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1.
Zhonghua Wai Ke Za Zhi ; 58(5): 397-400, 2020 May 01.
Artigo em Chinês | MEDLINE | ID: mdl-32393008

RESUMO

Liver transplantation is currently the only effective treatment for end-stage liver disease. The preservation of donor liver before transplantation is important. But both traditional static cold storage and machine perfusion are limited by the preservation time, so that the allotment space of donor liver is limited, which inevitably leads to the abandonment of part of donor liver.At present, to find a preservation technology that not only guarantees the quality of donor liver but also has a longer effective preservation time is the direction of joint efforts of all clinicians. Supercooling liver preservation(SLP) to find a preservation technology that not only guarantees the quality of donor liver but also has a longer effective preservation time is the direction of joint efforts of all clinicians. SLP, a new method based on using cryoprotectants to keep donor liver under -6 ℃ and recovering the graft with subnormothermic machine perfusion that enables long-term transplantation survival following 4 days of liver preservation, made a revolutionary breakthrough in the field of liver preservation, carved out a new field for the research of liver preservation. This article reviews the latest experimental research progress of SLP in the field of liver transplantation.


Assuntos
Criopreservação/métodos , Doença Hepática Terminal/cirurgia , Transplante de Fígado , Fígado , Preservação de Órgãos/métodos , Criopreservação/tendências , Humanos , Preservação de Órgãos/tendências , Perfusão/métodos
2.
Cir. pediátr ; 33(1): 25-29, ene. 2020. tab, ilus
Artigo em Espanhol | IBECS | ID: ibc-186134

RESUMO

Introducción: La transposición ovárica es una técnica quirúrgica que permite alejar los ovarios de la zona de irradiación en pacientes que van a recibir radioterapia abdominal o pélvica. Se han descrito buenas tasas de conservación de función. Sin embargo, no existen estudios en pacientes pediátricos. Presentamos nuestra serie de pacientes intervenidas en nuestro centro. Material y métodos: Estudio retrospectivo de pacientes a las que se le realizó transposición ovárica en nuestra unidad de cirugía oncológica pediátrica entre los años 2008 y 2018. La técnica empleada dependió de la edad, la zona de la irradiación y de la asociación o no con la cirugía del tumor primario. Resultados: Durante el periodo de estudio se realizaron un total de 21 transposiciones ováricas en 13 pacientes (8 bilaterales, 4 izquierdas y 1 derecha). En todos los casos se realizó criopreservación de corteza ovárica dentro del programa de preservación de fertilidad. Once de las 13 intervenciones fueron por laparoscopia, seccionándose el ligamento tubo-ovárico en 16 unidades y en 5 se realizó transposición ovárica sim-ple. La estancia hospitalaria media fue de 2,4 días sin registrarse ninguna complicación en el postoperatorio. Actualmente 9 pacientes continúan seguimiento en sus unidades de oncología pediátrica de referencia, sin haberse notificado ninguna complicación. Conclusión: La transposición ovárica es una técnica segura y reproducible. Estas pacientes requieren un seguimiento prolongado para conocer el estado de los ovarios tras el tratamiento oncológico


Introduction: Ovarian transposition is a surgical procedure allow-ing gonadal mobilization from a radiation spotlight to a safer, radiation therapy-free place in patients receiving abdominal or pelvic radiation therapy. And these patients can be managed using minimally invasive surgery. Although some authors have reported good results in fertility preservation with this technique, there are no long-term studies in the pediatric population. We present our results with this procedure in our oncological patients from the last decade. Materials and methods: Retrospective review of medical reports of patients who underwent laparoscopic ovarian transposition in our pediatric oncological surgery unit from 2008 to 2018. The technique varied depending on age, irradiation zone, and concomitant oncologi-cal resections. Results: A total of 21 ovarian transpositions were successfully per-formed in 13 patients. Eight were bilateral, four were left and only one was right. An ovarian cortex cryopreservation was simultaneously carried out in all patients. Eleven procedures were completed laparoscopically, and the suspensory ovarian ligament was divided in sixteen cases. The Fallopian tube was divided in one case, and a simple ovarian transposition was conducted in five cases. Mean hospital stay was 2.4 days, and no complications in the immediate postoperative period were noted. Conclusion: Ovarian transposition is a feasible, safe technique. These patients require an extended follow-up to assess ovarian function after oncological treatment


