiver at the age of 3 weeks and Experimenter-Caregiver at the age of 5 weeks, while dogs showed preference to the caregiver in three tests; conspecific Pup-Caregiver and Experimenter-Caregiver at the age of 4 weeks and dog-caregiver at the age of 5. Compared to wolves, dogs tended to display more communicative signals that could potentially facilitate social interactions, such as distress vocalization, tail wagging, and gazing at the humans' face. In contrast to dog puppies, wolf pups showed aggressive behavior toward a familiar experimenter and also seemed to be more prone to avoidance. Our results demonstrate that already at this early age--despite unprecedented intensity of socialization and the comparable social (human) environment during early development--there are specific behavioral differences between wolves and dogs mostly with regard to their interactions with humans.

Age Factors↗
PubMed1998

Binding of human urokinase-type plasminogen activator to its receptor: residues involved in species specificity and binding.

Urokinase-type plasminogen activator (UPA), particularly when bound to its receptor (UPAR), is thought to play a major role in local proteolytic processes, thus facilitating cell migration as may occur during angiogenesis, neointima and atherosclerotic plaque formation, and tumor cell invasion. To facilitate understanding of the need and function of the UPA/UPAR interaction in cell migration and vascular remodeling, we changed several amino acid residues in UPA so as to interfere with its interaction with its receptor. The receptor-binding domain of UPA has been localized to a region in the growth factor domain between residues 20 and 32. Since the binding of UPA to UPAR appears to be species specific, we used the differences in amino acid sequences in the growth factor domain of UPA between various species to construct a human UPA variant that does not bind to the human UPAR. We substituted Asn22 for its mouse equivalent Tyr by site-directed mutagenesis. This mutant UPA had similar plasminogen activator characteristics as wild-type UPA, including its specific activity and interaction with plasminogen activator inhibitor-1. However, no UPA/UPAR complexes could be observed in cross-linking experiments using DFP-treated 125I-labeled mutant UPA and lysates of various cells, including U937 histiocytic lymphoma cells, phorbol myristate acetate-treated human ECs, and mouse LB6 cells transfected with human UPAR cDNA. In direct binding experiments, DFP-treated 125I-labeled mutant UPA could not bind to phorbol myristate acetate-treated ECs, whereas wild-type UPA did bind. Furthermore, a 25-fold excess of wild-type UPA completely prevented the binding of DFP-treated 125I-labeled wild-type UPA to the human receptor on transfected LB6 cells, whereas an equal amount of mutant UPA had only a very small effect. In ligand blotting assays, very weak binding of mutant UPA to human UPAR could be observed. Changing Asn22 into the other amino acid residues alanine or glutamine had no effect on binding to UPAR on human ECs. The functional integrity of the growth factor domain in the non-receptor binding Asn22Tyr mutant is suggested by the fact that binding of this mutant to a murine UPAR can be restored after additional mutations in the growth factor domain, Asn27,His29,Trp30 to Arg27,Arg29,Arg30. We conclude that Asn22 and Asn27,His29,Trp30 in human UPA are key determinants in the species-specific binding of the enzyme to its receptor and that changing Asn22 into Tyr results in a UPA mutant with strongly reduced binding to UPAR.

Amino Acid Sequence↗
PubMed1988

Reaction of monoclonal antibodies with species specific determinants in Leptospira interrogans outer envelope.

A set of 24 monoclonal antibodies (MABs) was produced against an outer envelope preparation from Leptospira interrogans serovar copenhageni. The MABs reacted in enzyme immunoassay with species-specific determinants of an antigen in the leptospiral outer envelope (OE) of pathogenic but not of saprophytic species of Leptospira. The MABs did not agglutinate whole leptospires, nor could they opsonise homologous leptospires for phagocytosis by mouse macrophages or protect new-born guinea-pigs against lethal infection. The MABs reacted by Western blotting with a 35 x 10(3)-mol-wt band in OE separated on SDS-polyacrylamide gels, and also reacted with other bands to a lesser extent. The determinants to which the MABs were directed were localised in the leptospiral OE by immunogold labelling techniques.

Agglutination Tests↗
PubMed1979

Species specific grass pollen sensitivity: diagnosis and treatment with single grass species Allpyral vaccines.

