Gibbons and their territorial songs.

Discovery of the great call of the Javan gibbon and finding an enclave of the agile gibbon in Kalimantan permit for the first time a comparison of vocalizations among all major taxa of Hylobates. The songs are stereotyped, constant throughout the interrupted areas of distribution of each taxon, and are sexually divocal.

Regulation of expression of a sheep metallothionein 1a-sheep growth hormone fusion gene in transgenic mice.

Transgenic mice containing a sheep metallothionein 1a-sheep growth hormone fusion gene exhibited low, tissue-specific basal levels of transgene mRNA expression, resulting in slightly elevated levels of circulating growth hormone that did not lead to a detectable increase in growth. After zinc stimulation, high levels of transgene mRNA expression were induced in a number of tissues; these levels correlated with increased levels of circulating growth hormone, resulting in growth increases of up to 1.5 times the levels of controls and unstimulated transgenic mice. After removal of the zinc stimulus, transgene expression and circulating growth hormone concentrations returned to basal levels. Additional evidence from the pattern of developmental expression of the transgene suggests that zinc is the main regulator of this promoter in mice. The demonstrated regulation and low basal level of expression of the sheep metallothionein 1a promoter make it a candidate for use in other mouse transgenic studies and for use in transgenic livestock, in which regulation of expression is essential.

Cloning and sequencing of a cDNA encoding an ovine oestrus-associated oviducal protein.

Ovine oestrus-associated oviducal glycoprotein (oEGP) is synthesized and secreted specifically by the ampullary region of the ovine oviduct during the peri-ovulatory stages of the oestrous cycle. A cDNA that encodes oEGP was isolated and sequenced. Isolation of oEGP was achieved using the polymerase chain reaction (PCR) with primers based on a bovine oestrus-associated oviducal glycoprotein cDNA (bOGP) sequence. A 1599-bp cDNA encodes, in part, a deduced 519-amino acid sequence of mature protein which carries two potential N-linked glycosylation sites. The deduced amino acid sequence is more than 95% identical to that of bOGP and more than 74% identical to the first 491 amino acids of human oestrogen-dependent oviducal glycoprotein (hOGP). Northern blot hybridizations of RNA from several sheep tissues detected mRNA (2.4 kb) only in an ampulla oviduct sample.

Production of transgenic merino sheep by microinjection of ovine metallothionein-ovine growth hormone fusion genes.

Seven transgenic Merino sheep have been produced by the technique of pronuclear microinjection. Two different Sheep Metallothionein-1a-Sheep Growth Hormone fusion genes were used. Four of the transgenic sheep, all of which contained the gene MTsGH5, did not express the transgene. The remaining three sheep carrying the second fusion gene, MTsGH9, expressed the gene at high levels in a variety of tissues and had elevated blood levels of sheep growth hormone.

Blastocyst trophectoderm biopsy and preimplantation genetic diagnosis for familial monogenic disorders and chromosomal translocations.

OBJECTIVE: Modern in vitro fertilization practices involve transfer of embryos as blastocysts, when anabolic metabolism is well established and pregnancy rates can be maintained while transferring embryos singly to avoid multiple pregnancies. Embryo biopsy for preimplantation genetic diagnosis (PGD), however, is generally performed on day 3, when the embryo comprises just 6 to 8 cells, one or two of which are removed for testing. Implantation rates and clinical pregnancy rates have remained relatively low and a harmful effect from losing one or more cells from such early embryos has not been excluded. METHODS: We performed a sequential study involving 399 egg retrievals and 1879 embryo biopsies for patients undergoing PGD to avoid a serious monogenic disease or an unbalanced chromosomal translocation. We compared implantation and viable pregnancy rates after biopsies taken on day 3 (cleavage-stage biopsy) with biopsies delayed until day 5 or 6, when the embryo is a blastocyst and 5 or more cells can be sampled from the trophectoderm while the inner cell mass, from which the fetus develops, remains intact. All embryos were transferred as blastocysts. RESULTS: Despite fewer blastocysts than cleavage embryos biopsied and tested (3.6 compared to 6.6), implantation rates per embryo transferred were 43.4% if biopsied at the blastocyst stage and 25.6% if biopsied at the cleavage stage (P < 0.01), with ongoing or live-birth pregnancy rates per egg retrieval of 34.2% (average transfer number 1.1) for blastocyst biopsies and 25.5% (transfer number 1.6) for cleavage stage biopsies (P < 0.05, 1-tailed). The multiple pregnancy rate for monogenic disease exclusion fell from 16.7% to 2% (P = 0.04, 1-tailed). CONCLUSIONS: For exclusion of genetic disease, day 5-6 blastocyst-stage biopsies are more likely to be followed by implantation and singleton births than is the case after PGD performed on day 3.

The role of preimplantation genetic diagnosis in the management of severe rhesus alloimmunization: first unaffected pregnancy: case report.

