Cryopreservation Research Articles

Dying tumor cells-inspired vaccine for boosting humoral and cellular immunity against cancer.

Whole tumor cells can act as effective antigen depots and have been regarded as prospective candidate for cancer vaccines. However, the clinical outcomes of whole tumor cell vaccine were restricted by the poor immunogenicity and potential in vivo oncogenicity risks. Herein, a simple and effective cancer vaccine named frozen dying tumor cells (FDT) was constructed to initiate a cascade of immune attacks against cancer. By introducing immunogenic dying tumor cells and integrating cryogenic freezing technology, FDT was endowed with high immunogenicity, good in vivo safety, and long-time storage superiority. In syngeneic mice with malignant melanoma, FDT primed the polarization of follicular helper T cells and the differentiation of germinal center B cells in lymph nodes, and promoted the infiltration of cytotoxic CD8+ T cells in the tumor microenvironment, triggering the dual activation of humoral and cellular immunity. Of note, when combined with cytokines and immune checkpoint inhibitors, the FDT vaccine achieved 100% eradication of pre-existing tumors in mice, as demonstrated in the peritoneal metastasis model of colorectal carcinoma. Taken together, our work suggests an efficient cancer vaccine inspired by dying tumor cells and provides an alternative treatment candidate for cancer.

Kinetics of Austenite Decomposition in 54SiCr6 Steel during Continuous Slow Cooling Conditions

In this study, dilatometry and metallography were used to investigate the effect of silicon and copper alloying on the decomposition kinetics of 54SiCr6 steel during continuous slow cooling. It is different from the published literature for using the approach of the local activation energy of the austenite decomposition Ef and the local Avrami exponent n of the volume fraction of the transformed phase f to study the kinetics of austenite-pearlitic transformation in cooled 54SiCr steel at slow cooling rates. The Johnson–Mehl–Avrami equation was used to determine the dependence of the local activation energy for austenite decomposition Ef and the local Avrami exponent n on the volume fraction of the transformed phase f. The mechanism of the austenite decomposition was analysed based on the calculated values of n. Both the local and average activation energies were used to evaluate the alloying effect, and the results were compared with those obtained from other methods. The type of microstructure formed as a result of cooling at rates of 0.5 K/s, 0.3 K/s, 0.1 K/s and 0.05 K/s was determined. The effects of changes in the cooling rate and the content of silicon (1.5–2.5 wt.%) and copper (0.12–1.47 wt.%) on the dimension of nucleation and growth kinetics of the transformed phase were studied. It was revealed that the pearlite microstructure was formed predominantly in 54SiCr6 steel as a result of continuous cooling at slow cooling rates. It was also found that alloying this steel with copper led to a significant decrease in the value of Ef, as well as to a change in the mechanism of the kinetics of the austenite-pearlite transformation, which was realised in predominantly two- and three-dimensional nucleation and growth at a constant nucleation rate. At the same time, alloying this steel with silicon led only to a slight change in Ef. The results of the study of 54SiCr steel presented the dependence of the activation energy of transformation and the local Avrami exponent on the volume fraction of the transformed phase at a given cooling rate at different copper and silicon contents. In addition, the study provides insight into the mechanism of kinetics in cooled 54SiCr steel as a function of the cooling rate.

Prestressed compressive strength model of engineered cementitious composite subjected to freeze–thaw damage in cryogenic freezing state

Engineered cementitious composite (ECC) is characterized by excellent strain-hardening properties and strong crack-control abilities. However, in the freezing-thawing environment of some cold regions, the concrete materials, including ECC, are subject to varying degrees of freeze–thaw (FT) damage. This paper investigates the impacts of the cryogenic freezing environment on the evolutionary mechanism of the pore structure and compressive properties of ECC. A nuclear magnetic resonance (NMR) T2 spectrum experiment and uniaxial compression experiment at −18 °C were conducted on ECC with 0 to 300 freeze–thaw cycles (FTs). The results indicated that, as the number of FTs increased, the distribution curves of the NMR T2 spectrum exhibited three peaks. Meanwhile, the drop rate of the compressive stress–strain curves in the freezing state was less than that in the thawing state, and the compressive strength, elastic modulus, and peak strain in the freezing state were higher than those in the thawing state, respectively. Based on the elastic mechanics theory and experimental analysis, the pore frost heave stress equivalence and cryogenic freezing strength equivalence were proposed, and the prestressed compressive strength model of ECC with FT damage in the cryogenic freezing state was developed. It was found that the model prediction results were well consistent with the experimental values.

