Erythrocyte antigens in Norwegian goats: serological and genetic studies.

Goat alloantisera and bovine blood typing reagents were used to characterize eight erythrocyte antigen specificities in Norwegian goats by cluster analysis, absorption and family studies. Most of the goat sera were produced by injecting dams once or twice with blood cells or blood from their own kids. The characterized specificities were designated N1-N8. The two specificities N5 and N8 were recognized both by goat alloantisera and by reagents against the bovine factors E'1 and E'2 (N5) and I (N8), which are allelic factors in the bovine B-system. In goat families, the two specificities also behaved as alleles. Consequently, the locus or gene system coding for these specificities was called the B-system of goats. The six other erythrocyte antigens were provisionally assigned to six separate loci. In addition, a bovine anti-sheep R factor reagent reacted with cells from 3.3% of the goats tested, whereas a monoclonal antibody against the Forssman antigen reacted with all the goats tested.     

A comparison of lymphocyte antigen specificities in Norwegian and Swiss goats

In all, 363 alloantireagents were tested in Berne, Switzerland and in Oslo, Norway against lymphocytes from 1679 goats of different breeds. The same lymphocytotoxicity test was used at both laboratories. The test data were pooled, and correlation coefficients for pairs of sera were used to group the sera in clusters. Twelve clusters were accepted as defining lymphocyte antigen specificities believed to be coded by genes within the major histocompatibility complex (MHC). The specificities defined by these clusters were designated Eu1-Eu12. Two clusters defined specificities which were not coded from loci within the MHC. These were designated GLY-1.1 and GLY-2.1, and the loci GLY-1 and GLY-2, respectively. GLY-1.1 was also located on erythrocytes.     

A comparison of lymphocyte antigen specificities in Norwegian and Swiss goats

Summary. In all, 363 alloantireagents were tested in Berne, Switzerland and in Oslo, Norway against lymphocytes from 1679 goats of different breeds. The same lymphocytotoxicity test was used at both laboratories. The test data were pooled, and correlation coefficients for pairs of sera were used to group the sera in clusters. Twelve clusters were accepted as defining lymphocyte antigen specificities believed to be coded by genes within the major histocompatibility complex (MHC). The specificities defined by these clusters were designated Eu 1-Eu12. Two clusters defined specificities which were not coded from loci within the MHC. These were designated GLY-1.1 and GLY-2.1, and the loci GLY-1 and GLY-2, respectively. GLY-1.1 was also located on erythrocytes.     

Molecular identification of human Ia antigens coded for by a gene locus closely linked toHLA-DR locus

Human Ia(-like) specificities controlled by gene loci other thanHLA-DR were searched for at the molecular level in cells of human B-cell-type cell lines which carrytwo established DR specificities. Chevalier cells of DRw3 and 7 and U698M cells of DRw2 and 4 were used. Their Ia molecules were partially purified, radioiodinated and analyzed for Ia specificities by the direct binding and sequential binding assays with a selected panel of human Ia alloantisera. It was possible in both the cell lines to define a third subset of Ia molecules carrying a new specificity in addition to two Ia subsets carrying the established DR specificities. The new specificity was detected by putative anti-DRw4 and anti-DRw7 antisera and was closely associated with DRw4 and DRw7 at population level. It was thus designated provisionally as BR4X7. These results suggest that the BR4X7 specificity is coded for by a separateIa locus closely linked toHLA-DR locus. The determinant(s) responsible for BR4X7 was located on the small subunit of Ia molecules.     

Joint Report of the Fourth International Workshop on Lymphocyte Alloantigens of the Horse, Lexington, Kentucky, 12–22 October 1985

Summary. The workshop consisted of 12 monthly cell exchanges of full-sibling families among the 10 participating laboratories. A total of 33 parents, 52 offspring and five unrelated horses were typed by each laboratory using local antisera. The raw data were submitted for central analysis before any identification of the animals was revealed.
Confidence derived from the consistent agreement between the laboratories on the assignment and segregation of the first 10 ELA-W specificities led to the removal of the W (workshop) notation and acceptance of full status as locus A antigens. The seemingly supertypic W11 specificity, however, remained unchanged.
Ten additional specificities were seen to segregate with the ELA system, suggesting either splits of previously described specificities or products of linked loci. The workshop (W) notation was given to the 10 specificities W12-W21, befitting their status as specificities under study.
The previously described ELY-1.1 specificity, characterized by segregation independent from the ELA system, was confirmed along with a new specificity, ELY-1.2, which behaves as an allele of ELY-1.1. For informative families, the two specificities showed codominant expression and appeared to constitute a closed, autosomal system.
The ELY-2.1 specificity was confirmed to segregate independently from the ELA-A and ELY-1 loci.     