Skin test titrations and nasal provocation tests in sixty patients with hay fever showed specific reactions to extracts of individual grass species. There was, however, no correlation between skin and nasal sensitivity. Repeat testing after treatment with Allpyral vaccines consisting of only the grass species to which the nasal reaction was most severe, or only one of several pollens to which reactions were equally severe, showed marked diminution of skin and nasal sensitivity not only to the single pollen used for immuno-therapy but to all five common pollens used in the Allpyral grass mix. Clinical results seemed much improved as compared with results in the same year for Allpyral five grass mix vaccines, especially in the case of patients treated with Timothy, rye, or cocksfoot. It was concluded that these three grassees were to be preferred for treatment in England, and that these grasses contain common allergens.

Dose-Response Relationship, Immunologic↗
PubMed2005

Species-specific variation in the B30.2(SPRY) domain of TRIM5alpha determines the potency of human immunodeficiency virus restriction.

Retroviruses encounter dominant postentry restrictions in cells of particular species. Human immunodeficiency virus type 1 (HIV-1) is blocked in the cells of Old World monkeys by TRIM5alpha, a tripartite motif (TRIM) protein composed of RING, B-box 2, coiled-coil, and B30.2(SPRY) domains. Rhesus monkey TRIM5alpha (TRIM5alpha(rh)) more potently blocks HIV-1 infection than human TRIM5alpha (TRIM5alpha(hu)). Here, by studying chimeric TRIM5alpha proteins, we demonstrate that the major determinant of anti-HIV-1 potency is the B30.2(SPRY) domain. Analysis of species-specific variation in TRIM5alpha has identified three variable regions (v1, v2, and v3) within the B30.2 domain. The TRIM5alpha proteins of Old World primates exhibit expansion, duplication, and residue variation specifically in the v1 region. Replacement of three amino acids in the N terminus of the TRIM5alpha(hu) B30.2 v1 region with the corresponding TRIM5alpha(rh) residues resulted in a TRIM5alpha molecule that restricted HIV-1 nearly as efficiently as wild-type TRIM5alpha(rh). Surprisingly, a single-amino-acid change in this region of TRIM5alpha(hu) allowed potent restriction of simian immunodeficiency virus, a phenotype not observed for either wild-type TRIM5alpha(hu) or TRIM5alpha(rh). Some of the chimeric TRIM5alpha proteins that are >98% identical to the human protein yet mediate a strong restriction of HIV-1 infection may have therapeutic utility. These observations implicate the v1 variable region of the B30.2(SPRY) domain in TRIM5alpha(rh) antiviral potency.

Amino Acid Sequence↗
PubMed2006

Species-specific secondary metabolite production in marine actinomycetes of the genus Salinispora.

Here we report associations between secondary metabolite production and phylogenetically distinct but closely related marine actinomycete species belonging to the genus Salinispora. The pattern emerged in a study that included global collection sites, and it indicates that secondary metabolite production can be a species-specific, phenotypic trait associated with broadly distributed bacterial populations. Associations between actinomycete phylotype and chemotype revealed an effective, diversity-based approach to natural product discovery that contradicts the conventional wisdom that secondary metabolite production is strain specific. The structural diversity of the metabolites observed, coupled with gene probing and phylogenetic analyses, implicates lateral gene transfer as a source of the biosynthetic genes responsible for compound production. These results conform to a model of selection-driven pathway fixation occurring subsequent to gene acquisition and provide a rare example in which demonstrable physiological traits have been correlated to the fine-scale phylogenetic architecture of an environmental bacterial community.

Micromonosporaceae↗
PubMed1986

Identification of Brugia malayi in vectors with a species-specific DNA probe.

We evaluated the potential value of a cloned sequence of genomic DNA of Brugia malayi as a species-specific probe. Clone pBm 15 reacted with all stages of 8 different geographic isolates of B. malayi and cross-hybridized with microfilariae of B. timori. It did not hybridize with Wuchereria bancrofti or with B. pahangi, W. kalimantani, Dirofilaria repens, Breinlia booliati or Cardiofilaria species, animal filariids that can be sympatric with B. malayi. P32-labeled clone pBm 15 correctly identified mosquitoes infected even with 1 infective larva of B. malayi. This specific DNA probe should be an invaluable tool to monitor control programs of Brugian filariasis.

