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1.
Postepy Biochem ; 69(4): 291-297, 2024 01 30.
Artigo em Polonês | MEDLINE | ID: mdl-39012696

RESUMO

The problem of regeneration of damaged peripheral nerves is an ongoing topic and has long been the subject of intensive research worldwide. This study examined the morphological and functional evaluation of the regeneration process within the damaged sciatic nerve, a mouse animal model. The effect of impaired expression of the TSC-1 gene on the process of nerve regeneration was evaluated, depending on the mode of damage. The research object consisted of 48, 2-month-old male TSC lines. The test group consisted of animals that underwent damage to the sciatic nerve by crushing, freezing and electrocoagulation, while the control group includes mice whose sciatic nerve was not damaged. Behavioral tests were conducted to evaluate the functional return of the limb, after 3,5,7 and 14 days. The first changes in the regeneration process of the damaged neurite are observed as early as day 3 after the injury, while on day 14 after the injury the functional return of the damaged limb was noted.


Assuntos
Modelos Animais de Doenças , Eletrocoagulação , Regeneração Nervosa , Nervo Isquiático , Animais , Camundongos , Regeneração Nervosa/fisiologia , Nervo Isquiático/lesões , Masculino , Eletrocoagulação/métodos , Congelamento/efeitos adversos , Compressão Nervosa/métodos
2.
Cryobiology ; 115: 104892, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38593909

RESUMO

Refreezing the remaining genetic resources after in vitro fertilization (IVF) can conserve genetic materials. However, the precise damage inflicted by repeated freezing and thawing on bovine sperm and its underlying mechanism remain largely unexplored. Thus, this study investigates the impact of repeated freeze-thaw cycles on sperm. Our findings indicate that such cycles significantly reduce sperm viability and motility. Furthermore, the integrity of the sperm plasma membrane and acrosome is compromised during this process, exacerbating the advanced apoptosis triggered by oxidative stress. Additionally, transmission electron microscopy exposed severe damage to the plasma membranes of both the sperm head and tail. Notably, the "9 + 2" structure of the tail was disrupted, along with a significant decrease in the level of the axonemal protein DNAH10, leading to reduced sperm motility. IVF outcomes revealed that repeated freeze-thaw cycles considerably impair sperm fertilization capability, ultimately reducing the blastocyst rate. In summary, our research demonstrates that repeated freeze-thaw cycles lead to a decline in sperm viability and motility, attributed to oxidative stress-induced apoptosis and DNAH10-related dynamic deficiency. As a result, the utility of semen is compromised after repeated freezing.


Assuntos
Apoptose , Criopreservação , Fertilização in vitro , Congelamento , Estresse Oxidativo , Preservação do Sêmen , Motilidade dos Espermatozoides , Espermatozoides , Animais , Masculino , Bovinos , Criopreservação/veterinária , Criopreservação/métodos , Preservação do Sêmen/veterinária , Preservação do Sêmen/métodos , Espermatozoides/fisiologia , Fertilização in vitro/veterinária , Congelamento/efeitos adversos , Membrana Celular , Sobrevivência Celular , Acrossomo
3.
Biochem Pharmacol ; 197: 114950, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35143754

RESUMO

We previously reported that 2,5-dimethylcelecoxib (DM-C), a derivative of celecoxib, lacks cyclooxygenase-2 inhibitory effects and suppresses cardiac remodeling by activating glycogen synthase kinase-3 (GSK-3). However, it remains unclear whether DM-C attenuates fibroblast-to-myofibroblast transformation (FMT), which plays a key role in cardiac fibrosis. Therefore, we evaluated the effect of DM-C on FMT using a cryoinjury-induced myocardial infarction (CMI) mouse model. We found that DM-C attenuated the deterioration of left ventricular ejection fraction after CMI by decreasing cardiac fibrosis. Analysis of the expression level of α-smooth muscle actin (α-SMA), a marker for myofibroblasts, indicated that DM-C decreased FMT at the cardiac injury site. To investigate the mechanism by which DM-C attenuated FMT, fibroblasts obtained from the heart were stimulated with TGF-ß to induce FMT, and the effect of DM-C was analyzed. DM-C suppressed the expression of α-SMA and the phosphorylation levels of Smad 2/3 and GSK-3, indicating that DM-C suppressed α-SMA expression by inhibiting the transforming growth factor (TGF)-ß signaling pathway via activation of GSK-3. DM-C decreased the expression of collagen, connective tissue growth factor (CTGF) and Snail, which are also known to accelerate cardiac fibrosis. These results suggested that DM-C attenuated cardiac fibrosis by suppressing FMT at the injured site after CMI by inhibiting the TGF-ß signaling pathway via activation of GSK-3. Thus, DM-C has potential against cardiac disease as a novel anti-fibrotic agent.