Genetics of gliadins coded by the group 1 chromosomes in the high-quality bread wheat cultivar Neepawa

Summary The inheritance and biochemical properties of gliadins controlled by the group 1 chromosomes of the high-quality bread wheat cultivar Neepawa were studied in the progeny of the cross Neepawa x Costantino by six different electrophoretic procedures. Chromosome 1B of Neepawa contains two gliadin loci, one (Gli-B1) coding for at least six - or -gliadins, the other (Gli-B3) controlling the synthesis of gliadin N6 only. The map distance between these loci was calculated as 22.1 cM. Amongst the chromosome 1A gliadins, three proteins are encoded at the Gli-A1 locus whereas polypeptides N14-N15-N16 are controlled by a remote locus which recombines with Gli-A1. Six other gliadins are controlled by a gene cluster at Gli-D1 on chromosome 1D. Canadian wheat cultivars sharing the Gli-B1 allele of Neepawa were found to differ in the presence or absence of gliadin N6. The electrophoretic mobilities of proteins N6 and N14-N15-N16 were unaffected by the addition of a reducing agent during two-dimensional sodium dodecyl sulphate polyacrylamid-gel electrophoresis, suggesting the absence of intra-chain disulphide bonds in their structure.Research supported by a grant from the Commission of the European Communities, ECLAIR programme, Contract AGRE 0052     

Allotypic specificities of murine IgD and IgM recognized by monoclonal antibodies

Hybridomas generated from mice immunized with allotype and H-2-incompatible spleen cells were screened by flow cytometry. Monoclonal antibodies (MAb) to four of the five known specificities of IgD were identified. The location of these specificities on the IgD molecule was determined by the ability of a given MAb to bind to cells stripped of the Fab fragment by trypsin proteolysis. Igh-5.1 (present on IgDa and IgDe) and Igh-5.5 (unique to IgDe) were both located on the Fab fragment. Results from cross-blocking experiments suggest that, except for Igh-5.3, MAb which recognize the same specificity bind to the same antigenic determinant. Both the trypsin digest and blocking studies indicate that Igh-5.3 is composed of at least two antigenic determinants. AF6-78.25, a MAb specific for IgM of the b, d, and n haplotypes, was also identified; this MAb defines a new specificity, Igh-6.6. On the basis of the reactivities of these MAb, it appears that although the Igh-Ce and Igh-Cd haplotypes are virtually identical at the Igh-3(gamma 2b), Igh-1(gamma 2a), and Igh-2(alpha) loci, they are highly divergent at the Igh-5(delta) and Igh-6(mu) loci. This indicates that the Igh-Cd haplotype may have resulted from a recombination between Igh-Ce and another haplotype.     

Fourteen polymorphic microsatellite loci characterized in the house sparrow Passer domesticus (Passeridae,Aves)

We characterized 14 polymorphic microsatellite loci in the house sparrow Passer domesticus. Four loci were isolated from house sparrow genomic libraries and 10 loci were identified by testing 100 loci that had been originally isolated in other passerine species. Loci were characterized in 37–54 unrelated sparrows from British and Norwegian populations. Each locus displayed between two and 31 alleles, with the observed heterozygosity ranging between 0.30 and 0.91.     