Effect of heat-reinforcing needling on the inflammation and necrotizing apoptosis of synovial cells in synovial tissues of knee joint in rabbits with cold syndrome rheumatoid arthritis

To observe the effect of heat-reinforcing needling (HRN) on inflammatory factors and necrotizing apoptosis of synovial cells in synovial tissues of knee joint in rabbits with cold syndrome rheumatoid arthritis (RA), so as to explore its underlying mechanisms in treating RA. By using the random number table method, 40 New Zealand rabbits were randomly divided into normal, model, antagonist(AG), twist-reinforceing needling (TRN) and HRN groups, with 8 rabbits in each group. The model of cold syndrome RA was established by ovalbumin induction combined with Freund's complete adjuvant injection and cryogenic freezing method. In the AG group, the antagonist TAK-632 (25 mg/kg) was administered intragastrically, once every 2 days, for a total of 7 times. Rabbits of TRN and HRN groups were treated with corresponding acupuncture techniques on bilateral "Zusanli" (ST36) for 30 min, once a day for 14 days. After intervention, the changes of knee skin temperature and circumference were measured. Color Doppler ultrasound was used to observe the joint cavity effusion, synovial thickness and internal blood flow signal. The histomorphological changes of synovial tissues were observed after HE staining. ELISA was used to detect the contents of tumor necrosis factor (TNF)-α, interleukin (IL)-1β and IL-6 in serum. Transmission electron microscope was used to observe the ultrastructure, necrosis and apoptosis of synovial cells. Western blot was used to detect the protein expressions of receptor-interacting protein kinase1 (RIPK1), RIPK3, mixed lineage kinase domain-like protein (MLKL), and phosphorylation （p）-MLKL in synovial tissues. Compared with the normal group, the synovial was diffusely hyperplasia, joint cavity effusion and abnormal blood flow signal were obvious, inflammatory cells were clustered, arranged closely and disordered in the model group. The findings of transmission electron microscopy showed disruption of cell membrane integrity, swollen or ruptured mitochondria, obviously ruptured nucleus, condensed and pyknotic chromatin and nucleolus in the model group. Also, the skin temperature of the knee joint was significantly decreased (P<0.01), while the circumference of the knee joint, the contents of TNF-α, IL-1β and IL-6 in serum, the protein expressions of RIPK1, RIPK3, p-MLKL and MLKL in synovial tissues were significantly increased (P<0.01) in the model group. Compared with the model group, synovial tissue hyperplasia, joint cavity effusion, abnormal blood flow signals, synovial cell proliferation, inflammatory cell infiltration, disruption of cell membrane integrity, cell swelling, cell rupture, and nuclear pyknosis were reduced to different degrees in the AG, TRN and HRN groups. Additionally, the skin temperature of the knee joint was increased (P<0.01, P<0.05), while the circumference of the knee joint, the contents of TNF-α, IL-1β and IL-6 in serum, the expressions of RIPK1, RIPK3, p-MLKL and MLKL in synovial tissues were decreased (P<0.01, P<0.05) in the AG, TRN and HRN groups. The effects of HRN and AG were notably superior to that of TRN in up-regulating skin temperature of the knee joint, and down-regulating the circumference of the knee joint, the contents of TNF-α, IL-1β and IL-6 in serum, the expressions of RIPK1, RIPK3, p-MLKL and MLKL in synovial tissues (P<0.01, P<0.05). HRN can reduce synovial inflammation of knee joint in rabbits with cold syndrome RA, which may be related to its function in inhibiting the necrotizing apoptosis of synovial cells.

