Effects of temperature on virus-host relationships and on activity of the noninclusion virus of citrus red mites, Panonychus citri

The nonoccluded virus of citrus red mite retained full infectivity when exposed to 40.5°C for 24 hr within intact mite bodies but was inactivated at 46°C for 6 hr and 60°C for 1 hr. Exposures to 38°C for 28 days failed to destroy infectivity. Virus inoculated mites exposed to different temperature regimens had shortened periods of lethal infection at high temperatures and greatly lengthened periods of lethal infection at cool temperatures suggesting that failures in mite control by virus in the early spring and late fall may be due to previously unrecognized temperature relationships.     

Protection of Measles Virus by Sulfate Ions Against Thermal Inactivation.

Rapp, Fred (Baylor University College of Medicine, Houston, Tex.), Janet S. Butel, and Craig Wallis. Protection of measles virus by sulfate ions against thermal inactivation. J. Bacteriol. 90:132-135. 1965.-The infectivity of measles virus in water is rapidly destroyed at temperatures of 37 C and above. More than 50% of the infectivity is lost after 1 hr at 25 C, and almost 90% loss of infectivity occurs within 24 hr at 4 C. Magnesium chloride enhances the inactivation of the virus at all temperatures tested. Addition of either magnesium or sodium sulfate protects the virus against thermal inactivation. The stabilizing effect is demonstrable at temperatures ranging from 4 to 56 C, but is especially pronounced through 45 C. Prolonged storage (up to 6 weeks) of the virulent virus at 4 C in 1 m magnesium sulfate permits retention of substantial infectivity, whereas storage at 4 C in either water or 1 m magnesium chloride results in a loss of infectivity approximating 99% after 2 weeks. Magnesium chloride also enhances inactivation of the attenuated vaccine strain of measles virus. The attenuated virus, however, is strongly protected by magnesium sulfate against thermal inactivation, and retention of infectivity for long periods of time at 4 C seems feasible when the virus is kept in 1 m magnesium sulfate.     

Environmental factors controlling the time of flower-opening in Pharbitis nil

Pharbitis nil, strain Violet, subjected to various photoperiods(24-hr cycle at 24?C) bloomed about 10 hr after light-off whenthe light period was 10 hr or longer, and about 20 hr afterlight-on when the light period was shorter. The higher the temperature(20–30?C) during the dark period, the later the time offlower-opening, with the temperature during the last half ofthe dark period having a stronger effect than that during thefirst half. In continuous dark or light, flower buds of Pharbitis openedabout every 24 hr at all temperatures tested between 20 and28?C, which suggests the participation of a circadian rhythmin determining the time of flower-opening. A light pulse given6–12 or 28–36 hr after the onset of the dark periodgreatly advanced the phase of this rhythm (8–10 hr). Phasedelay of this rhythm could not be obtained by light pulses givenat any time. (Received September 29, 1979; )     

In Vitro Chick Embryo Cell Response to Strain MC29 Avian Leukosis Virus

Strain MC29 avian leukosis (myelocytomatosis) virus induced infection, elaboration of virus, and morphological alteration in chick embryo cells in vitro. Virus liberation began within 18 hr, morphological change was detectable at about 40 hr, and the cultures could be completely altered within 80 hr after infection. Altered cells were about half the volume and grew at approximately twice the rate of uninfected elements. The output of virus estimated by electron microscopy was about 140 particles per cell per hr. Deoxyribonucleic acid remained constant, but ribonucleic acid increased in both infected and control cells in adjustment to culture environment. The rates of uptake and incorporation of ³H-uridine and the incorporation of ³H-thymidine increased in the infected cells with onset of morphological change but were unaffected by processes of infection and virus elaboration per se. Incorporation of a ¹⁴C-amino acid mixture was slightly greater in the infected than in control cells. The speed of continuity of infection and massive morphological alteration constitute a unique response to avian tumor viruses, and the system gives promise of singular value for detailed studies of the processes of infection and morphological change.     

Electron Microscopic Study of the Morphogenesis of Vesicular Stomatitis Virus

Except for the rate, vesicular stomatitis virus (VSV) grows as well at 25 C as at 37 C in primary chick embryo fibroblast cells and in a pig kidney cell line [PK(H13)]. Maximal yields were reached at about 28 hr at 25 C and 10 hr at 37 C in these cells. Morphogenesis, as observed by electron microscopy, was similar at the two temperatures. The main feature was accumulation of virus in intracytoplasmic vacuoles. Mode of release of VSV has been controversial; both budding (as displayed by myxoviruses) and maturation at membranes of cytoplasmic vacuoles (as with arboviruses) have been claimed. Our observations support the latter view, and the apparent dichotomy in interpretation is discussed.     

