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Gas fracturing technology for enhancing rock permeability is an area with considerable potential for development. However, the complexity and variability of underground conditions mean that a variety of rock physical parameters can affect the outcome of gas fracturing, with temperature being a critical factor that cannot be overlooked. The presence of a temperature field adds further complexity to the process of gas-induced rock fracturing. To explore the effects of temperature fields on gas fracturing technology, this paper employs numerical simulation software to model the extraction of shale gas under different temperature conditions using gas fracturing techniques. The computer simulations monitor variations in the mechanical characteristics of rocks during the process of gas fracturing. This analysis is performed both prior to and following the implementation of a temperature field. The results demonstrate that gas fracturing technology significantly improves rock permeability; temperature has an impact on the effectiveness of gas fracturing, with appropriately high temperatures capable of enhancing the fracturing effect. The temperature distribution plays a crucial role in influencing the results of gas fracturing. When the temperature is low, the fracturing effect is diminished, resulting in a lower efficiency of shale gas extraction. Conversely, when the temperature is high, the fracturing effect is more pronounced, leading to a higher shale gas production efficiency. Optimal temperatures can enhance the efficacy of gas fracturing and consequently boost the efficiency of shale gas extraction. Changes in the parameters of the rock have a substantial impact on the efficiency of gas extraction, and selecting suitable rock parameters can enhance the recovery rate of shale gas. This paper, through numerical simulation, investigates the influence of temperature on gas fracturing technology, with the aim of contributing to its improved application in engineering practices.

Nanomaterials with unique structural and properties can be synthesized by rapid transition of the thermodynamic state. One promising method is through electrical explosion, which possesses ultrafast heating/quenching rates (dT/dt~109 K/s) of the exploding conductor. In this study, experiments were performed with fine metallic wire exploding in liquid nitrogen (liq N2, 77 K) under different applied voltages. For the first time in the literature, the physical image of the electrical explosion dynamics in liq N2 is depicted using electro-physical diagnostics and spatial-temporal-resolved photography. Specifically, the pulsation and collapse processes of the vapor bubble (explosion products) have been carefully observed and analyzed. As a comparison, an underwater electrical explosion was also performed. The experimental results suggest that the vapor bubble behavior in liq N2 differs from that in water, especially in the collapse phase, characterized by secondary small-scale bubbles in liq N2, but multiple bubble pulses in water; correspondingly, the products’ characteristics are discrepant.

To investigate the effect of liquid nitrogen on the granite failure process, the deterioration effect of liquid nitrogen on heated granite was investigated from experimental and theoretical perspectives. The mechanical properties of heated granite (25, 100, 200, 300, and 400 °C) after different cooling treatments (air cooling and liquid nitrogen cooling) were investigated by uniaxial compression tests. The damage evolution analysis was performed by a statistical damage constitutive model and the dissipation energy ratio was newly defined. The results show that there is an increase in the uniaxial compressive strength of heated granite before 200 °C, which is due to the competitive relationship between the thermal cracking and crack closure. Liquid nitrogen cooling can deteriorate the mechanical properties of heated granite in terms of strength and deformability. At 400 °C, the reduction rates of compressive strength and stiffness between air cooling and liquid nitrogen cooling reached 32.36% and 47.72%, respectively. Liquid nitrogen cooling induces greater initial thermal damage and, consequently, leads to a greater degree of total damage before the peak stress and makes rock easier to be damaged. At 400 °C, the total damage at the peak stress increased from 0.179 to 0.587 after the liquid nitrogen cooling. The difficulty of damage can be quantified by the dissipation energy ratio. In addition, the deterioration of liquid nitrogen on granite is positively related to temperature. This study confirmed the deterioration effect of liquid nitrogen and promoting effect of temperature, providing a theoretical approach to the degradation mechanism of liquid nitrogen.

An experimental bench was constructed for liquid nitrogen spray cooling in an open loop system, and the combined effect of the spray height, heat sink surface diameter, and spray mass flow on the heat transfer characteristics of liquid nitrogen spray cooling was studied by comparing 50 sets of experimental data. In general, the heat transfer performance decreased with an increase in the spray height and heat sink surface diameter, with the latter’s influence being more dominant. In the experiment, when the heat sink surface diameter increased to 24 mm, the influence of the spray height on the heat transfer performance decreased. The maximum critical heat flux and maximum heat transfer coefficient were 191.90 W·cm−2 and 16.67 W·cm−2·K−1.

