The effect of incubation temperature on the maintenance of rat palatal mucosa in organ culture

The effect of incubation temperature on the behavior of neonatal rat palatal mucosa maintained in a chemically defined medium in organ culture for periods up to 7 days was investigated. Explant survival was optimal at 37 degrees C with increasing mortality at temperatures of 34 degrees C and 30 degrees C. There was a transient increase in the epithelial mitotic activity at all temperatures, but at all time intervals mitotic activity was greatest at 37 degrees C. While the mitotic activity at 37 degrees C after 5 hr in vitro was comparable with previously described in vivo values, it was subsequently increased, only returning to values approximating those at the start of the experiment at 6 days. At 30 degrees and 34 degrees C the epithelial mitotic activity increased more slowly than at 37 degrees C; then it followed a similar pattern with time and after 5 days in vitro had fallen to values approximating initial values. At the cut edges of the explants, the rate of epithelial migration and subsequent keratinization increased with increasing temperature. It is suggested that survival of neonatal rat palatal mucosa is optimal in this organ culture system when maintained at 37 degrees C.     

Cell proliferation in vivo and in vitro in visceral organs of the adult newt, Triturus cristatus carnifex

The mitotic and labelling incidence of intestine, liver, spleen and pancreas cells of Triturus cristatus carnifex adults kept at 15°C, 20°C, 25°C and 30°C were examined. Intestine mitotic and labelling incidences were highest at 25°C and lowest at 30°C. There was no significant difference between 15°C and 20°C. No such relationship could be shown for liver, spleen or pancreas, which had very much lower mitotic and labelling incidences. In culture, intestine mitotic and labelling incidences fell significantly within the first four hours, and maintained these low levels for the next five days. In contrast, liver mitotic and labelling incidences rose for 9–11 days, and then began to fall, while pancreas mitotic and labelling incidences reached peak values at day 5, and were kept in good condition for up to 14 days.     

The effects of temperature on development of the large narcissus fly (Merodon equestris)

Eggs, larvae, pupae and adults of the large narcissus fly (Merodon equestris) were reared at a series of constant temperatures between 9–24°C. Egg development required from 37 days at 9°C to 7 days at 21.5°C. The low-temperature threshold for development was 6.7°C. Larvae reared at 1424°C were fully-grown after 18 weeks, but it took much longer for such insects to pupate, and adult flies emerged only after about 45 weeks of development. Large narcissus flies enter diapause during the larval stage and overwinter as fully-fed larvae, forming pupae in the following spring. Post-winter pupation and pupal development took from 169 days at 10°C to 36 days at 21.5°C. Of this, pupal development required from 91 days at 10°C to 19 days at 21.5°C. The low-temperature threshold for post-winter pupation and pupal development was 7.1°C, and for pupal development alone, 7.2°C. Females maintained at or below 19°C laid few eggs, whereas some females kept at or above 21.5°C laid more than 100 eggs (mean 69 ± 36). Approximately 50% of females maintained at or above 21.5°C laid less than 10 eggs during their lifetime. The mean egg-laying time was 6 to 9 days. Although temperatures at or below 19°C inhibited mating, once a female had mated, such temperatures did not prevent oviposition.     

Effects of microwave exposure and temperature on survival of mice infected with Streptococcus pneumoniae

Female CD-1 mice were injected with an LD₅₀ dose of Streptococcus pneumoniae and then exposed to 2.45 GHz (CW) microwave radiation at an incident power density of 10 mW/cm² (SAR = 6.8 W/kg), 4 h/d for 5 d at ambient temperatures of 19 °C, 22 °C, 25 °C, 28 °C, 31 °C, 34 °C, 37 °C and 40 °C. Four groups of 25 animals were exposed at each temperature with an equal number of animals concurrently sham-exposed. Survival was observed for a 10-d period after infection. Survival of the sham-exposed animals increased as ambient temperature increased from 19 °C–34 °C. At ambient temperatures at or above 37 °C the heat induced in the body exceeded the thermoregulatory capacity of the animals and deaths from hyperthermia occurred. Survival of the microwave-exposed animals was significantly greater than the shams (～20%) at each ambient temperature below 34 °C. Based on an analysis of the data it appears that the hyperthermia induced by microwave exposure may be more effective in increasing survival in infected mice than hyperthermia produced by conventional methods (ie, high ambient temperature). Microwave radiation may be beneficial to infected animals at low and moderate ambient temperatures, but it is detrimental when combined with high ambient temperatures.     

