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Identification of a serum protein polymorphism via two-dimensional electrophoresis. Family and population studies in two genetically isolated groups: North American Hutterites and Australian aborigines.

We report the identification and initial family and population studies of a previously undescribed serum protein polymorphism with two allelic forms. It was discovered in Hutterites, a reproductively isolated religious sect, and is also present in Australian aborigines and a sample of Chicago residents. A two-allele model is consistent with the segregation pattern observed in five kindreds within our initial study group. This polymorphism, provisionally designated SPPM-158, appears as a horizontal (charge-based) doublet in silver-stained ISO-DALT high-resolution two-dimensional electrophoresis gels. It is a low-concentration polypeptide (approximately 1 mg/dL) that has an apparent MWSDS of 43.6 kD and an isoelectric point of approximately 5.5. We infer that it circulates as a multimer or in a high-molecular-weight (greater than 200 kD) complex with other proteins because it is not observed in normal body fluids derived from physiologically ultrafiltered plasma such as amniotic fluid, urine, or cerebrospinal fluid; however, it is present in urine of patients with glomerular proteinuria. The high heterozygosity rates imply utility of this new serum protein marker for both forensic and population studies.

Australia

Inverted Y chromosome polymorphism in the Gujerati Muslim Indian population of South Africa.

An inverted Y chromosome has been found at a very high frequency in a Muslim Indian community living in the Johannesburg-Witwatersrand area of the Transvaal Province of South Africa: 8 of 141 (5.7%) retrospectively identified Indian males had an inv(Y)(p11.2q11.23) and all were of the Muslim faith. The inversion was found in 22 of 72 (30.5%) prospectively studied normal Muslim Indian males. All the carriers of the inversion were Gujarati-speakers whose families migrated to the Transvaal from the Gujerat Province of India during the first half of this century. The origins of the ancestors of the individuals with inv(Y) were traced to a small village, Kholvad, near the city of Surat, and some neighbouring villages. The polymorphic frequency of the inv(Y) has probably been produced through random genetic drift in a reproductively isolated community, maintained by strict endogamous marriage customs based on religious and linguistic affiliations. There was no indication that the inverted Y was associated with any reproductive disadvantages.

Black People

The Cajuns of Southern Alabama: morphology and serology.

A survey was conducted of 324 members of the Cajun isolate of Southern Alabama. Tradition and appearance suggest that this population of about 3,000 are not entirely White, Black, or Indian but constitute a triracial community somewhat reproductively isolated and inbred. The earliest American settlement in the area, along the banks of the Mobile and Tombigbee Rivers, lay between Spaniards to the South and Indian tribes on the other sides: Creek, Choctaw, and Cherokee. Physical measurements are reported for 71 adults, plus color of skin, eyes, and hair. X-rays were taken of wrist and ankle bones of some 253 children. Red blood samples were typed on adults and children, and haptoglobin, Gm, and Gc types were determined from serum. History and physical examinations were also made. Physical measurements and observations suggest predominantly White ancestry, and D2 analysis confirms this, with least similarity to Indians. Analysis of serological traits implies almost 70% White, almost 30% Black, and very little Indians genes. Few defects of clear genetic etiology were discovered. Growth patterns judged from X-rays appeared normal. All genetic loci testable were in Hardy-Weinberg equilibrium except Gc. While history and some common surnames suggest endogamy in the past, the medical and serological findings, plus some additional surnames, indicate that the isolate has already been largely diluted or dissolved.

Adolescent

Purification and properties of cyclic-3',5'-GMP-dependent protein kinase from the nematode Ascaris suum.

A cyclic-3',5'-GMP-dependent protein kinase was purified 7400-fold from the reproductive tract of female ascarids to a specific activity of 718 nmol min-1 mg-1 (histone as substrate). The yield of the preparation was 25%. The enzyme protein obtained was homogeneous as judged by isoelectrofocusing and polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The native enzyme behaved as a dimer of two 82-kDa subunits in gel permeation chromatography on Superose 12. The protein kinase was inactive in the absence of cyclic purine nucleotides. Half-maximum velocity was obtained in the presence of 18 nM cGMP, whereas 400-fold higher concentrations of cAMP were required for the same activity. The enzyme underwent autophosphorylation in first-order kinetics (rate constant 0.054 min-1), leading to maximum incorporation of 0.96 phosphate per subunit. The autophosphorylation led to a 4-fold increase in Vmax, while the Km remained almost unchanged. In an extract from the reproductive tract, cGMP-stimulated phosphorylation was primarily observed in five proteins (molecular masses of 66, 60, 43, 30, and 25 kDa). These proteins also incorporated phosphate when isolated reproductive tracts were incubated in the presence of [32P]phosphate. The phosphate content in cellular proteins was enhanced when the incubation was performed in the presence of 10(-4) M of either octyl-cAMP or octyl-cGMP. In addition to the proteins mentioned above, however, six more electrophoretic bands containing radioactive phosphate were identified after in situ labeling of reproductive tracts with radioactive phosphate.