Shale gas is a promising new energy source stored in shale. This research aims to study the laws of rock crack initiation and propagation under the low-temperature fracturing of liquid nitrogen, explore the influencing factors of the shale reservoir fracturing effect, and identify the method that achieves the best fracturing effect and obtains the highest economic benefits. Herein, a visualized physical experiment of the liquid nitrogen effect is carried out, and the fracture process of a numerical model under cold shock is simulated to analyze the effect of homogeneity on shale crack propagation. The results show that two different crack development modes could be observed in the field test. The first one was the horizontal plane radial crack caused by longitudinal thermal shrinkage, and the other one was the vertical tensile crack caused by circumferential shrinkage. A certain interval length was frequently found between the horizontal cracks. The crack propagation of the specimens with different homogenization degrees was basically distributed in the direction perpendicular to the liquid nitrogen contact surface. When the homogenization degrees were m = 2 and 5, the crack surface was rough and the microfracture was disordered and dotted around the crack tip. When m ≥ 10, the dotted damage around the crack tip did not appear, and the crack propagation was close to the results obtained from using the homogeneous materials. Finally, this work simulates the fracture process of a circular hole plane model under cold shock, analyzes the influences of heat transfer coefficient, in situ stress and other parameters on shale temperature, minimum principal stress distribution, and crack propagation, and discusses the optimal method to improve the heat transfer coefficient. The results show that increasing the heat transfer coefficient can increase the tensile stress value and influence the range of the contact boundary, making the rock more prone to cracking and resulting in greater crack development and a better crack initiation effect. The lateral stress coefficient affects the propagation direction of the cracks, and the propagation depths of low-temperature cracks were found to be deeper in the direction of larger principal stress.

In the present study, a flow visualization experiment in the clearance between a clear floating-ring and a rotating journal was carried out in order to clarify the effects of the recess geometry on the flow characteristics. Five types of the floating-rings with different recess geometries, i.e., the plane rhombus, the plane circle, the revise circular Young Leaf Mark, the dome and the revise dome geometries, were operated at a rotational speed of up to 40,000 rpm using liquid nitrogen as the working fluid to observe the interaction between source flow from the recess and rotating flow induced by the journal rotation in the clearance. The cavitation cloud inside the clearance was induced by viscous frictional heating and the pressure drop. The influence of the sweepback angle (θs) of the recess leading edge on the flow characteristics was investigated by comparing flow coefficient (Cf) and the cavitation cloud area ratio (Ac) obtained from the visual image. Based on the experimental results, it may be confirmed that the effects of the recess geometry on the flow characteristics of the cryogenic hybrid journal bearings are mainly caused by the influence of the θs value on the flow condition.

Cooling in liquid nitrogen is a typical service condition of high-temperature superconducting wire, and the variation of boiling stages on the wire protective layers such as the brass layers could be crucial for the quench behavior of superconducting devices. In this study, the influence of brass surface morphology (parameters of surface roughness and fractal dimension) on the Leidenfrost effect (including the wall superheat at critical heat flux and the wall superheat at Leidenfrost point, which are respectively characterized by the temperatures of ΔTCHF and ΔTLP) was studied. The surfaces of brass samples were polished by sandpaper to obtain different morphologies, which were characterized by using white light interferometer images, and the boiling curves were recorded and analyzed by Matlab with lumped parameter method. The experimental results demonstrated that the surface morphology of brass samples could influence the ΔTLP significantly, but had no clear relationship with the ΔTCHF. Moreover, the multi-scaled analysis was carried out to explore the influencing mechanism of surface microstructure, the relationship between ΔTLP and scale was more clear when the scale was small, and the fractal dimension was calculated and discussed together with surface roughness. The findings of this study could be instructive for surface treatment of superconducting wires to suppress quench propagation.