Joint Report of the Fifth International Workshop on Lymphocyte Alloantigens of the Horse, Baton Rouge, Louisiana, 31 October-1 November 1987

Six laboratories participated in the Fifth International Workshop on Lymphocyte Alloantigens of the Horse, testing 132 alloantisera against lymphocytes of 880 horses chosen to represent different families and breeds. Most of the alloantisera were produced by lymphocyte immunization between horses matched at the ELA-A locus. All horses were also tested with antisera contributed to the workshop by participating laboratories which identified ELA specificities A1-A10 and W12-W21. Previously identified workshop specificities ELA-W14, W15 and W19 were accepted as products of the ELA-A locus based on family and population studies by the workshop. Their designations were changed to ELA-A14, ELA-A15 and ELA-A19, respectively. Two new specificities were identified, namely ELA-W22 (W22) and ELA-W23 (W23). Population and family studies indicated that W22 and W23 as well as W13 are products of an ELA locus other than ELA-A. The presence of these specificities was correlated with the presence of certain ELA-A locus specificities, e.g. W13 with A3, W22 with A2 and W23 with A5. However, the association was not complete and W13, W22 and W23 also segregated with other ELA-A specificities in some families. Evidence for recombination was found between the ELA-A locus and the locus or loci encoding these specificities resulting in seven recombinant haplotypes found among the data presented in this workshop. Further studies are required for definitive assignment of the specificities to a class I or class II locus.     

Organization of the gene for human factor XI

Factor XI (plasma thromboplastin antecedent) is a plasma glycoprotein that participates in the early phase of blood coagulation. The gene for the human protein has been isolated from two different lambda phage genomic libraries. Four independent recombinant lambda phage carrying overlapping DNA inserts that coded for the entire gene for factor XI were isolated and characterized by restriction mapping, Southern blotting, and selective DNA sequencing to establish the number and location of the intron-exon boundaries. The gene for human factor XI was 23 kilobases in length and consisted of 15 exons (I-XV) and 14 introns (A-N). Exon I coded for the 5' untranslated region, and exon II coded for the signal peptide. The next eight exons (III-X) coded for the four tandem repeats of 90 or 91 amino acids that were present in the amino-terminal region of the mature protein. Each of these tandem repeats was coded by two exons that were interrupted by a single intron, and these introns were located in essentially the same position within each of the four tandem repeats. The carboxyl-terminal region of the protein, which contained the catalytic chain, was coded by five exons (XI-XV) that were interrupted by four introns. The last four introns were located in the same positions as those in the genes for human tissue plasminogen activator and human urokinase.     

缅甸英雄岛始新世-上新世浮游有孔虫及其古环境意义

英雄岛位于印度洋缅甸西海岸,前人在该区所做的工作较少,生物地层研究程度较低.围绕该区石油勘探的进行,作者分析了RM19-3-1井的29件钻井岩屑样品,根据所含浮游有孔虫识别出7次有孔虫生物事件.考虑到研究区处于低纬度地区,故采用Bolli和Saunders(1985)的分带标准建立了7个浮游有孔虫化石带:P12-P17带;P18/19-P21/N2带;P22/N3带,N4-N7带;N8-N10带;N11-N14带和N15-N20?带,其延限时代为始新世-上新世.根据所含有孔虫动物群的演替规律,可将研究区的沉积环境变化由下而上划分为6个阶段,总体具有大陆架-陆坡的沉积环境特征.     

Multipoint genetic mapping of the Xq26-q28 region in families with fragile X mental retardation and in normal families reveals tight linkage of markers in q26–q27

Summary The q26–q28 region of the human X chromosome contains several important disease loci, including the locus for the fragile X mental retardation syndrome. We have characterized new polymorphic DNA markers useful for the genetic mapping of this region. They include a new BclI restriction fragment length polymorphism (RFLP) detected by the probe St14-1 (DXS52) and which may therefore be of diagnostic use in hemophilia A families. A linkage analysis was performed in fragile X families and in large normal families from the Centre d'Etude du Polymorphisme Humain (CEPH) by using seven polymorphic loci located in Xq26-q28. This multipoint linkage study allowed us to establish the order centromere-DXS100-DXS86-DXS144-DXS51-F9-FRAX-(DXS52-DXS15). Together with other studies, our results define a cluster of nine loci that are located in Xq26-q27 and map within a 10 to 15 centimorgan region. This contrasts with the paucity of markers (other than the fragile X locus) between the F9 gene in q27 and the G6PD cluster in q28, which are separated by about 30% recombination.     