Aedes↗
PubMed1999

Species-specific alternative splicing of transgenic RNA in the mammary glands of pigs, rabbits, and mice.

Gene-farming techniques provide an effective tool for the production of recombinant proteins in livestock. Transgenes consisting of genomic DNA sequences are as a rule more efficiently expressed than those in which the product of interest is encoded by a cDNA. However, the processing of pre-mRNA from genomic constructs may yield unexpected messenger RNAs and subsequently protein variants. We describe the appearance of different alternative mRNA splice patterns of a gene construct in which a mutant human growth hormone (hGH-N) gene is transcriptionally controlled by 2.5 kb of mouse whey acidic protein (WAP2) regulatory sequences in the mammary gland of different livestock species. Compared to the transcription products in transgenic mice harboring the same gene construct and to cell transfection experiments, expression analysis in transgenic pigs and rabbits revealed different mRNA splice patterns with regard to the proportion of the processed transcripts. Apart from already-known physiological mRNA splice products, previously undescribed processed hGH transcripts were observed in these species. Sequence analysis of the transgenes suggests that the species-specific hGH mRNA patterns may be caused by species- and tissue-specific differences in trans-acting splice factors.

Alternative Splicing↗
PubMed2004

Validation of a simplified field-adapted procedure for routine determinations of methyl mercury at trace levels in natural water samples using species-specific isotope dilution mass spectrometry.

A field-adapted procedure based on species-specific isotope dilution (SSID) methodology for trace-level determinations of methyl mercury (CH(3)Hg(+)) in mire, fresh and sea water samples was developed, validated and applied in a field study. In the field study, mire water samples were filtered, standardised volumetrically with isotopically enriched CH(3) (200)Hg(+), and frozen on dry ice. The samples were derivatised in the laboratory without further pre-treatment using sodium tetraethyl borate (NaB(C(2)H(5))(4)) and the ethylated methyl mercury was purge-trapped on Tenax columns. The analyte was thermo-desorbed onto a GC-ICP-MS system for analysis. Investigations preceding field application of the method showed that when using SSID, for all tested matrices, identical results were obtained between samples that were freeze-preserved or analysed unpreserved. For DOC-rich samples (mire water) additional experiments showed no difference in CH(3)Hg(+) concentration between samples that were derivatised without pre-treatment or after liquid extraction. Extractions of samples for matrix-analyte separation prior to derivatisation are therefore not necessary. No formation of CH(3)Hg(+) was observed during sample storage and treatment when spiking samples with (198)Hg(2+). Total uncertainty budgets for the field application of the method showed that for analyte concentrations higher than 1.5 pg g(-1) (as Hg) the relative expanded uncertainty (REU) was approximately 5% and dominated by the uncertainty in the isotope standard concentration. Below 0.5 pg g(-1) (as Hg), the REU was >10% and dominated by variations in the field blank. The uncertainty of the method is sufficiently low to accurately determine CH(3)Hg(+) concentrations at trace levels. The detection limit was determined to be 4 fg g(-1) (as Hg) based on replicate analyses of laboratory blanks. The described procedure is reliable, considerably faster and simplified compared to non-SSID methods and thereby very suitable for routine applications of CH(3)Hg(+) speciation analysis in a wide range of water samples.

Environmental Monitoring↗
PubMed1995

An improved method for the species-specific assessment of mycobacteria in routinely formalin-fixed and paraffin-embedded tissues.

A polymerase chain reaction (PCR) assay for the rapid and species-specific diagnosis of mycobacterial infections in paraffin-embedded clinical specimens was developed using oligonucleotide primers to amplify a fragment of the DNA coding for the ribosomal 16S RNA of mycobacteria. The oligonucleotide primers amplified DNA from all 14 species of mycobacteria tested. By means of a reamplification protocol, as few as one to two mycobacteria could be detected in the presence of human DNA. The method of DNA isolation and amplification was applied on sections of routinely formalin-fixed and paraffin-embedded tissues. PCR for the beta-actin gene served as a control for successful DNA isolation. Mycobacterial DNA could be detected in cases of mycobacterial infections. The mycobacterial species was determined by additional sequencing of the PCR fragment. This PCR method may be a powerful tool for the diagnosis of mycobacterial infections from histopathological material and for the assessment of those mycobacteria that cannot readily be cultured, such as Mycobacterium leprae.