Assuntos
Humanos , Feminino , Criança , Adolescente , Preservação da Fertilidade/métodos , Preservação da Fertilidade/tendências , Atenção Terciária à Saúde , Criopreservação/métodos , Ovariectomia/métodos , Estudos Retrospectivos , Complicações Pós-Operatórias , Epidemiologia Descritiva , Ovário/cirurgia , Radioterapia/métodos , Laparoscopia
3.
Vox Sang ; 114(8): 876-883, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31625187

RESUMO

BACKGROUND AND OBJECTIVES: There is no standard methodology for post-thaw sample preparation for viability analysis of umbilical cord blood units (CBU). A common challenge faced by CB bank is for their product to meet the post-thaw cell viability threshold for CD45+ cells set at 40% by NetCord-FACT. The objective of this work was to improve the post-thaw staining method to maximize CD45+ cell viability so that clinically valuable samples meet the NetCord-FACT threshold criteria for CD45+ and CD34+ cell viabilities. MATERIALS AND METHODS: Samples of CBU buffy coats and CBU segments were thawed and taken for staining. Various parameters were evaluated on CD45+ and CD34+ cell viability as measured by 7-actinomycin D (7-AAD) staining. RESULTS: The results revealed that initiating the staining at 20 min post-thaw instead of 30, shortening the red cell lysis treatment, or performing lysis on ice and removing this step all together, all improved the viability of CD45+ cells. Using CBU segments, it was shown that the most effective approach in increasing the viability of CD45+ cells was the complete omission of red cell lysis step. However, removal of the lysis step can create technical artefacts during flow cytometry acquisition that results in an underestimation of the viability of CD34+ cells. This can be avoided and CD34+ cell viability restored with additional thresholding on CD45 signal. CONCLUSION: CB CD45+ cells are sensitive to red cell lysis treatment post-thaw; omission of this step provides the best viability and ultimately better reflects the quality of cells used for transplantation.


Assuntos
Criopreservação/métodos , Sangue Fetal/citologia , Antígenos Comuns de Leucócito/metabolismo , Antígenos CD34/genética , Antígenos CD34/metabolismo , Buffy Coat/citologia , Buffy Coat/metabolismo , Sobrevivência Celular , Criopreservação/normas , Sangue Fetal/metabolismo , Humanos , Antígenos Comuns de Leucócito/genética
4.
Med Sci Monit ; 25: 7908-7913, 2019 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-31638104

RESUMO

BACKGROUND Dimethyl sulfoxide (DMSO) cryoprotectant can effectively alleviate the damage to single tissue during cryopreservation and restore its physiological activity after rewarming. However, studies have not been successful for preserving large tissue. This study aimed to investigate the application conditions of DMSO in large composite-tissue by performing femoral artery perfusion and soaking in a rabbit hind-limb model. MATERIAL AND METHODS A microdialysis-freezing point osmometer was used to detect the minimum time required for effective perfusion of 10% v/v perfusion and 20% v/v perfusion group. Magnetic resonance spectroscopy (MRS) was used to detect the area under the spectrum peak of DMSO in perivascular, intramuscular, subcutaneous areas, and compare the area under the spectrum peak in the 20% vascular perfusion group and other whole immersion groups. RESULTS The minimum time required for effective perfusion of muscle in the 10% v/v group was 30 minutes, the DMSO concentration was stable at 5% v/v; In the 20% v/v group the minimum time was at 20 minutes, stable at 12% v/v. There was a statistically difference of the area under the spectrum peak in the 10% group and the 20% v/v group after vascular perfusion in different tissue locations (P<0.05). The 20% vascular perfusion group and the different concentration of DMSO in the large tissue soaking group were statistically different (P<0.05). There was a significant difference in the 20% v/v vascular perfusion group compared to the low concentration immersion group, but no difference compared to the 50% immersion group. CONCLUSIONS The effect of blood perfusion on cryopreservation in large tissue by using DMSO was slightly better than overall soaking, especially in preservation of skin and subcutaneous tissue.