Assuntos
Fibroblastos/efeitos dos fármacos , Congelamento/efeitos adversos , Infarto do Miocárdio/tratamento farmacológico , Miofibroblastos/efeitos dos fármacos , Pirazóis/uso terapêutico , Transdução de Sinais/efeitos dos fármacos , Sulfonamidas/uso terapêutico , Animais , Células Cultivadas , Fibroblastos/enzimologia , Fibroblastos/patologia , Fibrose , Quinase 3 da Glicogênio Sintase/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Infarto do Miocárdio/enzimologia , Infarto do Miocárdio/etiologia , Infarto do Miocárdio/patologia , Miofibroblastos/enzimologia , Miofibroblastos/patologia , Nitrogênio/toxicidade , Pirazóis/farmacologia , Ratos , Ratos Endogâmicos Lew , Transdução de Sinais/fisiologia , Sulfonamidas/farmacologia
4.
Front Endocrinol (Lausanne) ; 12: 709648, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34630326

RESUMO

Background: With the development of embryo freezing and warming technology, frozen-thawed embryo transfer (FET) has been widely utilized. However, studies investigating the association between cryopreservation duration and FET outcomes are limited and controversial, and previous studies did not conduct stratification analyses based on demographic or clinical characteristics. Methods: This multicenter retrospective study included 17,826 women who underwent their first FET following the freeze-all strategy during the period from January 2014 to December 2018. Duration of cryopreservation was categorized into five groups: 3-8 weeks, 8-12 weeks, 12-26 weeks, 26-52 weeks, and >52 weeks. Modified Poisson regression and multivariate logistic regression were used to assess the association between cryostorage time of vitrified embryos and transfer outcomes. Moreover, further stratification analyses were performed according to variables with p <0.05 in multivariate models. Results: In this large multicenter study, we observed that storage duration was inversely associated with the possibility of pregnancy and live birth (p <0.001), but not with the risk of ectopic pregnancy and miscarriage. Stratification analyses based on maternal age, the number of oocytes retrieved, and condition of embryo transferred indicated that the inverse correlation was significant in the subpopulation with characteristics: (1) less than 40 years old, (2) more than 3 oocytes retrieved, and (3) only high-quality blastocysts transferred. Conclusion: The results of this large, multicenter, retrospective study suggested that prolonged cryopreservation was inversely associated with the probability of pregnancy and live birth. Therefore, for patients who adopt a freeze-all strategy, early FET might achieve a better outcome.


Assuntos
Aborto Espontâneo/epidemiologia , Criopreservação/estatística & dados numéricos , Transferência Embrionária , Embrião de Mamíferos/patologia , Congelamento/efeitos adversos , Nascido Vivo/epidemiologia , Indução da Ovulação/métodos , Aborto Espontâneo/etiologia , Adulto , Coeficiente de Natalidade , China/epidemiologia , Técnicas de Cultura Embrionária , Feminino , Seguimentos , Humanos , Prognóstico , Estudos Retrospectivos , Vitrificação
5.
Bull Exp Biol Med ; 171(5): 596-600, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34617171

RESUMO

We studied the effect of xenon on the survival rate of the spermatozoa of the common frog Rana temporaria during slow freezing with saturation of the suspension with xenon at a pressure of up to 1.2 bar. The cryoprotective properties of xenon were analyzed in comparison with nitrogen. No specific cryoprotective effect of xenon was revealed. Viability of spermatozoa pretreated with xenon at atmospheric pressure (0 bar) or under excess pressure of 0.6 bar and frozen in a cryoprotective medium with dimethylformamide, sucrose, and BSA did not differ significantly. The use of overpressure of xenon of 1.0 or 1.2 bar in the pretreatment and freezing process significantly impaired viability of the biomaterial.


