The effect of temperature at which slow cooling is terminated and of thawing rate on the survival of one-cell mouse embryos frozen in dimethyl sulfoxide or 1,2-propanediol solutions

We investigated the slow freezing of one-cell mouse embryos with either dimethyl sulfoxide (Me2SO) or 1,2-propanediol (PROH) as the cryoprotectant. One-cell embryos, collected from superovulated C57BL/6J x CBA/Ca females were exposed to 1.5 M solutions of either Me2SO or PROH. The embryos were cooled at 0.3 degrees C/min to temperatures between -10 degrees and -80 degrees C before being plunged into LN2 and then warmed at either 20 degrees C/min or 450 degrees C/min. Survival was expressed as the percentage of hatching or hatched blastocysts per frozen-thawed embryo. When the slow cooling was in 1.5 M PROH, the temperature at which survival rates after slow thawing began to increase was -35 degrees C (52.6 +/- 5.2% survival). For slow cooling in 1.5 M Me2SO this temperature was -50 degrees C (45.0 +/- 2.9% survival). The addition of sucrose to the 1.5 M PROH solution raised the temperature at which survival rates after slow thawing began to increase to -30 degrees C (54.8 +/- 3.7% survival). If slow cooling was stopped at high subzero temperatures, embryos survived better after rapid thawing than slow thawing. If slow cooling was stopped at low subzero temperatures, the survival rate was not dependent on the thawing rate if freezing was done in 1.5 M PROH. When freezing was in Me2SO solutions and to subzero temperatures of -60 degrees and -80 degrees C, slow thawing gave better survival than rapid thawing. The addition of sucrose to the Me2SO freezing solution restored the survival rates at -60 degrees and -80 degrees C. These results indicate that high rates of survival may be obtained from one-cell mouse embryos by a rapid or a slow thawing procedure, as has been found for other developmental stages. The results also indicate that PROH provides superior protection compared to Me2SO against freezing-thawing damage and that the addition of sucrose to the freezing solutions prior to freezing improves the overall survival rates. Embryos that survived freezing and developed in culture implanted and formed normal fetuses at rates similar to those of nonfrozen control embryos (60% vs 68% and 53% vs 58%, respectively.     

A comparative assessment of cryosurgical devices: application to prostatic disease

OBJECTIVES: To determine the comparative freezing ability of the Cryotech (CT) and AccuProbe (CMS) cryosurgical systems. METHODS: Four conditions designed to model clinical situations were produced: (1) Single-probe performance in water at 17 degrees C; (2) five-probe performance in water at 17 degrees C; (3) single-probe performance in gel at 22 degrees C; and (4) single-probe performance in bovine liver. Parameters evaluated included temperatures at various time points (rates to and final low temperature), configuration of a freeze zone, and shaft freezing characteristics. In addition, isotherms were measured at predetermined distances from the center of the freeze zone. RESULTS: Both systems provided freezing of various media under operational conditions. In water, the CMS 3-mm probe delivered more rapid freezing temperature rates than the 3-mm CT probe, with a 110 degrees C difference in probe surface temperature. In gel, the CMS probe increased freeze volume fourfold versus a twofold increase for the CT probe. In bovine liver, there was nearly equivalent performance with respect to geometry of the freeze ball. Extrapolation of the CT cooling curve indicated temperature equivalence at 30 minutes. A larger shaft diameter 4.9-mm CT probe produced results similar to the CMS probe in all the tested media. In addition, the freeze configuration of the CMS probe was spherical; the CT configuration was more cylindrical. CMS probe (equivalent diameter) tip temperatures were on average 100 degrees C lower. CONCLUSIONS: Our tests demonstrated differences between the CMS and CT probe. The major differences are in the configuration of the freeze zone and shaft freezing. In equivalent conditions, the CMS 3-mm probe delivered more rapid cooling rates, a more spherical freeze ball, and lower absolute temperatures than the CT 3-mm probe. The larger CT probe produces equivalent freezing temperatures to the CMS probe, albeit with a more spherical shape. However, these in vitro systems may not adequately reflect varied prostate morphology. Further research is under way to determine if these differences affect relative efficacy of cryotherapy of the prostate.     

