Ethylene production from propanal

Tracer studies using a model system consisting of Cu²⁺ and ascorbate indicate that carbons 2 and 3 of propanal are converted to ethylene, and carbon 1 is converted to formic acid and CO₂. A mechanism accounting for this reaction is described. In apple tissue, methionine but not propanal is readily incorporated into ethylene. It is therefore concluded that propanal is not a precursor of ethylene in this fruit.     

Hemoglobin tacoma — A β-chain variant

A hitherto unknown inherited hemoglobin variant, Hb Tacoma, was discovered in three healthy members of a family of European extraction.Hybridization experiments with canine hemoglobin indicated a structural abnormality in the -chain. The variant was therefore designated as Hb _{2 2} ^Tacoma. Separation of Hb Tacoma from Hb A could only be clearly achieved by starch grain electrophoresis in TEB buffer at pH 8.6–9.0, where Hb Tacoma moved more rapidly towards the anode than Hb A; gradient column chromatography on DEAE cellulose and CM-Sephadex achieved partial separation. The proportion of abnormal hemoglobin in the heterozygote amounted to 43 per cent of the total hemoglobin. Hb Tacoma was less heat resistant and became more rapidly denatured in 8 M urea solution than Hb A, Hb C, or Hb S.On thin-layer starch gel electrophoresis in TCB buffer at pH 8.6 Hb Tacoma was associated with an electrophoretically slow, benzidine-positive component, Hb Tacoma-slow. The exact biochemical nature of this minor component could not be determined, although a polymeric product of Hb Tacoma is suspected.Heterozygosity for Hb Tacoma is associated with a raised Hb A₂ level without evidence of -thalassemia.A preliminary report of Hb Tacoma was presented at the Annual Meeting of the American Society of Human Genetics, Boulder, Colorado, August, 1964, and at the Seventh Annual Meeting of the American Society of Hematology, November, 1964, Seattle, Washington (Blood, 24, 848 (1964)).     

A Sample-Grouping Technique for Paraffin Embedments

A technique is described which facilitates histological preparation of multiple tissue specimens for light microscopy. The procedure enables the investigator to separate and label identifiable subgroups from a larger number of specimens in one histological section. After standard fixation, murine esophagi were arranged longitudinally and secured within segments of murine intestine. Markers such as plant fibers and human hairs were threaded alongside the esophagi within each intestinal casing. After standard dehydration and infiltration, several segments of intestine were arranged parallel to each other and at right angles to the intended plane of sectioning and were embedded together in one paraffin block. This method made it possible to assemble onto one microscope slide cross sections of 42 individual esophagi in 6 identifiable subgroups, each containing 7 esophagi.     

The relative efficiency of the Hardy-Weinberg equilibrium-likelihood and the conditional on parental genotype-likelihood methods for candidate-gene association studies.

Selecting a control group that is perfectly matched for ethnic ancestry with a group of affected individuals is a major problem in studying the association of a candidate gene with a disease. This problem can be avoided by a design that uses parental data in place of nonrelated controls. Schaid and Sommer presented two new methods for the statistical analysis using this approach: (1) a likelihood method (Hardy-Weinberg equilibrium [HWE] method), which rests on the assumption that HWE holds, and (2) a conditional likelihood method (conditional on parental genotype [CPG] method) appropriate when HWE is absent. Schaid and Sommer claimed that the CPG method can be more efficient than the HWE method, even when equilibrium holds. It can be shown, however that in the equilibrium situation the HWE method is always more efficient than the CPG method. For a dominant disease, the differences are slim. But for a recessive disease, the CPG method requires a much larger sample size to achieve a prescribed power than the HWE method. Additionally, we show how the relative risks for the various candidate-gene genotypes can be estimated without relying on iterative methods. For the CPG method, we represent an asymptotic power approximation that is sufficiently precise for planning the sample size of an association study.     

Human osteoblasts from younger normal and osteoporotic donors show differences in proliferation and TGFβ-release in response to cyclic strain

Mechanical stimulation of bone tissue by physical activity stimulates bone formation in normal bone and may attenuate bone loss of osteoporotic patients. However, altered responsiveness of osteoblasts in osteoporotic bone to mechanical stimuli may contribute to osteoporotic bone involution. The purpose of the present study was to investigate whether osteoblasts from osteoporotic patients and normal donors show differences in proliferation and TGFβ production in responses to cyclic strain. Human osteoblasts isolated from collagenase-treated bone explants of 10 osteoporotic patients (average age 70 ± 6 yr) and 8 normal donors (average age 54 ± 10 yr) were plated into elastic rectangular silicone dishes. Subconfluent cultures were stimulated by cyclic strain (1%, 1 Hz) in an electromechanical cell stretching apparatus at three consecutive days for each 30 min. The cultures were assayed for proliferation, alkaline phosphatase activity and TGFβ release in each three parallel cultures. In all experiments, osteoblasts grown in the same elastic dishes but without mechanical stimulation served as controls. Significant differences between stimulated cultures and unstimulated controls were determined by a paired two-tailed Wilcoxon test. In comparison to the unstimulated controls, osteoblasts from normal donors significantly increased proliferation (p = 0.025) and TGFβ secretion (p = 0.009) into the conditioned culture medium. In contrast, osteoblasts from osteoporotic donors failed to increase both proliferation (p > 0.05) and TGFβ release (p > 0.05) in response to cyclic strain. Alkaline phosphatase activity was not significantly affected (p > 0.05) in normal as well as osteoporotic bone derived osteoblasts.