A novel method to pack cryopreserved amniotic membrane for wound dressing—the pathway through validation of a new biological product

Amniotic membrane (AM) has been widely used as a biological dressing for many pathologies and illnesses worldwide, and products derived from this tissue have been commercially available in several countries. In Brazil, regulatory agencies have recently authorized its clinical use as a non-experimental therapy for burns, diabetic and venous stasis ulcers, and intrauterine adhesions. In this study, we present our pathway through validating the first available service in the country of AM cryopreservation, with a protocol for long-term storage in high-efficiency nitrogen cryogenic freezers and a specific way of packing the tissue for optimal clinical handling and efficient storage space utilization while preserving live cells and the tissue’s biological properties. Using gauze as support, cryoprotectant dimethyl sulfoxide and product presentation as a multilayer roll exhibited the best cell viability results and maintained the tissue integrity and presence of stem/progenitor cells. Essential proteins involved in tissue regeneration and immune and antimicrobial control were detected from the secretome of cryopreserved tissue similar to fresh tissue. Furthermore, immunogenic markers, such as human leukocyte antigens, were detected at very low levels in the tissue, confirming their low immunogenicity. Finally, we demonstrate that the tissue can be kept under refrigerated conditions for up to 7 d for further use, maintaining sterility and considerable cell viability. Our cryopreservation and storage protocol kept the AM viable for at least 20 months. In conclusion, this study enabled us to determine a novel efficient protocol for long-term AM preservation for future clinical applications.

Dynamic response characteristics of moraine-soil slopes under the combined action of earthquakes and cryogenic freezing

Freeze-thaw and earthquake are significant factors for the stability and failure of moraine soil slopes in high-altitude cold and strong earthquake area. The cryogenic freezing (CF) and heat thawing (HT) of moraine soil slopes were completed by using a self-designed constant-temperature test chamber, and several shaking table tests under different seismic intensity were carried out. In these tests, the displacement and acceleration responses were recorded by the non-contact measurement system and accelerometers, and the slope dynamic responses were clarified by the displacement, acceleration and energy. The results showed that: (1) the seismic responses of the CF slope are larger than that of the HT slope. (2) The failure process of the CF slope under earthquakes could be summarized into three phases: elastic deformation phase with seismic magnitude <0.4 g, brittle failure phase with seismic magnitude from 0.4 g to 0.8 g, and failure slip phase with seismic magnitude >0.8 g. (3) The failure mode of the HT slope is characterized by the overall collapse of loose moraine soil, and the particle size of the collapsed moraine soil redistributes during the sliding process. The failure mode of CF slope is a frozen block landslide with the soil-ice interface as the sliding surface. (4) The peak acceleration of the input wave, slope surface and elevation have amplification effect on the slope dynamic response, and the top of slope surface is the main deformation and failure area. (5) Reinforcement measures such as soil improvement of slope surface, weight reduction at slope top and backpressure at slope toe, heat pipes are recommended for moraine soil slopes in cold and strong earthquake area. Such findings may provide scientific basis for the damage state identification and seismic reinforcement of moraine soil slopes in alpine and strong earthquake areas.

Best Washing Method for Oligoasthenospermic Patients by Evaluating DNA Integrity by using Comet Assay Study