Assuntos
Congelamento/efeitos adversos , Espermatozoides/efeitos dos fármacos , Xenônio/farmacologia , Animais , Sobrevivência Celular/efeitos dos fármacos , Criopreservação/métodos , Crioprotetores/farmacologia , Masculino , Rana temporaria , Análise do Sêmen/veterinária , Preservação do Sêmen/efeitos adversos , Preservação do Sêmen/métodos , Preservação do Sêmen/veterinária , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatozoides/fisiologia
7.
Eur J Pharm Biopharm ; 169: 44-51, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34534655

RESUMO

Lyophilization can extend protein drugs stability and shelf life, but it also can lead to protein degradation in some cases. The development of safe freeze-drying approaches for sensitive proteins requires a better understanding of lyophilization on the molecular level. The evaluation of the freezing process and its impact on the protein environment in the nm scale is challenging because feasible experimental methods are scarce. In the present work we apply pulse EPR as a tool to study the local concentrations of the solute in the 20 nm range and of the solvent in the 1 nm range for a spin labeled 27 kDa monomeric green fluorescent protein, mEGFP, and the 172 Da TEMPOL spin probe, frozen in different water/glycerol-d5 mixtures. For average glycerol volume fractions, φgly-d5avg, ≥ 0.4 we observed transparent glassy media; the local concentration and the 1 nm solvent shell of TEMPOL and the protein correspond to those of a uniform vitrified glass. At φgly-d5avg ≤ 0.3 we observed partial ice crystallization, which led to ice exclusion of glycerol and TEMPOL with freeze-concentration up to the glycerol maximal-freeze local volume fraction, φgly-d5loc, of 0.64. The protein concentration and its shell behavior was similar except for the lowest φgly-d5avg (0.1), which showed a 4.7-fold freeze-concentration factor compared to sevenfold for TEMPOL, and also a smaller φgly-d5loc. We explain this behavior with an increased probability for proteins to get stuck in the ice phase during fast freezing at higher freeze-concentration and the related large-scale mass transfer.


Assuntos
Liofilização/métodos , Conformação Proteica , Estabilidade Proteica , Proteínas/farmacologia , Proteólise , Cristalização , Estabilidade de Medicamentos , Congelamento/efeitos adversos , Humanos , Preparações Farmacêuticas , Soluções/química , Soluções/farmacologia , Água/química , Água/farmacologia
8.
PLoS One ; 16(9): e0256836, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34473780

RESUMO

The study is aimed at the frosting problem of the air source heat pump in the low temperature and high humidity environment, which reduces the service life of the system. First, the frosting characteristics at the evaporator side of the air source heat pump system are analyzed. Then, a new defrost technology is proposed, and dimensional theory and neural network are combined to predict the transfer performance of the new system. Finally, an adaptive network control algorithm is proposed to predict the frosting amount. This algorithm optimizes the traditional neural network algorithm control process, and it is more flexible, objective, and reliable in the selection of the hidden layer, the acquisition of the optimal function, and the selection of the corresponding learning rate. Through model performance, regression analysis, and heat transfer characteristics simulation, the effectiveness of this method is further confirmed. It is found that, the new air source heat pump defrost system can provide auxiliary heat, effectively regulating the temperature and humidity. The mean square error is 0.019827, and the heat pump can operate efficiently under frosting conditions. The defrost system is easy to operate, and facilitates manufactures designing for different regions under different conditions. This research provides reference for energy conservation, emission reduction, and sustainable economic development.


