Possible cytogenetic mechanisms of direct paternal influence on the human twinning tendency and their consequences: a hypothesis

Certain cytogenetic mechanisms are suggested to explain the puzzling cases of the direct male influence on the repeated twin births in mammals including humans. The hypothesis is based on the peculiarities of female oogenesis and meiosis, the peculiarities of fertilization and on the established facts of the occurrence of true viable chimaeras produced by separate fertilization of two meiotic products of oogenesis. We postulate that definite genetic factors are transferred from the paternal side whose products become active in male gametes and promote penetration of two spermatozoa (polyspermy) or appearance of two male pronuclei in the egg cytoplasm. The results of such events may be twinning and occurrence of chimaeric or heteroploid individuals. The appearance of viable twins produced by male-dependent polyspermy may be considered as a fortunate outcome of various possible cytogenetic anomalies of fertilization, meiosis, and cleavage divisions. The existence of non-canonical cases of twins, except mono-and dizygotic ones is postulated, according to the hypothesis. Twins pairs produced by two paternal and one maternal genomes may be called "one and halfzygotic or sesquizygotic". The different types of twins may be classified in an order, according to the degree of genetic similarity; monozygotic, chimaeric, sesquizygotic chimaeric, sesquizygotic and dizygotic. This gives an opportunity to explain the appearance of 2 to 3% of "doubtful cases" in mass classification of twin pairs into mono- and dizygotic. The verification of the hypothesis involves the special thorough genetic and cytogenetic analysis of all twin sibs and their parents in families with the direct paternal influence on twin births.(ABSTRACT TRUNCATED AT 250 WORDS)     

Emergency queen cell production in the honey bee colony

Summary Emergency queen cell production was examined in honey bee colonies of mixed European races. Thirteen colonies were dequeened and followed on a daily basis until after queen emergence. Observations were made on the number of cells, the temporal sequence of queen cell construction, cell location within the nest, the age of larvæ selected for queen rearing, mortality of immature queens and the scenting behavior of workers in queenless colonies.Queen loss was detected within 6–12 hours and was first indicated by an increase in scenting behavior (on colony disturbance) and queen cup construction. The number of scenting workers reached a peak in 12–24 hours and then declined, as queen cell numbers increased. The time of queen cell initiation varied from 12–48 hours in different colonies. Emergency queen cells were usually started over worker larvæ less than 2 days of age (64.7%), but cells were built over 3 (25.3%) and 4 (10.0%) day old larvæ. Only 2 of 268 cells (0.8 %) were started over eggs; one survived and developed into a drone larva. In 6 colonies emergency queen cells were started over drone larvæ but these were destroyed immediately before or shortly after capping. The overall rate for queen cell construction over drone larvæ was 9.3%.The rate at which new queen cells were started after queen loss was high for two to four days, but then declined although new queen cells were started as late as eight or nine days after queen removal. The number of cells produced by a colony usually peaked by the third or fourth day and then leveled. Slight declines in total cell number often occurred because of cell mortality. The number of queen cells started by colonies varied from 11–49 with a mean of 20.4; cell mortality averaged 39.1%. Queen cells were well distributed throughout the brood nest but placement was biased toward the bottom of the frames and away from the entrance.

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Production de cellules royales après orphelinage accidentel dans des colonies d'abeilles à miel