Assuntos
Criopreservação/métodos , Dimetil Sulfóxido/química , Perfusão/métodos , Animais , Crioprotetores , Dimetil Sulfóxido/metabolismo , Artéria Femoral/patologia , Membro Posterior/patologia , Modelos Animais , Coelhos
6.
Turk J Pediatr ; 61(1): 1-6, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31559714

RESUMO

Konak M, Minici M, Tarakçi N, Altunhan H, Toker A, Örs R. Effects of the storage of breast milk at different temperatures on total antioxidant capacity, total oxidant status, and paraoxonase-1 level. Turk J Pediatr 2019; 61: 1-6. Breast milk is a well-balanced ideal nutritional source with high bioavailability for infants. As being a fresh, biological and dynamic product, changes in the breast milk during these storage periods have been the subject of ongoing research. This study aims to evaluate total antioxidant capacity (TAC), total oxidant status (TOS), and paraoxonase-1 (PON-1) levels of fresh and freezestored breast milk. Ten cc of breast milk was obtained from the mothers as the days between 10 and 15 in the morning within a 1-hour period. TAC, TOS, and PON-1 levels were evaluated in the fresh breast milk. Collected breast milk samples were divided into two groups for storage at -20°C or -80°C. Stored samples were tested for TAC, TOS, and PON-1 levels after 72 hours. The highest TAC level was detected in fresh breast milk (p < 0.05). The TOS levels of fresh breast milk showed a statistically significant reduction in rate after storage. The TOS levels at -20°C and -80°C were significantly lower at -80°C (p < 0.05). Our study results show that oxidant and antioxidant activities are at the maximum level in the fresh breast milk. In terms of antioxidant status the effect of freezing temperatures hasn`t been determined. We conclude that it is more convenient to store the breast milk at -80°C than to store at -20°C in terms of preserving the storage TOS level.


Assuntos
Antioxidantes/análise , Arildialquilfosfatase/análise , Temperatura Baixa , Criopreservação/métodos , Leite Humano/química , Oxidantes , Adulto , Antioxidantes/metabolismo , Arildialquilfosfatase/sangue , Biomarcadores/sangue , Extração de Leite , Feminino , Humanos , Lactente , Recém-Nascido , Recém-Nascido Prematuro , Masculino , Oxidantes/sangue , Estresse Oxidativo
7.
Int J Mol Sci ; 20(18)2019 Sep 17.
Artigo em Inglês | MEDLINE | ID: mdl-31533312

RESUMO

Mammalian sperm must undergo a set of structural and functional changes collectively termed as capacitation to ensure a successful oocyte fertilization. However, capacitation can be compromised by cryopreservation procedures, which alter the proteome and longevity of sperm. To date, how the protein changes induced by cryopreservation could affect the acquisition of sperm fertilizing potential remains unexplored. The present study investigated the protein profile of ram sperm during in vitro capacitation before and after cryopreservation to elucidate the impact of cryopreservation on sperm capacitation at a molecular level. Fresh and cryopreserved ram sperm were incubated under capacitating (CAP) and non-capacitating (NC) conditions for 240 min. The sperm proteome of these four treatments was analyzed and compared at different incubation times using reverse phase liquid chromatography coupled to mass spectrometry (RP-LC-MS/MS). The comparison between fresh and cryopreserved sperm suggested that cryopreservation facilitated an apoptosis-stress response and redox process, while the comparison between sperm incubated in CAP and NC conditions showed that capacitation increased those biological processes associated with signaling, metabolism, motility, and reproductive processes. In addition, 14 proteins related to mitochondrial activity, sperm motility, oocyte recognition, signaling, spermatogenesis, and the apoptosis-stress response underwent significant changes in abundance over time when fresh and cryopreserved sperm incubated in CAP and NC conditions were compared. Our results indicate that disturbances in a ram sperm proteome after cryopreservation may alter the quality of sperm and its specific machinery to sustain capacitation under in vitro conditions.


