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The effect of refrigeration on the osmolality and pH of nonpreserved artificial tears containing carboxymethylcellulose.

INTRODUCTION: Nonpreserved artificial tears (NPAT) are a recommended treatment for dry eye. The manufacturers' instructions state to discard the container after initial opening and use. Some clinicians advocate the use and storage of NPAT in a zip-lock bag in a refrigerator for up to 12 h. The purpose of this study was to evaluate whether refrigeration of opened NPAT over a 12-h period had any effect on the pH or osmolality. METHODS: Forty individual carboxymethylcellulose NPAT samples were used in this study. The initial osmolality and pH of each sample were measured with a vapor pressure osmometer and electronic pH meter. The samples were refrigerated (4 degrees C) in closed zip-lock plastic bags for 12 h. After storage, the pH and osmolality of the samples were measured. The data were statistically analyzed for significant differences using a paired t-test. RESULTS: The mean initial pH and osmolality before refrigeration were 6.46 pH units and 304.10 mmol/kg, respectively. After refrigeration, the mean pH was 6.44 units, and mean osmolality was 305.87 mmol/kg. Paired t-tests revealed a nonsignificant difference (p > 0.05) for both pH and osmolality. CONCLUSION: Refrigeration of opened carboxymethylcellulose NPAT stored in closed zip-lock plastic bags does not have a significant effect on the osmolality or pH of the solution. Storage of NPAT containing carboxymethylcellulose is an acceptable practice with regards to stability of pH and osmolality.

Carboxymethylcellulose Sodium↗

Chondrocyte viability in refrigerated osteochondral allografts used for transplantation within the knee.

PURPOSE: To evaluate cell viability and matrix characteristics of refrigerated osteochondral allografts implanted up to 44 days after harvest. METHODS: Sixteen refrigerated allografts underwent histologic and ultrastructural examination and fluorescence excitation analysis prior to implantation. The average size of the graft implanted was 6.2 cm(2) (+/-3.4 cm(2)). Refrigerated allografts averaged 30 days (range, 17 to 44 days) from donor expiration to implantation. Nine specimens underwent cell viability testing. The percent viability of refrigerated allografts prior to implantation averaged 67%. RESULTS: No significant correlations were noted between histologic score, electron microscopy score, matrix staining percent (MSP) score, and viability. When time to implantation was assessed, an inverse correlation was noted with MSP score (r =.539) (P < 0.05), indicating less matrix staining in grafts refrigerated longer after harvest. CONCLUSION: The current data indicate that refrigerated osteochondral allografts can be maintained for up to 44 days with average chondrocyte viability of 67%.

Cartilage, Articular↗

[Factors affecting the temperature of domestic refrigerators].

A survey was carried out in France in 1999 in order to know the air temperature in domestic refrigerators and the factors which may effect this temperature. Temperatures were recorded at three levels (top, middle and bottom of the refrigerator compartment). A questionnaire was filled to acquire the following information: characteristic of family (number of family members, age, profession, income...), characteristic of refrigerator (trade, type, age, temperature setting, refrigerating type...) and the use condition (room temperature, near by heat source, built in, door opening frequency...). The average temperature of the 119 surveyed refrigerators was 6.6 degrees C. Descriptive analysis and multi dimensional analysis of factors effecting refrigerator temperature were carried out. The classification tree and the segmentation confirm the influence of the use condition (frequency of door opening, temperature setting, near by heat source and built in). There is no direct effect of one factor but the combination of all of them.

Equipment Design↗

Medications not to be refrigerated.

A survey of selected drug labelers was conducted to generate a list of drug products that should not be refrigerated. Letters asking for information on products adversely affected by refrigeration were mailed to 109 drug product labelers. A second letter was sent to nonresponders and to labelers providing incomplete information. Responses were received from 97 labelers, 43 of whom stated that none of their products would be harmed by refrigeration. Eleven labelers were unable to provide conclusive data or a list of specific drug products that refrigeration would harm. Lists of drug products not to be refrigerated were provided by 43 labelers, some including explanations of the adverse effects of refrigeration. Pharmacists may find the survey's data useful in their patient education activities.