This study experimentally explores the thermofluidic performance of a cryogenic micro-pin fin cooler with two-phase liquid nitrogen flows. The liquid nitrogen cooling system is introduced to investigate the performance of the micro-pin cooler in a cryogenic condition. The result reveals that the nominal value of the base heat transfer coefficients of the micro-pin fin cooler with liquid nitrogen flows, 240 kW/m2-K at a mass flow rate of 2.23 g/s, is an order of magnitude greater than that with FC-72 flows. The result also demonstrates that the base heat transfer coefficient of the micro-pin fin cooler is nearly three times greater than that of the micro-gap cooler, not containing any fins. This study shows the feasibility of the cryogenic micro-pin fin cooler for thermally controlling very high heat density devices such as high-power laser diode bars, of which the heat density can reach 2000 kW/m2.

In this research work, the machinability of turning Hastelloy X with a PVD Ti-Al-N coated insert tool in dry, wet, and cryogenic machining environments is investigated. The machinability indices namely cutting force (CF), surface roughness (SR), and cutting temperature (CT) are studied for the different set of input process parameters such as cutting speed, feed rate, and machining environment, through the experiments conducted as per L27 orthogonal array. Minitab 17 is used to create quadratic Multiple Linear Regression Models (MLRM) based on the association between turning parameters and machineability indices. The Moth-Flame Optimization (MFO) algorithm is proposed in this work to identify the optimal set of turning parameters through the MLRM models, in view of minimizing the machinability indices. Three case studies by considering individual machinability indices, a combination of dual indices, and a combination of all three indices, are performed. The suggested MFO algorithm’s effectiveness is evaluated in comparison to the findings of Genetic, Grass-Hooper, Grey-Wolf, and Particle Swarm Optimization algorithms. From the results, it is identified that the MFO algorithm outperformed the others. In addition, a confirmation experiment is conducted to verify the results of the MFO algorithm’s optimal combination of turning parameters.

A simple process to synthesize metal nitride particles was proposed using submerged arc discharge plasma in liquid nitrogen. Gibbs standard free energy was considered for the selection of the nitride-forming materials. In this study, titanium (Ti) and aluminum (Al) electrodes were used as raw materials for nitride particle preparation. Liquid nitrogen acted as a dielectric medium as well as a nitridation source in this process. A copper electrode was also used as a non-reactive material for comparison with the reactive Ti and Al electrodes. As the operating conditions of the experiments, the arc discharge current was varied from 5 A (low-power mode) to 30 A (high-power mode). The formation of titanium nitride (TiN) and aluminum nitride (AlN) was confirmed in the particles prepared in all experimental conditions by X-ray powder diffraction (XRD). The observation using a field emission scanning electron microscope (FE-SEM) and a field emission transmission electron microscope (FE-TEM) indicated that the synthesized TiN particles showed a cubic morphology, whereas AlN particles containing unreacted Al showed a spherical morphology. The experiments using different metal electrode configurations showed that the anode generated most of the particles in this process. Based on the obtained results, a particle formation mechanism was proposed.

In this paper, the extended finite element method (XFEM) is first applied to account for the weak discontinuity of the axisymmetric electrostatic field. Firstly, the interface between two materials in an element is described by the level set method. The enrichment function is used to modify the shape function of enrichment elements. Secondly, to illustrate the feature of the enrichment function, the distribution diagrams of enrichment functions in sub-elements are drawn. The 3D field can be simplified to an axisymmetric field, which can reduce the difficulty of calculation. Finally, models with bubbles in liquid nitrogen in the axisymmetric field are used to prove the reliability of XFEM. Compared with the conventional finite element method (CFEM), XFEM costs lower computing resources with almost the same computational accuracy.