Assuntos
Criopreservação , Proteoma , Capacitação Espermática , Espermatozoides/metabolismo , Animais , Apoptose , Criopreservação/métodos , Ensaio de Imunoadsorção Enzimática , Congelamento , Masculino , Mitocôndrias , Capacitação Espermática/genética , Motilidade Espermática
8.
Transplant Proc ; 51(8): 2514-2519, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31473005

RESUMO

BACKGROUND: Hypothermic machine perfusion (HMP) has become a standard method of preservation for kidneys procured from expanded-criteria donors and donors after cardiac death. There are different systems and approaches to the HMP preservation period, with cold storage prior to HMP sometimes taking several hours. This study evaluated whether the time at which kidneys receive HMP had any influence on the outcomes of kidney transplantation. METHODS: In this analysis, patient and graft survival were evaluated over a 1-year post-transplantation period. Patients who received HMP kidneys (n = 379) were divided into 2 groups: those who received kidneys with a cold ischemia time (CIT) prior to HMP <295 minutes (group G1; n = 254) and those who received kidneys with CIT prior to HMP >295 minutes (group G2; n = 125). RESULTS: Delayed graft function was observed in 31.8% (81/254) of patients in group G1 vs 46.4% (58/125) of patients in group G2 (P = .007). One-year graft survival was statistically higher in the group G1 (93.2%; 233/254) vs group G2 (86.5%; 105/125, P = .029). Mean 1-year estimated glomerular filtration rate was significantly better in the group G1. CONCLUSIONS: In conclusion, introduction of HMP up to 295 minutes from procurement led to better early and 1-year graft results. Kidneys should receive HMP as soon as possible after retrieval, preferably during procurement.


Assuntos
Isquemia Fria/efeitos adversos , Criopreservação/métodos , Transplante de Rim/efeitos adversos , Rim , Preservação de Órgãos/efeitos adversos , Perfusão/efeitos adversos , Adulto , Isquemia Fria/métodos , Morte , Função Retardada do Enxerto/etiologia , Feminino , Taxa de Filtração Glomerular , Sobrevivência de Enxerto , Humanos , Hipotermia Induzida , Transplante de Rim/métodos , Masculino , Pessoa de Meia-Idade , Preservação de Órgãos/métodos , Perfusão/métodos , Fatores de Tempo , Doadores de Tecidos , Resultado do Tratamento
9.
Biomed Res Int ; 2019: 5397804, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31531356

RESUMO

For human infertility both male and female factors may be equally important. Searching for molecular biomarkers of male infertility, neglected for decades, and the attempts to explain regulatory mechanisms of fertilization become thus extremely important. Apart from examination of the structure and function of male gametes, also the possible importance of seminal plasma components should be considered. In this article we discuss data that indicate for the substantial significance of active seminal plasma components for conception and achievement of healthy pregnancy. Seminal plasma impact on the storage and cryopreservation of human and animal sperm and regulatory role of glycodelin on human sperm capacitation as well as hypothesized course of female immune response to allogenic sperm and conceptus has been discussed. The possible involvement of carbohydrates in molecular mechanism of fetoembryonic defense has been also mentioned.