Background: Sperm is more than just a mere transporter of paternal genetic information, although serving purposes other than conception. DNA integrity (DI) is regarded as a crucial aspect of semen quality and has great significance in predicting male fertility. Washing sperm is a form of preparation of sperm which is essential before processing intra-uterine-insemination since it removes chemicals from semen that might cause adversarial reactions in the uterus. During the process of washing, sperm gets separated from seminal fluid, which gradually improves the fertilizing capability of sperm, and numerous existing methods have faced complications with the determination of effective integrity in washing methods. The objective of the present study was to find the DI of the sperms through three washing methods that, include the density gradient (DG) method, swim up (SU) method, and magnetically activated cell sorting (MACS) method after the cryo-preservation (CP) for 3 months. Methodology: Semen samples were obtained from 40 oligoasthenospermic patients with progressive motility &lt;32%, who underwent assisted reproductive technology procedure in Sumathi fertility center in Madurai between October 2021 and December 2021. Sperm washing is performed through three different mentioned techniques. The grades of the comet reflected five different classes from class 0 to class 4. These data were analyzed using one-way ANOVA test, and multiple comparisons were performed between these groups using Tukey’s test. Results: The results showed a high level of normal sperm (68%) in the DG method than the SU (47%) and MACS (42%) sperm preparation techniques that specified by significant elevation (P &lt; 0.001) of class 0 comet tails. Conclusion: It was observed that the DG method has the best and success rates for intracytoplasmic sperm injection and in vitro fertilization was relatively high.

Effects of cryogenic freezing upon lithium-ion battery safety and component integrity

Recycling capacity for lithium-ion batteries (LIBs) has not kept pace with the increase in battery manufacturing throughout the early 21st century. Cost-effective recycling practices must be developed to accommodate the pending influx of battery waste over the coming decades as the first generation of LIBs reach their end-of-life (EOL). Cryogenically freezing LIBs can passivate them against abusive conditions, and may therefore enable LIBs to be granted exemptions for certain hazardous material transportation requirements, significantly reducing the cost of their transportation to recycling facilities. This work aims to identify potential risks of a cryogenic transportation scenario to inform the development of standards and practices thereof. Results are presented from freeze/thaw experiments using liquid nitrogen to freeze LIBs to −197 °C. Cells are opened after thawing to assess structural damage to the cell components inherent to the freezing process. Additionally, nail penetration tests are performed on cells as they thaw to room temperature. LIBs appear undamaged after cryogenic freezing. Nail penetration experiments indicate that thermal runaway is forestalled at low temperatures, but will ensue after sufficient thawing. No thermal response was detected in cells penetrated at or below −80 °C, which approximates the melting point of the electrolyte, suggesting that LIBs are inert while the electrolyte remains frozen.

Cryoprotective Characteristics of L-Carnosine Dipeptide (β-Alanyl-L-Histidine)

The dipeptide (β-alanyl-L-histidine) is found in significant amounts in the muscles and brain of mammals, especially in the olfactory structures. L-Carnosine exhibits many protective effects when exposed to various cytotoxic factors on cells. We used slices of the rat olfactory cortex to study the cryoprotective characteristics of L-carnosine during cryopreservation (CP). Changes in the activity of N-methyl-D-aspartate receptors (NMDAR) were analyzed during registration of NMDA potentials induced by electrical stimulation of the lateral olfactory tract. Brain slices were preincubated with L-carnosine (20 mM) in solution, frozen (−10°C), and after a long CP (30 days) they were warmed up to 37°C and changes in the amplitudes of NMDA potentials were determined. It was found that the dipeptide optimized the pH of the freezing solution after CP and retained the activity of NMDAR, determined by the amplitude of NMDA potentials. L-Carnosine after CP contributed to the dehydration of excess free water from the slices. The dipeptide inhibited the development of glutamate excitotoxicity in brain slices during CP and maintained normal NMDAR functioning. The data obtained prove that L-carnosine exhibits the properties of an endogenous cryoprotector in the nervous tissue.