Life history parameters of the cattle long‐nosed sucking louse,Linognathus vituli

Cattle sucking lice, Linognathus vituli (L.) (Phthiraptera: Linognathidae), were obtained from naturally infected cattle and maintained within ‘arenas’ affixed to the backs of cattle confined in controlled environment chambers maintained at a constant temperature of 15 °C. Temperatures measured within the arenas at an ambient temperature of 15 °C were constant at about 34 °C and only slightly above the temperature on nearby skin. The effect of temperature on egg development was determined using a gradient of temperatures between 25 °C and 41 °C. Eggs did not develop at temperatures of < 26 °C or > 39 °C. Survival of eggs was highest at temperatures of 30 °C and 35 °C. The earliest hatch was observed at 5 days post‐oviposition (at 33–35 °C). Development was extended to as long as 13 days at the lower temperatures. Kaplan–Meier survival probabilities were compared for lice kept at two densities in the arenas and showed there to be no effect of density on louse survival. Similarly, the mean number of eggs/louse/day over an 8‐day period was not influenced by louse density.     

Maintaining mosquito cell lines at high tempetatures: Effects on the replication of flaviviruses

Summary The upper thermal limit for maintenance of eleven mosquito cell lines was studied. Although most cell lines could be grown at 32°C to 34°C,Anopheles stephensi cell line could be maintained at 37°C. At higher temperatures initial growth rate was higher, but yield of cells after about a week of incubation was lower than at the standard temperature (28°C). Replication of several flaviviruses inAedes albopictus cell cultures adapted to 34.5°C was faster, and viral titers were higher than at 28°C.     

Increased salinity improves the thermotolerance of mesophilic nitrification

Nitrification is a well-studied and established process to treat ammonia in wastewater. Although thermophilic nitrification could avoid cooling costs for the treatment of warm wastewaters, applications above 40 °C remain a significant challenge. This study tested the effect of salinity on the thermotolerance of mesophilic nitrifying sludge (34 °C). In batch tests, 5 g NaCl L^?1 increased the activity of aerobic ammonia-oxidizing bacteria (AerAOB) by 20–21 % at 40 and 45 °C. For nitrite-oxidizing bacteria (NOB), the activity remained unaltered at 40 °C, yet decreased by 83 % at 45 °C. In a subsequent long-term continuous reactor test, temperature was increased from 34 to 40, 42.5, 45, 47.5 and 50 °C. The AerAOB activity showed 65 and 37 % higher immediate resilience in the salt reactor (7.5 g NaCl L^?1) for the first two temperature transitions and lost activity from 45 °C onwards. NOB activity, in contrast to the batch tests, was 37 and 21 % more resilient in the salt reactor for the first two transitions, while no difference was observed for the third temperature transition. The control reactor lost NOB activity at 47.5 °C, while the salt reactor only lost activity at 50 °C. Overall, this study demonstrates salt amendment as a tool for a more efficient temperature transition for mesophilic sludge (34 °C) and eventually higher nitrification temperatures.     

Selection of extraction solvent and temperature effect on stability of the algicidal agent prodigiosin

An organic solvent for extracting prodigiosin from culture broth was selected and a test to determine the long-term stability of prodigiosin was performed to develop prodigiosin as a biological control agent against Chattonella antiqua, a harmful alga that can cause red tides. Prodigiosin was extracted using nine solvents, and the extracts were analyzed by liquid chromatography-mass spectroscopy. Acetone was selected as the best organic solvent because of its high extraction efficiency and less processing time. Stability tests for prodigiosin were performed at various temperatures, and algicidal activity against C. antiqua was also tested. Ultimately, > 98% stability was sustained after 30 days at 4°C, whereas < 30% stability was maintained after 30 days at 37°C. Although prodigiosin was kept for 30 days in an optimum organic solvent, its stability was safely maintained and algicidal activity was sustained at 4°C. These results indicate that acetone is a very useful extraction and storage solvent for prodigiosin.     