Efficient RNA extraction methods are needed to study transcript regulation. Such methods must lyse the cell without degrading the genetic material. For cyanobacteria this can be particularly challenging because of the presence of the cyanobacteria cell envelope. The great breath of cyanobacterial shape and size (unicellular, colonial, or filamentous multicellular) created a variety of cell lysis methods. However, there is still a lack of reliable techniques for nucleic acid extraction for several types of cyanobacteria. Here we designed and tested 15 extraction methods using physical, thermic or chemical stress on the filamentous cyanobacteria Planktothrix agardhii. Techniques based on the use of beads, sonication, and heat shock appeared to be too soft to break the Planktothrix agardhii cell envelope, whereas techniques based on the use of detergents degraded the cell envelope but also the RNA. Two protocols allowed to successfully obtain good-quality RNA. The first protocol consisted to manually crush the frozen cell pellet with a pestle and the second was based on the use of high-intensity ultra-sonication. When comparing these two, the high-intensity ultra-sonication protocol was less laborious, faster and allowed to extract 3.5 times more RNA compared to the liquid nitrogen pestle protocol. The high-intensity ultra-sonication protocol was then tested on five Planktothrix strains, this protocol allowed to obtain >8.5 μg of RNA for approximatively 3.5 × 108 cells. The extracted RNA were characterized by 260/280 and 260/230 ratio > to 2, indicating that the samples were devoid of contaminant, and RNA Quality Number > to 7, meaning that the integrity of RNA was preserved with this extraction method. In conclusion, the method we developed based on high-intensity ultra-sonication proved its efficacy in the extraction of Planktothrix RNA and could be helpful for other types of samples. [ABSTRACT FROM AUTHOR]

Chronic wound infections are increasingly recognized to be dynamic and polymicrobial in nature, necessitating the development of wound models which reflect the complexities of infection in a non-healing wound. Wound slough isolated from human chronic wounds and transferred to mice was recently shown to create polymicrobial infection in mice, and there is potential this tool may be improved by cryogenic preservation. The purpose of this study was to investigate the application of cryogenic preservation to transferring polymicrobial communities, specifically by quantifying the effects of cryopreservation and wound microbiome transplantation. Slough from an established murine polymicrobial surgical excision model and five patients were subjected to three preservation strategies: refrigeration until infection, freezing in liquid nitrogen, or freezing in liquid nitrogen with glycerol solution prior to infection in individual mice. Four days following inoculation onto mice, wound microbiota were quantified using either culture isolation or by 16s rRNA gene community profiling and quantitative PCR. Cryogenic preservation did not significantly reduce bacterial viability. Reestablished microbial communities were significantly associated with patient of origin as well as host context (i.e., originally preserved from a patient versus mouse infection). Whereas preservation treatment did not significantly shape community composition, the transfers of microbiomes from human to mouse were characterized by reduced diversity and compositional changes. These findings indicated that changes should be expected to occur to community structure after colonization, and that compositional change is likely due to the rapid change in infection context as opposed to preservation strategy. Furthermore, species that were present in higher relative abundance in wound inoculate were more likely to colonize subsequent wounds, and wound inoculate with higher bacterial load established wound communities that were more compositionally similar. Results inform expectations for the complementation of chronic wound in vivo modeling with cryogenic preservation archives. [ABSTRACT FROM AUTHOR]

Cryopreservation is the most common method for long-term cell storage. Successful cryopreservation of cells depends on optimal freezing conditions, freezer storage and a proper thawing technique to minimize the cellular damage that can occur during the cryopreservation process. These factors are especially critical for sensitive stem cells with a consequential and significant impact on viability and functionality. Until now, slow-freezing has been the routine method of cryopreservation but, more recently rapid-cooling techniques have also been proposed. In this study, an ultra-rapid cooling technique [] was performed for the first time on human mesenchymal stem cells and the effectiveness evaluated in comparison with the conventional slow-freezing procedure. A thin nylon-membrane carrier was used combined with different cryoprotective agents: dimethyl sulfoxide, ethylene glycol and/or trehalose. Various aspects of the low cryoprotective doses and the ultra-rapid cooling procedure of the human mesenchymal stem cells were examined including: the physical properties of the nylon-support, cells encumbrance, viability, proliferation and differentiation. The expression of cell surface markers and apoptosis were also investigated. The study used an ultra-rapid cooling/warming method and showed an overall cell integrity preservation (83–99%), with no significant differences between dimethyl sulfoxide or ethylene glycol treatment (83–87%) and a substantial cell viability of 68% and 51%, respectively. We confirmed a discrepancy also observed by other authors in cell viability and integrity, which implies that caution is necessary when assessing and reporting cell viability data. [ABSTRACT FROM AUTHOR]