Assuntos
Simulação por Computador , Equipamentos e Provisões Elétricas , Congelamento/efeitos adversos , Calefação/instrumentação , Aprendizado de Máquina , Modelos Teóricos , Redes Neurais de Computação , Ar , Temperatura Alta , Umidade/efeitos adversos , Água/química
9.
Biol Open ; 10(7)2021 07 15.
Artigo em Inglês | MEDLINE | ID: mdl-34125176

RESUMO

Ozobranchus jantseanus is the largest metazoan known to survive in liquid nitrogen without pretreatment to date; however, the mechanism underlying this tolerance remains unclear. In this study, the first analyses of histological and morphological changes in normal, frozen, and dehydrated states were performed. Adults survived after direct placement in liquid nitrogen for 96 h, with a survival rate of approximately 86.7%. The leech could withstand rapid desiccation and its survival rate after rehydration was 100% when its water loss was below about 84.8%. After freezing, desiccation, and ethanol dehydration, the leech immediately formed a hemispherical shape. Particularly during drying, an obvious transparent glass-like substance was observed on surface. Scanning electron microscopy revealed many pores on the surface of the posterior sucker, creating a sponge-like structure, which may help to rapidly expel water, and a hemispherical shape may protect the internal organs by contraction and folding reconstruction in the anterior-posterior direction. A substantial amount of mucopolysaccharides on the surface and acid cells and collagen fibers in the body, all of which contained substantial polysaccharides, may play a key protective role during freezing. Our results indicate that the resistance of leeches to ultra-low temperatures can be explained by cryoprotective dehydration/vitrification strategies. This article has an associated First Person interview with the first author of the paper.


Assuntos
Adaptação Fisiológica/fisiologia , Congelamento/efeitos adversos , Sanguessugas/fisiologia , Animais , Microscopia Eletrônica de Varredura
10.
Cryo Letters ; 42(1): 53-58, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33973993

RESUMO

BACKGROUND: Several obstacles must be overcome before ovarian tissue cryopreservation can be used as a standard procedure. OBJECTIVE: To carry out a morphologic and functional study of the effect of cryopreservation on mouse follicles and stroma cells. MATERIALS AND METHODS: Female mice were divided into three groups (control, fresh graft and cryopreserved graft). Ultrastructural features of follicles and stroma cells were evaluated using transmission electron microscopy. After autologous transplantation, micro-vessel densities of grafts were examined. RESULTS: Vacuoles in granulosa cells and stromal cells are significantly greater than that of oocytes. The microvessel density of fresh grafts is significantly higher than that in frozen-thawed grafts. CONCLUSION: Granusola and stroma cells, rather than oocytes, are vulnerable to cryoinjury. Injuries to granulosa cells and stromal cells could be the critical part of ovarian damage caused by cryopreservation.


Assuntos
Criopreservação , Congelamento/efeitos adversos , Folículo Ovariano , Células Estromais , Animais , Feminino , Camundongos , Oócitos/patologia , Folículo Ovariano/fisiopatologia , Ovário , Células Estromais/patologia
11.
Cryo Letters ; 42(2): 87-95, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33970985

RESUMO

BACKGROUND: The role of oxidative stress during cryoprotectant treatment has received little attention. OBJECTIVE: To assess the effects of different cryoprotectants and discover relationships between cryodamage and oxidative stress parameters on Awassi ram sperm. MATERIALS AND METHODS: The sperm samples diluted with Salamon's tris-citrate (TRIS) containing 20% centrifuged egg yolk and 0.5, 1.0 or 1.5 M Glycerol (Gly), methanol (M), 2-methoxyethanol (2-ME), dimethylacetamide (DMA) and 1.2 propanediol (PR). After 2 h of equilibration at +4 ºC, the sperm samples were frozen in liquid nitrogen vapour and stored. RESULTS: The best post-thaw motility (43.3%, 41.7%) of sperm was achieved when protected with 0.5 and 1.0 M glycerol. Arylesterase and ceruloplasmin parameters were significantly different after equilibration, whereas sulfhydryl groups were significantly different after freezing in their respective groups (P < 0.05). CONCLUSION: The increased use of glycerol caused greater loss of motility. The role of oxidative stress in freezing was also found to be limited.


