Thawing In Water Bath Research Articles

Effect of Water Bath versus Refrigerator Thaw on Cryoprecipitate Fibrinogen and Factor VIII Content Using a Pre-Pooled Plasma Experimental Approach

<b><i>Introduction:</i></b> Originally developed as a form of factor VIII concentrate, cryoprecipitate’s primary clinical use has changed to treat fibrinogen deficiency as highlighted by recent approval of pathogen-reduced cryoprecipitated fibrinogen concentrates. The methodology by which frozen plasma is thawed during cryoprecipitate manufacturing is not standardized. This study compared plasma thawing techniques on cryoprecipitate fibrinogen and factor VIII levels. <b><i>Methods:</i></b> A matched pairwise experimental design was employed across three experiments to compare plasma thawing approaches (water bath or 24–48 h refrigerator). Each experiment involved the creation of 10 sets of ten homogenous frozen plasma pools which were then used to manufacture 10 pairs of cryoprecipitate pools differing only by assigned plasma thawing method. Total cryoprecipitate fibrinogen and factor VIII content between plasma thawing methods were compared using matched <i>t</i>-testing within each experiment. <b><i>Results:</i></b> Compared to water bath thawing, 24-h refrigerator thawing led to significantly higher cryoprecipitate fibrinogen content (2,554 mg vs. 1,824 mg; <i>p</i> < 0.001) and significantly lower cryoprecipitate factor VIII content (601 IU vs. 709 IU; <i>p</i> < 0.001). Longer refrigerator thaw times (36 and 48 h) led to significantly higher cryoprecipitate fibrinogen content than 24-h refrigerator thaw (3,180 mg vs. 2,956 mg and 2,893 mg vs. 2,483 mg, respectively; <i>p</i> = 0.01–0.03). <b><i>Conclusion:</i></b> Using homogenous frozen plasma units in a matched pairwise experimental design, refrigerator plasma thawing led to superior cryoprecipitate fibrinogen yields and inferior cryoprecipitate factor VIII yields. When maximizing cryoprecipitate fibrinogen yields, refrigerator plasma thawing, and in particular longer thawing times (36–48 h), should be considered.

Effects of different thawing methods on physical and physicochemical properties of frozen dough and quality of corresponding steamed bread

The thawing method is critical for the final quality of products based on the frozen dough. The effects of ultrasound thawing, proofer thawing, refrigerator thawing, water bath thawing, ambient thawing, and microwave thawing on the rheology, texture, water distribution, fermentation characteristics, and microstructure of frozen dough and the properties of steamed bread were investigated. The results indicated that the ultrasound thawing dough had better physicochemical properties than other doughs. It was found that ultrasound thawing restrained the water migration of dough, improved its rheological properties and fermentation capacity. The total gas volume value of the ultrasound thawing dough was reduced by 21.35% compared with that of unfrozen dough. The ultrasound thawing dough displayed a thoroughly uniform starch-gluten network, and an enhanced the specific volume and internal structure of the steamed bread. In conclusion, ultrasound thawing effectively mitigated the degradation of the frozen dough and enhanced the quality of steamed bread.

Use of Powdered Milk in Semen Cryopreservation Protocols for Fish: A Systematic Review.

This systematic review provides an overview of the history and current status of cryopreservation of fish sperm and a detailed evaluation of cryoprotocols using powdered milk. A literature search was performed in PubMed, Scopus, Web of Science, and SciELO databases. Twenty-nine articles were selected after excluding duplicate articles or articles that did not meet the eligibility criteria. Rhamdia quelen and Danio rerio were the most studied species. Slow freezing method, dry-shipper, freezing rate of -35.6°C/min, thawing in water bath (35.93°C ± 10°C), and 0.25 and 0.5 mL plastic straws were the main approaches evaluated. Methanol was the most used permeable cryoprotectant in combination with powdered milk, yielding the best results at 10% concentration. Motility rate was the main analysis performed after cryopreservation in virtually all studies, being subjectively evaluated by most authors. Powdered milk at 15% promoted the best results in the analyzed studies. For motility rate, the gains with the addition of powdered milk were observed in the orders Perciformes (Oreochromis mossambicus), Siluriformes (Pangasius pangasius, Pseudoplatystoma corruscans, and Pseudoplatystoma mataense), and Cypriniformes (Tor soro and Barbonymus gonionotus). For fertilization, gains were observed in the order Siluriformes (P. mataense) and Cypriniformes (T. soro). Sperm viability gains were observed in the orders Siluriformes (P. pangasius), Characiformes (Piaractus brachypomus), and Cypriniformes (B. gonionotus). The scientific evidence we present in this study may contribute and serve as a starting point for new and more refined studies to be developed in the field.

