Actin- and tubulin-dependent functions during Saccharomyces cerevisiae mating projection formation.

Several conditional-lethal mutant alleles of the single-copy Saccharomyces cerevisiae beta-tubulin and actin genes were used to evaluate the roles of microtubules and actin filaments in the pheromone-induced extension of mating projections. Mutants defective in tubulin assembly form projections indistinguishable in appearance from those formed by wild-type cells. However, the tubulin mutants are unable to move their nuclei into the projections and to orient the spindle pole body associated with each nucleus toward the projection tip. Actin mutants are defective in spatial orientation of cell-surface growth required for formation of normal mating projections. Migration of nuclei into mating projections and Spa2p segregation to projection tips are also defective in actin mutants. Studies with abp1 null mutants showed that the function of the Abp1p actin-binding protein is either not required for projection formation or there are other proteins in yeast with similar functions. Our findings demonstrate that actin is required to restrict cell-surface growth to a defined region for pheromone-induced morphogenesis and suggest that nuclear position and orientation in mating projections depend on direct or indirect interaction of microtubules with actin filaments.     

Studies on the Thermal Destruction of Escherichia coli in Milk and Milk Products

Escherichia coli (ATCC no. 9637) was shown to have mean D values (slope of destruction rate curve) of 28.2, 5.1, 1.3, 0.00195, 0.00100, 0.00055, 0.00029, and 0.00016 min in milk at temperatures of 125, 130, 135, 168, 170, 172, 174, and 176 F, respectively. The mean D values for this organism in chocolate milk were 32.2, 10.4, 2.6, 0.00265, 0.00133, 0.00069, 0.00035, and 0.00028 min at respective temperatures of 125, 130, 135, 168, 170, 172, 174, and 176 F. Mean D values of 34.4, 10.0, 3.5, 0.00093, 0.00080, 0.00068, 0.00043, and 0.00036 min were found in cream for respective temperatures of 125, 130, 135, 168, 170, 172, 174, and 176 F. In ice cream mix the organism was found to have mean D values of 39.3, 15.2, 5.1, 0.00147, 0.00120, 0.00078, 0.00070, and 0.00053 min for temperatures of 125, 130, 135, 170, 172, 174, 176, and 178 F, respectively. The slopes of the thermal death time curves were found to be 10.2, 10.2, 10.0, and 10.3 F for this organism in milk, chocolate milk, 40% fat cream, and ice cream mix, respectively.     

HK/LK polymorphism and its genetic determination in Speke's gazelle

Concentrations of potassium (K) and sodium (Na) were determined in the erythrocytes of 28 members of an interbreeding herd of Speke's gazelle. The distribution of K concentrations and Na/K concentration ratios suggested the presence of the high-K/low-K (HK/LK) polymorphism known in erythrocytes of domestic bovids. The pedigree of the herd of gazelles is known completely, permitting examination of the inheritance of HK/LK polymorphism by overlaying the distribution of phenotypes on the pedigree. Statistical analyses clearly indicate a strong genetic component in the phenotypic variability that is entirely consistent with a single autosomal locus, two-allele mode of inheritance, with the LK allele being dominant. This is the first demonstration of HK/LK polymorphism in a wild bovid species. The evidence indicates that HK/LK polymorphism is of considerable evolutionary age, is of monophyletic origin, and is maintained by selection.     

Iron depletion: Possible cause of tumor cell cytotoxicity induced by activated macrophages

The experiments reported here provide a possible molecular mechanism for the activated macrophage cytotoxic effect. Tumor cells that develop cytostasis and inhibition of mitochondrial respiration in response to cocultivation with activated macrophages release a significant fraction of their intracellular iron-59 content. Kinetic studies show that specific release of iron-59 from target cells begins 4–6 hours after initiating cocultivation which is the time point that inhibition of DNA synthesis is first detected. Treatment of tumor cells with metabolic inhibitors causing inhibition of respiration, protein synthesis, RNA synthesis, and DNA synthesis to a similar or greater extent than that caused by activated macrophages does not induce release of intracellular iron-59. It is significant that mitochondrial respiration and DNA replication, both strongly inhibited in target cells by activated macrophages, are metabolic pathways with enzymatic activity vulnerable to inhibition by depletion of intracellular iron.     