Development ofDiatraea Saccharalis [Lep.: Pyralidae] at constant temperatures

At constant temperatures between 15.6 and 32°C the incubation time of eggs ofDiatraea saccharalis (F.) was reduced by each increase in temperature. At 34°C the time decreased. Highest (98.6%) and lowest (9.9%) egg hatch occurred at 26 and 34°C, respectively. Larvae completed development at temperatures ranging from 22 to 34°C; however, only 4.4% of the larvae pupated at 34°C. Duration of the larval stage at 30°C (♂=18.1 days; ♀=19.1 days) was ca. 14 days shorter than at 22°C. Maximum rate of development in the pupal stage occurred at 28°C (ca. 6.8 days), and a higher temperature increased developmental time and mortality. Adult longevity and egg production generally were reduced with increasing temperatures and egg production was highest at 24°C (729.8 eggs/ moth). As many as 7 larval stages occurred; but most larvae completed development in 5 stages, and none completed development in less than 5 stages. The female larval stage was ca. 1 day longer than that of males, and this difference occurred primarily in the 5th stage.     

The influence of temperature upon polysomes of spermatids of rat testes

Incorporation of [³H]phenylalanine into protein by a reconstituted lysate subcellular system (ribosomes plus high-speed supernatant) from rat spermatids was measured at 34°C after 5 minutes preincubation of one component at 0°C while the other component was incubated at temperatures from 30°C to 40°C. Preincubation at temperatures above 34°C inhibits the ribosomal activity but not the high-speed supernatant activity. The incubation of lysate above 34°C results from a dissociation of polysomes to monosomes. These results indicate that ribosomes are the most sensitive component to the increased temperature on protein synthesis in lysate cell free system by spermatids and that the inhibition of protein synthesis in spermatids above 34°C is at least partly explained by the breakdown of polysomes in these cells.     

Glutamine synthetase in cultured whole retinas from the embryonic chick: Enhancement of basal and induced activity by 4 °C storage

Storage of whole retinas from the embryonic chick for 24 h at 4 °C resulted in increased basal levels of glutamine synthetase (GS) during subsequent incubation at 37 °C in the absence of cortisol. GS levels in these retinas maintained initially at 4 °C (CS), in many cases, exceeded GS levels in cortisol-induced whole retinas incubated solely at 37 °C. The increase in basal GS activity is seen within 48 h of the transfer of the retinas from 4 to 37 °C. If cortisol (0.001 μg/ml = 2.8 nm or 0.01 μg/ml = 28 nm) is added during the last 24 h of culture to CS retinas subsequently transferred to 37 °C, levels of GS are attained that are higher than those in the corresponding retinas cultured continually at 37 °C. However, the activity ratios (GS specific activity in cortisol-treated retinas/GS specific activity in retinas not exposed to cortisol) are similar for CS retinas and those maintained at 37 °C throughout. Monolayers of retinal cells display similar basal and cortisol-induced levels of GS independent of treatment. Retinal monolayers maintained at 4 °C for 24 h and subsequently incubated at 37 °C do not exhibit increases in either basal or cortisol-induced levels of GS over those in monolayers maintained at 37 °C throughout. The CS-promoted increase in the basal and cortisol-induced GS activity of whole retinas is eliminated by enzymatic dispersion of the retina just prior to 37 °C culture of the cells as monolayers. Both basal and cortisol-induced GS levels in the latter monolayers resemble those in retinal cells kept as monolayers throughout.     