Cryopreservation and culture of human corneal keratocytes

PURPOSE: To assess the effects of two different concentrations of albumin in a cryoprotective solution and two freezing methods on human corneal keratocyte ctyopreservation. METHODS: Isolated keratocytes were used for cryopreservation. Solutions of 10% dimethylsulfoxide with either 2% or 10% human albumin were used as cryoprotective agents. Cells either were transferred directly into a -80 degrees C freezer (freezing rate, 2 degrees C/min) or were cooled in a programmed freezer (1 degrees C/min until -40 degrees C and then 10 degrees C/min), which resulted in four different cryopreservation protocols. Cells were stored at -80 degrees C, then were thawed at 37 degrees C, and subsequently were cultured. Keratocytes were studied by means of trypan blue staining, growth assay, apoptosis assays, transmission electron microscopy, and immunochemistry. RESULTS: The percentage of cells that were alive after thawing ranged from 80% to 99% by trypan blue staining and from 45% to 60% by flow cytometry. The ratio of the number of living cells at the end of primary culture after cryopreservation to that before cryopreservation was significantly (P=0.04) higher after direct transfer into the -80 degrees C freezer than after controlled-rate freezing, whereas the albumin concentration had no significant influence on this ratio (P=0.45). The percentage of apoptotic cells was significantly higher after cryopreservation than in the control group of noncryopreserved cells; more than 5% 24 hours after thawing. Cryopreservation did not modify the keratocyte ultrastructure. Fibroblast growth factor dramatically decreased the serum-induced cell expression of alpha smooth muscle actin, whereas cryopreservation had no influence on this cell expression. CONCLUSIONS: A freeze-thaw trauma, which was related to cryopreservation-induced cell apoptosis, was revealed during primary culture after thawing. Direct transfer into the -80 degrees C freezer resulted in better postcryopreservation growth in the culture than controlled-rate freezing. A change in albumin concentration from 2% to 10% did not affect the results.     

Effect of freezing method, thawing temperature and post-thaw dilution/washing on motility (CASA) and morphology characteristics of high-quality human sperm

Sixteen semen samples, 12 donor and four patient samples of high initial quality, were processed to compare the effect of two freezing methods, two thawing temperatures and the effect of dilution and washing on sperm motility and morphology characteristics. Sperm samples were divided in two equal parts and frozen either by fast vapour freezing or by slow computer-controlled freezing. For each freezing method, half of the straws were thawed at room temperature (22 degrees C), the other half were thawed at 37 degrees C. From each freeze-thawing treatment, one straw was evaluated immediately post-thawing; another straw was washed to remove the cryoprotectant solution. In this way, each semen sample was subjected to eight freeze-thawing treatments. No effect of the freezing method and thawing temperature was observed on motility characteristics evaluated by computer-assisted semen analysis, nor on light-microscopical morphology parameters. Post-thaw dilution and washing, however, exerted a deleterious effect on sperm motility, by reducing percentage motility by 50% compared to unwashed thawed specimens. Linearity and percentage of morphologically normal spermatozoa were obviously impaired, while percentage of abnormal tails and beat cross frequency increased significantly. In general, freeze-thawing was most successful when rapid vapour freezing was followed by 37 degrees C thawing, and when slower computer-controlled freezing was combined with 22 degrees C thawing, causing significant interactions between the freezing method and the thawing temperature. For semen samples of high initial quality, vapour and computer-controlled freezing were equally effective in terms of recovery of morphologically normal, motile spermatozoa.     