Analysis of the fine specificities of sheep major histocompatibility complex class II-specific monoclonal antibodies using mouse L-cell transfectants

The fine specificities of two panels of monoclonal antibodies (mAbs) for sheep major histocompatibility complex (MHC) class II molecules were determined using five mouse L-cell transfectaents, each expressing a defined sheep DQ or DR MHC class II A/B gene pair. Using the transfectants in an indirect fluorescence antibody assay, previous immunochemical characterization of the mAbs was confirmed for 16 of 23 mAbs tested. The MHC class II subtype specificity ( DQ or DR ) of each mAb was assigned without interference from the products of other expressed class II loci. This allowed the identification of both cross-locus specificities as well as defining fine specificities of mAbs previously only partially characterized by immunochemical techniques.     

Null alleles for gliadin blocks in bread and durum wheat cultivars

Summary Wheat gliadin proteins are coded by clusters of genes (complex loci) located on the short arms of chromosomes of homoeologous groups 1 and 6 in bread (6x) and durum (4x) wheats. The proteins expressed by the various complex loci have been designated gliadin blocks. In a survey of accessions from the Germplasm Institute (C.N.R., Bari, Italy) collection, several different accessions have been found that lack particular blocks of proteins (null alleles). In some bread wheat accessions, seeds do not express gliadins that are coded by chromosomes 1D and 6A in normal cultivars. Similarly, some durum wheat accessions lack -gliadin components coded for by genes on chromosomes 1A and 1B. The missing proteins do not result from the absence of whole chromosomes, but may be the consequence of partial deletion of these genes at a complex locus or result from their silencing.     

Structure of recombinant N-terminal globule of type VI collagen alpha 3 chain and its binding to heparin and hyaluronan.

A large portion of the N-terminal globule of human collagen VI was prepared from the culture medium of stably transfected human embryonic kidney cell clones. The recombinant product corresponds to sequence positions 1-1586 of the alpha 3 (VI) chain that consists of eight homologous approximately 200 residue motifs (N9 to N2) being similar to the A domain motif of von Willebrand factor. By ultracentrifugation fragment N9-N2 showed a molecular mass of 180 kDa and an asymmetric shape. Elongated structures that consist of eight small globes (diameter approximately 5 nm) were demonstrated by electron microscopy. The data indicate that each A domain motif represents a separate folding unit which are connected to each other by short protease-sensitive peptide segments. Circular dichroism studies demonstrated about 38% alpha helix, 14% beta sheets and 17% beta turns. Fragment N9-N2 showed binding to heparin which could be abolished by moderate salt concentrations. Heparin binding was assigned to domains N9, N6 and N3 which were obtained after partial proteolysis. Domains N7, N5 and N4 lacked affinity for heparin. In addition, N9-N2 showed strong binding to hyaluronan that required exposure to 6 M urea for full dissociation. Ligand binding studies indicated some affinity of N9-N2 for the triple helical region of collagen VI suggesting a role of the N-terminal globule in the self-assembly of microfibrils. No or only little binding was, however, observed to fibril-forming collagens I and III, several basement membrane proteins and other extracellular proteins. Fragment N9-N2 was also an inactive substrate for cell adhesion.     

The swine histo compatibility system SLA: serological studies in the common Swiss and Danish swine breeds

Alloantisera were produced in common Swiss and Danish swine breeds by immunization with leucocytes or skin-grafts. Out of the 126 antisera, 66 were chosen for further study based on titrations employing lymphocytes from unrelated pigs typed with French SLA antisera (Vaiman, Chardon & Renard, 1979). The 66 selected antisera and the French reagents defining 26 SLA specificities were used to type lymphocytes from 595 unrelated pigs of the common Swiss and Danish breeds. The reaction-patterns of the French, Swiss and Danish antisera were adequately correlated for the French SLA specificities Nos FJ 2, 3, 6, 7, 8, 9, 11, 14, 19, 20 and 24 and for the haplotypes FJ 15.1.18 and FJ 5.4. In addition, a cluster of correlated Danish and Swiss antisera characterized a new specificity, provisionally designated CPH 31. This specificity was frequent in the Danish Landrace pigs.
Using the reagents identified in this report, the segregation of SLA markers was studied. Back-cross families demonstrated segregation of 15 distinct SLA haplotypes of which 14 are common in French Landrace or Large White. Differences were found in haplotype frequencies in both the Swiss and the Danish Landrace and Large White breeds.     