Study on the Flavor Compounds of Fo Tiao Qiang under Different Thawing Methods Based on GC-IMS and Electronic Tongue Technology.

“Fo Tiao Qiang” is a famous dish with Chinese characteristics. It is delicious, rich in materials, and high in nutritional value. Through physical and chemical analysis, electronic tongue, gas chromatography–ion mobility spectroscopy, and other technologies, the present study explored the quality characteristics and flavor differences of Fo Tiao Qiang by using different thawing methods (natural thawing, ultrasonic thawing, microwave thawing, and water bath thawing). The results show that the protein content was slightly higher in Fo Tiao Qiang with ultrasonic thawing than others. The fat content of the microwave-thawed Fo Tiao Qiang was significantly lower than the other three kinds of samples. After ultrasonic thawing, the number of free amino acids in the samples were the highest and the umami taste was the best. Compared with natural thawing, most of the flavor substances decreased in ultrasonic thawing, microwave thawing, and water bath thawing. However, several substances increased, such as alpha-terpineol, beta-phenylethyl alcohol, phenylacetaldehyde, cis-rose oxide, isobutyl acetate, and 2–3-pentanedione. This study revealed the changing laws of different thawing methods on the quality characteristics and flavor characteristics of Fo Tiao Qiang. It provides theoretical guidance for the industrial production and quality control of Fo Tiao Qiang.

Fe3O4 Nanoparticles with Carboxylic Acid Functionality for Improving the Structural Integrity of Whole Vitrified Rat Kidneys

The successful rewarming of cryopreserved organs has always been a big challenge for the cryopreservation technology aimed at improving the shortage of available organs for transplantation. The traditional water bath rewarming produces more obvious devitrification and thermal stress damages, resulting in negative effects on the organ structure and physiological function. Nanowarming technology via induction heating of iron oxide nanoparticles for rewarming large-volume frozen biosamples is a promising strategy to overcome related problems. In this study, Fe3O4 nanoparticles modified with carboxylic acid are used for nanowarming to rewarm the whole frozen kidney. The key steps including loading and elution of the cryoprotectant and magnetic nanoparticles (mNPs), vitrification of large-volume samples, and nanowarming of the whole kidney are explored in detail to achieve whole kidneys with a more integrated structure. Compared with water bath thawing, the nanowarming method could reduce the maximum thermal stress of the whole kidney by 2 orders of magnitude and has enough rewarming rate to avoid the devitrification phenomenon, so as to obtain a lower cell apoptosis rate, a more integrated vascular network, and a lower residual amount of mNPs inside the kidneys after elution. The optimization of the nanowarming method for the whole kidney could provide effective guidance for the large organ cryopreservation in clinical transplantation.

Adherent cell thawing by infrared radiation

Cryopreservation of adherent cells is crucial for commercial cell therapy technology, including effective distribution and storage. Fast thawing has been shown to increase cell recovery in vitrified samples. Previously, radiofrequency (RF) has been investigated as a heating source on large samples, either with or without magnetic particles. Also, laser heating with the aid of dye or nanoparticles has been utilized on sub–millimeter samples successfully. For slow freezing cryopreservation methods, the influence of rate of thawing on viability is less clear. Cryopreservation of surface adhered cells result in many cases in detachment from the surface. We illustrate how intense infrared radiation from a focused halogen illuminator accelerates thawing. We show that two epithelial cell lines, retinal pigment epithelium cells and heterogeneous human epithelial colorectal adenocarcinoma cells, can be effectively cryopreserved and recovered using a combination of slow freezing and fast thawing under infrared illumination. We were able to successfully thaw samples, of 2–4 mm thick, including the media, on the order of a second, providing a heating rate of thousands of Kelvin per minute. Under optimal conditions, we observed higher post–thawing cell viability rates and higher cell adhesion with infrared thawing than with water bath thawing. We suggest that bulk warming with infrared radiation has an advantage over surface warming of surface–attached cells, as it alleviates cell stress during the process of thawing. These findings will pave the way for novel approaches to treating substrate–adhered cells and 3D scaffolds with cells and organoids. This technology may serve as a crucial component in lab–on–chip systems for medical testing and therapeutic use.