Subzero nonfreezing preservation in a murine limb replantation model

This study was designed to examine the most effective temperature for hypothermic storage, without freezing, to prolong ischemic tolerance in an amputated murine hindlimb model. We measured freezing points in the calf muscle and the subcutaneous tissue of the foot in the amputated limbs of Fisher 344 strain male inbred rats. The highest freezing point was -1.5 degrees C, which was recorded in the calf muscle. To prevent freezing in any of the tissues in the amputated limb, the temperature for the lowest nonfreezing preservation was defined as -1 degrees C. The amputated limbs were preserved at subzero nonfreezing temperature (-1 degrees C) and at 4 degrees C for 4, 8, 12, 24, 48, and 72 h, and were then transplanted to other inbred rats by microsurgical techniques. We evaluated the vascular patency of the anastomoses by direct observation and performed histological examinations on the seventh day after replantation. Subzero nonfreezing preservation of a limb at -1 degrees C for 72 h was significantly superior to hypothermic preservation at 4 degrees C for 72 h in terms of anastomotic patency rates (P < 0.05). The histology of skeletal muscles preserved at -1 degrees C for 8 h showed greater similarity to the normal situation than the histology of those preserved at 4 degrees C for 8 h. Bone viability with osteoblastic activity was maintained in grafted limbs preserved at -1 degrees C for 72 h, but in the limbs preserved at 4 degrees C for 72 h the bone was not viable, showing no osteoblastic activity. Clinically, the period of ischemia in major limb replantation at normal ambient temperatures is limited to about 6 h. In this study, the maximum ischemia time for replantation of a limb containing muscle tissue was prolong to 8 h at -1 degrees C, but the maximum ischemia time at 4 degrees C could not be prolonged to 8 h. Our results suggest that, in the major replantation of a limb containing muscular tissue, hypothermic preservation at -1 degrees C would be more useful than preservation at 4 degrees C.     

Incubation temperatures affect adherence to plastic of Candida albicans by changing the cellular surface hydrophobicity

The influence of cellular surface hydrophobicity on the adherence capacity to plastic of Candida albicans was investigated at two culture temperatures (37 and 22 degrees C). The majority of the 42 strains studied were hydrophobic at 22 degrees C and hydrophilic at 36 degrees C. The hydrophobic cells showed a consistent adherence capacity which was absent from the hydrophilic strains. The culture temperatures affect adherence to plastic of C. albicans by changing the cellular surface hydrophobicity.     

Freezing temperature of finger skin

In 45 subjects, 154 frostnips of the finger were induced by cooling in air at -15 degrees C with various wind speeds. The mean supercooled skin temperature at which frostnip appeared was -9.4 degrees C. The mean skin temperature rise due to heat of fusion at ice crystallization was 5.3 degrees C. The skin temperature rose to what was termed the apparent freezing point. The relation of this point to the supercooled skin temperature was analyzed for the three wind speeds used. An apparent freezing point for a condition of no supercooling was calculated, estimating the highest temperature at which skin freezes at a given wind speed. The validity of the obtained differences in apparent freezing point was tested by an analysis of covariance. Although not statistically significant, the data suggest that the apparent freezing point with no supercooling decreases with increasing wind velocity. The highest calculated apparent freezing point at -15 degrees C and 6.8 m/s was 1.2 degrees C lower than the true freezing point for skin previously determined in brine, which is a statistically significant difference.     

Shear stress effects on growth and activity of Lactobacillus delbrueckii subsp. bulgaricus

