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At least 55 records · Page 3Linked to original sources
PubMed2004

Cold-temperature stability of five beta-lactam antibiotics in bovine milk and milk extracts prepared for liquid chromatography-electrospray ionization tandem mass spectrometry analysis.

The stability of five major beta-lactam antibiotics (amoxicillin, ampicillin, cloxacillin, oxacillin, and penicillin G) in fortified milk and in milk extracts prepared for LC-ESIMS/MS analysis was studied at varying cold temperatures (4, -20, and -76 degrees C). Storage of milk samples at 4 degrees C resulted in measurable losses of all beta-lactams after 6 days (>50% in most cases). Slow degradation of penicillin G, cloxacillin, and oxacillin was observed in milk stored at -20 degrees C, but no losses were recorded at -76 degrees C over 4 weeks. All antibiotics showed good stability at all temperature tested in milk extracts prepared for LC-ESIMS/MS analysis. The results of this study emphasize adherence to adequate sample handling and storage protocols as to reflect residue levels at the time of sample submission.

Animals↗
PubMed1999

Cold-temperature plastic resin embedding of liver for DNA- and RNA-based genotyping.

The standard practice of tissue fixation in 10% formalin followed by embedding in paraffin wax preserves cellular morphology at the expense of availability and quality of DNA and RNA. The negative effect on cellular constituents results from a combination of extensive cross-linking and strand scission of DNA, RNA, and proteins induced by formaldehyde as well as RNA loss secondary to ubiquitous RNase activity and negative effects of high temperature exposure during paraffin melting, microscopic section collection, and tissue adherence to glass slides. An effective strategy to correlate cellular phenotype with molecular genotype involves microdissection of tissue sections based on specific histopathological features followed by genotyping of minute representative samples for specific underlying molecular alterations. Currently, this approach is limited to short-length polymerase chain reaction amplification (<250 bp) of DNA, due to the negative effects of standard tissue fixation and processing. To overcome this obstacle and permit both cellular morphology and nucleic acid content to be preserved to the fullest extent, we instituted a system of cold-temperature plastic resin embedding based on the use of the water-miscible methyl methacrylate polymer known as Immunobed (Polysciences, Warminster, PA). The system is simple, easy to adapt to clinical practice, and cost-effective. Immunobed tissue sections demonstrate a cellular appearance equivalent or even superior to that of standard tissue sections. Moreover, thin sectioning (0.5-1.0 microm thickness) renders ultrastructural evaluation feasible on plastic-embedded blocks. Tissue microdissection is readily performed, yielding high levels of long DNA and RNA for genomic and transcription-based correlative molecular analysis. We recommend the use of Immunobed or similar products for use in molecular anatomical pathology.

Cold Temperature↗
PubMed2002

An investigation into the changed physiological state of Vibrio bacteria as a survival mechanism in response to cold temperatures and studies on their sensitivity to heating and freezing.

AIMS: To induce pathogenic Vibrio bacteria into a changed physiological state, in response to cold temperatures in sea water, and assess their sensitivity to heating and freezing, as compared with normal cells. METHODS AND RESULTS: Cells of exponential phase Vibrio vulnificus, V. cholerae and V. parahaemolyticus were washed and inoculated into flasks of sea water, which were stored at 20 and 4 degrees C. Cells stored at 20 degrees C could be recovered after 60 d on non-selective agar (heart infusion agar; HIA) and on the selective agar (thiosulphate citrate bile salts agar) which is used in most Vibrio detection methodology. At 4 degrees C cells became non-culturable on both agars over time. The non-culturable cells appeared to be metabolically active and maintained their membrane integrity, whilst undergoing a change in morphology from rod-shaped to coccoid cells. Resuscitation was possible, in some cases, by an upshift in temperature before plating and the addition of catalase to HIA plates was found to increase recovery. Studies were carried out to assess the sensitivity of the non-culturable cells to heating and freezing compared with the normal cells. Vibrio organisms, whether culturable or in the non-culturable form, were not inactivated by freezing to -20 degrees C. Heating studies showed that V. parahaemolyticus was very heat resistant at low temperatures. However, a pasteurization regime of 2 min at 70 degrees C was found to be effective against all three strains. Experiments showed that the non-culturable cells of all three strains were similar in their heat resistance or, in some cases, were more heat sensitive than cells in the normal form. CONCLUSIONS: Cells in the changed physiological form would not be detected in fish or seafood products by the current Vibrio detection methods. Freezing had no effect in reducing cell numbers. Vibrio parahaemolyticus was very heat resistant in the low temperature pasteurization studies. The higher pasteurization regime of 70 degrees C for 2 min was effective against all three pathogens. Non-culturable cells had similar heat sensitivity or were more heat sensitive than cells in the normal state. SIGNIFICANCE OF IMPACT OF THE STUDY: The study has highlighted a need for the development of better Vibrio detection methods. The low temperature pasteurization of oysters, which has been recommended in the USA, would not be adequate against the strain of V. parahaemolyticus used in this study. Heating regimes which were found to control cells in the normal form will also be effective for the control of the cells with changed physiology.