Nitrogen transformations revealed by isotope dilution in an organically fertilized hybrid poplar plantation

Measuring nitrogen (N) transformations from organic fertilizers can help in selecting applications rates that provide sufficient soluble N to promote tree growth in short-rotation plantations. The objective of this study was to determine how organic fertilizers (papermill biosolids, liquid pig slurry) affected microbially-mediated N transformations in soils. Soil samples were collected from a hybrid poplar plantation before fertilization, 1 month after fertilizer application and at the end of the growing season. Net N mineralization and nitrification were evaluated during a 28 d laboratory incubation, while gross N transformations were assessed using a ¹⁵N isotope dilution technique. Pig slurry application increased soil ammonium (NH₄-N) and nitrate (NO₃-N) concentrations within 1 month, while papermill biosolids increased soil NH₄-N and NO₃-N concentrations at the end of the growing season. Gross N consumption rates were greater than gross N production rates. The NH₄-N and NO₃-N consumption rates were positively correlated with labile carbon and microbial biomass. The gross nitrification rate was 18 to 67% of the gross mineralization rate but 30% or less of the gross NH₄-N consumption rate, indicating that NH₄ consumption was overestimated by the isotope dilution technique. We conclude that N cycling in this hybrid poplar plantation was characterized by rapid consumption of plant-available N following N mineralization and nitrification.     

The swine histocompatibility system SLA: serological studies in the common Swiss and Danish swine breeds

Alloantisera were produced in common Swiss and Danish swine breeds by immunization with leucocytes or skin-grafts. Out of the 126 antisera, 66 were chosen for further study based on titrations employing lymphocytes from unrelated pigs typed with French SLA antisera (Vaiman, Chardon & Renard, 1979). The 66 selected antisera and the French reagents defining 26 SLA specificities were used to type lymphocytes from 595 unrelated pigs of the common Swiss and Danish breeds. The reaction-patterns of the French, Swiss and Danish antisera were adequately correlated for the French SLA specificities Nos FJ 2, 3, 6, 7, 8, 9, 11, 14, 19, 20 and 24 and for the haplotypes FJ 15.1.18 and FJ 5.4. In addition, a cluster of correlated Danish and Swiss antisera characterized a new specificity, provisionally designated CPH 31. This specificity was frequent in the Danish Landrace pigs. Using the reagents identified in this report, the segregation of SLA markers was studied. Back-cross families demonstrated segregation of 15 distinct SLA haplotypes of which 14 are common in French Landrace or Large White. Differences were found in haplotype frequencies in both the Swiss and the Danish Landrace and Large White breeds.     

Design of a specific phenyllactate dehydrogenase by peptide loop exchange on the Bacillus stearothermophilus lactate dehydrogenase framework.

Restriction sites were introduced into the gene for Bacillus stearothermophilus lactate dehydrogenase which enabled a region of the gene to be excised which coded for a mobile surface loop of polypeptide (residues 98-110) which normally seals the active site vacuole from bulk solvent and is a major determinant of substrate specificity. Oligonucleotide-overlap extension (using the polymerase chain reaction) was used to obtain double-stranded DNA regions which coded for different length and sequence loops and which also contained the same restriction sites. The variable length and sequence loops were inserted into the cut gene and used to synthesize hydroxyacid dehydrogenases with altered substrate specificities. Loops which were longer and shorter than the original were made. The substrate specificities of enzymes with these new loops were considerably altered. For many poor enzyme-substrate pairs, the effect of fructose 1,6-bisphosphate on the steady-state kinetic parameters suggested that the substrate was mainly bound in a nonproductive mode. With one longer loop construction (BL1), activity with pyruvate was reduced one-million-fold but activity with phenylpyruvate was largely unaltered. A switch in specificity (kcat/KM) of 390,000-fold was achieved. The 1700:1 selectivity of enzyme BL1 for phenylpyruvate over pyruvate is that required in a phenyllactate dehydrogenase to be used in monitoring phenylpyruvate in the urine of patients with phenylketonuria consuming an apparently phenylalanine-free diet.