Glycosylation and immunocytochemistry of binucleate cells in pronghorn (Antilocapra americana, Antilocapridae) show features of both Giraffidae and Bovidae.

Although the pronghorn (Antilocapra americana) resembles an antelope, its nearest relatives are the giraffe and okapi. In this study we have examined the placentae of 6 pronghorns using lectin- and immunocytochemistry to identify giraffid and bovid features. Binucleate cells (BNC) of the placenta exhibited features intermediate between those of the giraffe and bovine; Dolichos biflorus agglutinin binding - strong in the bovine BNC and absent in the giraffe - was evident in only a subpopulation of BNC while binding to blood vessels, as in the giraffe. Binding of Phytolacca americana agglutinin resembled that of the giraffe and okapi whereas many other glycans were found in all four clades. PAG antigens were similar to bovine and okapi but not giraffe. In summary, although the pronghorn outwardly resembles an antelope, placental BNC show giraffid features. Although each clade has its own individual characteristics, there are far more similarities than differences between them, emphasizing the common ancestry of all four clades.

Multiple G proteins and phospholipase C isoforms in human myometrial cells: implication for oxytocin action.

Although a physiological role for oxytocin during parturition is well accepted, the mechanisms by which it activates myometrial contractility during labor have not been completely elucidated. We have previously shown the presence of Gq and two pertussis toxin (PT) substrates of the Gi family in human myometrial cells. In the present study, we have identified by Western blotting the G protein and phospholipase C (PLC) isoforms present in these cells and investigated their implication in oxytocin signaling by measuring the formation of inositol phosphates (IPs) and mobilization of intracellular calcium. We found G protein subunits alpha(q), alpha(11), alpha(i1), alpha(i2), alpha(i3), alpha(z), and two splice variants of alpha(s)- and beta-subunits. We have also detected the presence of five PLC isoforms: beta 1, beta 2, beta 3, gamma 1, and gamma 2. Oxytocin-induced IPs formation and intracellular Ca2+ mobilization were inhibited to approximately 50% after pretreatment of the cells with PT, suggesting that oxytocin activates PLC beta by interacting with at least two types of G proteins: a member of the Gq family (PT resistant) and a member of the Gi family (PT sensitive). The tyrosine phosphatase inhibitor pervanadate stimulated IPs formation in myometrial cells. Using the protein kinase inhibitors staurosporine, phenylarsine oxide, and Ro 31-8220 and the protein kinase C activator phorbol dibutyrate, we have shown that pervanadate and oxytocin activate PLC by different mechanisms. Furthermore, oxytocin did not activate tyrosine phosphorylation in human myometrial cells, as measured with an antiphosphotyrosine antibody, indicating that it does not activate a PLC gamma isoform. We conclude that oxytocin activates human myometrium by interacting with at least two G proteins and possibly three PLC beta isoforms.

Oxytocin signalling in human myometrium.

A physiological role for oxytocin in stimulating uterine contractions during labour is well accepted, but has not yet been well defined. Oxytocin activates phospholipase C to produce inositol 1,4,5-trisphosphate, which releases Ca2+ from intracellular stores. There is considerable evidence that G-proteins are involved in this signalling pathway. The objectives of the present study were to determine the mechanisms of action of oxytocin in human myometrium. We have measured the effect of oxytocin on the formation of inositol phosphates (InsPs) in cultured human myometrial cells labelled with [3H] inositol and on changes in intracellular free Ca2+ concentration ([Ca2+i]) in single cells using a dynamic calcium imaging system. Pertussis toxin was used to obtain information on the G-proteins involved. Oxytocin induced InsPs formation and [Ca2+i] mobilisation in a concentration-dependent manner in human myometrial cells. Our data suggest that two distinct types of G-proteins are involved in the oxytocin response: one most probably a member of the Gq family (pertussis toxin-resistant) and another of the Gi family (pertussis toxin-sensitive). Using Western blotting, we have found that the pertussis toxin-resistant G-proteins alpha(q), alpha(11) and alpha(2), and pertussis toxin-sensitive alpha(i1), alpha(i2), and alpha(i3) are expressed in these cells. We have also detected the phospholipase C isoforms beta(1), beta(2) and beta(3) which are regulated by G-proteins, and phospholipase C isoforms gamma(1) and gamma(2), regulated by receptor tyrosine kinase pathways. However, oxytocin does not stimulate tyrosine phosphorylation in myometrial cells. Extracellular Ca2+ does not play a direct role in the activation of phospholipase C by oxytocin. Protein kinase C causes a strong inhibitory feedback on the oxytocin pathway: protein kinase C activators abolish the response to oxytocin while inhibitors potentiate it. Oxytocin responsiveness is upregulated by incubating the cells in the presence of oestradiol. This effect is reversed by the anti-oestrogen tamoxifen. Oestrogens exert their effects on the oxytocin pathway at a postreceptor level, possibly by affecting the expression of G-proteins and/or phospholipase C isoforms.