Agar↗
PubMed2000

Extended alternating-temperature cold acclimation and culture duration improve pear shoot cryopreservation.

Meristems of many pear genotypes can be successfully cryopreserved following 1 week of cold acclimation, but an equal number do not survive the process or have very little regrowth. This study compared commonly used cold acclimation protocols to determine whether the cold acclimation technique used affected the cold hardiness of shoots or the regrowth of cryopreserved meristems. In vitro-grown pear (Pyrus L.) shoots were cold acclimated for up to 16 weeks, then either the shoot tips were tested for cold hardiness or the meristems were cryopreserved by controlled freezing. Cold acclimation consisted of alternating temperatures (22 degrees C with light/-1 degrees C darkness with various photo- and thermoperiods) or a constant temperature (4 degrees C with an 8-h photoperiod or darkness). Compared with nonacclimated controls, both alternating- and constant-temperature acclimation significantly improved postcryopreservation regrowth of P. cordata Desv. and P. pashia Buch. -Ham. ex D. Don meristems. Alternating-temperature acclimation combined with either an 8-h photoperiod or darkness was significantly better than constant-temperature acclimation. Alternating-temperature shoot acclimation for 2 to 5 weeks significantly increased postcryopreservation meristem regrowth, and recovery remained high for up to 15 weeks acclimation. Postcryopreservation meristem regrowth increased with 1 to 5 weeks of constant-temperature acclimation and then declined with longer acclimation. Shoot cold hardiness varied with the acclimation procedure. The LT(50) of shoots acclimated for 10 weeks with alternating temperatures was -25 degrees C; that with constant temperature was -14.7 degrees C; and that of the nonacclimated control was -10 degrees C. Less frequent transfer of cultures also improved acclimation of shoots. Shoots grown without transfer to fresh medium for 6-12 weeks had higher postcryopreservation recovery with shorter periods of acclimation than shoots with a 3-week transfer cycle.

Acclimatization↗
PubMed1975

Thermal responses of unclothed men exposed to both cold temperatures and high altitudes.

Six resting men were exposed to three temperatures (15.5, 21, 26.5 degrees C) for 120 min at three altitudes (sea level, 2,500 m, 5,000 m). A 60-min sea-level control at the scheduled temperature preceded the nine altitude episodes. Comparison of the base-line results at any one temperature showed no differences between rectal temperatures (Tre) or mean weighted skin temperatures (Tsk). After 120 min, Tre and Tsk not only depended on ambient temperature but also altitude. The initial rate of fall in Tre increased with altitude and equilibrium occurred earlier. At 15.5 degrees C, Tre was 0.3 degrees C lower at 5,000 m and 0.2 degrees C lower at 2,500 m than at sea level. Tsk was almost 2 degrees C higher at 15.5 degrees C at 5,000 m and 1 degrees C higher at 2,500 m than at sea level. Similar, smaller differences were observed at 21 degrees C. Mean weighted body temperature showed no change with altitude, but, since the gradient between core and shell was reduced, a shift of blood toward the periphery is implied.

Adolescent↗
PubMed1981

Responses of shorn and full-fleeced lambs given two levels of feed intake and exposed to warm and cold temperatures.

Studies on the effect of fleece length (5 and 85 mm) and feed intake (1 and 2 kg/day) were conducted in 9- to 10-month-old wether lambs kept at 2 environmental temperatures (-12.3 and 17.8 C). Digestion coefficients were lower (P less than 0.05) at the high level of intake and in cold-exposed lambs. Rectal temperature was lower (P less than 0.05) in lambs kept at - 12.3 C. Respiration rate increased (P less than 0.05) with increasing environmental temperature and higher feed consumption. Blood glucose and plasma urea nitrogen concentrations were not affected by feed intake level or fleece length. Glucose was higher (P less than 0.05) and plasma was lower (P less than 0.05) in lambs at low temperatures. Serum-free fatty acids were increased (P less than 0.05) in lambs given the low feeding level and kept at the low temperature. Plasma thyroxine and protein-bound iodine concentrations were lower (P less than 0.10) in full-fleeced lambs and were higher (P less than 0.05) at the low temperature.