Novas tecnologias combinadas com desidratação osmótica para aplicação na conservação de frutas: uma revisão

ABSTRACT: Osmotic dehydration (OD) is a technique used for the partial removal of water from foodstuff, including fruit and vegetables, with the aim of producing a desiccated product. The process involves placing the material in a hypertonic solution for several hours and allowing water to move from the cell compartment into the solution by osmosis. OD is influenced by various factors such as the concentration and composition of the osmotic solution, the solution temperature, the type of agitation and the time of exposure, as well as the size, shape and compactness of the food material. The main advantages of OD over conventional drying processes are the superior quality of the dried products and the minimization of shrinkage. In recent years, research effort has focused on the combination of OD with other technologies, such as ultrasound, cryogenic freezing with liquid nitrogen, pulsed electric field, gamma radiation and high hydrostatic pressure. The application of these methods prior to or concomitant with OD accelerates mass transfer and reduces the drying rate of fruit and vegetables by increasing the permeability of cell membranes. In this manner, combined processes tend to be more efficient and economical in comparison with conventional OD because they reduce operating times and; consequently, energy consumption. In addition, the dried products generated by such coupled processes typically exhibit improved nutritional and physicochemical characteristics. This review summarizes the basic principles and applications of OD in combination with other methods, with particular emphasis on the production of dried fruits.

Construction of ceRNA Regulatory Network in Human Oocytes.

Cryogenic freezing, often known as cryopreservation, is a technique for preserving human oocytes. In this study, differentially expressed long non-coding RNAs (lncRNAs) and messenger RNAs (mRNAs) were identified using the human frozen oocyte dataset GSE69768 from the Gene Expression Database (GEO). Subsequently, combined with the microRNA atlas database, the miRNAs combined with differentially expressed lncRNAs (DElncRNAs) were predicted, and the lncRNA-miRNA-mRNA interaction relationship and competitive endogenous (ceRNA) regulatory network were obtained. The results revealed that multiple DElncRNAs and DEmRNAs were involved in the ceRNA network of the human oocyte. Finally, GO functional annotation and KEGG pathway enrichment analysis were performed on the differentially expressed mRNA (DEmRNA) in the ceRNA network, and the biological processes and pathways that may be related to the ceRNA network in frozen oocytes were explored. In conclusion, in the ceRNA network for human oocyte, lncRNA, mRNA, and miRNA do not each operate via a distinct, independent mechanism. Not only does the RNA-RNA contact involve the ceRNA regulatory mechanism, but it also involves interactions between proteins that are encoded by genes. Furthermore, the negative effects of inter-individual variations and instability on the quality of high-throughput detection cannot be completely ruled out due to the paucity of human oocyte data containing both mRNA and lncRNA expression profiles.

Climate change risks and financial performance of the electric power sector: Evidence from listed companies in China

The frequency, intensity and duration of extreme weather events have seriously affected human life and production. The electric power sector is the foundation of economic activity as well as a core participant in the adaptation and mitigation of climate change. Therefore, in the context of climate change, it is crucial for the stable operation of the national economy for power sector to cope with different climate change risks and improve their adaptability to climate change. In this paper, listed power companies in China are selected as samples to analyse the impact of climate change risk on the financial performance of the power system's supply and transmission-distribution sides. The empirical results show a significant positive correlation between climate change risks and the financial performance of listed electric power companies. The rainfall index and drought index positively impact the financial performance of listed electric power companies. The cryogenic freezing index has a negative impact on the financial performance of listed electric power companies., which is further analysed and proved that a cryogenic freezing disaster will cause the regional breakdown of the power system. The operating cost ratio and the proportion of clean energy supply play a mediating effect on the correlation between the comprehensive climate risk index and the return on equity of companies. To increase electric power companies' ability to adapt to climate change, climate change risks should be integrated into the risk management framework, and the company's financial performance can be improved by optimizing the energy mix and constructing safe lines. The government can promote the transformation of electric power companies by launching green financial tools.