Human sample, from patients or healthy donors, are a valuable link between basic research and clinic. Especially in translational research, they play an essential role in understanding development and progression of diseases as well as in developing new diagnostic and therapeutic tools. Stored in biobanks, fast access to appropriate material becomes possible. However, biobanking in a clinical context faces several challenges. In practice, collecting samples during clinical routine does not allow to strictly adhere to protocols of sample collection in all aspects. This may influence sample quality to variable degrees. Time from sample draw to asservation is a variable factor, and influences of prolonged storage at ambient temperature of tissues are not well understood. We investigated whether delays between 5 minutes and 3 hours, and the use of RNAlater RNA-preserving reagent would lead to a relevant drop in sample quality, measured by quantitative mRNA expression analysis. Our findings suggest that even under ambient conditions, delays up to 3 hours do not have a major impact on sample quality as long as the tissue remains intact. [ABSTRACT FROM AUTHOR]

Flavonoids are secondary metabolites of plants that often have medical applications. The influences of different sample drying pretreatments on flavonoids and antioxidant activity of ferns have not studies. Dryopteris erythrosora leaves used to analyze flavonoid alterations resulting from drying pretreatments. The total flavonoid content of D. erythrosora leaves exposed to different pretreatments was significantly different. The total flavonoid content of samples initially air-dried in shade and then oven-dried at 75°C were the highest (7.6%), while samples initially dried at 75°C had the lowest content (2.17%). Antioxidant activities of D. erythrosora leaves with different pretreatments varied. Group B first air-dried in the shade and then oven-dried at 75°C and group C first air-dried in the sun and then oven-dried at 75°C, both showed relatively stronger antioxidant activity. The best pretreatment for preserving the flavonoids was to first dry the plant material in the shade and then complete the drying process in an oven at 75°C. It was tentatively identified 22 flavonoids among the four different pretreatments by HPLC-ESI-TOF-MS. [ABSTRACT FROM AUTHOR]

A theoretical model has been derived to examine the dissociation of shock-compressed liquid nitrogen. An important feature of the model is the introduction of a binding energy for the atomic state which leads to a volume-dependent dissociation energy. Calculated shock pressures and shock temperatures are in agreement with experiment. The model correctly predicts the highly unusual feature of a reflected shock point lying above the principal Hugoniot and of a decrease in the temperature of a reflected shock. The shock cooling is shown to be the consequence of the volume dependent dissociation energy. The overall transition is from a molecular to a metallic liquid. [ABSTRACT FROM AUTHOR]

In the present study, a flow visualization experiment in the clearance between a clear floating-ring and a rotating journal was carried out in order to clarify the effect of the sweepback angle (θs) of the leading edge of the recess on the flow characteristics. Four types of the floating-rings with different recess geometries, i. e., the Young Leaf Mark, the square and the non-recessed geometries, were operated at a rotational speed of up to 45,000 rpm using liquid nitrogen as the working fluid to observe the interaction between source flow from the recess and rotating flow induced by the journal rotation in the clearance. The cavitation cloud inside the clearance was induced by viscous frictional heating and the pressure drop. The influence of the θs value on the flow characteristics was investigated by comparing flow coefficient (Cf) and the cavitation cloud area ratio (Ac) obtained from the visual image. Based on the experimental results, the dependence of the flow characteristics on the rotational speed was confirmed to be affected by the sweepback angle of the leading edge of the recess.

Giant cell tumor (GCT) of bone is a benign aggressive bone lesion with significant recurrence rates following surgical curettage. Historically, these tumors were approached by performing an intralesional curettage of the tumoral tissue by filling the resulting cavity using morselized iliac bone autograft. The major problems of this therapy were the high recurrence rates of up to 40–50%. Several adjuvant treatments have been proposed in order to augment resection margins, including liquid nitrogen (LN), phenol, ethanol, hydrogen peroxide (H2O2) and bone cement (polymethyl methacrylate (PMMA)). LN can be used either to preserve tissues or for controlled necrosis depending on the cycles of freezing and thawing. Usually, a quick freeze followed by a slow thaw will lead to destruction of human cells. This article reviews the results of cryosurgery with LN associated with surgical resection and the additional use of PMMA in a small group of patients with a histopathological confirmation of bone GCT with different localizations (i.e., tibia, distal radius and iliac bone). Cryosurgery with LN of bone GCT proved to be an efficient tool to decrease the recurrence rate for this tumoral type. In our series of cases, there were no complications, oncological or otherwise, at the two-year minimum follow-up, with good and excellent functional results.

