Myocardial S-adenosylhomocysteine hydrolase is important for adenosine production during normoxia

(1) The coronary vasodilator adenosine can be formed in the heart by breakdown of AMP or S-adenosylhomocysteine (SAdoHcy). The purpose of this study was to get insight into the relative importance of these routes of adenosine formation in both the normoxic and the ischemic heart. (2) A novel HPLC method was used to determine myocardial adenosine and SAdoHcy. Accumulation of SAdoHcy was induced in isolated rat hearts by perfusion with L-homocysteine thiolactone or L-homocysteine. The release of adenosine, inosine, hypoxanthine, xanthine and uric acid was determined. Additional in vitro experiments were performed to determine the kinteic parameters of S-adenosylhomocysteine hydrolase. (3) During normoxia the thiolactone caused a concentration-dependent increase in SAdoHcy. At 2000 μM of the thiolactone an SAdoHcy accumulation of 0.49 nmol/min per g wet weight was found during normoxia. L-Homocysteine (200 μM) caused an increased of 0.37 and 4.17 nmol SAdony/soc per g wet weight during normaxia and ischemia, respectively. (4) The adenosine concentration in ischemic hearts was significantly lower when homocysteine was infused (6.2 vs. 115 nmol/g; P < 0.05). Purine release was increased 4-fold during ischemia. (5) The K_m for hydrolysis of SAdoHcy was about 12 μM. At in vitro conditions favoring near-maximal SAdoHcy synthesis (72 μM adenosine, 1.8 mM homocysteine), the synthesis rate in homogenates was 10 nmol/min per g wet weight. (6) From the combined in vitro and perfusion studies, we comclude that S-adenosylhomocysteine hydrolase can contribute significantly to adenosine production in normoxic rat heart, but not during ischemia.     

Saccharin induces morphological changes and enhances prolactin production in GH4C1 cells

Summary The artificial sweetener saccharin inhibits binding of epidermal growth factor (EGF) to cultured rat pituitary tumor cells (GH₄C₁ cells). Saccharin also causes morphological alterations in these cells, resulting in pronounced elongation, stretching, and firmer attachment of cells to the culture dishes. These alterations in cell shape are similar to those observed after treatment of GH₄C₁ cells with EGF and with thyrotropin-releasing hormone (TRH), both of which enhance prolactin (PRL) production in these cells. After assaying for PRL in saccharin-treated cultures, it was observed that this sweetener is also capable of stimulating PRL production two-to sixfold in a dose-dependent manner. Enhancement of PRL production can be observed at 0.5 mM saccharin, yet this is 10 times less than the saccharin concentration required to alter cell shape. These effects of saccharin on cell morphology and on PRL production are reversible in GH₄C₁ cell cultures. When added to cultures along with maximal concentrations of EGF or TRH, the effects of saccharin on PRL production are additive, suggesting that the actions of saccharin are mediated by a somewhat different pathway from that of the peptide hormones. Pulse labeling studies indicate that the enhancement of PRL production is highly specific inasmuch as saccharin was found to decrease the overall rate of protein synthesis in these cells. Saccharin also causes a decrease in the rate of DNA synthesis under these treatment conditions. Mitomycin C, which similarly inhibited DNA synthesis, had no effect on cell morphology or PRL production. This investigation was supported by a Faculty Research Grant from Wheaton College     

Foraminifera and meiofauna on an intertidal mudflat,Cornwall, England: Populations; respiration and secondary production; and energy budget

A rich and varied meiofauna inhabits a Cornish mudflat near the mouth of the Tamar River in southwestern England. Population densities range from 117 to 943 individuals · g^–1 (wet) sediment (1.4–11.4 × 10⁶ individuals · m^–2), with foraminifera, harpacticoid copepods and nematodes appearing in nearly equal numbers and comprising most of the meiofauna. Seasonally, meiofaunal numbers rise and fall with solar radiation and vary inversely with river discharge. Two species, the atestate allogromiid A and the calcareous Haynesina germanica (Ehrenberg), far outnumber other foraminifera; their population densities and growth rates reach maxima in spring and summer.Monthly rates of sediment respiration are locally variable, but clearly increase from winter (4.13 ml O₂ · m^–2 · h^–1 in December) to spring (38.87 ml O₂ · m^–2 · h^–1 in April). Experiments and calculations ascribe approximately 30% of this total to the meiofauna (including microfauna and microflora), 50% to bacteria and less than 20% to chemical oxidation. A tentative energy budget for the mudflat suggests that secondary production by meiofauna is small as compared with coastal environments elsewhere, and that meiofaunal production (426 Kcal · m^–2 · y^–1) is nearly twice meiofaunal respiration (252 Kcal · m^–2 · yr^–1).     

Biomass and production of Argyrodiaptomus furcatus,a tropical calanoid copepod in Broa Reservoir,southern Brazil

The biomass and the production of Argyrodiaptomus furcatus (Sars), the most abundant copepod in Broa Reservoir (São Carlos, São Paulo State), were estimated, determining in the laboratory the development time and the quantity of organic carbon and establishing the relationship between these two parameters. The daily production was calculated from P = B(1- e^gt) and the annual production was obtained by integrating daily production against time. The maximum production of Argyrodiaptomus furcatus in the reservoir depends on the region considered and on the period of the year. The maximum production was 45.15 mg C m^–3d^–1 in March, 1976 at station II, region of macrophytes and 6.74 mg C m^–3d^–1 at station IV, near the dam. The mean production for the year is 6.26 mg C m^–3d^–1 at station II and 1.43 mg C m^–3d^–1 at station IV.     