These findings suggest a different responsiveness to 1% cyclic strain of osteoblasts isolated from normal and osteoporotic bone that could be influenced by both the disease of osteoporosis and the higher average age of the osteoporotic patient group. While osteoblasts from osteoporotic donors failed to increase proliferation and TGFβ release under the chosen mechanical strain regimen that stimulated both parameters in normal osteoblasts, it is possible that some other strain regimen would provide more effective stimulation of osteoporotic cells.     

Oviposition and chemosensory stimulation of the root flies Delia radicum and D. floralis in response to plants and leaf surface extracts from resistant and susceptible Brassica genotypes

In Brassica crops differences in susceptibility to root fly attack can be largely attributed to antixenotic resistance. Plants of four genotypes (two swedes and two kales) with widely differing resistance in field trials, were compared in laboratory choice assays for their susceptibility to oviposition by the root flies Delia radicum (L.) and D. floralis (Fallen) (Diptera, Anthomyiidae). For both species the preference among the genotypes corresponded to the susceptibility of the genotypes in the field. The preference ranking in response to surrogate leaves treated with methanolic surface extracts of the four genotypes was identical to the preference among potted plants, demonstrating that chemical factors on the leaf surface mediate host preference for oviposition in these species.For both species of fly, glucosinolates are major oviposition stimulants and for D. radicum an additional, nonglucosinolate oviposition stimulant, presently called CIF, is known. We describe a procedure for chromatographic separation of glucosinolates from CIF in leaf surface extracts. In oviposition-choice assays with D. radicum, the CIF-fractions of the two swede genotypes applied to surrogate leaves received a 1.8 and 4.6 times higher proportion of eggs than the respective glucosinolate-fractions, confirming the major importance of CIF as an oviposition stimulant. The genotype of swede that was preferred by both fly species in tests with plants and methanolic leaf surface extracts, also stimulated oviposition more in tests with the glucosinolate-fractions or the CIF-fractions derived from the surface extracts, respectively. Thus, glucosinolates and CIF together account for the observed preference among the genotypes and may also be responsible for their susceptibility under field conditions. In the two kale genotypes the preference for plants or surface extracts differed from the preference among the corresponding glucosinolate- and CIF-fractions, indicating that additional, as yet unknown chemical factors may also be involved.For both groups of stimulants tarsal chemoreceptors allow electrophysiological monitoring of glucosinolate- and CIF-activity in fractionated surface extracts. For D. radicum the chemosensory activity of both glucosinolate- and CIF-fractions corresponded to the respective behavioural activity in the oviposition preference tests, suggesting that preference for oviposition among genotypes can be predicted from the electrophysiological activity of their fractions. The chemosensory response of D. floralis, in particular to the CIF-fractions, was less pronounced than the response of D. radicum, indicating interspecific differences in the perception of the major oviposition stimulants. We discuss the potential application of electrophysiological techniques in support of other screening methods used in breeding for root fly resistance in Brassica crops.     

Self-contained, tetracycline-regulated retroviral vector system for gene delivery to mammalian cells.

Retroviral vectors that contain the tetracycline-inducible (Tet) system were developed. The two components of the Tet system were organized within the vectors in a manner that stringently maintains tetracycline-dependent regulation. Regulated expression of an indicator gene inserted into the retroviral vectors was examined in several different cell types. In infected NIH 3T3 cells, levels of induction in the absence of tetracycline were observed to be as much as 336-fold higher than levels in the presence of tetracycline, which were extremely low. Tetracycline-dependent regulation was observed in all other transduced cell types and ranged from 24- to 127-fold. The generation of retroviral vectors containing regulatory elements that allow for the regulated expression of heterologous genes and that have the ability to infect virtually all dividing target cells should greatly facilitate the biochemical and genetic examination of a broad range of genes. Moreover, these inducible retroviral vectors should prove useful in gene therapy applications.     

Effects of population subdivision and catastrophes on the persistence of a land snail metapopulation

We modeled the dynamics of a metapopulation of the land snail Arianta arbustorum in north-eastern Switzerland to investigate the effect of population subdivision on the persistence of a land snail metapopulation and to analyze the interaction between spatial factors, population subdivision, and catastrophes. We developed a spatially structured, stochastic, age-structured metapopulation model with field data from previous studies on the metapopulation in Switzerland and experimental and meteorological data. The model incorporated distance-dependent dispersal through stream banks, correlated environmental fluctuations, and catastrophes resulting from heavy rains. The results point to various complex interactions among factors involved in metapopulation dynamics and suggest that in some cases population subdivision may act to decrease threats from environmental fluctuations and catastrophes.