Drug Labeling↗

Clinical and laboratory comparison study of refrigerated and cryopreserved bone marrow for transplantation.

Refrigerated storage for short-term preservation of bone marrow is an alternative to cryopreservation where chemotherapeutic regimens include drugs with short in vivo half-lives. We performed a clinical and laboratory comparison of bone marrow stored at 4 degrees C for up to 9 days to bone marrow cryopreserved at -90 degrees C for autotransplantation. After adjusting for the confounding effects of disease type or sex, no clinically meaningful variation in post-transplant course between refrigerated storage and cryopreserved was found. Therefore, the data presented in this study suggest that the clinical recovery indices following transplantation between the two storage groups are essentially equivalent. One potential advantage to refrigerated storage, however, is that it may provide an opportunity for extended exposure to growth factors and/or purging agents in vitro prior to transplantation. To prepare for an in vitro analysis of this hypothesis, we concentrated the stem cell population and compared the nucleated cell recovery, viability and colony forming potential following refrigerated storage of whole bone marrow and buffy coat to cryopreserved bone marrow stored for the same interval. While the nucleated cell recovery for cryopreserved marrow was significantly greater than for refrigerated storage, the viability and colony forming potential of the refrigerated storage was superior or equivalent, independent of prior processing.

Adolescent↗

Detection of Salmonella in dry foods using refrigerated pre-enrichment and enrichment broth cultures: interlaboratory study.

An interlaboratory study was performed in 11 laboratories to validate the use of pre-enrichment and tetrathionate brilliant green (TBG35) and selenite cystine (SC35) enrichment cultures refrigerated 72 h at 2-5 degrees C for greater analytical flexibility in the detection of Salmonella in dry foods. Productivities of refrigerated pre-enrichment and enrichment cultures were compared with that of the AOAC/Bacteriological Analytical Manual (BAM) procedure using 4 food types: whole egg powder, milk chocolate, animal feed, and instantized skim milk powder. Uninoculated and inoculated samples were included in each food group. There was complete agreement between the results obtained by the standard AOAC/BAM procedure and the 2 refrigeration procedures. Of 660 samples tested, the AOAC/BAM procedure identified 393 contaminated samples that were readily detected from the corresponding refrigerated pre-enrichment cultures and from the combined productivity of homologous refrigerated TBG35 and SC35 cultures. Refrigeration (72 h) of pre-enrichment or enrichment cultures for greater analytical flexibility and laboratory productivity in the examination of dry foods is under review for adoption by AOAC International.

Animal Feed↗

Analysis of volatiles in meat from Iberian pigs and lean pigs after refrigeration and cooking by using SPME-GC-MS.

The volatile compounds generated in meat from Iberian and lean pigs after four different treatments (raw, refrigerated, cooked, and refrigerated cooked meat) were analyzed. The different treatments showed different volatile profiles. Methyl alcohols and ketones (such as 2-ethyl-hexan-1-ol, 2-methyl-butan-1-ol, 3-methyl-butan-1-ol, and 3-hydroxy-butan-2-one) were the most representative in refrigerated meat because of the degradation of carbohydrates and proteins together with the Strecker degradation pathway. Lipid-derived volatiles were the most abundant in cooked meat and refrigerated cooked meat. Meat from different pig breeds presented different volatile profiles, probably due to different enzymatic and oxidative deterioration susceptibility. Otherwise, the fat content and its compositional characteristics also played an important role in the generation of volatiles. As compared to samples from lean pigs, muscles from Iberian pigs showed a higher content of heme iron that may have promoted the generation of higher content of total lipid-derived volatiles during the refrigeration of cooked meat. Despite that, the formation of volatiles with low thresholds and related to intense rancidity perception likely to be derived from polyunsaturated fatty acids was higher in lean pork than in meat from Iberian pigs. This might be expected to lead to a more intense development of a warmed over flavor during refrigeration of cooked samples from lean pigs.