Effects of acute heat exposure on respiratory metabolism and blood glucose in freshwater fishes, Prochilodus scrofa (curimbatá) and Cyprinus carpio (carp) acclimatized to tropical winter

Routine oxygen consumption and blood glucose were determined from freshwater fishes, Prochilodus scrofa and Cyprinus carpio, exposed at high temperatures for 1 hr. Prochilodus scrofa had a significantly higher rate of oxygen consumption at 30 degrees C than at 25 degrees C, and carp higher at 25 degrees C than at 30 degrees C. Blood glucose was significantly higher for Cyprinus carpio than for Prochilodus scrofa at 25 and 30 degrees C; however, after exposure to these temperatures for 1 hr blood glucose did not change significantly for both species. The results suggest that these interspecific variations may be linked to the differences between native and foreign fishes and their way of life.     

Reduction of ribonuclease by glutathione at elevated temperatures: the molecular mechanism

1. The mechanism of the reaction between ribonuclease and GSH at elevated temperatures has been studied by using N-(4-dimethylamino-3,5-dinitrophenyl)-maleimide to label the reduced ribonuclease. 2. After incubation for 2hr. at 35 degrees , enzymically active ribonuclease was recovered; at 50.8 degrees half of the initial ribonuclease was recovered as enzymically active ribonuclease and half as reduced labelled ribonuclease; at 55 degrees all of the initial ribonuclease was recovered in the labelled form. 3. It was inferred that the rate-limiting step was the reduction of the first disulphide bond in any one molecule. This was followed by rapid reduction of the other bonds in the same molecule.     

THE INFLUENCE OF TEMPERATURE AND DAYLENGTH ON SPOTTED WILT VIRUS DISEASE OF TOMATO

The responses of young tomato plants to infection with tomato spotted wilt virus were studied at constant temperatures. The plants were grown in nutrient solutions and given three daylength treatments.
The length of the incubation period was positively correlated with temperature and a regression equation for this relationship is given. The calculated temperature at which this period becomes infinite is 6.7°C. The temperature coefficient ( Q ₁₀) varied from 1.7 to 2.9.
Dry-weight determinations were made of stems, leaves and roots. In healthy plants the leaf/stem ratio generally fell with rise of temperature and the shoot/root ratio rose. Infection changed these ratios, which is interpreted as indicating that it interfered with supplies of carbohydrate to the growing regions.
Systemic symptoms are described for plants raised at 11.4°, 18.3°, 23.9°, and 29.4°C.
Changes in daylength did not affect the systemic symptoms or incubation period. With a 9 hr. day at 29.4°C., the virus content of sap was significantly lower than with 12 or 15 hr., and with a 12 hr. day was significantly higher than with a 15 hr. day.     

Interferon production in chick-embryo cells. The effect of puromycin and p-fluorophenylalanine

1. When chick-embryo cells were treated with ultraviolet-inactivated influenza virus (Melbourne strain), interferon was produced after a lag period of about 10hr. 2. The addition of small amounts of either puromycin or p-fluorophenylalanine immediately after the virus inhibited the subsequent production of interferon. Both inhibitors primarily affected protein synthesis, and it is concluded that interferon production involves new protein synthesis. 3. Results obtained by the addition of either inhibitor for short periods during the lag phase demonstrated a requirement for protein synthesis during the second half of the lag phase. 4. Addition of puromycin during the course of interferon production caused almost immediate inhibition, but interferon formation became insusceptible to the action of p-fluorophenylalanine at about 26hr. after infection. Possible explanations of this effect are discussed.     

Development of diurnal rhythm in some metabolic parameters in foals

1. The development of diurnal rhythm activity of FDPA, AspAT and A1AT and in levels of cortisol, T3 and T4 was observed in the blood serum of six foals. 2. The studies began when a foal was 7 days old and were repeated every month until foals reached 1 year of age. Blood samples were taken every 4 hr for one day each month. 3. As a control group four barren mares were used, kept and examined in the same conditions. 4. In mature mares, diurnal rhythms in activity of A1AT (acrophase at 2200 hr), AspAt (2400 hr) and cortisol (0630 hr) but in T3 only in summer months (acrophase at 0100 hr) were observed. 5. During the first 6 months of foal life, significantly higher mean levels of FDPA, A1AT, T3 and T4 than in control mares were found. 6. The cortisol level in foals was half as much as that of mature mares throughout the year. 7. In foals the diurnal rhythm in A1AT activity occurred in the 5th month and in AspAt--in the 12th month (acrophase at 2400 hr), but in cortisol levels it was developed already in the second month of foal life (acrophase at 0830 hr).     