Effects of heat stress in the leaf mitotic cell cycle and chromosomes of four wine-producing grapevine varieties

Grapevine varieties respond differentially to heat stress (HS). HS ultimately reduces the photosynthesis and respiratory performance. However, the HS effects in the leaf nuclei and mitotic cells of grapevine are barely known. This work intends to evaluate the HS effects in the leaf mitotic cell cycle and chromosomes of four wine-producing varieties: Touriga Franca (TF), Touriga Nacional (TN), Rabigato, and Viosinho. In vitro plants with 11 months were used in a stepwise acclimation and recovery (SAR) experimental setup comprising different phases: heat acclimation period (3 h—32 °C), extreme HS (1 h—42 °C), and two recovery periods (3 h—32 °C and 24 h—25 °C), and compared to control plants (maintained in vitro at 25 °C). At the end of each SAR phase, leaves were collected, fixed, and used for cell suspensions and chromosome preparations. Normal and abnormal interphase and mitotic cells were observed, scored, and statistically analyzed in all varieties and treatments (control and SAR phases). Different types of chromosomal anomalies in all mitotic phases, treatments, and varieties were found. In all varieties, the percentage of dividing cells with anomalies (%DCA) after extreme HS increased relative to control. TF and Viosinho were considered the most tolerant to HS. TF showed a gradual MI reduction from heat acclimation to HS and the lowest %DCA after HS and 24 h of recovery. Only Viosinho reached the control values after the long recovery period. Extrapolating these data to the field, we hypothesize that during consecutive hot summer days, the grapevine plants will not have time or capacity to recover from the mitotic anomalies caused by high temperatures.     

Temperature-dependent antimutagenic activity of acrolein in Escherichia coli

The effect of temperature on the antimutagenic activity of acrolein was investigated using UV-irradiated E. coli B. When incubated at lower temperatures (30°C or 37°C), acrolein greatly reduced the mutation frequency in WP2 (wild-type strain), but no such effect was observed with WP2s and ZA159 (excision repair-deficient strains). The antimutagenic activity of acrolein increased when cells were incubated at higher temperatures (40°C or 42°C). Particularly in excision repair-deficient strains, the antimutagenic activity was observed only at higher temperatures. In heat shock response-deficient background, however, the antimutagenic activity was observed at 30°C even in the excision repair-deficient strains.     

Corpora lutea in submerged organ cultures at low incubation temperatures

Summary Corpora lutea in submerged organ cultures of mouse ovary retained viability when incubated at 24°C. At harvest, whole corpora lutea appeared uniformly viable, and mitotic figures were occasionally seen among the luteal cells. At 24°C, the observed excellent cytological condition of the corpora lutea was in contrast to the necrosis seen when the explants were incubated at 34°C.     

Corpora lutea in submerged organ cultures at low incubation temperatures

Summary Corpora lutea in submerged organ cultures of mouse ovary retained viability when incubated at 24°C. At harvest, whole corpora lutea appeared uniformly viable, and mitotic figures were occasionally seen among the luteal cells. At 24°C, the observed excellent cytological condition of the corpora lutea was in contrast to the necrosis seen when the explants were incubated at 34°C.     

Effect of temperature on the life history and demographic parameters of Spalgis epius (Lepidoptera: Lycaenidae), a candidate biological control agent of mealybugs (Hemiptera: Pseudococcidae)

Many species of mealybugs (Hemiptera: Pseudococcidae) are serious pests of economically important crops worldwide. We evaluated the influence of constant temperatures: 14, 16, 18, 20, 22, 24, 26, 28, 30, 32 and 34°C on the life history and demographic parameters of Spalgis epius (Lepidoptera: Lycaenidae), a candidate biological control agent of various species of mealybugs. No eggs completed their development at 14 and 34°C. Egg-to-adult developmental time significantly decreased from 89.9 days at 16°C to 20.4 days at 32°C. The estimated lower temperature threshold of 10.2°C and 416.6 degree-days were required to complete egg-to-adult development. The mortality of immature stages was maximum at 16 and 32°C and minimum at 28°C. The highest lifetime fecundity was recorded at 28°C and it significantly decreased at 32°C. The longevity of adults was about three times more at 16°C than at 30 and 32°C. The net reproductive rate (R ₀) significantly increased with increased temperatures up to 28°C and significantly decreased at 32°C. The mean generation time (T) significantly decreased with increased temperature up to 30°C, but it significantly increased at 32°C. The intrinsic rate of population increase (r _m ) was highest at 30°C. The finite rate of increase (λ) was significantly greater at 30°C than at other temperatures. These data suggest that S. epius can develop, reproduce and survive in a wide range of temperatures and thus could be regarded a potential biological control agent of mealybugs.     