Animals

[A perfusion device for the survival of isolated organs of the reproductive system].

A perfusion device for life supporting in isolated reproductive organs is described. With the device, the following functions are possible: gas exchange in recirculating blood flow, perfusate replacement in the course of perfusion, blood flow generation in a circular circuit with simultaneous provision of hemodynamic parameters closely approximating native ones.

Animals

Differentiation of U.S. and European isolates of porcine reproductive and respiratory syndrome virus by monoclonal antibodies.

Monoclonal antibodies (MAbs) to two U.S. isolates of porcine reproductive and respiratory syndrome (PRRS) virus were prepared. Two MAbs specifically recognized a conserved epitope on the putative 15-kDa nucleocapsid protein of U.S. and European isolates of PRRS virus. Four other MAbs recognized epitopes on the 15-kDa protein of U.S. but not European isolates of PRRS virus. Collectively, this indicates that PRRS viruses contain both conserved and divergent epitopes on the 15-kDa viral protein.

Animals

Patterns of mortality in the the Old Order Amish. I. Background and major causes of death.

The major causes of death were studied in the Old Order Amish people in three settlements in Indiana, Ohio and Pennsylvania to determine if lifestyle and genetic isolation altered their mortality risk compared to neighboring non-Amish. The Amish are a conservative religious group who live in farm settlements, use horses for work and travel, exercise vigorously, and avoid cigarettes and alcohol. They are reproductively isolated and highly inbred. Death certificates and Amish censuses were used to determine mortality risks, which were summarized using age-adjusted mortality ratio (MRs). Amish mortality patterns were not systematically higher or lower than those of the non-Amish, but differed by age, sex, and cause. Amish males had slightly higher all-cause MRs as children and significantly lower MRs over the age of 40, due primarily to lower rates of cancer (MR = 0.44, age 40-69), and cardiovascular diseases (MR = 0.65, age 40-69). Amish females MRs for all causes of death were lower from age 10-39, not different from 40-69, and higher over age 69. MRs were not significantly different for all cancer sites combined in Amish women and they had higher cardiovascular mortality ratio aged 70 and over (MR =1.34). Other major causes of death were also examined. Because the Amish and other farming groups have similar mortality patterns, it is suggested that lifestyle may be the primary determinant of the overall mortality patterns in the Amish.

Adolescent

Counter current transfer and back transport of 3H-PGF2 alpha in the cow's broad ligament vasculature ipsilateral and contralateral to the corpus luteum.

Eight cows of similar age (5-7 years) were chosen for the experiment. Isolated reproductive tract was supplied with autologous oxygenated and heated (40 degrees C) blood through the uterine artery and ovarian artery. 3H-PGF2 alpha in total dose of 2 MBq (10(7) cpm) was injected into each of the uterine lumen of isolated organ. Blood samples were collected at 5 min intervals during 120 min of experiment using cannulae inserted into the branches of uterine arteries about 1 cm below the horns and from ovarian arteries inserted 0.5 cm below the ovaries. The concentration of 3H-PGF2 alpha found in blood plasma taken from uterine artery or from ovarian artery on the side with active corpus luteum (CL) was significantly lower (p less than 0.001) compare with contralateral side to active CL. Radioactive PGF2 alpha found in branches of uterine arteries on both ipsilateral and contralateral side to CL was significantly higher (p less than 0.001) compare to ovarian artery of the same side. It is concluded that absorption of 3H-PGF2 alpha from uterine lumen into venous blood as well as its counter current transfer in area of broad ligament vasculature were reduced on the side of uterine horn with active CL probably as an effect of estrogen:progesterone ratio on vascular constriction in area of uterine vasculature.

Adnexa Uteri

Cytoplasmic incompatibility. Giant steps sideways.

The horizontal transfer of a bacterial endosymbiont that is intimately associated with reproductive isolation in insects is now feasible and may, in principle, lead to new strategies for biological pest control.

Animals

Postcopulatory, prezygotic isolation: intraspecific and interspecific sperm precedence in Tribolium spp., flour beetles.