Turnover of coniferin in pine seedlings

Coniferin (coniferyl alcohol-β-d-glucoside) was not detected in pine seeds (Picea abies) but it accumulated in the stems and roots of pine seedlings. Pulse labeling experiments with l-phenylalanine-[U-¹⁴C] and 100-day-old pine seedlings in hydroponic solution showed a turnover of coniferin with a half life of about 60 hr. Pulse labeling with ¹⁴CO₂ and seedlings kept in soil gave a half life of about 120 hr for coniferin. The results indicate that coniferin could be an intermediate of lignin biosynthesis in pine seedlings.     

STUDIES ON THE EFFECT OF TEMPERATURE ON VIRUS MULTIPLICATION IN INOCULATED LEAVES

The rate at which the Rothamsted tobacco necrosis virus (RTNV) accumulates in inoculated French bean leaves increases with rising temperature to 22°C. and then decreases. Three days after inoculation, leaves at 22°C. contain 4000 times as much virus as at 10°C. and 1000 times as much as at 30°C. At all temperatures the rate of accumulation may depend on the balance between synthesis and inactivation of RTNV, but inactivation becomes increasingly important with rise of temperature above 22° C. and as the virus content of the leaves increases. Above 22°C. the rate of multiplication may increase but less rapidly than the rate of inactivation, and exposing inoculated leaves to ultra-violet radiation at various intervals after inoculation suggests that at 30°C. RTNV multiplies in and moves from the initially infected epidermal cells in slightly less than the 6 hr. needed at 22°C. Thirty hr. are needed at 10°C. Newly formed virus is rapidly inactivated at 30°C. Raising the ambient temperature also decreases the numbers of local lesions produced by RTNV, possibly by increasing the chances that the introduced virus particles will become inactivated. Increasing the virus content of the inoculum above the level giving one lesion per sq.cm. does not increase the subsequent virus content of inoculated leaves.
At temperatures of 30°C. and below, tomato aucuba mosaic virus produces necrotic lesions in leaves of tobacco and Nicotiana glutinosa whereas above 30°C. the lesions are chlorotic. In both hosts this virus multiplies more rapidly when the infected cells are killed.     

Rhythmic cortisol secretion in the equine: Analysis and physiological mechanisms

Abstract

Four Thoroughbred mares (no. 1–4) were maintained under constant temperature (24°C) and controlled light (L/D:12/12 with lights on at 06.00 hr) conditions. They were fed and watered ad libitum with fresh feed and water given at 09.00 hr. After a 45‐day pre‐conditioning period, blood samples were obtained by veinipuncture at 4‐hr intervals for 14 days to determine circadian and day‐to‐day variation. The horses exhibited a circadian rhythm with maximum values attained at about 12.00 hr, however, there are periods of days in which no rhythm is distinguishable. Ultradian rhythms with mean periods of 105 to 128 and 24 to 31 min are superimposed upon the circadian rhythm. The individual rhythms are quite variable from horse to horse and within the same horse. During periods of decline in plasma cortisol with metabolic half‐lives of approximately 70 min, secretion of cortisol was very low or had ceased. During periods of increasing plasma concentration, secretion was occurring at a faster rate than degradation. Rapid decreases in plasma concentration (metabolic half‐life of approximately 30 min) was accompanied by a rise in specific activity indicating cortisol with a high specific activity was entering the plasma pool from other storage pools.     

Different Administration Schedules of the Same Dose of 2,5-Hexanedione Influence the Development of Neuropathy and the Toxicokinetics

The same total dose (1.2 g/kg/week) of 2,5-hexanedione (2,5-HD) was administered subcutaneously at 100 mg/kg/12 hr, 200 mg/kg/24 hr, and 400 mg/kg/48 hr to three groups of Donryu rats. The peripheral neuropathy induced by 2,5-HD was confirmed by clinical observation every day, and neurophysiological measurements every 4 weeks. During the 15th week of this experiment, 2,5-HD concentrations in plasma 0.5 to 24 hours after injection were determined. It was found that the greater the dose of 2,5-HD per treatment injected, the earlier peripheral neuropathy developed. Toxicokinetic analysis showed that both the values of the area under the plasma concentration versus time curve and the half life of 2,5-HD were increased, but the excretion parameters (Ke) were decreased, in animals treated with 200 mg/kg/24 hr and 400 mg/kg/48 hr 2,5-HD.     

Photochlorophyllide (P650) turnover in dark-grown barley leaves

Laevulinate (LA) induced an increase in protochlorophyllide (P650) in dark-grown ageing barley leaves. The increase was due to a suppression of a P650 breakdown mechanism. The LA inhibition of P650 destruction allowed an estimate to be made of turnover of P650 in ageing etiolated leaves. The rate constant for P650 destruction in 8-day-old dark-grown leaves was 139 pmol/nmol/hr with a half life of 5 hr.     