Alcohols↗

Fault tree analysis for exposure to refrigerants used for automotive air conditioning in the United States.

A fault tree analysis was used to estimate the number of refrigerant exposures of automotive service technicians and vehicle occupants in the United States. Exposures of service technicians can occur when service equipment or automotive air-conditioning systems leak during servicing. The number of refrigerant exposures of service technicians was estimated to be 135,000 per year. Exposures of vehicle occupants can occur when refrigerant enters passenger compartments due to sudden leaks in air-conditioning systems, leaks following servicing, or leaks caused by collisions. The total number of exposures of vehicle occupants was estimated to be 3,600 per year. The largest number of exposures of vehicle occupants was estimated for leaks caused by collisions, and the second largest number of exposures was estimated for leaks following servicing. Estimates used in the fault tree analysis were based on a survey of automotive air-conditioning service shops, the best available data from the literature, and the engineering judgement of the authors and expert reviewers from the Society of Automotive Engineers Interior Climate Control Standards Committee. Exposure concentrations and durations were estimated and compared with toxicity data for refrigerants currently used in automotive air conditioners. Uncertainty was high for the estimated numbers of exposures, exposure concentrations, and exposure durations. Uncertainty could be reduced in the future by conducting more extensive surveys, measurements of refrigerant concentrations, and exposure monitoring. Nevertheless, the analysis indicated that the risk of exposure of service technicians and vehicle occupants is significant, and it is recommended that no refrigerant that is substantially more toxic than currently available substitutes be accepted for use in vehicle air-conditioning systems, absent a means of mitigating exposure.

Air Conditioning↗

Bacterial and fungal growth after freezing or refrigerating parenteral nutrient solutions.

Parenteral nutrient (PN) solutions were evaluated for growth of pathogenic organisms after refrigeration or freezing and then thawing. Sixteen bags of hypertonic dextrose and amino acid solutions were divided into two series (refrigerated and frozen), inoculated with Escherichia coli. Candida albicans, Staphylococcus aureus, or Streptococcus faecalis, and exposed to freezing or refrigeration. The inoculum concentration was greater than would likely occur with patient contamination of the solution. Microbial growth in the solutions was determined after warming to room temperature and at 17 or 18 hours after reaching room temperature. There was no increased growth of C. albicans in PN solutions that were frozen versus the refrigerated samples. Counts for all of the organisms in the frozen series, immediately after freezing and then thawing, decreased or stayed the same compared with baseline counts. Growth of E. coli, Staph. aureus, and Strep. faecalis increased in the frozen samples compared with the refrigerated samples after room-temperature storage, suggesting a possible increased risk of infectious complications if contaminated solutions are left at room temperature for extended periods. Since no increased risk of microbial growth is likely in frozen versus refrigerated PN solutions that are thawed and promptly infused, batch freezing may be an effective and convenient means of preparing PN solutions for home patients.

Bacteria↗

Effects of refrigeration on the biometry and development of Protophormia terraenovae (Robineau-Desvoidy) (Diptera: Calliphoridae) and its consequences in estimating post-mortem interval in forensic investigations.