Cells are frequently submitted to shear stresses during industrial processes. Shear stress can be either beneficial or detrimental to the cells depending on the organism and on the level of intensity. The present work was designed to study the effect of shear stress on cell activity of a widely used lactic acid bacterium, Lactobacillus delbrueckii subsp. bulgaricus (L. bulgaricus). A constant shear stress bioreactor, based on Couette device, was developed and used to control shear stress from 0 to 72 Pa during a 4-h cultivation of a supplemented whey permeate medium with L. bulgaricus at 42 degrees C. In order to reach high shear stresses and to perform experiments within the laminar flow range where hydrodynamic conditions are accurately defined, the medium was thickened with carboxymethylcellulose (CMC). pH, cell counts, optical density, lactose and lactic acid concentrations were monitored during culture, and cell activity was evaluated after culture and after a freezing treatment at -80 degrees C, using a standardized activity test based on optical density measurement. Cell metabolism was significantly improved by intermediate shear stress levels (36 and 54 Pa) during culture. Furthermore, biomass concentration, evaluated by optical density, was clearly higher at 36 Pa. Cell lengthening was observed, which was mainly related to the presence of CMC and partly to shear stress level, especially at 36 Pa. Hydrodynamic conditions during culture could affect the membrane permeability of the cell and its resistance to freezing. Cells cultivated at 72 Pa were certainly weakened by shearing forces, and these cultures exhibited lag times twice as long after freezing as those grown at 36 Pa. Furthermore, after freezing, cultures grown at 36 Pa had shorter lag times than at 0 Pa (1.1 and 1.3 h, respectively) and higher specific growth rates (1.24 and 0.99 h-1, respectively).     

MPP+ induced substantia nigra degeneration is attenuated in nNOS knockout mice

Differential scanning calorimetry was used to characterize thermal events associated with freezing and melting of suspensions and extracts of Panagrolaimus davidi, an Antarctic nematode which can survive intracellular freezing. Nematode suspensions produced a single freezing exotherm with a shoulder on the peak representing the freezing of the nematodes. A shoulder on the peak of melting endotherms indicates the melting of the nematodes and of the water surrounding them. Exotherms were also detected from individual nematodes mounted in liquid paraffin. The freezing of nematodes was very rapid and in marked contrast to that of freezing-tolerant insects and vertebrates, which take hours or days to freeze. Eighty-two percent of the nematodes' body water froze. High levels of survival were obtained in nematodes exposed to temperatures down to -40 degrees C. No additional thermal events were observed after the freezing event and before the melting of samples cooled to -40 degrees C, indicating no changes in the proportion of body water frozen. Ice nucleating activity is present in nematode suspensions but not in supernatants from nematode extracts. No thermal hysteresis activity was detected in nematode extracts.     

Comparison of the different strategies for cryopreserving and storage of the bone marrow CD34+ cells. Possibility of unprogrammed rate freezing and storage at -80 degrees C mechanical freezer

We have compared the efficiency of the different strategies allowing for a long term storage of a human CD34+ bone marrow cells. Accordingly, the aliquots of CD34+ cells isolated from bone marrow were frozen using: controlled rate freezing equipment, or freezer unprogrammed in a -80 degrees C mechanical freezer. After freezing, CD34+ cells were subsequently stored for one month in a liquid nitrogen tank at -196 degrees C or in mechanical freezer at -80 degrees C. We have found that both the viability and the recovery of clonogeneic progenitors of CD34+ cells samples stored at different temperature were similar. Therefore, regarding the costs and simplicity, we recommend the unprogrammed freezing and storage of human CD34+ cells at -80 degrees C in a mechanical freezer as a convenient, inexpensive, and reliable method for storing marrow for transplantation. This data also indirectly indicate that the aliquots of the CD34+ cells can be shipped frozen on dry ice (-80 degrees C), and that these cells will maintain viability under this conditions. Furthermore, in this study we have confirmed validity of our earlier observation that human CFU-Meg progenitors are more sensitive to cryopreservation.     