Assuntos
Criopreservação/veterinária , Congelamento/efeitos adversos , Estresse Oxidativo , Preservação do Sêmen/veterinária , Carneiro Doméstico , Animais , Crioprotetores/farmacologia , Glicerol , Masculino , Motilidade dos Espermatozoides , Espermatozoides
12.
Eur J Pharm Biopharm ; 163: 127-140, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33813056

RESUMO

In the pharmaceutical industry, cryoprotectants are added to buffer formulations to protect the active pharmaceutical ingredient from freeze- and thaw damage. We investigated the freezing and thawing of aqueous sodium citrate buffer with various cryoprotectants, specifically amino acids (cysteine, histidine, arginine, proline and lysine), disaccharides (trehalose and sucrose), polyhydric alcohols (glycerol and mannitol) and surfactants (polysorbate 20 and polysorbate 80). Hereby, we employed optical cryomicroscopy in combination with differential scanning calorimetry in the temperature range to -80 °C. The effect of cryoprotectants on the morphology of the ice crystals, the glass transition temperature and the initial melting temperature is presented. Some of the cryoprotectants have a significant impact on ice crystal size. Disaccharides restrict ice crystal growth, whereas surfactants and glycerol allow ice crystals to increase in size. Cysteine and mannitol cause dehydration after thawing. Either one or two glass transition temperatures were detected, where arginine, surfactants, glycerol, proline and lysine suppress the second, implying a uniform freeze-concentrated solution. The initial melting temperature of pure buffer solution can be shifted up by adding mannitol, both disaccharides and both surfactants; but down by glycerol, proline and lysine.


Assuntos
Crioprotetores/química , Soluções/química , Soluções Tampão , Varredura Diferencial de Calorimetria , Química Farmacêutica , Congelamento/efeitos adversos , Microscopia , Temperatura de Transição , Vitrificação
13.
J Assist Reprod Genet ; 38(6): 1293-1302, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33825118

RESUMO

PURPOSE: In vitro maturation (IVM) is an alternative to in vitro fertilization (IVF) for women at high risk of developing ovarian hyperstimulation syndrome (OHSS). This study determined the effectiveness and safety of a freeze-only strategy versus fresh embryo transfer (ET) after IVM with a pre-maturation step (CAPA-IVM) in women with a high antral follicle count (AFC). METHODS: This randomized, controlled pilot study (NCT04297553) was conducted between March and November 2020. Forty women aged 18-37 years with a high AFC (≥24 follicles in both ovaries) undergoing one cycle of CAPA-IVM were randomized to a freeze-only strategy with subsequent frozen ET (n = 20) or to fresh ET (n = 20). The primary endpoint was ongoing pregnancy resulting in live birth after the first ET of the started treatment cycle. RESULTS: The ongoing pregnancy rate in the freeze-only group (65%) was significantly higher than that in the fresh ET group (25%; p = 0.03), as was the live birth rate (60% versus 20%; p = 0.02). Clinical pregnancy rate was numerically, but not significantly, higher after frozen versus fresh ET (70% versus 35%; p = 0.06), while the number of day 3 or good quality embryos, endometrial thickness on the day of oocyte pick-up, implantation rate, and positive pregnancy test rate did not differ significantly between groups. No cases of OHSS were observed, and miscarriage and multiple pregnancy rates were similar in the two groups. CONCLUSIONS: These findings suggest that the effectiveness of CAPA-IVM could be improved considerably by using a freeze-only strategy followed by frozen ET in subsequent cycles. TRIAL REGISTRATION NUMBER: NCT04297553 ( www.clinicaltrials.gov ).


Assuntos
Congelamento/efeitos adversos , Técnicas de Maturação in Vitro de Oócitos , Oócitos/crescimento & desenvolvimento , Folículo Ovariano/crescimento & desenvolvimento , Adolescente , Adulto , Coeficiente de Natalidade , Criopreservação/métodos , Transferência Embrionária , Feminino , Humanos , Nascido Vivo/epidemiologia , Indução da Ovulação/métodos , Gravidez , Taxa de Gravidez , Adulto Jovem
15.
Molecules ; 26(5)2021 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-33652771