Prolong cryopreservation duration negatively affects pregnancy outcomes of vitrified-warmed blastocyst transfers using an open-device system: A retrospective cohort study

ObjectiveTo investigate the impact of cryopreservation (CP) duration on pregnancy outcomes of vitrified-warmed blastocysts transfers using an open-device liquid-nitrogen (LN2) system. MethodsThis retrospective cohort study was conducted on 6327 first vitrified-warmed single blastocyst transfer cycles with autologous oocytes from January 2015 to December 2020. The CP duration was initially divided into six groups: Group I: 0–3 months (n = 4309); Group II: 4–6 months (n = 1061); Group III: 7–12 months (n = 304); Group IV: 13–24 months (n = 113); Group V: 25–72 months (n = 466); Group VI: 73–120 months (n = 74). Multivariate logistic regression was performed to evaluate the independent effect of CP duration on pregnancy outcomes. To further examine the time limit of vitrification, propensity score matching (PSM) was applied to compare pregnancy outcome of patients with storage duration of 25–120 months to those of 0–24 months. After that, pregnancy outcomes were compared among the subgroups of Group I’: 0–24 months, Group II’: 25–48 months, Group III’: 49–72 months, Group IV’: 73–120 months. Stratification analysis based on embryo quality was also performed. Primary outcomes were clinical pregnancy rate and live birth rate. Secondary outcomes were implantation, biochemical pregnancy rate, ongoing pregnancy rate and early miscarriage rate. ResultsLogistic regression demonstrated that the odds of pregnancy outcomes were similar across Group I to IV. However, the implantation rate, chances of biochemical pregnancy, clinical pregnancy, ongoing pregnancy, and live birth significantly decreased as the storage duration increased up to 25 months, while miscarriage rate did not significantly differ between groups. Subgroup analysis confirmed a dramatical decrease of clinical pregnancy and live birth rate when cryopreserved for more than 24 months. After that, the slope was relatively steady between 25 and 72 months, then steeply decreased again as CP reached 73–120 months. In addition, there was a more remarkable decline of pregnancy outcomes in the average quality embryo transfers than in the high quality embryo transfers as cryopreservation storage increased. ConclusionProlonged cryopreservation of vitrified blastocysts in an open-device LN2 system up to 24 months might negatively affect pregnancy outcomes. This negative impact progresses as storage duration increases, especially when exceeds 72 months. Average quality embryo appears to be less sustainable with long-term cryo-storage.

Optimizing radiofrequency assisted cryogenic freezing to improve meat microstructure and quality

Radiofrequency-assisted nitrogen cryogenic freezing previously proposed was optimized to increase meat quality. Continuous cryogenic (CCF, for 2.5 min) and pulsed cryogenic (PCF, 3 s pulse with 10 s interval for 10 min) freezing were set up. CCF and PCF were then combined with radiofrequency (RF) pulses to set up RF-CCF (10 s 2 kV RF pulse with 20 s interval during 2.5 min of continuous N 2 flow) and RF-PCF (30 s 2 kV RF pulse with 60 s interval, during 3 s nitrogen pulse with 10 s interval for 10 min). These processes were compared with slow (SF) and blast (BF) freezing. Meat quality ( i.e. , microstructure, drip loss, color, firmness) decreased in the order RF-PCF ≥ PCF > RF-CCF > BF > SF > CCF, indicating that nitrogen pulsing considerably reduced tissue damages and its combination with RF was crucial to further preserve tissue integrity. RF-PCF could be adapted to different matrices by tuning processing conditions. • Radiofrequency (RF) nitrogen cryogenic freezing of meat was optimized by a pulsing approach. • Nitrogen pulsing during meat freezing reduced tissue damage. • RF pulsing was modulated to guarantee homogeneous cryogenic freezing of meat. • Combining nitrogen pulses with RF was crucial to preserve meat microstructure.