Takayasu''s arteritis in Mexico: a clinical review of 44 consecutive cases

Experiments were designed to determine the effects of sub-zero temperatures on the function of the noradrenergic innervation of a peripheral blood-vessel. The central ear-artery of the rabbit was used for this purpose. The ear was exposed to temperatures of -6, -9 or -18 degrees C in vivo for 15 min. After 1 day (24 h) or 6 days in vivo, the central ear-artery was dissected free, incubated in [3H]-noradrenaline (NA) and stimulated in vitro with high potassium (75 mM) for 5 min to evoke release of [3H]-NA. The release of [3H]-NA was Ca(2+)-dependent. One day after exposure to -6, -9 or -18 degrees C, increases of 45-57 and 44-72% and a reduction of 12-35% were observed, respectively, in three successive potassium-evoked NA-releases. After 6 days in vivo an increase of 30-34% was observed following exposure to -6 degrees C, while no alteration was observed after exposure to -9 degrees C. A reduction of 84-89% was recorded after exposure to -18 degrees C. Following this exposure to -18 degrees C, there was also a great reduction in the evoked release of [3H]-NA compared with the spontaneous release, whereas this correlation did not change after exposure to -6 and -9 degrees C. The total uptake of [3H]-NA was unchanged after freezing the tissue at -6 degrees C, but was substantially reduced after exposure to -9 and -18 degrees C. A short period of in vivo restoration (6 days, enhanced the uptake of [3H]-NA.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of anti-freeze protein (AFP) on the cooling and freezing of equine embryos as measured by DAPI-staining

Equine embryos recovered on Day 6 after ovulation were cooled to +4 degrees C, or frozen with AFP alone or together with glycerol. Twenty embryos (140-200 microm in diameter) were randomly assigned to 6 treatment groups. In the first 3 groups, the embryos were cooled from room temperature to +4 degrees C at a rate of 3 degrees C/min and warmed again at a rate of 32 degrees C/min in a programmable freezer. In the second 3 groups, the embryos were frozen using a standard protocol, stored in liquid nitrogen for 5-7 days and then thawed in a 37 degrees C waterbath. After cooling/warming or freezing/thawing all the embryos were stained with DAPI. The percentage of dead cell area was significantly lower in the cooling groups than in the freezing groups and no significant differences were apparent between the cryoprotectants used in the study.     

Kinetics of heat inactivation of Venezuelan equine encephalomyelitis virus

Thermal inactivation of Venezuelan Equine Encephalomyelitis Virus (VEEV) was studied at temperatures from 26 degrees to 55 degrees C. Inactivation of infectivity took place by two thermodynamically different reactions, one of which predominated at temperatures below 44 degrees C and the other at higher temperatures. The presence of 1 or 2 M NaCl stabilized the VEE virus at low temperatures but enhanced the inactivation at high temperatures. This latter effect at temperatures higher than 50 degrees C, is associated with the occurrence of two-component survival curves. The different effects of hypertonic NaCl concentrations at the two ranges of temperature, are related to different mechanisms of inactivation operating at each range (protein denaturation and nucleic acid-RNA breakdown). Different kinetics of thermal inactivation at 55 degrees C were observed between virus strains with different virulence. However, no significant correlations was found between the virulence of the eleven VEE virus strains studied and their thermostability at 37 degres and 55 degrees C.     

Changes in biogenic amines and microbiological analysis in albacore (Thunnus alalunga) muscle during frozen storage

Albacore specimens of extra quality were analyzed for their biogenic amine contents after 1, 3, 6, and 9 months of frozen storage at -18 degrees C or -25 degrees C. A high-performance liquid chromatography method involving a linear elution gradient was optimized for the identification and determination of putrescine, cadaverine, histamine, spermidine, and spermine in albacore tuna. Putrescine was the biogenic amine that showed the highest increase, reaching concentrations of 59.04 ppm (815% of the initial level) and 68.26 ppm (942% of the initial level) in the white muscle of albacore after 9 months of frozen storage at -18 and -25 degrees C, respectively. Cadaverine, histamine, and spermidine concentrations were below 3, 5, and 11 ppm, respectively, after 9 months of frozen storage, while spermidine underwent a significant decrease at both storage temperatures. Microbiological analysis confirmed the absence of species of Enterobacteriaceae in 75% of the albacore specimens after 9 months of frozen storage; coliforms were always below 3 CFU/g. The survival rate of the psychrotrophic microorganisms after 9 months of frozen storage at -25 degrees C was 4.6%, while 38.9 and 92.1% of the aerobic mesophiles present in the white muscle of albacore before freezing survived 9 months of storage at -18 and -25 degrees C, respectively.     