RESUMO

The aim of this research was to investigate the effect of the number of freeze-thaw cycles (0, 1, 3, 5, and 7) on porcine longissimus protein and lipid oxidation, as well as changes in heterocyclic aromatic amines (HAAs) and advanced glycation end products (AGEs) and their precursors. We analyzed the relationship among HAAs, AGEs, oxidation, and precursors and found the following results after seven freeze-thaw cycles. The HAAs, Norharman and Harman, were 20.33% and 16.67% higher, respectively. The AGEs, Nε-carboxyethyllysine (CEL) and Nε-carboxymethyllysine (CML), were 11.81% and 14.02% higher, respectively. Glucose, creatine, and creatinine were reduced by 33.92%, 5.93%, and 1.12%, respectively after seven freeze-thaw cycles. Norharman was significantly correlated with thiobarbituric acid reactive substances (TBARS; r2 = 0.910) and glucose (r2 = -0.914). Harman was significantly correlated to TBARS (r2 = 0.951), carbonyl (r2 = 0.990), and glucose (r2 = -0.920). CEL was correlated to TBARS (r2 = 0.992) and carbonyl (r2 = 0.933). These changes suggest that oxidation and the Maillard reaction during freeze-thaw cycles promote HAA and AGE production in raw pork.


Assuntos
Tecido Adiposo/metabolismo , Aminas/metabolismo , Compostos Heterocíclicos/metabolismo , Proteínas/metabolismo , Aminas/química , Animais , Galinhas , Culinária , Congelamento/efeitos adversos , Compostos Heterocíclicos/química , Humanos , Reação de Maillard , Carne/análise , Oxirredução , Carne de Porco/análise , Suínos , Substâncias Reativas com Ácido Tiobarbitúrico/química , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo
16.
Transfusion ; 61(4): 1202-1214, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33569783

RESUMO

BACKGROUND: The kinetics of hematopoietic recovery after autologous stem cell transplantation (ASCT) may be affected by laboratory procedures. The aim of this study was to evaluate the influence of characteristics of the cryopreserved units of peripheral blood stem cells (PBSC) on postthawing cell viability and engraftment outcomes after ASCT. STUDY DESIGN AND METHODS: This was a retrospective cohort study including individuals referred for ASCT. Cryopreservation was conducted at a single processing facility between 2014 and 2019, and patients received clinical care at six transplant centers. Covariates and outcome data were retrieved from participants' records. RESULTS: The study population comprised 619 patients (345 [55.7%] male). Median age was 53 years. Multiple myeloma was the most common diagnosis (62.7%). Higher preapheresis CD34+ cell count, lower nucleated cell (NC) concentration per cryobag, and composition of the cryoprotectant solution (5% dimethyl sulfoxide [DMSO] and 6% hydroxyethyl starch) were statistically significantly associated with higher postthawing cell viability. The linear regression model for time to neutrophil and platelet engraftment included the infused CD34+ cell dose and the composition of the cryoprotectant solution. Patients who had PBSC cryopreserved using 10% DMSO solution presented six times higher odds (odds ratio [OR] = 6.9; 95% confidence interval [CI]: 2.2-21.1; p = .001) of delayed neutrophil engraftment (>14 days) and two times higher odds (OR = 2.3, 95%CI: 1.4-3.7; p = .001) of prolonged hospitalization (>18 days). DISCUSSION: The study showed that mobilization efficacy, NC concentration, and the composition of the cryoprotectant solution significantly affected postthawing cell viability. In addition, the composition of the cryoprotectant solution significantly impacted engraftment outcomes and time of hospitalization after ASCT.


Assuntos
Transplante de Células-Tronco Hematopoéticas , Células-Tronco Hematopoéticas/fisiologia , Laboratórios , Células-Tronco de Sangue Periférico/fisiologia , Prática Profissional , Adulto , Idoso , Sobrevivência Celular , Estudos de Coortes , Criopreservação/normas , Feminino , Congelamento/efeitos adversos , Mobilização de Células-Tronco Hematopoéticas/normas , Transplante de Células-Tronco Hematopoéticas/normas , Células-Tronco Hematopoéticas/citologia , Humanos , Laboratórios/normas , Masculino , Pessoa de Meia-Idade , Mieloma Múltiplo/sangue , Mieloma Múltiplo/epidemiologia , Mieloma Múltiplo/terapia , Transplante de Células-Tronco de Sangue Periférico , Células-Tronco de Sangue Periférico/citologia , Prática Profissional/normas , Estudos Retrospectivos , Manejo de Espécimes/métodos , Manejo de Espécimes/normas , Transplante Autólogo , Resultado do Tratamento
17.
Lipids Health Dis ; 20(1): 21, 2021 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-33632254