The aim of this study was to simulate the low temperatures that insects could experience between the time being sampled from cadavers and their arrival in the laboratory. This was in order to investigate the effect of low temperature on development of maggots. At different stages of development, individuals of Protophormia terraenovae (Robineau-Desvoidy) reared at 24 degrees C were submitted to a temperature of 4.0+/-0.5 degrees C for a period varying from 1 to 10 days. Independent of the stage of development at which the insects were refrigerated, the treatment induced significant changes on the duration of development. The effect of low temperature on the developmental time between the return to 24 degrees C to adult emergence depended on the larval stage that was refrigerated. When first instar larvae and prepupae were refrigerated, the time to emergence at 24 degrees C decreased with an increase of duration of the refrigeration period. Time to emergence increased under the same conditions when second instar larvae and pupae were refrigerated. These results indicate that keeping larvae of P. terraenovae at 4 degrees C does not just simply lead to a cessation of metabolism but disturbs the regular development. Ten days of cooling induced an error in estimating post-mortem interval (PMI) of more than 6h.

Animals↗

Occupational exposure to fluorinated hydrocarbons during refrigeration repair work.

This study describes refrigeration repair workers' occupational exposures to halogenated refrigerants, focusing on difluorochloromethane (HCFC 22), tetrafluoroethane (HFC 134a) and a mixture of tri-, tetra- and pentafluoroethane (R404A) in 30 work operations. Unlike earlier reported studies, the present study includes working procedures involving welding in order to measure possible occupational exposure to decomposition products. The measurements included hydrogen fluoride (HF), hydrogen chloride (HCl), phosgene (COCl2) and volatile organic compounds (VOC). The exposures were assessed during work operations on small-scale cooling installations like refrigerators and freezers. The repair workers' occupational exposures to refrigerants were moderate, and the major part of the exposures were associated with specific working procedures lasting for relatively short periods of time (<20 min). During these exposure events the concentrations were occasionally high (up to 42434 mg m(-3)). Although welding operations lasted only for short periods of time, HF was detected in 9 out of 15 samples when HCFC 22, HFC 134a or R404A had been used. Hydrogen chloride was detected in 3 out of 5 samples in air polluted with HCFC 22. Phosgene was not detected. A large number of VOCs in various concentrations were found during welding. Except for the applied refrigerants, halogenated compounds were only found in one sample.

Adult↗

Low-temperature storage of bone marrow in nitrogen vapor-phase refrigerators: decreased temperature gradients with an aluminum racking system.

Large temperature gradients may exist in nitrogen vapor-phase refrigerators. Cryopreserved cells stored at higher levels may be considerably warmer than those stored closer to the liquid nitrogen reservoir. To decrease this temperature gradient, racking systems constructed of aluminum were placed in marrow storage refrigerators. The higher heat conductivity of aluminum resulted in a vapor-phase temperature gradient of only 5.9 degrees C at 22.5 inches above the liquid, as compared to the gradient of 86 degrees C seen with steel frames in a similar refrigerator. Temperature fluxes were minimal with lid opening or nitrogen filling. The thicker frame size and loss of the lowest storage level resulted in a storage capacity 63 percent of that achievable with steel frames and liquid immersion. Consumption of nitrogen was estimated to be 174 to 220 percent of the static usage in this model of refrigerator with 6 inches of nitrogen, but comparable to the consumption expected with full immersion of the racking system, regardless of frame construction. These data demonstrate the feasibility of achieving very low, stable, cryogenic temperatures in a vapor-phase refrigerator.

Aluminum↗

Impact of microbial ecology of meat and poultry products on predictions from exposure assessment scenarios for refrigerated storage.