Production of microsporidia pathogenic to the Colorado potato beetle (Leptinotarsa decemlineata) in alternate hosts

The microsporida Nosema gastroideae and N. equestris, which are highly pathogenic for Leptinotarsa, have been successfully produced in some other chrysomelid species, Gastrophysa polygoni and G. viridula. As the principal target host, Leptinotarsa is very susceptible to these pathogens, and death occurs before massive sporulation by the microsporidia. By contrast, the infected larvae of G. polygoni or G. viridula are able to develop until the adult stage when most of the tissues become filled with spores. In addition, the larvae and adults of these species can be reared in the laboratory on Polygonum aviculare and Rumex obtusifolius. These plants have longer vegetative periods and are better sources of food than potato leaves. In both species of Gastrophysa the yields of spores related to unit weight were about five times higher than in Leptinotarsa. In the adults of G. viridula there was up to 4.8 × 10⁶ spores mg^?1 body weight of N. gastroideae, or 9.1 × 10⁶ spores mg^?1 of N. equestris. The higher content of microsporidian spores facilitates their purification and isolation.     

Chronobiological aspects of the host-parasite relationships between Biomphalaria glabrata and Schistosoma mansoni: cercarial production and infectivity, and growth kinetics of the host

In the Biomphalaria glabrata-Schistosoma mansoni association, variations in cercarial production, in cercarial infectivity, and in the growth of infected snails are rhythmic. These chronobiological aspects are correlated with the dynamics of the intramolluscan larval stages of the parasite during the course of the infection. Rhythmic variations in the growth kinetics of infected snails are interpreted in terms of host-parasite metabolic exchanges.     

Direct analysis of growth factor requirements for isolated human fetal hepatocytes

Summary Hepatocytes were isolated from human fetal liver in order to analyze the direct effects of growth factors and hormones on human hepatocyte proliferation and function. Mechanical fragmentation and then dissociation of fetal liver tissue with a collagenase/dispase mixture resulted in high yield and viability of hepatocytes. Hepatocytes were selected in arginine-free, ornithine-supplemented medium and defined by morphology, albumin production and ornithine uptake into cellular protein. A screen of over twenty growth factors, hormones, mitogenic agents and crude organ and cell extracts for effect on the stimulation of hepatocyte growth revealed that EGF, insulin, dexamethasone, and factors concentrated in bovine neural extract and hepatoma cell-conditioned medium supported attachment, maintenance and growth of hepatocytes on a collagen-coated substratum. The population of cells selected and defined as differentiated hepatocytes had a proliferative potential of about 4 cumulative population doublings. EGF and insulin synergistically stimulated DNA synthesis in the absence of other hormones and growth factors. Although neural extracts enhanced hepatocyte number, no effect on DNA synthesis of neural extracts or purified heparin-binding growth factors from neural extracts could be demonstrated in the absence or presence of defined hormones, hepatoma-conditioned medium or serum. Hepatoma cell-conditioned medium had the largest impact on both hepatocyte cell number and DNA synthesis under all conditions. Dialyzed serum protein (1 mg/ml) at 10 times higher protein concentration had a similar effect to hepatoma cell-conditioned medium (100 μg/ml). The results suggest that hepatoma cell conditioned medium may be a concentrated and less complicated source than serum for purification and characterization of additional normal hepatocyte growth factors. This work was supported by NIH grant DK35310. Editor’s statement Many investigators have struggled with the special problems associated with culture of differentiated hepatocytes. In this paper attention is given to the specific growth factor requirements for fetal human hepatocytes. The observation that factors from hepatoma conditioned medium or neural extracts enhanced the growth of the cells may indicate that additional growth factors are to be identified that are important in the survival and proliferation of hepatocytes, and may also indicate that the malignant transformation of these cells may involve the production of autocrine growth stimulators.     

Disinhibition of oocyte growth in adult,virgin Periplaneta americana by corpus allatum denervation: Age dependency and relatedness to mating

Unilateral section of the nervi corporis allati I (NCA-1) of isolated, starved, adult, virgin Periplaneta americana disinhibited oocyte growth during a specific period following their adult emergence. The effect required that the corpus allatum (CA) be free of NCA-1 innervation for 4 days beyond the time the females were 7–8 days old. The onset of this sensitive period corresponds to when most isolated, starved virgins become sexually receptive. The results suggest that NCA-1 inhibition of CA activity, initiated about 7 days, is relieved by mating. When done on sexually receptive, starved virgins, unilateral NCA-1 section was as effective as insemination for stimulating growth and chorionation of the first generation of oocytes. Neural inhibition of juvenile hormone (JH) secretion by the CA may also explain diminished production of oocytes by isolated, fed virgins, for during 30 days following unilateral NCA-1 section they produced 2.6 to 5 times more oothecae than did controls with a single CA removed or after the sham operation. The number of oothecae deposited by fed virgins was similarly increased after bilateral NCA-1 section, but to a lesser extent than when the operation was done on fed, inseminated females of the same age. Specificity of the response of the CA to denervation was substantiated by experiments in which the CA were extirpated and reimplanted, by topically applying C₁₆JH, and by experiments in which the nervus corporis cardiaci 1 and 2 on the right or left side were severed.