RESUMO

BACKGROUND: PCSK9 monoclonal antibody lowers plasma PCSK9 and LDL-cholesterol levels. The manufacturers recommend drug storage at 2-8 °C, and not above 25 °C. This study aimed to investigate drug stability at various temperatures that this drug could be exposed to during medication handling and transportation in tropical countries. METHODS: Alirocumab and evolocumab were tested in 3 study conditions: room temperature (RT), cooler device with cold pack, and freeze-thaw for 9 and 18 h. Heated drugs were used as negative control. Free plasma PCSK9 levels from 9 hyperlipidemia subjects were measured with ELISA. RESULTS: Average subject age was 49.2 ± 18.4 years. Percent PCSK9 inhibition significantly declined in heated drugs compared to baseline. Average RT during the study period was 30.4 ±2.6 °C. Change in percent PCSK9 inhibition of PCSK9 mAb at RT from baseline was - 5.8 ± 4.4% (P = 0.005) and - 11.0 ± 8.9% (P = 0.006) for alirocumab at 9 h and 18 h, and - 9.7 ± 11.8% (P = 0.04) and - 15.1 ± 14.3% (P = 0.01) for evolocumab at 9 and 18 h, respectively. In contrast, there were no significant changes in percent PCSK9 inhibition from baseline when PCSK9 mAb was stored in a cooler. In freeze-thaw condition, changes in percent PCSK9 inhibition from baseline to 9 and 18 h were - 5.2 ± 2.9% (P = 0.001) and - 2.6 ± 4.9% (P = 0.16) for alirocumab, and - 1.8 ± 4.2% (P = 0.24) and 0.4 ± 6.1% (P = 0.83) for evolocumab. CONCLUSION: Proper drug storage according to manufacturer's recommendation is essential. Drug storage at RT in tropical climate for longer than 9 h significantly decreased drug efficacy; however, storage in a cooler device with cold pack for up to 18 h is safe.


Assuntos
Anticorpos Monoclonais/química , LDL-Colesterol/sangue , Estabilidade de Medicamentos , Pró-Proteína Convertase 9/química , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais Humanizados/química , Anticorpos Monoclonais Humanizados/farmacologia , Feminino , Congelamento/efeitos adversos , Humanos , Masculino , Pessoa de Meia-Idade , Inibidores de PCSK9 , Pró-Proteína Convertase 9/imunologia , Temperatura , Adulto Jovem
18.
Genes (Basel) ; 12(2)2021 02 03.
Artigo em Inglês | MEDLINE | ID: mdl-33546197

RESUMO

The cryoprotection of cell activity is a key determinant in frozen-dough technology. Although several factors that contribute to freezing tolerance have been reported, the mechanism underlying the manner in which yeast cells respond to freezing and thawing (FT) stress is not well established. Therefore, the present study demonstrated the relationship between DaMDHAR encoding monodehydroascorbate reductase from Antarctic hairgrass Deschampsia antarctica and stress tolerance to repeated FT cycles (FT2) in transgenic yeast Saccharomyces cerevisiae. DaMDHAR-expressing yeast (DM) cells identified by immunoblotting analysis showed high tolerance to FT stress conditions, thereby causing lower damage for yeast cells than wild-type (WT) cells with empty vector alone. To detect FT2 tolerance-associated genes, 3'-quant RNA sequencing was employed using mRNA isolated from DM and WT cells exposed to FT (FT2) conditions. Approximately 332 genes showed ≥2-fold changes in DM cells and were classified into various groups according to their gene expression. The expressions of the changed genes were further confirmed using western blot analysis and biochemical assay. The upregulated expression of 197 genes was associated with pentose phosphate pathway, NADP metabolic process, metal ion homeostasis, sulfate assimilation, ß-alanine metabolism, glycerol synthesis, and integral component of mitochondrial and plasma membrane (PM) in DM cells under FT2 stress, whereas the expression of the remaining 135 genes was partially related to protein processing, selenocompound metabolism, cell cycle arrest, oxidative phosphorylation, and α-glucoside transport under the same condition. With regard to transcription factors in DM cells, MSN4 and CIN5 were activated, but MSN2 and MGA1 were not. Regarding antioxidant systems and protein kinases in DM cells under FT stress, CTT1, GTO, GEX1, and YOL024W were upregulated, whereas AIF1, COX2, and TRX3 were not. Gene activation represented by transcription factors and enzymatic antioxidants appears to be associated with FT2-stress tolerance in transgenic yeast cells. RCK1, MET14, and SIP18, but not YPK2, have been known to be involved in the protein kinase-mediated signalling pathway and glycogen synthesis. Moreover, SPI18 and HSP12 encoding hydrophilin in the PM were detected. Therefore, it was concluded that the genetic network via the change of gene expression levels of multiple genes contributing to the stabilization and functionality of the mitochondria and PM, not of a single gene, might be the crucial determinant for FT tolerance in DaMDAHR-expressing transgenic yeast. These findings provide a foundation for elucidating the DaMDHAR-dependent molecular mechanism of the complex functional resistance in the cellular response to FT stress.