A novel extension of traditional growth models for exposure assessment of food-borne microbial pathogens was developed to address the complex interactions of competing microbial populations in foods. Scenarios were designed for baseline refrigeration and mild abuse of servings of chicken broiler and ground beef Our approach employed high-quality data for microbiology of foods at production, refrigerated storage temperatures, and growth kinetics of microbial populations in culture media. Simple parallel models were developed for exponential growth of multiple pathogens and the abundant and ubiquitous nonpathogenic indigenous microbiota. Monte Carlo simulations were run for unconstrained growth and growth with the density-dependent constraint based on the "Jameson effect," inhibition of pathogen growth when the indigenous microbiota reached 10(9) counts per serving. The modes for unconstrained growth of the indigenous microbiota were 10(8), 10(10), and 10(11) counts per serving for chicken broilers, and 10(7), 10(9) and 10(11) counts per serving for ground beef at respective sites for backroom, meat case, and home refrigeration. Contamination rates and likelihoods of reaching temperatures supporting growth of the pathogens in the baseline refrigeration scenario were rare events. The unconstrained exponential growth models appeared to overestimate L. monocytogenes growth maxima for the baseline refrigeration scenario by 1500-7233% (10(6)-10(7) counts/serving) when the inhibitory effects of the indigenous microbiota are ignored. The extreme tails of the distributions for the constrained models appeared to overestimate growth maxima 110% (10(4)-10(5) counts/serving) for Salmonella spp. and 108% (6 x 10(3) counts/serving) for E. coli O157:H7 relative to the extremes of the unconstrained models. The approach of incorporating parallel models for pathogens and the indigenous microbiota into exposure assessment modeling motivates the design of validation studies to test the modeling assumptions, consistent with the analytical-deliberative process of risk analysis.

Animals↗

Marrow storage techniques: a clinical comparison of refrigeration versus cryopreservation.

Fifty-three patients were evaluated for a comparison of the efficacy, safety, and cost efficiency of bone marrow (BM) transplanted after either refrigeration or cryopreservation. Thirty-eight patients had BM stored at 4 degrees C for an average of 3 days and 15 patients had cryopreserved BM stored for an average of 56 days. The average number of cells harvested was 3.8 x 10(8)/kg. The time to WBC recovery greater than 1 x 10(9)/l was 17 days refrigerated and 23 days for cryopreserved BM. The time to platelet recovery greater than 20 x 10(9)/l was 24 days for refrigeration storage and 51 days for cryopreserved BM. Four of 38 patients with refrigerated vs. 4/15 patients with cryopreserved BM experienced delayed engraftment (p less than 0.05). Refrigeration storage requires no special equipment, is cheaper than and presents a safe and viable alternative to cryopreserved BM in reconstituting hemopoiesis following high-dose chemo-radiotherapy.

Adolescent↗

Effect of centrifuge speed, refrigeration medium, and sperm washing medium on cryopreserved sperm quality after thawing.

Cryopreservation and subsequent thawing of semen for assisted reproductive procedures decreases sperm motility; motility further reduces when the cryoprotectant medium is removed because of the osmotic shock and centrifugation done to prepare the sperm. To compare motility and thus pregnancy rates, this study examined the effects of adding an additional refrigeration medium and three different centrifugation speeds for sperm preparation. Semen samples from 16 healthy normal volunteers were obtained by masturbation after 48 h of abstinence. Sperm motility and other motion characteristics were analyzed with a computer-assisted semen analyzer before freezing, after thawing, and after centrifugation. Each sample was divided into 6 aliquots and frozen using the liquid nitrogen vapor method. After thawing, human tubal fluid (HTF) with 5% human serum albumin was added to 3 aliquots, and refrigeration medium (identical to freezing medium without glycerol) was added to the remaining 3 tubes for each specimen. The tubes containing the two media were then washed by centrifugation at 100 g, 300 g, and 500 g for 10 min. Aliquots with refrigeration medium did not significantly differ from those with HTF for percent motility, curvilinear velocity, straight-line velocity, and amplitude of lateral head displacement at any centrifugation speed. Motile sperm count was significantly greater only at 100 g and 300 g for refrigeration medium (p = .02 and .01) and HTF (p = 0.001); at 300 g, average path velocity in refrigeration medium aliquots (p = .01) and linearity in HTF (p = .01) were greater. The results indicated that the reduction in motility and other motion characteristics probably cannot be overcome by changing factors such as the sperm preparation medium or centrifugation speed. More effective cryopreservation techniques or preparation methods that eliminate centrifugation need to be developed.

Centrifugation↗

Food safety knowledge of consumers and the microbiological and temperature status of their refrigerators.

