The ecology of the yeast flora in necroticOpuntia cacti and of associatedDrosophila in Australia

A survey was made of the yeast communities isolated from necrotic tissue of 4 species of prickly-pear cacti (Opuntia stricta, O. tomentosa, O. monacantha, andO. streptacantha) which have colonized in Australia. Yeast communities were sampled from a number of localities and at different times. Cactus specific yeasts accounted for 80% of the total isolates, and the 3 most common species contributed 63% of the total. Comparisons of the species compositions of the yeast communities indicated that the differences among communities were greater betweenOpuntia species than between different localities within a single cactus species, and also that differences between years were greater than average differences between localities within years. Multivariate statistical tests of association between yeast community and physical features of rots indicated that temperature, pH, and age of rot all exerted some influence on the structure of the yeast community. Similar analyses involvingDrosophila species inhabiting these cactus rots suggested the existence of complex associations betweenDrosophila community, yeast community, and physical and chemical attributes of the cactus necroses.     

Studies on metacercarial excystment in Himasthla leptosoma (Trematoda: Echinostomatidae) and newly emerged metacercariae

Metacercarial cysts of Himasthla leptosoma were subjected to a solution containing bile salts, trypsin and l cysteine at 41°C. The treatment induced intense metacercarial activity and after 20 min metacercariae burst through the cyst walls and emerged. Electron microscopy demonstrated that organisms burst through a small area of cyst wall which was devoid of a layer of lamellae present elsewhere towards the innermost surface. The appearance of ruptured cyst walls indicated that they had been softened by the excystment medium. Newly emerged metacercariae possessed a reniform collar of 29 cephalic spines and these were sometimes withdrawn into pits, presumably by action of a muscle complex in the head region. Sensory papillae were distributed in a bilaterally symmetrical arrangement around the anterior sucker and none were visible on the surface of the ventral sucker. Tegumental spines were found only from a point some distance behind the head collar to the region of the ventral sucker. The most anterior spines were simple and peg-like and they quickly merged posteriorly into more complex palmate forms.     

The occurrence of species of semi-aquatic Enchytraeidae (Oligochaeta) in Ireland

The Enchytraeidae are essentially terrestrial oligochaetes but many species have marked aquatic tendencies. Over two thirds of recorded Irish species were found in soils which were submerged or frequently flooded and 35% showed a distinct preference for these conditions. Relatively few species were living in soils subject to drought. Red blood was present in 28 species, all but one from soils with more than 55% water. Cognettia sphagnetorum and C. glandulosa developed red blood in very wet conditions. In a survey of Irish wetlands, samples were taken from bog, heath, marsh, fen, margins of lakes and rivers, and saltmarsh. The influence of various environmental parameters was determined using ordination techniques. Magnesium and pH were found to be the most important factors. A high level of magnesium distinguished coastal sites and pH 5.2 separated two clusters representing acid peat and marsh-fen-aquatic sites. Groups of indicator species characterized each of the three clusters. The ecological distribution of the indicator species is described, and their usefulness in classifying enchytraeid communities is discussed.     

Possible genotoxicity of melanin synthesis intermediates: tyrosinase reaction products interact with DNA in vitro

Summary The actual cellular target of the cytotoxic intermediates of melanin synthesis is not yet known. In the present paper it is shown that eukaryotic DNA binds in vitro to soluble reaction products of tyrosinase (EC 1.14.18.1) and is physically modified, as ascertained by the following criteria: (a) buoyant density in cesium chloride density gradients; (b) polyacrylamide gel electrophoresis: (c) deoxyribonuclease (EC 3.1.4.5) test; (d) electron microscopy. The results reported here support the view that DNA itself may be a target for the cytotoxic intermediates of melanin synthesis.     

Alteration of the acyl chain composition of free fatty acids,acyl coenzyme A and other liPids by dietary polyunsaturated fats

Dietary alterations were used to demonstrate selective handling of fatty acids during their redistributionin vivo. Differences in the mol Per cent of individual acyl chains in the non-esterified fatty acid, acyl-coenzyme A and PhosPholiPid fractions reflected a result of relative Precursor abundance combined with enzymic selectivities. Selective distributions were observed in the utilization of individual acyl chains between 16:0 and 18:0, 18:1 and 18:2, and among 20:3, 20:4 and 20:5, 22:6 by ligase(s), hydrolase(s) and acyl-transferases. The variations in the mol Per cent of linoleate Present in the acyl-coenzyme A fraction of liver relative to that in the non-esterified fatty acids suggested anin vivo regulation of the level of linoleoyl-coenzyme A that influenced the synthesis of both arachidonoyl-coenzyme A and lipids. The greater abundance of eicosaPentaenoic acid in the free fatty acid fraction relative to that in the acyl-coenzyme A fraction may increase the ability of dietary 20: 5n-3 to be an effective inhibitor of the synthesis of Prostaglandins derived from 20:4n-6.     