Assuntos
Congelamento/efeitos adversos , NADH NADPH Oxirredutases/genética , Saccharomyces cerevisiae/genética , Regulação Fúngica da Expressão Gênica/genética , Redes Reguladoras de Genes/genética , Poaceae/enzimologia , Estresse Fisiológico/genética , Fatores de Transcrição/genética
19.
Photosynth Res ; 149(1-2): 135-153, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-33033976

RESUMO

In Antarctica, multiple stresses (low temperatures, drought and excessive irradiance) hamper photosynthesis even in summer. We hypothesize that controlled inactivation of PSII reaction centres, a mechanism widely studied by pioneer work of Fred Chow and co-workers, may effectively guarantee functional photosynthesis under these conditions. Thus, we analysed the energy partitioning through photosystems in response to temperature in 15 bryophyte species presenting different worldwide distributions but all growing in Livingston Island, under controlled and field conditions. We additionally tested their tolerance to desiccation and freezing and compared those with their capability for sexual reproduction in Antarctica (as a proxy to overall fitness). Under field conditions, when irradiance rules air temperature by the warming of shoots (up to 20 °C under sunny days), a predominance of sustained photoinhibition beyond dynamic heat dissipation was observed at low temperatures. Antarctic endemic and polar species showed the largest increases of photoinhibition at low temperatures. On the contrary, the variation of thermal dissipation with temperature was not linked to species distribution. Instead, maximum non-photochemical quenching at 20 °C was related (strongly and positively) with desiccation tolerance, which also correlated with fertility in Antarctica, but not with freezing tolerance. Although all the analysed species tolerated - 20 °C when dry, the tolerance to freezing in hydrated state ranged from the exceptional ability of Schistidium rivulare (that survived for 14 months at - 80 °C) to the susceptibility of Bryum pseudotriquetrum (that died after 1 day at - 20 °C unless being desiccated before freezing).


Assuntos
Adaptação Fisiológica , Briófitas/fisiologia , Temperatura Baixa/efeitos adversos , Desidratação , Congelamento/efeitos adversos , Fotossíntese/fisiologia , Luz Solar/efeitos adversos , Regiões Antárticas
20.
Meat Sci ; 172: 108335, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33059179

RESUMO

The inhibitory effect of ice structuring protein (ISP) on the quality deterioration of quick-frozen pork patties subjected to multiple freeze-thaw (F-T) cycles was investigated. The inhibitory effect of ISP on patty quality deterioration was obvious after five F-T cycles (P < 0.05). The hardness and springiness of patties with 0.20% ISP were 3.84% and 10.61% higher than those of patties without ISP, and the thawing loss of patties with 0.20% ISP was 43.64% lower than that of patties without ISP (P < 0.05). In addition, ISP effectively restrained moisture migration and destruction of pork patty microstructure during F-T cycles. More importantly, thiobarbituric acid reactive substance levels and carbonyl contents in the patties with 0.20% ISP were 25% and 32% lower than those in the control group (no significant difference with patties with 0.30% ISP) after five F-T cycles. Therefore, these results illustrated the potential benefits of ISP in meat products.


Assuntos
Congelamento/efeitos adversos , Produtos da Carne/análise , Proteínas de Carne/química , Animais , Qualidade dos Alimentos , Gelo , Carbonilação Proteica , Suínos , Substâncias Reativas com Ácido Tiobarbitúrico/análise
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