Effects of different thawing methods on flavor compounds and sensory characteristics of raspberry

AbstractIn order to identify more suitable thawing method to preserve aroma, taste, and other sensory quality of raspberry, five thawing methods were used to treat frozen raspberry, namely water bath thawing, microwave thawing, ultrasonic thawing, room temperature thawing, and refrigeration thawing. Gas chromatography‐mass spectrometry was employed to study the quantitative and qualitative analysis of the volatile compounds, and the characteristic flavor components of raspberry were compared by relative odor activity value (ROAV). Electronic tongue technique and fuzzy comprehensive evaluation were used to ensure the changes in sensory qualities of raspberry. The results showed that the time of microwave thawing was 0.57 minutes, and drip loss was 4.40%. Both of them were the lowest among the five thawing methods. The volatile components of raspberry included aldehydes, esters, ketones, alcohols, fatty acids, terpenes, and other compounds. Among them, there were 19 kinds of volatile components in quick‐frozen raspberries, 18 volatile components in raspberries after treated by microwave thawing and refrigeration thawing, 16 volatile components after room temperature thawing, 12 volatile components after ultrasonic thawing, and 10 volatile components after water bath thawing. The characteristic flavor compounds of raspberry were identified as acetaldehyde, alpha‐ionone, beta‐ionone, raspberry ketone, alpha‐terpene, and alpha‐pinene. Microwave thawing was the most suitable way to preserve the characteristic flavor components of raspberry, but water bath thawing could destroy the most of compounds. After microwave thawing, raspberry tasted best and was most acceptable to consumers. Therefore, microwave thawing was the most appropriate method to preserve raspberry flavor and sensory quality among the five thawing methods.

Effect of Oral Administration of Selenium and Vitamin E on the Quality of Fresh, Refrigerated and Frozen Semen in French Bulldog Breed Dogs

Background: New methodologies have been developed seeking to maximize pregnancy rate in female dogs created in commercial kennels, and also in order to maintain the quality of canine semen after dilution, refrigeration or freezing. One of the main factors that generate damage to sperm is oxidative stress, to minimize sperm damage, selenium and antioxidants like vitamin E are administered, by oral administration, seeking to improve the quality of semen. The objective was to study the effect of vitamin E and selenium, by oral administration, in the quality of fresh, refrigerated and frozen semen in adult dogs French Bulldog breed.Materials, Methods & Results: Semen samples were collected from 5 adult dogs, French Bulldog breed, being 2 semen drawing before the daily oral supplementation with vitamin E and selenium (ESE®) and semen drawing at 20, 40 and 60 days after the beginning of oral supplement. The ejaculated samples were diluted in TRIS - fructose citric acid (3.28 g TRIS-hydroxy-methyl-amino-methane, 1.78 g of citric acid monohydrate and 1.25 g of D - fructose, dissolved in 100 mL of distilled water and added of 20% egg yolk and 6% of glycerol. The characteristics evaluated in fresh semen were: volume (mL), color, appearance, concentration (x106 / mL), sperm motility (%), sperm strength (1 to 5) and morphology (%). For refrigerated and frozen semen were analyzed: sperm motility (%), sperm strength (1-5) and morphology (%). Diluted semen samples were centrifuged at: 1500 g/10 min and “pellets” formed by sperm of each ejaculated, detached from the tube wall were diluted homogeneously in the diluent TRIS type up to the final volume of 1.5 mL. After that, packaged in 0.5 mL French straws, kept under refrigeration at 5ºC/4 h, placed in nitrogen vapor at -120ºC/15 min, and dipped in liquid nitrogen at -196ºC and then stored on identified rachis and stored in liquid nitrogen container until the time of thawing in water bath at 37°C/30 s for semen microscopic analysis. Data from fresh, refrigerated and frozen semen were statistically analyzed by analysis of variance and the average compared by 5% of Tukey test. Fresh semen sperm concentration differed (P < 0.05) between the samples, rising after 40 days after the beginning of oral supplementation with selenium and vitamin E. For the spermatic strength, better score (P < 0.05) was observed at collection 4, in 40 days after the beginning of oral supplementation to dogs. For fresh and refrigerated semen, the total defects, defects of head, acrosome and tail did not differ (P > 0.05) between the samples. Total sperm defects and minor head and tail defects did not differ (P > 0.05) between the samples in post-thawing. Regarding the acrosome defects after thawing, there was a significant reduction (P < 0.05) in samples performed 40 and 60 days after the beginning of oral supplementation with selenium and vitamin E.Discussion: Attention should be paid for what purpose the extenders within the refrigeration or freezing biotech will be used. The managed supplement, by oral administration, containing selenium and vitamin E, influenced beneficially raising the sperm concentration in fresh semen and decreasing the acrosome defects in frozen semen. Oral administration of supplementation with selenium and vitamin E is recommended for improving the quality of fresh and frozen semen in dogs.