Animals↗
PubMed2005

Cold temperature PM emissions measurement: method evaluation and application to light duty vehicles.

This work examines the methodology to sample and measure the number and size of motor vehicle particulate emissions (PM) at subambient temperatures. The study has two principal objectives. The first is to address the following question: which aspects of the particle sampling, dilution, and size measurement process must be made at the vehicle test temperature to obtain an accurate representation of the PM emissions? The second is to perform a preliminary overview of how subambient temperature operation affects PM emissions from the major classes of current model light duty vehicles. The principal findings are the following: (1) The temperature of the particle size instruments, test cell versus room temperature, has little effect on the measurements. (2) Once the engine has warmed, solid particle (soot) mode emissions in the cold test cell are similar to those at room temperature. The first finding simplifies cold temperature emissionstesting because it allows particle sizing instruments to be placed outside the cold test cell and operated at room temperature. The latter is consistent with the expectation that solid particles are formed in the engine and are therefore relatively unaffected by ambient conditions after engine warm-up. Use of cold dilution air in the room-temperature test cell increases the number and size of nuclei particles; however, the effect of dilution airtemperature was inconclusive in the cold test cell.

Air Pollutants↗
PubMed2002

Protective effects of cold temperature and surface-contact on acid tolerance of Salmonella spp.

AIMS: To determine the effect of cold-storage temperature and surface contact on the survival of Salmonella spp. during acid challenge. To determine the contribution of sigma transcriptional factor (encoded by rpoS) in surface contact-mediated acid tolerance. METHODS AND RESULTS: Salmonella serovar Typhimurium cells in exponential and stationary growth phase were subjected to acid challenge in planktonic and surface-associated states. Low temperatures offered protection against acid challenge to cells which were in stationary growth phase (but not to those in exponential growth phase). The cells from stationary, as well as logarithmic growth phase, acquired increased acid tolerance upon surface contact with various surfaces, such as fresh-cut apples, agar and polyethersulphone membranes. The alternative sigma transcription factor was not required to acquire surface contact-mediated acid tolerance. CONCLUSIONS: Salmonella spp. take advantage of low temperature and surface association to overcome acid challenge. Some of the acid tolerance mechanisms are independent of the sigma transcription factor regulon. SIGNIFICANCE AND IMPACT OF THE STUDY: Effective control measures during produce processing must take into account the different susceptibilities of planktonic vs surface-associated food-borne pathogens.

Agar↗
PubMed1978

Depolarization of the Electrogenic Transmembrane Electropotential of Zea mays L. by Bipolaris (Helminthosporium) maydis Race T Toxin, Azide, Cyanide, Dodecyl Succinic Acid, or Cold Temperature.

The transmembrane electrical potential of root cells of Zea mays L. cv. W64A in a modified 1x Higinbotham solution was partially depolarized by semipurified toxin obtained from Bipolaris (Helminthosporium) maydis race T. At a given toxin concentration depolarization of Texas cytoplasm cells was much greater than for normal cytoplasm cells. This observation correlated directly to the differential host susceptibility to the fungus. The time course and magnitude of depolarization were dependent on toxin concentration; at high concentration the electropotential difference change was rapid. Cortex cells depolarized more slowly than epidermal cells indicating that the toxin slowly permeated intercellular regions. Toxin concentrations which affected electropotential difference were of the same magnitude as those required to inhibit root growth, ion uptake, and mitochondrial processes.Azide, cyanide, and cold temperature (5 C) gave the same partial depolarization as did the toxin. Dodecyl succinic acid caused complete depolarization. These and other data indicate that one of the primary actions of the toxin is to inhibit electrogenic ion pumps in the plasmalemma.

Journal Article↗
PubMed2002

Cold temperatures reduce the sensitivity of stored platelets to disaggregating agents.