Measles Virus Ribonucleic Acid and Protein Synthesis: Effects of 6-Azauridine and Cycloheximide on Viral Replication

Cycloheximide and 6-azauridine were employed to study the time course of measles virus protein and nucleic acid syntheses in AV3 cells. Synthesis of ribonucleic acid (RNA) essential for infectivity was first detected at 6 hr and increased concurrently with the formation of essential protein. Maximum levels of virus-specific RNA and protein were present by 18 hr, a time when only 5% of progeny virus was detected. Essential RNA and protein syntheses preceded the formation of infectious virus by at least 10 to 12 hr. The time course of RNA and protein syntheses essential for the formation of complement-fixing (CF) antigen and salt-dependent agglutinin (SDA) was also determined. RNA synthesis essential for the formation of SDA was first detected at 2 hr and was present maximally by 6 hr, whereas SDA-protein increased concurrently with the protein essential for infectivity. This suggested that the last protein essential for infectivity may be SDA. RNA synthesis essential for the formation of CF antigen was first detected at 4 hr, while CF-protein increased at 5 hr and preceded SDA-protein and protein essential for infectivity by approximately 3 hr. Reversal of inhibition of protein synthesis by cycloheximide indicated that early protein synthesis (1 to 3 hr) was required for the formation of infectious virus. The data suggest that the relatively long eclipse period observed with measles virus is related to a long maturation period rather than to late formation of early proteins, viral RNA, or structural proteins.     

Sequential Formation of Vaccinia Virus Proteins and Viral Deoxyribonucleic Acid Replication

In vaccinia virus-infected cell cultures, cellular protein synthesis was inhibited 50% at 2 hr postinfection (PI) and 80 to 90% by 4 hr PI. Input virus was responsible for this inhibition. Five early proteins, coded for by the viral genome, could be detected at 2 to 3 hr PI. Normally, their synthesis did not continue beyond 6 hr PI, at which time synthesis of a different set of proteins began. When DNA replication was blocked, synthesis of these early proteins continued until 9 to 12 hr PI. The bulk of the proteins which were incorporated into mature virus were synthesized at 8 hr PI and thereafter. The time of their formation was close to the time at which virus maturation occurred. However, 15% of the protein found in mature virus was synthesized early in the infectious cycle. The quantity of “early viral protein” which was not incorporated into mature virus was almost as large as the quantity of viral protein which did appear in mature virus. The “early” and “late” proteins could be shown to have separate and distinct immunological properties. The role of this large quantity of “early” protein is discussed.     

Quantitative analysis of cell proliferation and differentiation in the cortex of the postnatal mouse cerebellum

The generation cycle of germinative cells (external matrix cells) in the external granular layer of the cerebellar cortex of the 10-to 11-day-old mouse was studied by radioautography following repeated injections of H³-thymidine. The generation time is 19 hr, presynthetic time 8.5 hr, DNA-synthetic time 8 hr, postsynthetic time 2 hr, and mitotic time 0.5 hr. These proliferating cells occupy the outer half of the external granular layer and make up the external matrix layer. Neuroblasts are differentiated from the external matrix cell, migrate out from the layer and accumulate in the inner half of the external granular layer to form the external mantle layer. The transit time of the neuroblasts in the external mantle layer is 28 hr. Thereafter, they migrate farther into the molecular layer and the internal granular layer. By means of long-term cumulative labeling, the rate of daily production of neuroblasts from the external matrix cell is studied in quantitative terms. It becomes clear that the entire population of the inner granule neurons arises postnatally in the external granular layer between 1 and 18 days of age and that 95% of them is produced between postnatal days 4 and 15. Finally, the fate of the cells in the external granular layer at its terminal stage was studied by marking the cells with H³-thymidine during 15–16 days of life and following their subsequent migration and developmental changes up to 21 days of life. Comparison of radioautographs taken before and after the migration disclosed that the external matrix cells give rise to a small number of neuroglia cells. This finding revealed their multipotential nature.     

Epizootiology of a nuclear polyhedrosis virus in European spruce sawfly (Gilpinia hercyniae): the rate of passage of infective virus through the gut of birds during cage tests.

The passage of a nuclear polyhedrosis virus (NPV) of the sawfly, Gilpinia hercyniae, through avian gut was studied during cage tests on Sturnus vulgaris (three individuals), Parus ater (one), Parus caerulus (five), and Parus major (one). Following brief infection feeds, polyhedral inclusion bodies of the virus could be detected in bird feces within 0.5 hr. Peak passage of polyhedra occurred in less than 1 hr and none were detected after 2.5 hr. The feces of all birds remained infective (in bioassay tests using first instar G. hercyniae larvae) to the end of the day of infection while those of nine birds remained infective to the next day and of six birds to the third day. One bird, P. major, was also infective on Days 4, 6, and 7. The infectivity of NPV in feces stored for 2 years at +3°C declined by half. Though the scale of their epizootiological contribution is unknown, the comparatively long retention and passage of infective virus suggests birds may be effective in short- and long-distance transport of baculoviruses.