Changes of sample temperature inside a cryo storage robot

Changes of sample temperature inside a cryo storage robot

Effect of heat-reinforcing needling on expression of P2X7R, NLRP3 and Caspase-1 in synovial tissues of knee joint in rabbits with cold syndrome rheumatoid arthritis

To observe the effects of heat-reinforcing needling on the expression of purinergic ligand-gated ion channel 7 receptor(P2X7R)，Nodlike receptor protein 3(NLRP3) and Caspase-1 in synovium tissues of knee joint of rabbits with cold syndrome rheumatoid arthritis(RA)，so as to explore the anti-inflammatory mechanism of heat reinforcing needling in the treatment of RA. Thirty male New Zealand white rabbits were randomly divided into normal group, model group, reinforcing-reducing needling group(RRG), heat-reinforcing needling group(HRG), and antagonist group(AG), with 6 rabbits in each group.The model of cold syndrome RA was established by ovalbumin combined with Freund's adjuvant and cryogenic freezing. Rabbits of RRG and HRG were treated with corresponding acupuncture techniques on both sides of "Zusanli"(ST36) for 30 min, once a day for 7 days; Rabbits of the AG was intraperitoneally injected with A438079(2.5 mg/kg), once a day for 7 days. After intervention, color Doppler ultrasound was used to observe the joint cavity effusion, synovial thickness and internal blood flow signal.The histomorphological changes of synovial tissues were observed by HE staining. Quantitative real-time PCR method was used to detect the mRNA expression levels of P2X7R, NLRP3, and Caspase-1 in synovial tissues. Western blot was used to detect the protein expression of interleukin(IL)-1β, IL-6 and tumor necrosis factor(TNF)-α in synovial tissues. Compared with the normal group, the synovial tissues in the model group was thickened, linear and punctate blood flow signals were increased, joint effusion was obvious, synovial coating cells were enlarged, the synovial matrix was severely hyperplasia, inflammatory cells infiltration was obvious.The mRNAs expression levels of P2X7R, NLRP3, Caspase-1 and IL-1β, IL-6, TNF-α proteins in synovial tissues were significantly increased(P<0.01). Compared with model group, abnormal blood flow signals, synovial thickness, joint effusion, proliferation of synovial matrix, inflammatory cell infiltration and synovial coating cells in the RRG, HRG and AG were improved. The mRNAs expression levels of P2X7R, NLRP3, Caspase-1 and IL-1β, IL-6, TNF-α proteins in synovial tissues were decreased in the RRG, HRG and AG (P<0.05, P<0.01). Compared with the RRG, the above indexes were lower in the HRG and AG (P<0.05, P<0.01).There was no significant difference in all indexes above between the HRG and AG (P<0.01, P<0.05). Heat reinforcing needling can improve synovial inflammation of RA, which may be related to regulating the expressions of P2X7R/NLRP3/Caspase-1 signaling pathway.

In Vitro versus Cryo-Induced Capacitation of Bovine Spermatozoa, Part 2: Changes in the Expression Patterns of Selected Transmembrane Channels and Protein Kinase A.

Since the molecular similarities and differences among physiological capacitation and cryocapacitation have not been studied in detail, this study was designed to assess the gene and protein expression levels of the Cation channel of sperm (CatSper) 1 and 2, sodium bicarbonate (Na+/HCO3−) cotransporter (NBC) and protein kinase A (PKA) in un-capacitated (control), in vitro capacitated (CAP) and cryopreserved (CRYO) bovine spermatozoa. All samples were subjected to motility evaluation using the computer assisted sperm analysis and chlortetracycline (CTC) assay for the assessment of the capacitation patterns. Furthermore, quantitative reverse transcription PCR (qRT-PCR) and Western blots were used to monitor the expression patterns of the selected capacitation markers. The results showed a significant reduction in the gene and protein expression levels of CatSper1 and 2 in the CRYO group when compared to the CAP group (p < 0.0001). In the case of NBC, the results were not significantly different or were inconclusive. While a non-significant down-regulation of PKA was found in the CRYO group, a significant reduction in the expression of the PKA protein was found in frozen-thawed spermatozoa in comparison to the CAP group (p < 0.05). In conclusion, we may hypothesize that while in vitro capacitated and cryopreserved spermatozoa exhibit CTC-patterns consistent with capacitation events, the molecular machinery underlying CTC-positivity may be different.