Assuntos
Fertilização/fisiologia , Sêmen/fisiologia , Espermatozoides/fisiologia , Animais , Criopreservação/métodos , Humanos , Infertilidade Masculina/fisiopatologia , Masculino , Preservação do Sêmen/métodos , Capacitação Espermática/fisiologia , Motilidade Espermática/fisiologia
10.
Reprod Domest Anim ; 54 Suppl 3: 53-56, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31512317

RESUMO

DNA fragmentation of cumulus cells could be used as an indicator of oocyte vitrification success as an indirect indicator of the quality of the oocyte. This study was designed to compare the DNA fragmentation of post-mortem equine cumulus cells before or after vitrification in the absence of permeable cryoprotectant agents. Cumulus-oocyte complexes (COCs; n = 56) were recovered from slaughterhouse ovaries and subjected to in vitro maturation (42 hr/38.2°C/5%CO2 ) before (control group) or after a permeable cryoprotectant-free vitrification method using 1 M sucrose (vitrification group). After in vitro maturation, COCs were denuded, and cumulus cells were washed and stored at -80°C until thawing. Cumulus cell samples were processed with the chromatin dispersion test (Ovoselect, Halotech DNA, Spain). Low, high and total DNA fragmentation percentages of cumulus cells were recorded and compared between the two groups by Student's t test. Results were expressed as mean ± SEM. The vitrified group resulted in significantly higher (p < 0.05) percentages for low (16.81 ± 1.62 vs. 6.63 ± 0.77) and total (21.14 ± 1.84 vs. 12.76 ± 1.48) DNA fragmentation of cumulus cells. There were no significant differences between groups for high DNA fragmentation of cumulus cells. In conclusion, permeable cryoprotectant-free vitrification of equine oocytes increased the total DNA fragmentation rate of cumulus cells but protected them against high DNA fragmentation rates. Further studies are needed to examine the relationship between DNA fragmentation of cumulus cells and the developmental competence of equine oocytes.


Assuntos
Criopreservação/veterinária , Células do Cúmulo , Fragmentação do DNA , Oócitos , Animais , Criopreservação/métodos , Feminino , Cavalos , Vitrificação
11.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi ; 33(9): 1102-1107, 2019 Sep 15.
Artigo em Chinês | MEDLINE | ID: mdl-31512450

RESUMO

Objective: To review the current status and progress of sterilization and preservation for allograft in anterior cruciate ligament reconstruction. Methods: The related literature about the sterilization and preservation of allografts in anterior cruciate ligament reconstruction was extensively reviewed and summarized. Results: There are many sterilization methods for allografts, the most commonly used method is γ-ray irradiation, but the optimal irradiation dose is still unclear. Electron beam irradiation is also available, but excessive dose is harmful to graft shaping. A combined sterilization method combining physics and chemistry methods is still being explored. Cryopreservation is the most commonly used method of preservation. In order to reduce the influence of crystals, the principle of "slow cooling and rapid rewarming" should be adhered to as far as possible. Conclusion: The processing methods of allograft can affect the effectiveness of anterior cruciate ligament reconstruction. The clinical doctors should consider the sterilization and preservation methods in practice.


Assuntos
Reconstrução do Ligamento Cruzado Anterior , Criopreservação , Esterilização , Transplante Homólogo , Aloenxertos , Lesões do Ligamento Cruzado Anterior/cirurgia , Reconstrução do Ligamento Cruzado Anterior/métodos , Criopreservação/métodos , Humanos , Esterilização/métodos , Transplante Homólogo/métodos
12.
Anim Reprod Sci ; 208: 106103, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31405464