Gas chromatographic detection of D-(-)-2,3-butanediol and butyric acid produced by sporeformers in cream-style corn and canned beef noodle soup: collaborative study.

A gas chromatographic method that identifies sporeformers as the cause of spoilage in swollen cans of low-acid foods was collaboratively studied in 2 stages. Two organic compounds produced by sporeformers, D-(-)-2,3-butanediol and butyric acid, are measured in the upper phase after centrifugation of the liquid portion of the can contents. Each sample is assayed on 2 packed columns designed for the assay of aqueous solutions of volatile fatty acids, using flame ionization detectors. For study 1, 16 duplicate inoculated cans of cream-style corn and beef noodle soup were sent to 9 collaborators. For study 2, 7 collaborators received 11 duplicate inoculated cans of the 2 foods. Duplicate uninoculated cans of each food served as negative controls. The inocula were 6 sporeforming organisms (4 Clostridium and 2 gas-forming Bacillus species) and 2 nonsporeformers. After the deletion of marginal samples, the percentages of correctly identified sporeformers and nonsporeformers in beef noodle soup were 83 (110/132) and 90 (54/60), respectively; corresponding percentages for cream-style corn were 80 (98/123) and 100 (35/35). The method has been adopted official first action.

The vital statistics method of estimating net migration by age cohorts.

The focus of this paper is the development and testing of a method of estimating deaths which occur during a decade to aging birth and death cohorts, so that it may be possible to estimate net migration by the vital statistics (VS) method for age cohorts. Until now the VS method has been used only in making estimates of total net migration.The results obtained by using the VS method for age cohorts show that (1) the average census survival rate (CSR) method generally yields algebraically lower estimates of net migration than does the VS method; but (2) there are some striking exceptions which are apparently associated with errors in census enumeration by age, sex, and color. Comparisons between the average CSR and the VS methods are shown, by age, for both the North Carolina and the coterminous United States populations.A cursory examination of these comparisons suggests that the exclusive use of the VS method in estimating net migration for age cohorts may lead to substantial error. Finally, the magnitude of these errors in estimating net migration, as well as in census enumeration, can be roughly approximated if it is assumed that the use of the CSR method yields reasonably accurate estimates of net migration.

Effect of census errors on the measurement of net migration.

This paper traces the history of the use of vital statistics, survival rates, and ratios in the estimation of net migration from one decade to another. Net migration studies by Hart (1921); Baker (1933) ; Hamilton (1934); Thornthwaite (1934); Lively and Taeuber (1939) ; Henderson (1943); Hamilton and Henderson (1943); Hamilton (1951); Siegel and Hamilton (1952); Lee and Bowles (1954); Price (1955); Lee, Miller, and others (1957); Hamilton (1959); Zachariah (1962); Tarver (1962); Shryock (1964); Eldridge (1965); Hamilton (1965); and the United States Census Bureau are cited as the principal users of various residual methods of estimating net migration. All these demographers have either implicitly or explicitly recognized that errors in census enumeration and in the registration of births and deaths have been reflected in errors of estimated net migration.The underlying characteristic of all the methods used by these demographers has been the estimation of net migration as a residual obtained by subtracting natural increase in an area during a decade from the population change during the same decade. This method has been most generally stated in the classic formula {fx394-1} This formula has been used both with total populations and with aging cohorts. The principal variations of the basic formula have involved the use of life table and census survival ratios as a means of measuring natural increase (B - D), or of estimating "expected" populations assuming no migration. The main points of controversy have involved life table v. censm survival ratios, assumptions regarding the similarity in national and state census enumeration errors, and ways and means of estimating the errors involved in estimates of migration and of migration rates by the various methods.Daniel O. Price (1955) and Zachariah (1962) made important mathematical contributions and attempted to evaluate the errors involved in the me of census survival rates. Eldridge (1965) discovered that, in the United States between 1950 and 1960, the use of the census survival rate method usually gave much lower estimates of net migration than did the classic vital statistic method. Hamilton (1965), using some suggestions by Hope T. Eldridge, developed a mathematical theory or explanation of not only why the CSR estimates were usually lower than the VS estimates but also why the CSR estimates would usually give closer estimates on the true net migration than would the EVS method, which itself is subject to errors of census enumeration and of underregistration of births and deaths. The author also discusses the effect of improvement in census enumeration between 1950 and 1960 on estimates of net migration and derives a generalized formula which takes the timing of migration into consideration.The author acknowledges with sincere appreciation important constructive suggestions made by Dr. Hope T. Eldridge, Population Studies Center, University of Pennsylvania, and the authors of the many papers used as original material. This paper is a revision of a paper read before the annual meeting ot the Population Association of America, Hotel Roosevelt, New York, New York, April 29-30, 1966. Contribution from the Departments of Sociology and Experimental Statistics, North Carolina Agricultural Experiment Station, North Carolina State University. Published with the approval of the Director of Research as Paper No. 2227 of the Journal series.