A Brief Overview on Female Infertility, It's Causes and Diagnostic Approaches

Female infertility is the major disorder which has altered the man kind foe lack of conception and reproducibility, stressful world, excess radiation, lack of biological food, genetically disorder ,changing life style, increased electronic discharge have resulted the female infertility. Infertility/ childlessness cause great personal suffering &amp; distress. Most of this agony &amp; misery is hidden from the public gaze. That is the reason this topic is not discussed about openly. The dismal ignorance &amp; neglect about the causes of childlessness and its treatment are main reason for the lack of public support for childless couple. Female fertility can be limited or diminished or destroyed in a number of ways. Women have a finite number of germs cells and follicles that are available for a limited period, from menarche to menopause, during their lifetimes. The process of ovulation is mediated by the interactions of hypothalamic, pituitary and ovarian hormones. Interference with ovulation can occur at any one or combinations of these sites. The oviducts can be distorted or blocked by the consequences of endometriosis or infection. The quality of the ova and spontaneous pregnancy decreases steadily with age. Drugs are available that stimulate ovulation and donor eggs can be used. The cryo preservation of ova or ovarian tissue is technique now receiving research attention. Diagnosis is straightforward when causes are severe and laparoscopy is still the preferred method for assessing for tubal factor infertility and endometriosis. IVF remains the dominant treatment, although traditional measures still have a major role. Internationally, IVF opportunities are limited in view of cost. About 10-15% of couples experience some difficulty with fertility. Remedies range from a visit to a primary physician, education and adjustments in timing attempts to conceive, to placing the entire reproductive process in the hands of specialist.The purpose of the study is to evaluate existing literature for possible associations between female infertility, infertility-associated diagnoses, and the following areas of disease: infertility and mental health significance of the study common factors ,diagnosis methods artificial fertility treatment methods ethical issues.

Identification of Carbides and Phase Transformations in Sintered Fe–Mo–Mn–C Alloys Produced under a Slow Continuous Cooling

Different sintered alloys were produced by sintering and slow cooling of powder compacts made from pre-alloyed Fe-0.50Mo-0.15Mn powder mixed with varied graphite powder contents (0.30–1.20 wt.% with 0.15% increment). According to microstructures, sintered alloys were divided into sintered hypo-eutectoid, near eutectoid, and hyper-eutectoid alloys. By using Groesbeck color tinting and X-ray diffraction technique, the common phase transformation products of these sintered alloys were found to include ferrite and carbides. The sintered hypo-eutectoid alloys had microstructures consisting of polygonal ferrite grains and two forms of ferrite + carbide mixtures, such as ferrite + M23C6 carbide and ferrite + M3C carbide. In sintered near-eutectoid alloys, only ferrite + M3C carbide mixtures occupied microstructures. In sintered hyper-eutectoid alloys, large proeutectoid M23C6 carbide formed first and followed by abnormal ferrite, degenerate ferrite + M23C6 pearlite, lamellar ferrite + M23C6 pearlite, Widmanstätten M3C carbide, inverse bainite (ferrite + M3C) and upper bainite (ferrite + M3C).

An analysis on the impacts of cryogenic freezing on raspberry quality

Counter-season supply of horticultural products is of increasing demand. Consumers are demanding annual supply of raspberries, which has historically been challenging due to their seasonal summer supply and characteristically high metabolism resulting in a short shelf life and limited period of availability. However, the development of freezing technologies for increasing the length of storage of raspberries offers an opportunity for continual supply of premium quality raspberries. We investigated berry quality after freezing whole fresh raspberries, comparing conventional freezing methods with a modern cryogenic freezing method over a period of six months. Significant increases in total soluble solids, titratable acidity, hue and chroma were found when raspberries were frozen compared to fresh raspberries. No overall difference in berry quality was observed between freezing methods for any parameter assessed. When assessed at time intervals, total soluble solids, pH, titratable acidity, chroma and hue were consistent between freezing methods for all durations of time frozen. These findings provide decision support for producers and distributors pursuing a novel counter season supply chain.