Base Sequence↗
PubMed1988

The amino-acid residues on the C-terminal side of the cleavage site of angiotensinogen influence the species specificity of reaction with renin.

The N-terminal sequences of human and canine angiotensinogen and two hybrid sequences were synthesized and used to determine whether the species specificity of renin is influenced by amino-acid residues adjacent to the cleavage site. kcat/Km for the generation of angiotensin I from the N-terminal tridecapeptide of human angiotensinogen by canine renin is 0.37% of that observed when the N-terminal tetradecapeptide from canine angiotensinogen is used as a substrate. Replacement of the valine residue at P'1 in the human tridecapeptide with the leucine residue from the canine sequence triples kcat and improves Km 4-fold. Replacement of isoleucine residue at P'2 with the valine residue from the canine sequence enhances Km 8-fold. Substitution of the histidine residue at P'3 with the tyrosine serine sequence of canine angiotensinogen increases kcat an order of magnitude. Results obtained with the synthetic substrate are similar to those observed with the protein substrates. Canine renin does not cleave human angiotensinogen. Also, kcat/Km of canine renin for its homologous substrate is about 6-times greater than the kcat/Km value for human renin acting on human angiotensinogen.

Amino Acid Sequence↗
PubMed1986

Cross-reacting and species-specific determinants on a major allergen from Dermatophagoides pteronyssinus and D. farinae: development of a radioimmunoassay for antigen P1 equivalent in house dust and dust mite extracts.

Two species of mites of the genus Dermatophagoides are common in house dust and make a major contribution to the allergen content of house dust. As judged by skin tests and the radioallergosorbent test, these mites, D. pteronyssinus and D. farinae, demonstrate extensive cross-reactivity. When the major allergen from D. pteronyssinus, antigen P1, was compared with the equivalent allergen from D. farinae, these two proteins were found to have both species-specific and common antigenic determinants. With specifically purified antibodies directed against the common determinants, we developed a radioimmunoassay for these antigen P1 equivalent proteins in mite extracts as well as house dust extracts. The quantity of allergen in dust from 63 houses (255 samples) was measured, and values ranged from less than 100 to greater than 100,000 ng/gm of fine dust. The correlation between antigen P1 equivalent (ng/gm) and the number of mites per gram of dust (identified by microscopy) was very good (r = 0.74; p less than 0.001). Furthermore, this correlation was not affected by the species of mite in the dust sample. Of 9243 mites identified, 95% were of the genus Dermatophagoides, and of these, 16.4% and 18% could be positively identified as D. farinae and D. pteronyssinus, respectively. When the assay for antigen P1 equivalent was applied to isolated components of mite cultures of both species, significant allergen was found in the mite bodies, cuticles, and their excreta (fecal particles). For the house-dust samples, less than 10% of the allergen measured could be explained by the allergen content of mite bodies. Our results demonstrate that a radioimmunoassay for antigen P1 equivalent can provide an accurate and simple means of assessing the quantity of mite-derived allergen in dust-mite extracts, house-dust extracts, or house dust.

Allergens↗
PubMed1990

Pathfinding during spinal tract formation in the chick-quail chimera analysed by species-specific monoclonal antibodies.

In order to analyse the spinal tract formation at early stages of development in avian embryos, chick-quail spinal cord chimeras were prepared and species-specific monoclonal antibodies (MAb) were developed. MAbs CN, QN and CQN uniquely stained chick, quail, and both chick and quail nervous tissues, respectively. All three antibodies appeared to bind to the same membrane molecule, but to different epitopes. Cord reversal revealed the features of axonal growth of both cord interneurons and dorsal root ganglion cells. Quail cord interneurons grew along an originally ventral marginal layer in the quail cord transplanted in a reversed position, then turned toward the ventral side at the boundary between the graft and the host, and grew along the host chick ventral marginal layer. Central axons of dorsal root ganglia were restricted to the ventrolateral region of the cord which originally formed the dorsal funiculus. These results suggest that cord interneurons and dorsal root ganglion cells actively select to grow along specific regions of the cord and that spinal tract formation appears to be determined by cord cells, and not by sclerotome cells.