Effect of temperature on growth and activity of a methanogenic culture utilising acetate

Studies with a methanogenic culture enriched for use of acetic acid showed that this culture had an optimum growth temperature of 35 degrees C, with only small differences for other temperatures between 30 and 40 degrees C. The optimum temperature was the same when determined on the basis of biomass production rate during the exponential (log) phase of growth (0.08-0.09 day-1, at 35 degrees C), amount of biomass present at the end of the log phase (100 mg/l), activity of the biomass (rate of conversion in millimoles per day per milligram (dry wt.) biomass present, 0.08 at end of log phase), or biomass yield (mg (dry wt.) biomass produced per millimole acetic acid converted, 1.0-1.1). Temperatures outside the range 30 to 40 degrees C caused marked reductions in the above parameters. The maximum temperature for growth was 42-44 degrees C; the minimum, below 15 degrees C, the lowest temperature studied. Acetic acid conversion to methane was 0.8-1.0 mol/mol, and was independent of temperature.     

Survival and growth of Campylobacter jejuni after artificial inoculation onto chicken skin as a function of temperature and packaging conditions

Campylobacter jejuni is one of the major causes of food poisoning in humans. C. jejuni is also widespread in food animals, and meat and meat products derived from food animals are the most common vector of bacterial transmission to humans. To determine the role of packing and storage conditions on the replication of C. jejuni on chicken, the virulent strain C. jejuni 81116 was artificially inoculated onto chicken skin pieces (1 cm2) and stored at different temperatures and under various packaging conditions. C. jejuni 81116 remained viable at -20 and -70 degrees C and was able to replicate at 4 degrees C and at ambient room temperature. C. jejuni 81116 was also inoculated onto chicken skin and subjected to repeated freeze thawing and the viability of the inoculum was quantified. C. jejuni 81116 could withstand repeated freeze thawing similar to that which may occur in the domestic home. Under all freezing conditions, C. jejuni 81116 retained a high level of viability and quickly replicated to levels which exceeded Australian food authorities' permitted bacteria level on raw food products after the sample was thawed.     

Overexpression and mutations of p53 in metastatic malignant melanomas

Heterocypris incongruens (RAMD.) was selected as an intermediate host, having emerged as the main host for Diorchis stefanskii Czapl. in experimental conditions. It was found that the index of infective activity for larvae of D. stefanskii was high on the first day, amounting to 50-54%, with an extensity of infection of 100%. The infective activity of larvae declined slowly but steadily at the low temperature (5 degrees C). Oncospheres retained their infective activity for more than 2.5 months. In contrast, in culture at 18-20 degrees C and 25 degrees C, the infective activity of larvae had declined rapidly only a few days into the experiment. The irrevocable loss of tapeworm infectivity occurred at temperatures of -5 degrees C and +38 degrees C, at which 100% of oncospheres died.     

Physiopathology of bedsores

Listeria monocytogenes is a food-borne pathogenic bacterium that can be found in soft cheese. At the beginning of cheese ripening, the pH is about 4.85-4.90. The aim of this work was to study the influence of temperature, preincubation temperature (temperature at which the inoculum was cultivated) and initial bacterial concentration on the survival of L. monocytogenes (strain Scott A) at pH 4.8. It was demonstrated in an earlier study that these factors did influence growth kinetics. Survival studies of L. monocytogenes were done in a laboratory broth simulating cheese composition. Four test temperatures (2, 6, 10 and 14 degrees C) and two preincubation temperatures were studied (30 degrees C or the test temperature). Listeria monocytogenes (strain Scott A) was unable to grow at pH 4.8 under all conditions tested. The time for 10% survival was about 11 and 2 d, at 2 degrees C with preincubation at 2 degrees C and 30 degrees C, respectively; 9 d at 6 degrees C with preincubation at 6 degrees C; 4 d at 6 degrees C with preincubation at 30 degrees C; and 1 d at 14 degrees C with preincubation at 14 degrees C or at 30 degrees C. The results show that survival of L. monocytogenes (strain Scott A) at pH 4.8 is not dependent on initial bacterial concentration but on both the test and preincubation temperatures.     