RESUMO

The purpose of this study was to evaluate the effect of different concentrations of CoQ10 in soybean lecithin (SL) or egg yolk (EY) extenders on ram semen cryopreservation. Semen samples were collected from five rams, twice a week, then diluted in the extenders (SL and EY) containing different concentrations of CoQ10 as follows: extender containing SL: 0 µM (control, SL/Q0), 1 µM (SL/Q1), 2 µM (SL/Q2), 5 µM (SL/Q5) and 10 µM (SL/Q10) CoQ10; extender containing EY: 0 µM (control, EY/Q0), 1 µM (EY/Q1), 2 µM (EY/Q2), 5 µM (EY/Q5) and 10 µM (EY/Q10) CoQ10. Sperm motion characteristics, membrane integrity, abnormal morphology, viability, apoptotic-like changes, mitochondria active potential, acrosome integrity and lipid peroxidation were evaluated after freeze-thaw process. The SL/Q1, SL/Q2, EY/Q1 and EY/Q2 resulted in greater (P ≤ 0.05) sperm total motility, progressive motility, membrane integrity and mitochondria active potential compared to the other groups. Acrosome integrity in the SL/Q0, SL/Q1, SL/Q2, EY/Q0, EY/Q1 and EY/Q2 groups was greater (P ≤ 0.05) than in the SL/Q5, SL/Q10, EY/Q5 and EY/Q10 groups. The SL/Q2 and EY/Q2 treatment groups had greater (P ≤ 0.05) sperm viability rates and less apoptotic-like changes and lipid peroxidation. The CoQ10 compound could be explored as a novel potential antioxidant for cryopreservation of ram semen because with used of this compound in the present study there was an improved post-thawed sperm quality.


Assuntos
Gema de Ovo , Lecitinas/farmacologia , Preservação do Sêmen/veterinária , Ovinos , Espermatozoides/efeitos dos fármacos , Ubiquinona/análogos & derivados , Animais , Antioxidantes/farmacologia , Criopreservação/métodos , Criopreservação/veterinária , Congelamento , Peroxidação de Lipídeos , Masculino , Sêmen/efeitos dos fármacos , Preservação do Sêmen/métodos , Ubiquinona/farmacologia
13.
J Assist Reprod Genet ; 36(9): 1823-1835, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31376104

RESUMO

PURPOSE: Vitrification is a well-accepted fertility preservation procedure for cryopreservation of oocytes and embryos but little is known regarding ovarian tissue, for which slow freezing is the current convention. The aim of the present study was to assess the efficiency of non-equilibrium vitrification compared to conventional slow freezing for ovarian cortex cryopreservation. METHODS: Using prepubertal sheep ovaries, the capacity of the tissue to sustain folliculogenesis following cryopreservation and in vitro culture was evaluated. Ovarian cortex fragments were cultured in wells for 9 days, immediately or after cryopreservation by conventional slow freezing or non-equilibrium vitrification in straws. During culture, follicular populations within cortex were evaluated by histology and immunohistochemistry for PCNA and TUNEL. Steroidogenic activity of the tissue was monitored by assay for progesterone and estradiol in spent media. RESULTS: No significant differences in follicle morphology, PCNA, or TUNEL labeling were observed between cryopreservation methods at the initiation of culture. Similar decreases in the proportion of primordial follicle population, and increases in the proportion of growing follicles, were observed following culture of fresh or cryopreserved ovarian tissue regardless of cryopreservation method. At the end of culture, PCNA and TUNEL-positive follicles were not statistically altered by slow freezing or vitrification in comparison to fresh cultured fragments. CONCLUSIONS: Overall, for both cryopreservation methods, the cryopreserved tissue showed equal capacity to fresh tissue for supporting basal folliculogenesis in vitro. Taken together, these data confirm that both non-equilibrium vitrification and slow-freezing methods are both efficient for the cryopreservation of sheep ovarian cortex fragments.


Assuntos
Criopreservação/métodos , Folículo Ovariano , Ovário/fisiologia , Animais , Estradiol/metabolismo , Feminino , Preservação da Fertilidade/métodos , Folículo Ovariano/citologia , Folículo Ovariano/fisiologia , Progesterona/metabolismo , Antígeno Nuclear de Célula em Proliferação/metabolismo , Puberdade , Ovinos , Técnicas de Cultura de Tecidos , Vitrificação
14.
Pediatrics ; 144(3)2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31383814

