On the genetics of transplantation in the lizardCnemidophorus tigris

A total of 195 allografts were analyzed among 14 wild individuals of the whiptail lizardCnemidophorus tigris. The cumulative time for rejection of all allografts by each individual varied in a uniformly graded sequence ranging from 502 to 2263 days. One individual did not reject 9 of 13 allografts. A comparison of rejection rates revealed that the relative degree of antigenicity of a donor was more or less proportional to its immune response, i. e., the strongest rejectors tending to elicit the strongest immune response and the weakest rejectors the weakest response. Such a relationship as well as the graded sequence of rejection suggest that neither the many weak transplantation antigen hypothesis, nor the strong locus hypothesis adequately explain the present results. Probable genetic models are discussed and a summary of transplantation studies in lower vertebrates is presented.     

Inhibition of nuclear protein kinases by adenosine analogues

Two cyclic AMP-independent protein kinases from rat liver nuclei were inhibited competitively by adenosine and a variety of its analogues: cardycepin, tubercidin, 6-mercaptopurine riboside, 6-methylaminopurine riboside, 6-dimethylaminopurine riboside, and 2'-deoxyadenosine. Neither enzyme was inhibited by 6-methoxypurine riboside. Kinase NI was more sensitive to cordycepin, tubercidin, 6-methylaminopurine riboside,, 2'-deoxyadenosine, and adenosine than was kinase NII. Although the effects of all analogues tested were reversed by increasing the concentration of ATP, kinases NII and NI exhibited marked differences in their affinities for adenosine. The results presented here suggest that 1) differences in the catalytic properties of nuclear protein kinases can be detected by inhibitor studies and 2) modifications in the purine ring and sugar moiety of an adenosine analogue can alter its ability to inhibit nuclear protein kinases.     

Membrane trafficking along the phagocytic pathway

Phagosome maturation involves extensive remodelling of the phagosomal membrane as a result of intracellular transport events. Newly formed phagosomes exchange membrane-associated and soluble proteins with early endosomes by fusion. Budding of vesicles from the phagosome and fusion with Golgi-derived vesicles may also contribute to the remodelling of the phagosomal compartment. As a consequence of changes in membrane composition, phagosomes acquire the ability to fuse with late endocytic compartments. In vitro reconstitution and other studies suggest that the trafficking events underlying phagosome maturation require several GTP-binding proteins, including Rab5 and Galphas', NSF-SNAP-SNARE complexes and coatomers.     

Genetic diversity and ecological distribution ofPhaseolus vulgaris (Fabaceae) in northwestern South America

Our goal was to investigate in more detail wild and cultivated common bean (Phaseolus vulgaris) accessions from northwestern South America (Colombia, Ecuador, and northern Peru) because prior research had shown this region to be the meeting place of the two major gene pools (Middle American and Andean) of common bean. Explorations were conducted in these countries to collect additional materials not represented in germplasm collections. It was possible to identify wild common bean populations in Ecuador and northern Peru, where they had never been described before. In addition, we were able to extend the distribution of wild common bean in Colombia beyond what was known prior to this study. In all areas, the wild common bean habitat had suffered severely from destruction of natural vegetation. In Colombia, wild common beans were found on the Eastern slope of the Andes (in continuation of its distribution in Venezuela), whereas in Ecuador and northern Peru they were found on the western slope of this mountain range. This geographic distribution was correlated with an ecological distribution in relatively dry environments with intermediate temperatures (known as “dry mountain forest”). Isozyme andphaseolin seed protein analyses of the northern Peruvian and Ecuadoran wild populations showed that they were intermediate between the Middle American and Andean gene pools of the species. Phaseolin analyses conducted on landraces of the Upper Magdalena Valley in Colombia showed that Andean domesticates were grown at a higher altitude than Middle American domesticates suggesting that the former are adapted to cooler temperatures. Our observations and results have the following consequences for the understanding and conservation of genetic diversity in common bean and other crops: 1) Our understanding of the distribution of the wild relative of common bean (and other crops) is imperfect and further explorations are needed to more precisely identify and rescue wild ancestral populations; 2) For crops for which the wild ancestor has not yet been identified, it may be worthwhile to conduct additional explorations in conjunction with genetic diversity studies at the molecular level to guide the explorations; 3) Our study shows the benefit for more efficient germplasm conservation which can be derived from the dynamic interplay between field explorations (and other conservation operations) and molecular analyses to determine genetic distances and diversities; 4) The intermediate materials identified in northern Peru and Ecuador may have basic importance to understand the origin of the common bean and an applied role as a bridge between the Middle American and Andean gene pools; and 5) The differential adaptation to temperature of the two major cultivated gene pools may help breeders select genotypes based at least partially on their evolutionary origin.     

DISTRIBUTION AND SYSTEMATICS OF THE FRESHWATER GENUS SIRODOTIA(BATRACHOSPERMALES,RHODOPHYTA) IN NORTH AMERICA1

Multivariate morphometrics and image analysis were used to determine the number of well-delineated infrageneric taxa of Sirodotia in North America. Three groupings were distinguished from 25 populations examined from Newfoundland and Quebec in the north to central Mexico in the south. These groupings were statistically related to 10 type specimens, and the following species were recognized: Sirodotia huillensis (Welwitsch ex W. et G. S. West) Skuja (syn. S. ateleia Skuja), S. suecica Kylin (syn. S. acuminata Skuja ex Flint and S. fennica Skuja), and S. tenuissima (Collins) Skuja ex Flint. These species are differentiated on the basis of whorl shape and degree of separation at maturity (S. suecica, rounded and appressed; S. huillensis and S. tenuissima, truncated apex and separated), the density of spermatangia (S. huillensis, dense clusters; S. suecica and S. tenuissima, sparsely aggregated), and the mode of germination of the gonimoblast initial (S. suecica and S. tenuissima, from the nonprotuberant side of the fertilized carpogonium; S. huillensis from the protuberant side). Sirodotia huillensis was found only in the desert-chaparral, whereas S. suecica and S. tenuissima occurred from south-temperate to boreal regions in cool (temperature 8–18° C), low ion (specific conductance 10–99 μS · cm^?1), and mildly acidic to neutral (pH 5.7–7.3) waters.     