Resume La production de cellules royales après orphelinage accidentel fut examinée dans des colonies d'abeilles de différentes races européennes. Treize colonies ont été quotidiennement placées dans un orphelinat expérimental après l'apparition d'une nouvelle reine. Des observations ont été faites sur le nombre de cellules, le timing de la reconstruction des cellules royales, l'emplacement des cellules à l'intérieur du nid, l'âge des larves sélectionnées en vue de l'élevage des reines, le taux de mortalité des cellules et le phénomène d'exhibition de la glande de Nassanoff des ouvrières dans les colonies orphelines.On a pu détecter la perte d'une reine après 6 à 12 heures; celle-ci fut tout d'abord indiquée par le fait qu'un certain nombre d'abeilles exhibent leur organe odorant lors de l'ouverture de la ruche, et l'élaboration de la cupule royale. Le nombre des ouvrières exposées a atteint son record entre 12 et 24 heures puis s'est mis à décroître, alors que les cellules royales augmentaient. Le temps requis pour l'initiation des cellules royales a varié entre 12 et 48 heures, selon les colonies. Les cellules royales de remplacement ont commencé ordinairement à se former sur des larves d'ouvrières de moins de 2 jours (64,7%), mais des cellules se sont développées sur des larves âgées de 3 (25,3%) à 4 jours (10,0%). Sur 268 cellules, 2 étaient uniquement formées à partir d'ufs, dont un seul survivait et devenait une larve mâle. Dans six des colonies, des cellules royales se sont développées à partir de larves mâles, mais celles-ci furent immédiatement détruites soit avant, soit juste après l'operculation. Le taux de développement de cellules royales était de 9,3% par rapport aux cellules mâles.Le taux de développement de nouvelles cellules royales après la perte d'une reine a été assez élevé pendant une période de 2 à 4 jours, mais s'est mis à décroître bien que de nouvelles cellules royales se formaient entre 8 et 9 jours après le début de l'orphelinage. Nous avons noté un taux record de cellules produites par une colonie vers le 3^e ou 4^e jour, qui s'est ensuite réparti de façon plus égale. Le taux de mortalité des cellules a alors provoqué la baisse du nombre total des cellules. Le nombre des cellules royales des colonies a varié entre 11 et 49, c'est-à-dire une moyenne de 20,4; le taux de mortalité des cellules s'est avéré de 39,1%. Les cellules royales étaient bien distribuées dans tout le nid à couvain, mais surtout vers le fond du cadre, et loin de l'entrée de la ruche.

     

Extracellular lumazine from aggregating Dictyostelium discoideum cells. Influence of pH on its fluorescence

Lumazine has been demonstrated to be one of the two main compounds responsible for the extracellular fluorescence linked with the aggregation ability of Dictyostelium discoideum, strain Ax-2. The other compound, also having lumazine properties, is, however, different from the 7-hydroxylumazine proposed previously. The influence of pH on the fluorescence of lumazine was studied. The possible use of extracellular pteridines as pH markers was stressed and a method for the determination of the amount of "lumazine equivalents" in the extracellular medium of aggregating D. discoideum cells is elaborated.     

Effect of dissolved oxygen on nitrogen fixation by A. vinelandii. I. Free cell cultures

As part of an investigation into the use of biological nitrogen fixation for fertilizer ammonia production, continuous culture studies of respiration and nitrogen fixation in the aerobic bacteria Azotobacter vinelandii under oxygen-limited conditions were conducted. Respiration and growth rates followed Monod forms with respect to dissolved oxygen concentration. However, specific nitrogen fixation rate and nitrogenase activity exhibited maximum values at dissolved oxygen concentrations of ca. 0.02 mM (10% of air saturation). These results suggest careful control of oxygen in the environment is necessary to optimize fixed nitrogen production by this organism.     

Thermal stabilization of amylolytic enzymes by covalent coupling to soluble polysaccharides

The immobilization of pullulanase and beta-amylase on soluble polysaccharides (dextrans and amylose) has been carried out. The method used for coupling the enzymes to the carbohydrate support involves limited periodate oxidation of the polysaccharide followed by reductive alkylation with sodium cyanoborohydride or borohydride. The influence of the degree of functionalization of the carbohydrate, the incubation time, the nature of the reducing agent and, for the dextrans studied, their molecular weight, on the properties of the conjugate were studied. We have observed an apparent correlation between the molecular weight of the glycoprotein conjugates formed and their thermal stability, resistance to urea denaturation and their kinetic parameters. By selecting the proper experimental conditions leading to conjugates with maximum thermal stabilities, it has also been shown that beta-amylase conjugates can hydrolyze starch at a temperature 20 degrees C higher than the corresponding value for the native enzyme. This result demonstrates that conjugation may result in modified enzymes leaving a high operational stability at elevated temperatures. We suggest that the immobilization method presented in this article represents an approach to the stabilization of enzymes employed at an industrial level, which may be of general application.     