Revisiting crucial steps of an encapsulation/desiccation based cryopreservation process: importance of thawing method in the case of Pelargonium meristems

This study questioned (1) the need for progressive osmotic desiccation (2) the efficiency of a new thawing method using 0.75 M sucrose liquid medium (3) the effect of various evaporative desiccation rates, in an encapsulation/desiccation cryopreservation process established for Pelargonium meristems. Whether progressive or not, osmotic desiccation in presence of high sucrose concentrations (up to 1 M) was equally efficient in the induction of meristem tolerance to further evaporative desiccation. The latter step was necessary for meristems to reach a water content window compatible with cryopreservation. This window was dependent on the thawing regime. Indeed, when frozen at relatively high water content (0.56 g g −1 DW), high meristem survival rates (30–54%) were recorded only after sucrose-thawing. Moreover, at relatively lower water content (0.30 g g −1 DW or lower), sucrose-thawing resulted in significantly higher survival rates than the traditional ambient- and 40 °C water bath-thawing. Irrespective of evaporative desiccation rates, meristem viability after dehydration to an approximate water content of 0.30 g g −1 DW, was similar. Additional cryopreservation of such dehydrated meristems did not affect their survival. The beneficial effect of sucrose-thawing on residual freezable water behaviour (detected via differential scanning calorimetry (DSC)) is discussed.

Microwave technology for the rapid thawing of frozen blood components.

Based on a continuing need to provide a more rapid response to requests for thawed fresh frozen plasma, the authors evaluated plasma thawing with the use of a microwave oven and compared it with conventional 37 degrees C waterbath thawing methods. Their results indicate that microwave-thawed plasma contains precipitated denatured protein (mainly albumin and fibrinogen) and that there is a significant reduction of coagulation Factors IX, X, XI, and fibrinogen compared with fresh plasma. They also measured levels of di-ethyl hexyl phthalate after microwave thawing and found its rate of accumulation similar to that of the 37 degrees C waterbath. More importantly, fundamental principles of microwave heating preclude uniform temperatures being maintained throughout the thawing of plasma; hence, the denaturation of plasma proteins is expected to occur under even low heating conditions.

Microwave thawing of tissue culture cells

A comparison is made between microwave and water-bath thawing on the survival of tissue culture cells which have been frozen to −79 °C, with DMSO and HES protective agents (freezing rate: 20 °C per min). The microwave thawing rates, selected in the range 100–300 °C/min, are comparable to those achieved with a water bath (115 °C/min). Microwave thawing is stopped at various temperatures between 0 and 40 °C, and survival, determined by cell plating indicated that the microwave thawing is acceptable. Survival rates tend to be highest when the rapid microwave thawing terminated at temperatures just above 0 °C.