Rhesus (Rh) D alloimmunization may cause haemolytic disease of the fetus and newborn if the fetal Rh blood type is positive. Although the incidence of severe RhD alloimmunization has decreased with prophylactic anti-D immunoglobulin administration during and after pregnancy, sensitization still occurs in a small group of women. In such women, Rh disease will continue to be significant problem and for their babies who may be affected. Preimplantation genetic diagnosis (PGD) may be utilized to avoid materno-fetal blood group incompatibility in an RhD-sensitized woman. Biopsy of a single cell from early cleavage-stage embryos screening for RhD-negative embryos allows the transfer of only RhD-negative embryo(s) into the uterus. This avoids any complications related to haemolytic disease of the fetus and newborn. This article describes the first reported case of an unaffected pregnancy using PGD for Rh disease. IVF and embryo transfer resulted in a clinical pregnancy and the birth of a healthy girl confirmed to be blood type RhD negative. PGD in couples with a heterozygous RhD-positive male partner provides an option for avoiding haemolytic disease of the newborn in RhD alloimmunized mothers.

Production of transgenic sheep.

The production of transgenic sheep has proven difficult compared to the mouse and lower animals. The work load is far greater and the rates of success far less by most criteria. However, the benefits to human and animal health and agricultural productivity are potentially enormous (Ward and Nancarrow, Chapter 5) and support for the continuation of the work is assured. Unfortunately, the low rate of transgenesis for sheep, at about 1% of injected, transferred embryos, means that investigation of the regulation of expression of the transgenes, their phenotypic effects, and optimization of the fusion gene constructs, all of utmost importance to the agricultural industry, can seldom be addressed. We know now that the mouse may not be a good model for the sheep, an example being the ovine metallothioneinovine growth hormone fusion gene, GH9, for which expression and phenotypic effects were quite different for sheep and mice. In sheep, pronuclear microinjection of several hundred copies of the foreign gene into embryos is the only published method used to regularly produce transgenics and it will be the standard by which future methods for incorporation of the transgene are judged.

A Milk-Like Beverage from Neutralized Direct-Acid-Set Cottage Cheese Whey 1.

An imitation milk was formulated by combining 4 parts of neutralized, direct-acid-set, cottage cheese whey with 6 parts of whole milk, and fortifying with .5% nonfat dry milk solids (NFDM). Whey collected from a commercial plant was neutralized, clarified, blended with milk, fortified with NFDM, pasteurized, homogenized, and packaged in ½-gal. paper cartons in the Kansas State University (KSU) dairy processing plant. Calculated raw material costs were compared for 2.0% low fat milk, 3.25% milk, and for imitation milks with those same milkfat concentrations. The 2.0 and 3.25% imitation milk could be formulated with savings of 35 and 25 cents per gallon, respectively. Coded samples of the product were compared with regular KSU whole milk by 112 persons. Of these, 45% identified the KSU milk samples, 35% thought the experimental product was the KSU milk, and 20% could tell no difference. The milk sample was preferred for flavor by 42% of the consumers, 32% had no preference, and 26% preferred the imitation milk. The imitation milk, containing 11.5% total solids and 2.4% protein, was .5% lower in total solids and 1.0% lower in protein than KSU whole milk.

Prevalence of leprosy among slum dwellers.

Distribution of leprosy among slum dwellers in two areas of Bangalore City were studied. Out of a random sample of 483 it was found that 150 were suffering from leprosy while 74 tuberculoid and 25 lepromatous cases were detected, the remaining showed up as borderline and polyneuritic types. The important socio economic features and the consequences of this chronic disease have been discussed.

Flavor and Texture of Cottage Cheese Made by Direct-Acid-Set and Culture Methods 1.

Consumers were asked to indicate a flavor and texture preference for one of two creamed and salted small-curd cottage cheeses made by direct-acid-set or conventional short-set culture methods. When a commercial starter distillate flavor was added to direct-acid-set cheese, its flavor and texture were preferred over cheese made by the culture method. Conversely, when flavor was not added to the direct-acid-set cheese, consumers preferred the flavor and texture of the cultured product. Differences among mean flavor or texture scores of three age groups were not significant (< 20, 20-40, > 40). Preferences for texture appeared to be biased by flavor.

Chromosomes and DNA of Mus: the karyotypes of M. fulvidiventris and M. dunni.

The chromosomes of the Asian mice, Mus fulvidiventris (booduga?), are typical of the Mus in general, viz., 40 telocentric chromosomes. The centromeric heterochromatin does not fluoresce brightly. The G band pattern of the euchromatin is the same as that of M. musculus. The diploid number of M. dunni is also 40, but each autosome possesses a short, heterochromatic second arm. The X chromosome is a long submetacentric, whose entire short arm and the terminal segment of the long arm are heterochromatic. The Y is a long telocentric and is heterochromatic. The G band pattern of the long arms of M. dunni involved only the addition of C bands. Mus dunni and M. booduga are sympatric in many localities in India, but they can be separated by karyological and subtle morphological differences.

Zygote viability in gene transfer experiments.

To learn why some zygotes remain viable after gene transfer while others lyse, we injected DNA into fertilized eggs and compared those that lysed within 1 h of injection with others that retained a normal appearance. Using scanning electron microscopy, we found open holes on the surface of lysed eggs, indicating failure of the plasma membrane to reseal after microinjection. No holes were seen in unlysed eggs, but many of them had membrane alterations suggestive of healed punctures. We also examined aspects of the gene transfer procedure that might influence survival such as the size of injection pipettes and their taper relative to zygote diameter, possible toxicity of the injection medium, the timing of injection, and immediate vs. delayed pipette withdrawal. The only factors that significantly affected cell viability were pipette size and taper, and timing of injection in relation to first cleavage. This suggests that zygote viability correlates inversely with the size of the hole produced by the injection pipette and that damage to the membrane is less successfully repaired as the fertilized egg readies itself for division.