Agriculture has always been an important part of human evolution. Traditionally, farming is changing and developing with regard to challenges it faces. The major challenges of modern agriculture are food and nutrition safety for the growing world population. Promoting species and genetic diversity in agriculture appears to be an important approach to dealing with those challenges. Gene banks all around the world play a crucial role in preserving plant genetic resources for future crop improvements. The plant germplasm can be preserved in different ways, depending on the species or form of stored plant tissue. This review focuses on a special preservation method—cryopreservation. Cryopreservation is an effective technique for storing living systems at ultra-low temperatures, usually in liquid nitrogen or its vapor phase. This conservation method is crucial for plants that do not produce seeds or that produce non germinating seeds, as well as for plants that propagate vegetatively. Moreover, based on the cryopreservation method, a novel plant biotechnology tool for pathogen eradication called cryotherapy has been developed. The use of liquid nitrogen eliminates plant pathogens such as viruses, phytoplasmas, and bacteria. Our article reviews recent advances in cryo-biotechnologies such as cryopreservation and cryotherapy, with special focus on studies concerning fruit plants.

In the present study, two types of nine-recess hybrid journal bearings (HJBs) with different recess geometries, i.e., the Young Leaf Mark and the square, were operated at a rotational speed of up to 67,000 rpm using the cryogenic journal bearing test apparatus at JAXA to clarify influences of the journal rotating direction on the cryogenic HJB. Liquid nitrogen was supplied to the test bearings with a pressure of up to 4 MPa. The test bearings were 60 mm in diameter and 25 mm in width, with a 0.055 mm clearance and a recess area ratio of 0.186. Based on the experimental results the dependence of the rotor vibration on the rotational speed was confirmed to be affected by the journal rotating direction. The test HJBs with the positive sweepback angle of the recess leading edge showed the smallest rotor vibration in the high-rotational-speed operation where the hydrodynamic effect was predominant.

We present sample transfer instrumentation and integrated protocols for the preparation and atom probe characterization of environmentally-sensitive materials. Ultra-high vacuum cryogenic suitcases allow specimen transfer between preparation, processing and several imaging platforms without exposure to atmospheric contamination. For expedient transfers, we installed a fast-docking station equipped with a cryogenic pump upon three systems; two atom probes, a scanning electron microscope / Xe-plasma focused ion beam and a N2-atmosphere glovebox. We also installed a plasma FIB with a solid-state cooling stage to reduce beam damage and contamination, through reducing chemical activity and with the cryogenic components as passive cryogenic traps. We demonstrate the efficacy of the new laboratory protocols by the successful preparation and transfer of two highly contamination- and temperature-sensitive samples—water and ice. Analysing pure magnesium atom probe data, we show that surface oxidation can be effectively suppressed using an entirely cryogenic protocol (during specimen preparation and during transfer). Starting with the cryogenically-cooled plasma FIB, we also prepared and transferred frozen ice samples while avoiding significant melting or sublimation, suggesting that we may be able to measure the nanostructure of other normally-liquid or soft materials. Isolated cryogenic protocols within the N2 glove box demonstrate the absence of ice condensation suggesting that environmental control can commence from fabrication until atom probe analysis. [ABSTRACT FROM AUTHOR]