Teratogenic activity of trichloroacetic acid in the rat

Trichloroacetic acid (TCA) is a by-product of the chlorine disinfection of water containing natural organic material. It is detectable in finished drinking water at levels comparable to the trihalomethanes (30-160 micrograms/L). TCA is also formed in vivo after ingestion of hypochlorite and has been identified as a major metabolite of chlorinated hydrocarbons such as trichloroethylene. The developmental effects of TCA were evaluated in the pregnant Long-Evans rat. Animals were dosed by oral intubation on gestation days 6-15 (plug = 0) with 0, 330, 800, 1,200, or 1,800 mg/kg/day. The vehicle control was distilled water. Maternal observations included clinical signs, weight change, and gross evaluation of organ weights and uterine contents at necropsy (day 20). Live fetuses were examined for external, skeletal, and soft tissue malformation. There were no maternal deaths associated with toxicity prior to sacrifice. Weight gain during treatment was reduced at 800, 1,200, and 1,800 mg/kg. Spleen and kidney weights were increased in a dose-related manner. The mean percent of resorbed implants per litter was 34, 62, and 90 at 800, 1,200, and 1,800 mg/kg, respectively. Live fetuses showed dose-dependent reductions in weight and length. The mean frequency of soft tissue malformations ranged from 9% at the low dose to 97% at the high dose. These were principally in the cardiovascular system (interventricular septal defect, levocardia). Skeletal malformations were found only at 1,200 and 1,800 mg/kg and were mainly in the orbit. Based on these observations TCA was considered to be developmentally toxic in the pregnant rat at doses of 330 mg/kg and above.     

Structure of subnucleosomal particles. Tetrameric (H3/H4)2 146 base pair DNA and hexameric (H3/H4)2(H2A/H2B)1 146 base pair DNA complexes

The tetrameric (H3/H4)2 146 base pair (bp) DNA and hexameric (H3/H4)2(H2A/H2B)1 146 bp DNA subnucleosomal particles have been prepared by depletion of chicken erythrocyte core particles using 3 or 4 M urea, 250 mM sodium chloride, and a cation-exchange resin. The particles have been characterized by cross-linking and sedimentation equilibrium. The structures of the particles, particularly the tetrameric, have been studied by sedimentation velocity, low-angle neutron scattering, circular dichroism, optical melting, and nuclease digestion with DNase I, micrococcal nuclease, and exonuclease III. It is concluded that since the radius of gyration of the DNA in the tetramer particle and its maximum dimension are very close to those of the core particle, no expansion occurs on removal of all the H2A and H2B. Nuclease digestion results indicate that histones H3/H4 in the tetramer particle protect a total of 70 bp of DNA that are centrally located within the 146 bp. Within the 70 bp DNA length, the two terminal regions of 10 bp are, however, not strongly protected from digestion. The optical melting profile of both particles can be resolved into three components and is consistent with the model of histone protection of DNA proposed from nuclease digestion. The structure proposed for the tetrameric histone complex bound to DNA is that of a compact particle containing 1.75 superhelical turns of DNA, in which the H3 and H4 histone location is the same as found for the core particle in chromatin by histone/DNA cross-linking [Shick, V. V., Belyavsky, A. V., Bavykin, S. G., & Mirzabekov, A. D. (1980) J. Mol. Biol. 139, 491-517]. Optical melting of the hexamer particle shows that each (H2A/H2B)1 dimer of the core particle protects about 22 base pairs of DNA.     

Hypothesis-driven science in large-scale studies: the case of GWAS

This is a response to the nine commentaries on our target article “Unlimited Associative Learning: A primer and some predictions”. Our responses are organized by theme rather than by author. We present a minimal functional architecture for Unlimited Associative Learning (UAL) that aims to tie to together the list of capacities presented in the target article. We explain why we discount higher-order thought (HOT) theories of consciousness. We respond to the criticism that we have overplayed the importance of learning and underplayed the importance of spatial modelling. We decline the invitation to add a negative marker to our proposed positive marker so as to rule out consciousness in plants, but we nonetheless maintain that there is no positive evidence of consciousness in plants. We close by discussing how UAL relates to development and to its material substrate.