Heat and cold denaturation of beta-lactoglobulin B.

The thermal denaturation of bovine beta-lactoglobulin B was investigated by high-sensitivity differential scanning microcalorimetry between pH 1.5 and 3.0 in 20 mM phosphate buffer. The process was found to be a reversible, two-state transition. Progressive addition of guanidine hydrochloride at pH 3.0 leads to the appearance of a low-temperature calorimetric endotherm, corresponding to the cold renaturation of the protein. Circular dichroism experiments have confirmed the low and high temperature denaturation processes, and have shown some structural differences between both denatured states of beta-lactoglobulin B.     

Transport of phagosomal components to an endosomal compartment.

The participation of phagosomes in interorganellar protein and membrane exchange is important to the maturation of phagosomes into phagolysosomes. To investigate this process, we have developed an assay to measure protein transport from phagosomes to other vesicle populations. J774-E clone macrophages phagocytosed 125I-anti-dinitrophenol IgG-coated Staphylococcus aureus for 3 min followed by chase for intervals of 0-30 min. Following cell fractionation, the intracellular distribution of radioiodinated protein was assayed. We observed a time-dependent increase radioiodinated protein in a non-phagosome vesicle fraction which displayed endosome characteristics. Concomitantly, radioiodinated protein within phagosomes decreased over the chase period. As assessed via Percoll density gradient fractionation, the phagocytosed radioiodinated protein migrated to both heavy (lysosome density) and light (endosome density) vesicle populations. Characterization of the fusogenic properties of the transport vesicles demonstrated that they are capable of in vitro fusion with early endosomes. Furthermore, this fusion event shares many of the biochemical requirements identified for phagosome-endosome and endosome-endosome fusion. Morphological analysis of phagosome maturation provides additional evidence for phagosome to endosome transport. These results suggest phagocytosed material is transferred from phagosomes to endosomes and then recycled out of the cell.     

PKC cellular distribution in TPA activated human peripheral blood mononuclear cells, treated with an anti-HLA class I monoclonal antibody

Cytosolic and Particulate Protein Kinase C has been studied in Peripheral Blood Mononuclear Cells activated with 12-O-Tetradecanoyl phorbol 13-acetate and treated with the anti-HLA Class I Monoclonal Antibody 01.65. No effects on the cellular distribution of PKC activity nor to the proliferative response has been found. In phytohemagglutinin stimulated PBMC cultures treated with MoAb 01.65 total PKC activity depletion and 3H-Thymidine incorporation inhibition has been found. In PBMC cultures activated with both PHA and TPA, the proliferative response was similar to cultures activated with PHA alone, while the PKC cellular distribution was similar to the one detected in TPA stimulated cultures. Addition of the MoAb 01.65 was ineffective on both PKC activity and 3H-Thymidine incorporation. These data indicate that anti-HLA Class I MoAb induced 3H-Thymidine incorporation inhibition may be related to low levels of PKC activity.     

DNA polymorphisms and linkage relationship of the human complement componentC6,C7, andC9 genes

In this report we describe the linkage between genes encoding human complement componentsC6,C7, andC9. Polymorphisms have been described at the DNA level for theC7 andC9 genes. We have studied 20 individuals by Southern blot analysis with fourC6 cDNA subclones to detect restriction fragment length polymorphisms (RFLPs). We have found a Taq I polymorphism defined by two alleles of 8.0 (C6 H) and 6.0 (C6 L) kilobases (kb). RFLP segregation for theC6, C7, andC9 loci in informative families allowed us to estimate the maximum Lod scores at a recombination fraction of =0.0 (C6–C7), =0.0 (C7–C9), and =0.0 (C6–C9). Significant linkage disequilibrium was found betweenC6 andC7 and betweenC7 andC9 loci in directly determined haplotypes of unrelated parents. Data from this study show that the genes encoding the human terminal complement componentsC6, C7, andC9 define a cluster in the short arm of chromosome 5. We propose that the clusters involving theC8A andC8B and theC6, C7, andC9 genes be referred to as MACI and MACH, respectively.     

Isolation and characterization of a mouse subtelomeric sequence

A mouse subtelomeric sequence, ST1, was generated from genomic DNA of the mouse HR9 (129/Sv origin) cell line by the polymerase chain reaction (PCR) using a single telomeric primer. ST1 was cloned and characterized: it is composed of 670 bp of novel DNA sequence flanked on each end by inverted telomeric hexanucleotide repeats (TTAGGG)_n. PCR amplification from BALB/c mouse DNA using this single primer gave the same major product. Southern analysis and PCR using internal ST1 primers confirmed that the ST1 sequence is present in mouse genomic DNA. In situ hybridization to metaphase chromosomes of SJL origin mapped ST1 to many, if not every, mouse telomere. PCR experiments using different combinations of the telomeric, minor satellite, and ST1 primers indicated that some ST1 copies are adjacent to minor satellite sequences, that telomeric and ST1 sequences are not generally interspersed with minor satellite sequences,and that ST1 and the minor satellite have a consistent and specific orientation relative to each other and to the telomere.by H.F. Willard