Background: Cryopreservation of leukocytes isolated from the cervicovaginal and colorectal mucosa is useful for the study of cellular immunity (see Hughes SM et al. PLOS ONE 2016). However, some questions about mucosal biology and sexually transmitted infections are better addressed with intact mucosal tissue, for which there is no standard cryopreservation protocol. Methods and findings: To find an optimal preservation protocol for mucosal tissues, we tested slow cooling (1°C/min) with 10% dimethylsulfoxide (designated “cryopreservation”) and fast cooling (plunge in liquid nitrogen) with 20% dimethylsulfoxide and 20% ethylene glycol (“vitrification”). We compared fresh and preserved human cervicovaginal and colorectal tissues in a range of assays, including metabolic activity, human immunodeficiency virus infection, cell phenotype, tissue structure by hematoxylin-and-eosin staining, cell number and viability, production of cytokines, and microbicide drug concentrations. Metabolic activity, HIV infectability, and tissue structure were similar in cryopreserved and vitrified vaginal tissues. However, vitrification led to poor cell recovery from the colorectal mucosa, with 90% fewer cells recovered after isolation from vitrified colorectal tissues than from cryopreserved. HIV infection rates were similar for fresh and cryopreserved ectocervical tissues, whereas cryopreserved colorectal tissues were less easily infected than fresh tissues (hazard ratio 0.7 [95% confidence interval 0.4, 1.2]). Finally, we compared isolation of cells before and after cryopreservation. Cell recoveries were higher when cells were isolated after freezing and thawing (71% [59–84%]) than before (50% [38–62%]). Cellular function was similar to fresh tissue in both cases. Microbicide drug concentrations were lower in cryopreserved explants compared to fresh ones. Conclusions: Cryopreservation of intact cervicovaginal and colorectal tissues with dimethylsulfoxide works well in a range of assays, while the utility of vitrification is more limited. Cell yields are higher from cryopreserved intact tissue pieces than from thawed cryopreserved single cell suspensions isolated before freezing, but T cell functions are similar. [ABSTRACT FROM AUTHOR]

Cryopreservation of biological materials is predominantly done using liquid nitrogen, and its application involves high maintenance costs and the need for periodical refilling of liquid nitrogen. Stable storage in mechanical freezers at −80°C would eliminate these issues and allow for shipment of frozen specimens using dry ice. In this work, the possibility of increasing the storage temperature of cryopreserved samples to −80°C by using combinations of DMSO and sucrose has been studied. Preservation efficacy was studied by measuring stability of liposomes encapsulated with carboxyfluorescein during storage at −150, −80 and −25°C for up to three months. Thermal and molecular mobility properties of the different DMSO-sucrose formulations were measured using differential scanning calorimetry, whereas hydrogen bonding interactions of the formulations were probed by Fourier transform infrared spectroscopy. It was found that addition of sucrose to DMSO solutions increases the Tg, and decreases molecular mobility in the glassy state at a particular temperature. Although it was expected that storage above or close to Tg at −80°C would affect liposome stability, stability was found to be similar compared to that of samples stored at −150°C. Higher molecular mobility in the glassy state could not be associated with faster CF-leakage rates. Distinct differences in storage stability at −25°C, far above Tg, were found among the sucrose/DMSO formulations, which were explained by the differences in permeability of sucrose and DMSO resulting in different levels of osmotic stress in the formulations. [ABSTRACT FROM AUTHOR]

Genetic resource banks (GRB) are a valuable tool for maintaining genetic variability and preserving breeds from pathogens or catastrophe, enabling us to assess and correct breeding schemes, minimizing the impact of genetic drift and facilitating diffusion. This study tests their efficiency in re-establishing two extinct populations of a synthetic rabbit line selected for daily weight gain, using vitrified embryos from two generations (18th and 36th) separated by 15 years of genetic selection. The effect of long-term storage of vitrified embryos in liquid nitrogen was also evaluated. A total of 516 vitrified embryos using the same protocol were transferred into 54 recipients. The embryos had been maintained in liquid nitrogen during 2 different periods, (i) 1 year (301 embryos and 26 transfers, 36th generation) and (ii) 15 years (259 embryos and 28 transfers, 18th generation). A total of 80.0% (8/10 to 18th) and 60.0% (9/15 to 36th) of the foundational sire families were eventually re-established. Over approximately one year, animals within each population were crossed to produce the next generation and re-establish the original population size. Our study demonstrated that our GRB of embryos vitrified 15 years ago is a successful strategy to re-establish rabbit populations to continue the breeding programme. [ABSTRACT FROM AUTHOR]