Human bronchial epithelial cells with integrated SV40 virus T antigen genes retain the ability to undergo squamous differentiation

Human bronchial epithelial cells transformed by either DNA virus infection (SV40 or Adenovirus 12-SV40 hybrid virus) or transfection with the SV40 large T antigen gene were studied for their ability to undergo squamous differentiation when exposed to 12-O-tetradecanoylphorbol-13 acetate (TPA), transforming growth factor-beta 1 (TGF-beta 1), or fetal bovine serum (FBS), agents that induce the squamous differentiation of normal human bronchial epithelial cells. Squamous differentiation occurred in all ten T-antigen-positive cell cultures when they were exposed to either FBS or TGF-beta 1, but none differentiated when exposed to TPA. From one cell line, designated BEAS-2B, two subclones were isolated, one of which was induced to undergo squamous differentiation by FBS, and a second that failed to undergo squamous differentiation and was mitogenically stimulated when exposed to serum. These phenotypically different subclones provide a new in vitro cellular system for delineating the mechanism(s) of human bronchial epithelial cell squamous differentiation in response to FBS or TGF-beta 1.     

Liposome-entrapped tyrosinase: a tool to investigate the regulation of the Raper-Mason pathway

The effect of the entrapment of mushroom tyrosinase (EC 1.14.18.1) within liposomes on the enzyme activity and Km vs. L-3,4-dihydroxyphenylalanine is reported in the present work; the effect of cholesterol insertion within liposome membranes on the enzyme activity has also been studied. The oxidation rates of various monophenols and diphenols by free and liposome-integrated mushroom tyrosinase were measured and the oxidation latencies vs. different substrates investigated. The different substrates are apparently oxidized according to the properties of the substituents as electron donors or acceptors; the Km values vs. L-3,4-dihydroxyphenylalanine calculated on measuring O2 consumption are higher than those calculated on measuring the dopachrome production rates. It is interesting that natural substrates of tyrosinase are oxidized according to a negative catalysis by the liposome-entrapped enzyme; this point is discussed in relation to the well known cytotoxicity of some intermediates of the Raper-Mason pathway.     

Early developmental exposure to benzodiazepine ligands alters brain levels of thiobarbituric acid-reactive products in young adult rats

Levels of thiobarbituric acid (TBA)-reactive material were measured in brain regions of 3–4 monthold rats following prenatal exposure to several benzodiazepine (BDZ) receptor ligands over gestational days 14–20. Prenatal exposure to diazepam (DZ) at 1.0 mg/kg/day markedly elevated levels of brain TBA-reactive material while exposure to a higher dose (2.5 mg/kg) induced a significant increase only in the hippocampus. Early exposure to the central-type BDZ agonist clonazepam as well as to the central-type antagonist Ro 15-1788 also increased brain levels of TBA-reactive material. Concurrent exposure to the higher dose of DZ partially attenuated the effect of Ro 15-1788. Prenatal exposure to the peripheral-type BDZ ligand PK11195 produced a profound increase in TBA-reactive products in all regions, and concurrent DZ exposure did not attentuate this effect, except in the basal ganglia. Measurement of TBA-reactive material.from birth to 3 months indicated that the effect of prenatal exposure to DZ was not apparent until after 8 weeks of age. Acute in vitro exposure of adult and fetal tissue to DZ had no effect on TBA-reactive material. The results suggest an interference in the organization of cellular metabolism in the brain by developmental exposure to BDZ ligands.     

Identification of developmental toxicants using the Frog Embryo Teratogenesis Assay-Xenopus (FETAX)

Because growth and development are processes sensitive to the action of many chemicals, bioassays that screen for developmental toxicants may be more indicative of chronic effects than acute toxicity assays. FETAX is a 96 h whole embryo static renewal test employing the embryos of the frog Xenopus laevis. Endpoints are mortality, malformation and growth. Because of the frog's fecundity, its extensive use in basic research and the ability to obtain embryos year-round, it is an ideal organism to use in screening for developmental toxicants. By validating using known mammalian teratogens and the use of rat liver microsomes to stimulate mammalian metabolism, we have extended the use of the system for the prescreening of human developmental toxicants. In past validation work, we have correctly identified the teratogenicity of 15 to 17 compounds used in validation for a predictive accuracy of approximately 88%. In the present study, the ability of FETAX to detect developmental toxicants in groundwater samples taken from an industrial waste dump was evaluated. FETAX showed that it was sensitive enough to detect developmental toxicants in samples without prior concentration. In some samples, less than half the LC50 concentration was required to cause significant malformation. In some cases, a dose-response curve was not obtainable but the test results nonetheless indicated some developmental toxicity. The results of this study indicate that it is necessary to routinely screen for developmental toxicants when establishing water quality criteria for the preservation of species and for human health.     

Effects of monensin on ATP levels and cell functions in rat liver and lung in vitro

Summary Effects of the proton-alkali cation-exchanging ionophore, monensin, on aspects of cellular metabolism and ionic exchanges have been studied in rat tissues in vitro. Incubation of liver slices at 38°C with 0.1 m monensin induced timedependent vesiculation, initially in the Golgi region, reduction of ATP content and of protein synthesis. At 1 m, monensin also reduced net, active movements of K⁺, Na⁺, Cl^– and water in liver slices and inhibited state 3 respiration in isolated mitochondria. The respiratory inhibitor, amytal, similarly reduced ATP content and protein synthesis at concentrations lower than those inhibiting ion transport in slices. Low concentrations of monensin (0.1–1.0 m) had similar effects on ATP and ion transport in slices of adult lung. By contrast, late-fetal liver and lung were much less sensitive to monensin; in these tissues, glycolysis sustained substantial levels of ATP. Monensin also induced vesiculation of the Golgi apparatus in fetal lung cells. It is concluded that by lowering ATP levels, monensin can markedly alter various metabolic activities in those cells which depend primarily on oxidative phosphorylation for their metabolic energy.