Postcesarean endometritis. Clinical risk factors predictive of positive blood cultures

OBJECTIVE: To identify peripartum risk factors that are predictive of positive blood cultures in patients with postcesarean endometritis. STUDY DESIGN: A retrospective review of 179 patients diagnosed with postcesarean endometritis was conducted. Patients with positive and negative blood cultures obtained at the time of diagnosis were compared. Patient's charts were reviewed for intrapartum, intraoperative and postpartum factors. Chi-square and nonpaired Student's tests were used when appropriate, with P < .05 considered significant. RESULTS: During this period, 179 (20%) postcesarean patients developed endometritis. One hundred sixty-eight (94%) of those patients had blood cultures. Eleven (6.5%) were positive; however, one of these grew a skin contaminant and was disregarded. When patients with positive blood cultures were compared to those with negative blood cultures, length of labor, number of vaginal examinations, postoperative day when the diagnosis was established, estimated blood loss at the time of cesarean delivery, presence of intrapartum chorioamnionitis, number of hours of ruptured membranes, white blood cell count at the time of diagnosis, use of prophylactic antibiotics, development of wound infection or other infectious etiologies were not shown to be predictive. There were no positive blood cultures among patients with a temperature < 38.5 degrees C. At a temperature < 38.8 degrees C, 1/126 (0.79%) had a positive blood culture. At a temperature > or = 38.8 degrees C, 9/42 (21.4%) had a positive blood culture (P < .001). Approximately $5,890 was spent on obtaining positive blood cultures in patients with temperatures < 38.8 degrees C. In contrast, $218 was spent per positive blood culture obtained from patients with a temperature > or = 38.8 degrees C. CONCLUSION: The traditional practice of obtaining blood cultures at a temperature > or = 38.0 degrees C is not justified but elevating the threshold to 38.8 degrees C is equally effective and less costly.     

Identification of murine p120 isoforms and heterogeneous expression of p120cas isoforms in human tumor cell lines

Heat stress is a common problem for cattle. General consequences of heat stress include increased body temperatures and reduced feed intakes. As a measure of heat stress, core body temperatures of unshaded feedlot steers (crossbred Bos taurus) were monitored from mid-June to early November in Nebraska using transmitters implanted in the peritoneum of 10 steers (initially 10 mo of age). Steers were fed at 0630 and 1430 using a finishing diet of 1.52 NEg Mcal/kg with 13% protein and 4% roughage per day and housed in two open lots with stocking densities of 15.2 or 19.3 m2/steer. Core body temperatures, ambient temperature, relative humidity, and wind speed were measured at 3-min intervals and mathematically filtered to produce 120 readings/ d. For 94 usable daily records, body temperature means (39.04 +/- .12 degrees C), maxima (39.89 +/- .21 degrees C at 1836 +/- .73 h), minima (38.33 +/- .29 degrees C at 0823 +/- .38 h), and patterns were similar among steers. As daily maximum ambient temperatures increased, minimum body temperatures decreased slightly (.04 degree C per 5 degrees C; P < .01). After daily maximum ambient temperatures reached a threshold of 25.6 degrees C, daily maximum body temperatures increased linearly with maximum ambient temperatures (.42 degree C per 5 degrees C; P < .01). Sharp peaks in body temperature were often seen in the late evening (approximately 2200) after ambient temperature had decreased to well below maximum values. These evening peaks occurred on an average of 25% of the days, had amplitudes ranging from .7 to 3.5 degrees C relative to mean daily temperatures and lasted for 1.5 h. From a practical standpoint, we suggest that producers monitor meteorological forecast of peak ambient temperatures and make special efforts, such as spraying animals, when exceptionally hot weather is predicted.