The objectives of this study were to examine domestic food safety knowledge levels of consumers, establish the levels and incidence of bacterial contamination and operational temperatures in domestic refrigerators, and identify areas in which consumer food safety education is necessary in Ireland. A food safety knowledge questionnaire applied to a representative sample of households (n = 1,020) throughout the island of Ireland found the gaps in consumer food safety knowledge. Analysis of swab samples (n = 900) recovered from the domestic refrigerators in these households showed average total viable counts of 7.1 log CFU/cm2 and average total coliform counts of 4.0 log CFU/cm2. Analysis of swab samples also detected the incidence of Staphylococcus aureus (41%), Escherichia coli (6%), Salmonella enterica (7%), Listeria monocytogenes (6%), and Yersinia enterocolitica (2%). Campylobacter jejuni and E. coli O157:H7 were not detected in domestic refrigerators. The temperature profiles of a subset of the sampled refrigerators (100) were monitored for 72 h, and 59% were found to operate, on average, at temperatures above the recommended 5 degrees C. Knowledge and temperature survey results varied considerably, but consumers who scored better in terms of basic food safety knowledge had reduced levels of bacterial contamination in their refrigerators and reported a reduced incidence of food-associated illnesses. This study confirms the effect of basic food hygiene knowledge on hygienic practice and identifies specific areasfor emphasis in the development and delivery of effective food safety risk communication messages to consumers.

Adolescent↗

Comparison of a rapid ATP bioluminescence assay and standard plate count methods for assessing microbial contamination of consumers' refrigerators.

The feasibility of using an ATP bioluminescence assay for assessing microbial contamination of home refrigerators was evaluated and compared with the standard culture methods. Samples of refrigerator surfaces were collected from 123 households by swabbing an area of 100 cm2 on three locations in the refrigerator with premoisturized sterile swabs. Microbial contaminations were determined by aerobic plate count (APC; incubated at 35 degrees C for 48 h) and psychrotrophic plate count (PPC; incubated at 7 degrees C for 10 days) on plate count agar. The results were compared to the readings from the microbial ATP (mATP) bioluminescence assay. The correlation coefficient (r) between mATP and PPC (r = 0.851) was slightly higher than that between mATP and APC (r = 0.823). Our results indicated a potential discrepancy in the population of mesophilic and psychrotrophic bacteria in the refrigerator samples. Nevertheless, mATP appeared to be a reliable indication of the average of APC and PPC (r = 0.895). The mATP bioluminescence assay would provide a rapid and convenient test for researchers in field studies to assess microbial contamination in refrigerators.

Adenosine Triphosphate↗

Effect of refrigeration on in vitro penetration of Salmonella enteritidis through the egg yolk membrane.

Internally contaminated eggs have been implicated as leading sources of transmission of Salmonella Enteritidis (SE) to humans. Although SE is not often deposited inside the nutrient-rich yolks of naturally contaminated eggs, penetration through the vitelline membrane to reach the yolk contents could result in rapid bacterial multiplication. In previous studies, such penetration has been observed occasionally at warm temperatures during experiments with in vitro egg contamination models. The present study was conducted to determine whether refrigeration affects the frequency of in vitro SE penetration of the egg yolk membrane. After inoculation of small numbers of SE onto the outside of the vitelline membranes of intact yolks, immediate refrigeration of contaminated samples prevented the penetration of SE into the egg yolk contents during 24 h of storage. However, SE penetrated inside the yolk contents in 4% of contaminated egg samples refrigerated after 2 h of storage at 30 degrees C, 15% of samples refrigerated after 6 h of storage at 30 degrees C, and 40% of samples stored at 30 degrees C for 24 h (48 samples per treatment group). These results highlight the value of prompt refrigeration for restricting the opportunities for SE to multiply to high numbers inside the yolks of contaminated eggs.

Animals↗