In this study, we compared the effect of signal transduction inhibitors on fibrinogen binding, aggregation, the activation state of GPIIb-IIIa, and cytosolic calcium levels in cold and room temperature-stored platelets. Cold-stored platelets have a higher sensitivity to agonist-induced aggregation when compared to room temperature-stored platelets. We also found that cold-stored platelets had a significantly higher aggregation response to ADP and epinephrine, while platelets stored at room temperature responded poorly to these agonists (mean values of 61 vs. 18%, n = 14). Four inhibitors were selected to target various signaling pathways. Cold-stored platelets were more resistant to disaggregation by promethazine, prostaglandin D2, yohimbine, and echistatin. The effects of cold temperatures on stored platelets are targeted to activation pathways as there was no spontaneous aggregation or spontaneous fibrinogen binding as measured in this study. PAC-1 binding was not inhibited to the same degree as aggregation or fibrinogen binding responses, suggesting that the disaggregation was not caused by a change in the conformation of GPIIb-IIIa. Cytosolic calcium levels did not decrease in cold-stored platelets after inhibitor addition. The inhibitors are likely acting after the establishment of the GPIIb-IIIa activation state and may affect the post-occupancy signaling by the fibrinogen-occupied integrin. Differences between aggregation and disaggregation responses of cold- and room temperature-stored platelets suggest that cold-stored platelets may have different mechanisms to stabilize platelet aggregates during their formation.

Blood Platelets↗
PubMed1992

[Influence of environmental temperature (cold exposure) on nasal resistance].

Changes in nasal resistance were examined in 33 healthy subjects in order to investigate the physiological reactions of nasal mucosa to cold exposure. Each examinee was exposed to changes in room temperature from 25 degrees C to 15 degrees C with the same humidity in an artificial atmosphere chamber, regulated by an environmental control unit. Nasal resistance was measured by means of the active-anterior method using a rhinomanometer MPR-2200. The same experiments were repeated on different dates in all 18 subjects. The results can be summarized as follows. 1) The average value of nasal resistance significantly increased after room temperature was decreased from 25 degrees C to 15 degrees C. 2) Patterns of nasal resistance responses to cold exposure were classified into two types. One type was increased nasal resistance after cold exposure and the other was maintenance of resistance despite cold exposure. Each individual manifested a different pattern in experiments carried out on different dates. 3) The nasal resistance value in the chamber at 25 degrees C was significantly higher in winter than in summer, and the increase in nasal resistance in response to cold exposure was significantly larger in summer than in winter. This result was assumed to be due to cold adaptation of the nasal mucosa in winter. 4) Heart rate decreased significantly with cold exposure. However, changes of heart rate in response to cold exposure did not correlate with changes in nasal resistance. No correlation between changes in nasal resistance in response to cold exposure and orthostatic loading, which was carried out after the cold exposure experiment, was detected. 5) A nasal cycle was observed in 7 (14%) of the 33 cases. In 6 of these total nasal resistance was maintained despite cold exposure.

Adult↗
PubMed2000

Adaptive response to cold temperatures and characterization of cspA in Salmonella typhimurium LT2.

Salmonella typhimurium is a major foodborne microbial pathogen which primarily contaminates poultry products causing salmonellosis in humans. S. typhimurium LT2 cultures, when transferred from 37 degrees C to 5 degrees C or 10 degrees C, showed an initial lag period in growth with an approximate generation time of 10-25 h. Western blot assay using E. coli CS7.4 antibody and analysis of radiolabeled total cellular proteins from S. typhimurium cultures after exposure to 10 degrees C or 5 degrees C showed elevated expression of a major cold shock protein, CS7.4. Identification of a decreased level of CS7.4 at 37 degrees C suggests that the expression of this protein may require a large temperature downshift. Putative regulatory protein binding segment on the 5'-untranslated region referred as 'Fragment 7' in S. typhimurium exhibited a 90.6% and a 56.25% nucleotide sequence identity when compared with the Fragment 7 of E. coli and S. enteritidis, respectively. The differences in the nucleotide sequence within the Fragment 7 between S. typhimurium and S. enteritidis may explain the differential expression of CspA at 37 degrees C. The nucleotide sequence of the open reading frame of S. typhimurium cspA gene showed a single base difference at 816 bp position from a G to a C which altered the amino acid residue from a glycine to an alanine. In addition to CspA, an elevated expression of a 105 kDa, and decreased expression of 6 proteins were evidenced when cultures of S. typhimurium were exposed to 10 degrees C or 5 degrees C. Differential expression of the CspA and other proteins in S. typhimurium following exposure to cold temperatures suggest that adaptation and continued growth and survival at cold temperatures in this pathogen may be aided by these cold-responsive proteins.

Adaptation, Physiological↗