We investigated intraspecific and interspecific patterns of paternity in Tribolium castaneum, the flour beetle, by mating females either to pairs of conspecific males or to one conspecific and one heterospecific male of the closely related species, T. freemani. Females of both species store sperm in the spermatheca after copulation and postcopulatory, prezygotic reproductive isolation has been reported between this pair of species. When conspecific males of contrasting genotype are mated simultaneously to T. castaneum females, we observed extremely high levels of variation among females in the pattern of sperm precedence as shown by the offspring genotypes. In contrast, T. castaneum females mated simultaneously to a conspecific and a heterospecific male produced over 99 per cent conspecific progeny. When conspecific males were mated sequentially to T. castaneum females, within 3-7 days, all offspring were sired by the second male. In contrast, when a conspecific male was replaced with a T. freemani male, most females continued to produce only first-male, conspecific offspring for the next 10 days. When a heterospecific male was replaced with a conspecific male, T. castaneum females changed from producing hybrid to conspecific offspring within 3 days. We discuss the evolutionary implications of these findings.

Animals

Wolbachia and cytoplasmic incompatibility in mycophagous Drosophila and their relatives.

Bacterial symbionts belonging to the genus Wolbachia are associated with postzygotic reproductive incompatibility in a number of insect species. Using polymerase chain reaction (PCR) amplification of bacterial gene sequences, strains from 10 species belonging to the closely related quinaria, testacea and tripunctata groups of Drosophila were screened for the presence of Wolbachia in their reproductive tissues. Those screened included the mycophagous species D. falleni, D. recens, D. phalerata, D. testacea, D. neotestacea, D. orientacea, D. putrida and D. tripunctata, and the nonmycophagous species D. palustris and D. quinaria. Two species, D. recens and D. orientacea, were found to be infected with Wolbachia. Subsequent tests of four additional strains of D. recens found all to be infected with the bacteria. It was established that these bacteria cause partial cytoplasmic incompatibility in D. recens by antibiotic curing followed by crosses between cured and uncured strains. Curing was confirmed by a PCR assay. Although most species of insects shown to be infected with Wolbachia are cosmopolitan and/or have undergone recent range expansion in association with human activity, D. recens and D. orientacea are endemic species with specialized ecological habits. Preliminary molecular phylogenetic analysis indicates that, among the species we examined, D. quinaria is most closely related to D. recens. To determine whether the bacteria are involved in reproductive isolation between these two species, reciprocal crosses were carried out between D. quinaria and both infected and uninfected (cured) strains of D. recens. Although these species did mate with each other, all interspecific crosses failed to yield hybrid progeny, indicating that the bacteria are not responsible for reproductive incompatibility between these species.

Animals

The heterochromatin of grasshoppers from the Caledia captiva species complex. I. Sequence evolution and conservation in a highly repeated DNA family.

The restriction enzyme TaqI digests 0.2% of the genomic DNA from the grasshopper Caledia captiva to a family of sequences 168 bp in length (length of consensus sequence). The sequence variation of this "Taq family" of repeat units was examined among four races from C. captiva to assay the pattern of evolution within this highly repeated DNA. The Taq-family repeats are located in C-banded heterochromatin on at least one member of each homologous pair of chromosomes; the locations range from centromeric to telomeric. Thirty-nine cloned repeats isolated from two population 1A individuals along with 11 clones from seven populations taken from three of the races demonstrated sequence variation at 72 positions. Pairwise comparisons of the cloned repeats, both within an individual and between different races, indicate that levels of intraspecific divergence, as measured by reproductive incompatibility, do not correlate with sequence divergence among the 168-bp repeats. A number of subsequences within the repeat remain unchanged among all 50 clones; the longest of these is 18 bp. That the same 18-bp subsequence is present in all clones examined is a finding that departs significantly (P less than 0.01) from what would be expected to occur at random. Two other cloned repeats, from a reproductively isolated race of C. captiva, have sequences that show 56% identity with this 18-bp conserved region. An analysis showed that the frequency of occurrence of an RsaI recognition site within the 168-bp repeat in the entire Taq family agreed with that found in the cloned sequences. These data, along with a partial sequence for the entire Taq family obtained by sequencing uncloned repeats, suggest that the consensus sequence from the cloned copies is representative of this highly repeated family and is not a biased sample resulting from the cloning procedure. The 18-bp conserved sequence is part of a 42-bp sequence that possesses dyad symmetry typical of protein-binding sites. We speculate that this may be significant in the evolution of the Taq family of sequences.

Animals

Olfactory discrimination as an isolating mechanism in speciating mole rats.

Olfactory discrimination was tested in two chromosome forms of the speciating fossorial rodent, Spalax ehrenbergi, in Israel. Females of the chromosome forms 2n = 52 and 2n = 58 were tested for male odour discrimination, the source of odour being either case litter or urine. Estrous females of both forms preferred homochromosomal odurs, whereas diestrous females showed no discrimination. These results suggest that olfactory discrimination may serve as a reproductive isolating mechanism in the speciation of mole rats.

Animals

Sexual isolation in Drosophila melanogaster: a possible case of incipient speciation.