Animals↗
PubMed1995

Species-specific and mating type-specific DNA regions adjacent to mating type idiomorphs in the genus Neurospora.

Mating type idiomorphs control mating and subsequent sexual development in Neurospora crassa and were previously shown to be well conserved in other Neurospora species. The centromere-proximal flanks of the A and a idiomorphs, but not the distal flanks from representative heterothallic, pseudohomothallic, and homothallic Neurospora species contain apparent species-specific and/or mating type-specific sequences adjacent to the well-conserved idiomorphs. The variable flank is bordered by regions that are highly homologous in all species. The sequence of approximately 1 kb immediately flanking the conserved idiomorphs of each species was determined. Sequence identity between species ranged from 20% (essentially unrelated) to > 90%. By contrast, the mt-A1 gene shows 88-98% identity. Sequence and hybridization data also show that the centromere-proximal flanks are very different between the two mating types for N. intermedia, N. discreta, and N. tetrasperma, but not for N. sitophila and N. crassa. The data suggest a close evolutionary relationship between several of the species; this is suppported by phylogenetic analysis of their respective mt-A1 genes. The origin of the variable regions adjacent to the evolutionarily conserved mating type idiomorphs is unknown.

Base Sequence↗
PubMed2004

[Allele frequencies and species specificity of six short tandem repeat loci in Chinese population].

OBJECTIVE: To develop a set of new markers for forensic application, the authors have chosen 6 short tandem repeat(STR) loci to study the allele frequencies and species specificity in Chinese Han population in Chengdu. METHODS: One hundred and ten EDTA-blood samples were collected from the unrelated individuals in Chengdu city, Sichuan province. DNA was extracted by Chelex-100 and amplified by the polymerase chain reaction(PCR). Polyacrylamide gel electrophoresis (PAGE) and silver staining were used to analyze the PCR products. RESULTS: The polymorphisms of all 6 STR loci have been obtained in Chinese Han population in Chengdu, the alleles of D4S2366, D4S2367, D6S474, D6S1281, D2S1396 and D20S601 being 7, 7, 6, 7, 5, 7, the observed heterozygosity of them being 0.802, 0.708, 0.770, 0.627, 0.542, 0.672, the discrimination power of them being 0.887, 0.828, 0.849, 0.848, 0.794, 0.865; and the power of exclusion of them being 0.602, 0.441, 0.544, 0.325, 0.227, 0.386. Evaluated by comparison with the data from 14 different animals as controls, the 6 STR loci contain good specificity of human beings. CONCLUSION: The 6 STR loci are highly polymorphic and can play a key role in species identification. They are new candidate markers for forensic personal identification and paternity testing.

Animals↗
PubMed2002

Development of a monoclonal antibody detection assay for species-specific identification of abalone.

Species identification based on biochemical and molecular techniques has a broad range of applications. These include compliance enforcement, the management and conservation of marine organisms, and commercial quality control. Abalone poaching worldwide and illegal trade in abalone products have increased mainly because of the attractive prices obtained and caused a sharp decline in stocks. Alleged poachers have been acquitted because of lack of evidence to correctly identify species. Therefore, a robust method is required that would identify tissue of abalone origin to species level. The aim of this study was to develop immunologic techniques, using monoclonal and polyclonal antibodies, to identify 10 different abalone species and subspecies from South Africa, the United States, Australia, and Japan. The combination of 3 developed monoclonal antibodies to South African abalone (Haliotis midae) enabled differentiation between most of the 10 species including the subspecies H. diversicolor supertexta and H. diversicolor diversicolor. In a novel approach, using antibodies of patients with allergy to abalone, the differentiation of additional subspecies, H. discus discus and H. discus hannai, was possible. A field-based immunoassay was developed to identify confiscated tissue of abalone origin.

Journal Article↗