Improving germination and vigour of aged and stored onion seeds by matriconditioning

Germination and vigour of accelerated aged (AA) and naturally stored onion seeds were examined. Accelerated ageing was conducted at 40 °C and 100 % relative humidity (RH). Non aged seeds were stored for 34 months at 3 or 15 °C and 40, 60 or 90 % RH. To restore seed viability, stored and aged seeds were matriconditioned with Micro-Cel E. A distinct loss of germination was observed after 5 days of accelerated ageing. Naturally stored seeds maintained high viability for 34 months, when stored at 3 °C and 40, 60 and 90 % RH or at 15 °C and 40 %. An increase of RH to 60 and 90 % at 15 °C caused loss of germination and vigour. Matriconditioning improved germination and increased endogenic ethylene release and in vivo ACC oxidase activity of both aged and stored seeds.     

Evaluation of functional stability of quercetin as a raw material and in different topical formulations by its antilipoperoxidative activity

The present study evaluates the antioxidant activity of the flavonol quercetin, and its functional stability as a raw material and when added in formulations. The iron-chelating activity was determined using the bathophenanthroline assay, and the functional stability was evaluated with the antilipoperoxidative assay. Raw material presented concentration-dependent antilipoperoxidative and iron-chelating activities. The initial antilipoperoxidative activity of the raw material, cream and gel-cream were 63%, 78%, and 69%, respectively. There was no detectable loss of activity during 182 days (6 months) of storage at all tested temperatures (4°C, room temperature [RT], 37°C, and 45°C) for the raw material. Considering the method variability of 10%, activity loss greater than 10% for nonionic cream was detected after 126 days at 4°C (20.1%), decreasing thereafter to 22.2% after 182 days. At 45°C, the loss of activity started after 182 days (13.2%). For the anionic gel-cream, activity loss started after 84 days (28.4%, 45°C), decreasing after 182 days to 40.3% at 45°C. At 37°C, activity loss was detected after 182 days (12%). In conclusion, the results suggest that the activity of quercetin depends on iron chelation, and its posible usefulness as a topical antioxidant to prevent oxidative stress-induced skin damage depends on maintaining its antilipoperoxidative activity stored at RT, which avoids special storage conditions. Published: February 3, 2006     

Evaluation of efficacy of stabilizers on the thermostability of live attenuated thermo-adapted <Emphasis Type="Italic">Peste des petits ruminants</Emphasis> vaccines

In this study, thermo-adapted (Ta) PPR vaccines were assessed for their stability at 25, 37, 40, 42 and 45°C in lyophilized form using two extrinsic stabilizers {lactalbumin hydrolysate-sucrose (LS) and stabilizer E} and in reconstituted form with the diluents (1 mol/L MgSO₄ or 0.85% NaCl). The lyophilized vaccines showed an expiry period of 24–26 days at 25°C, 7–8 days at 37°C and 3–4 days at 40°C. LS stabilizer was superior at 42°C with a shelf-life of 44 h, whereas in stabilizer E, a 40 h shelf-life with a comparable half-life was observed. At 45°C, the half-life in stabilizer E was better than LS and lasted for 1 day. Furthermore, the reconstituted vaccine maintained the titre for 48 h both at 4°C and 25°C and for 24–30 h at 37°C. As both the stabilizers performed equally well with regard to shelf-life and half-life, the present study suggests LS as stabilizer as a choice for lyophilization with 0.85% NaCl diluent, because it has better performance at higher temperature. These Ta vaccines can be used as alternatives to existing vaccines for the control of the disease in tropical countries as they are effective in avoiding vaccination failure due to the breakdown in cold-chain maintenance, as this vaccine is considerably more stable at ambient temperatures.