It is generally believed that Drosophila melanogaster has no closely related species with which it can produce the viable and fertile hybrids that are essential for the genetic analysis of speciation. Following the recent report of molecular differentiation between a Zimbabwe, Africa, population and two United States populations, we provide evidence that strong sexual isolation exists between the D. melanogaster population in Zimbabwe and populations of other continents. In the presence of males of their own kind, females from most isofemale lines of Zimbabwe would not mate with males from elsewhere; the reciprocal mating is also significantly reduced, but to a lesser degree. The genes for sexual behaviors are apparently polymorphic in Zimbabwe and postmating reproductive isolation between this and other populations has not yet evolved. Whole chromosome substitutions indicate significant genetic contributions to male mating success by both major autosomes, whereas the X chromosome effect is too weak to measure. In addition, the relative mating success between hybrid and pure line males supports the interpretation of strong female choice. These observations suggest that we are seeing the early stages of speciation in this group and that it is driven by sexual selection. The genetic and molecular tractability of D. melanogaster offers great promise for the detailed analysis of this apparent case of incipient speciation.

Animals

Genetic distance and species formation in evolving populations.

We compare the behavior of the genetic distance between individuals in evolving populations for three stochastic models. In the first model reproduction is asexual and the distribution of genetic distances reflects the genealogical tree of the population. This distribution fluctuates greatly in time, even for very large populations. In the second model reproduction is sexual with random mating allowed between any pair of individuals. In this case, the population becomes homogeneous and the genetic distance between pairs of individuals has small fluctuations which vanish in the limit of an infinitely large population. In the third model reproduction is still sexual but instead of random mating, mating only occurs between individuals which are genetically similar to each other. In that case, the population splits spontaneously into species which are in reproductive isolation from one another and one observes a steady state with a continual appearance and extinction of species in the population. We discuss this model in relation to the biological theory of speciation and isolating mechanisms. We also point out similarities between these three models of evolving populations and the theory of disordered systems in physics.

Biological Evolution

Hybridization between Aedes simpsoni and Aedes woodi with observations on the genetic basis of morphological differences.

The public health importance of the genus Aedes makes it imperative that a better understanding of the genetic differences and isolating barriers between the species be developed. Aedes simpsoni and Aedes woodi from near Dar es Salaam, Tanzania, were used in this investigation. Analysis of F(1), F(2), and backcross progeny obtained from forced matings indicates that single factors form the genetic basis of some of the differences between the two species. There is evidence that Ae. simpsoni and Ae. woodi are separated by both premating and postmating reproductive isolating mechanisms.

Aedes

Sequence analysis of open reading frames (ORFs) 2 to 4 of a U.S. isolate of porcine reproductive and respiratory syndrome virus.

The sequence of ORFs 2 to 4 of a U.S. isolate of porcine reproductive and respiratory syndrome virus (PRRSV), ATCC VR2385, was determined by analysis of a cDNA lambda library. The cDNA clones containing PRRSV specific sequences were selected using a VR2385 ORF 4 specific PCR probe and sequenced. The ORFs 2, 3 and 4 overlapped each other and encoded polypeptides with predicted M(r) of 29.5 kDa (ORF 2), 28.7 kDa (ORF 3) and 19.5 kDa (ORF 4), respectively. No overlap was found between ORFs 4 and 5, and instead there was a 10 bp sequence which separated these two ORFs. The nucleic acid homology with corresponding ORFs of the European PRRSV isolate Lelystad virus (LV) was 65% for ORF 2, 64% for ORF 3 and 66% for ORF 4. Comparison of the ORF 4 sequences of VR2385 with that of another U.S. isolate MN-1b revealed only 86% amino acid sequence homology and the presence of deletions in the ORF 4 of MN-1b. Our results further strengthen the observation that there is sequence variation between US and European PRRSV isolates.

Amino Acid Sequence

Mhc class II DNA polymorphisms within and between chromosomal species of the Spalax ehrenbergi superspecies in Israel.

Restriction fragment length polymorphisms (RFLPs) of two major histocompatibility class II genes (P alpha 1 and Q beta) were studied in 13 populations of four chromosomal species (2n = 52, 54, 58 and 60) of the mole rat, Spalax ehrenbergi superspecies in Israel. A substantial frequency of allelic fragments was found in both genes for all populations, including a desert isolate. In the P alpha 1 gene, one allelic fragment is a result of a deletion mutation which is diagnostic of the 2n = 52 chromosomal species. All other ten allelic variants are the result of point mutations. All mutations are located in a short region flanking the 3' end of the gene. Based on Mhc polymorphisms we confirm earlier evidence that gene flow does not occur between the older chromosomal species (2n = 52, 54, 58), and that reproductive isolation decreases, progressively from the oldest to the youngest species (2n = 60).

Animals