Background: Placental dysfunction is related to many pregnancy complications, but collecting placental specimens for investigation in large scale epidemiologic studies is often infeasible. Standard procedures involving immediate collection after birth and snap freezing are often cost prohibitive. We aimed to collect pilot data regarding the feasibility and precision of a simpler approach, the collection of tissue samples following 24 hours of refrigeration of whole placentae at 4°C, as compared to the “gold standard” of snap freezing excised tissue within 40 minutes of delivery for the assessment of inflammatory cytokines. Methods: Placentae were collected from 12 women after delivering live-born singleton babies via uncomplicated vaginal delivery. Two placentae were utilized to establish laboratory tissue processing and assay protocols. The other 10 placentae were utilized in a comparison of three tissue collection conditions. Specifically, key inflammatory cytokines were measured in 3 sections, representing three collection conditions. Sections 1 (full thickness) and 2 (excised prior to freezing) were obtained within 40 minutes of delivery and snap frozen in liquid nitrogen, and section 3 (full thickness) was obtained after refrigerating the placenta at 4°C for 24 hours. Results: IL-6, IL-10, and IL-8 all had comparable concentrations and variability overall in all three section types. Levels of tumor necrosis factor alpha (TNF-α) were too low among samples to reliably measure using immunoassay. Conclusions: Refrigeration of placentae prior to processing does not appear to compromise detection of these cytokines for purposes of large scale studies. These findings provide a framework and preliminary data for the study of inflammatory cytokines within the placenta in large scale and/or resource-limited settings. [ABSTRACT FROM AUTHOR]

This is a tale of two scientists who, in their younger days, had their scientific judgement clouded by the promise of a big discovery. Two years later, they found that their conclusions had been considerably exaggerated. They were lucky, though, as their later work would prove to be significant. Now, more than 30 years after those events, they met again and put in writing their understanding of what went wrong. [ABSTRACT FROM AUTHOR]

Emerging technologies have enabled the isolation and characterization of rare circulating tumor cells (CTCs) from the blood of metastatic cancer patients. CTCs represent a non-invasive opportunity to gain information regarding the primary tumor and recent reports suggest CTCs have value as an indicator of disease status. CTCs are fragile and difficult to expand in vitro, so typically molecular characterization must be performed immediately following isolation. To ease experimental timelines and enable biobanking, cryopreservation methods are needed. However, extensive cellular heterogeneity and the rarity of CTCs complicates the optimization of cryopreservation methods based upon cell type, necessitating a standardized protocol. Here, we optimized a previously reported vitrification protocol to preserve patient-derived CTC cell lines using highly conductive silica microcapillaries to achieve ultra-fast cooling rates with low cryoprotectant concentrations. Using this vitrification protocol, five CTC cell lines were cooled to cryogenic temperatures. Thawed CTCs exhibited high cell viability and expanded under in vitro cell culture conditions. EpCAM biomarker expression was maintained for each CTC cell line. One CTC cell line was selected for molecular characterization, revealing that RNA integrity was maintained after storage. A qPCR panel showed no significant difference in thawed CTCs compared to fresh controls. The data presented here suggests vitrification may enable the standardization of cryopreservation methods for CTCs. [ABSTRACT FROM AUTHOR]

Our aim was to compare the process of bone formation after reconstruction of the vertebral body using a titanium cage with either a liquid nitrogen-treated (frozen) bone autograft or non-treated fresh bone autograft. Twelve canine beagles underwent anterior reconstruction of the 5th lumbar vertebrae using a titanium cage and bone autograft. Bone formation was compared across four experimental groups: fresh bone autograft groups, with animals sacrificed at either 8 or 16 weeks post-reconstruction, and liquid nitrogen-treated (frozen) bone autograft groups, with animals again sacrificed at either 8 or 16 weeks post-reconstruction. Bone formation was evaluated histologically by calculating the proportion of ‘reaction’ and ‘mature bone’ regions at the ends of the cage, its center, and ventral/dorsal sides. The reaction region contained osteocytes with a nucleus and osteoblasts accumulated on the surface of an osteoid, while a laminar structure was visible for mature bone regions. For fresh bone autografts, the reaction and mature bone regions significantly increased from 8 to 16 weeks post-reconstruction. By comparison, for frozen autografts, the reaction bone region did not significantly increase from 8 to 16 weeks post-reconstruction, while the mature bone region did increase over this time period. The proportion of reaction bone was higher at the ends and dorsal side of the cage at 8 weeks, for both graft types, with greater bone formation at the center of the cage at 16 weeks only for the fresh bone autograft. Therefore, bone formation in the anterior spinal reconstruction site tended to be delayed when using a frozen bone autograft compared to a fresh bone autograft. The bone formation process, however, was similar for both groups, beginning at the ends and dorsal side of the cage adjacent to the surrounding vertebral bone. [ABSTRACT FROM AUTHOR]