Occurrence of intramitochondrial Ca2+ granules in a hypertrophied heart exposed to adriamycin

Left ventricular hypertrophy was produced in rabbits by narrowing the abdominal aorta in the subdiaphragmatic region. Six weeks after the surgery, sham control as well as hypertrophied animals were treated with adriamycin. Myocardial cell damage resulting from a total cumulative dose of 5 mg/kg of adriamycin was seen only in hypertrophied hearts. Alterations in muscle cells of these hearts included prominent "contraction bands" and perinuclear edema. Mitochondria were characterized by swelling and accumulation of electron-opaque granules. Energy-dispersive x-ray analysis of the mitochondria revealed the presence of calcium in these granules. The study confirms that the hypertrophied heart is more vulnerable to adriamycin-induced cell damage and this may be due to an increased susceptibility of these hearts to the occurrence of Ca2+ overload in the cell.     

Differential effects of positive and negative proliferative stimuli on murine cytolytic and helper T-cell clones

Studies were performed to determine whether substances could be identified which exhibited differential regulatory effects--either positive or negative--on the growth of murine alloreactive cytolytic (Tc) and helper (Th) cloned T-cell lines. The following lines of evidence suggested that Tc and Th proliferate in response to the same growth factor (GF). (1) When GF-containing fluids from cultures of phorbol myristic acetate (PMA)-activated EL4 thymoma were fractionated by a variety of biochemical techniques. Tc and Th eluted together. (2) Absorption of GF-containing supernatants with either cloned Tc or cloned Th depleted GF activity for each to a similar extent, and GF eluted from either Tc or Th to which it had adsorbed supported the proliferation of Tc and Th equally well. (3) Lectin-depleted supernatants from cultures of concanavalin A (Con A)-activated Th stimulated the proliferation of Th as well as Tc. (4) Recombinant human interleukin (IL-2) supported the growth of Tc and Th with equal efficiency. On the other hand, the following observations indicated that Tc and Th differed in their responses to inhibitors of GF-driven proliferation. (1) Con A at greater than or equal to 0.3 micrograms/ml inhibited the GF-driven proliferation of each of three Th lines but not either of two Tc lines. To the contrary, Con A enhanced GF-dependent proliferation of Tc. (2) Like Con A, allogeneic splenocytes selectively depressed GF-driven proliferation of Th but not Tc. (3) A substance generated during the acid elution of GF from cells, possibly a modified fetal calf serum component, greatly reduced the GF-driven proliferation of Tc but not Th. These results suggest that differential control of the proliferation of Tc and Th in cellular immune responses may be achieved via negative regulatory signals and raise the possibility that substances which can selectively depress the proliferation of specific T-cell subsets might be found which would be of therapeutic value.     

Wheat germ agglutinin-resistant variant of EL4 containing altered oligosaccharides as a target cell for cytotoxic T cells

A lectin-resistant variant of the murine EL4 lymphocytic leukemia cell line was selected in the presence of wheat germ agglutinin for low levels of cell-surface sialic acid. H-2Kb was the major internally radiolabeled H-2b molecule on the cell-surface of WD1, and it was not sialylated, as determined by two-dimensional gel analysis. Endo-beta-N-acetylglucosaminidase H treatment of the WD1 membrane fractions suggested that the oligosaccharides on the cell-surface H-2Kb molecule were complex, but nonsialylated. Monoclonal antibody inhibition of the allogeneically primed cell-mediated cytotoxicity (CMC) reaction indicated that the T cells (BALB/c anti-EL4; H-2d anti-H-2b) were specific only for the H-2Kb target cell antigen. These WD1 variant cells were used as targets in the CMC assay using anti-H-2Kb T cells and compared with the parent EL4 in vitro line. The change in the cell-surface oligosaccharide did not affect the susceptibility to lysis by the cytotoxic T lymphocytes even though there were 2.5-fold more H-2Kb antigens on the WD1 variant cell (1.5 X 10(5) sites/cell) than on the parent EL4 in vitro cell (5.9 X 10(4) sites/cell). It was possible to isolate highly purified preparations of H-2Kb from either the EL4 or the WD1 line using a monoclonal antibody affinity column. Interestingly, the variant WD1 cell would no longer grow in the peritoneal cavity of the syngeneic C57BL/6 mouse.