RESUMO

BACKGROUND: Fertility preservation enables patients undergoing gonadotoxic therapies to retain the potential for biological children and now has broader implications in the care of transgender individuals. Multiple medical societies recommend counseling on fertility preservation before initiating therapy for gender dysphoria; however, outcome data pre- and posttreatment are limited in feminizing transgender adolescents and young adults. METHODS: The University of Pittsburgh Institutional Research Board approved this study. Data were collected retrospectively on transgender patients seeking fertility preservation between 2015 and 2018, including age at initial consultation and semen analysis parameters. RESULTS: Eleven feminizing transgender patients accepted a referral for fertility preservation during this time; consultation occurred at median age 19 (range 16-24 years). Ten patients attempted and completed at least 1 semen collection. Eight patients cryopreserved semen before initiating treatment. Of those patients, all exhibited low morphology with otherwise normal median semen analysis parameters. In 1 patient who discontinued leuprolide acetate to attempt fertility preservation, transient azoospermia of 5 months' duration was demonstrated with subsequent recovery of spermatogenesis. In a patient who had previously been treated with spironolactone and estradiol, semen analysis revealed persistent azoospermia for the 4 months leading up to orchiectomy after discontinuation of both medications. CONCLUSIONS: Semen cryopreservation is a viable method of fertility preservation in adolescent and young adult transgender individuals and can be considered in patients who have already initiated therapy for gender dysphoria. Further research is needed to determine the optimal length of time these therapies should be discontinued to facilitate successful semen cryopreservation.


Assuntos
Criopreservação/métodos , Preservação da Fertilidade/métodos , Disforia de Gênero/terapia , Sêmen , Adolescente , Aconselhamento , Estradiol/uso terapêutico , Estrogênios/uso terapêutico , Feminino , Disforia de Gênero/psicologia , Hormônio Liberador de Gonadotropina/agonistas , Humanos , Masculino , Estudos Retrospectivos , Espironolactona/uso terapêutico , Adulto Jovem
15.
Blood Transfus ; 17(4): 263-273, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31385799

RESUMO

BACKGROUND: Sex hormone intake in blood donors may affect the quality of red blood cell (RBC) products via modulation of RBC function and predisposition to haemolysis during cold storage. The aims of this study were to evaluate the association between female sex hormone intake and RBC storage outcomes, and to examine possible mechanisms by which sex hormones interact with RBCs. MATERIALS AND METHODS: Sex hormone intake by race/ethnicity and menopausal status, and association analyses between hormone intake and donor scores of storage, osmotic or oxidative haemolysis, were evaluated in 6,636 female donors who participated in the National Heart, Lung and Blood Institute's RBC-Omics study. A calcium fluorophore, Fluo-3AM, was used to define RBC calcium influx in response to exogenous sex hormones or transient receptor potential cation (TRPC) channel drugs. RESULTS: Sex hormone intake was more prevalent in premenopausal women from all racial groups (18-31%) than in postmenopausal women (4-8%). Hormone intake was significantly (p<0.0001) associated with reduced storage haemolysis in all females, reduced osmotic haemolysis in postmenopausal donors (23.1±10.2% vs 26.8±12.0% in controls, p<0.001), and enhanced susceptibility to oxidative haemolysis in premenopausal women. In vitro, supraphysiological levels of progesterone (10 µmol/L), but not 17ß-oestradiol or testosterone, inhibited calcium influx into RBC and was associated with lower spontaneous haemolysis after 30 days of cold storage (0.95±0.18% vs 1.85±0.35% in controls, p<0.0001) or in response to a TRPC6 activator. CONCLUSIONS: Sex hormone intake in female donors is associated with changes in RBC predisposition to haemolysis. Menstrual status and the type of hormone preparation may contribute to differences in haemolytic responses of female RBCs to osmotic and oxidative stress. Progesterone modulates calcium influx into RBC via a mechanism that may involve interactions with membrane TRPC6 channels.


Assuntos
Doadores de Sangue , Eritrócitos/metabolismo , Hormônios Esteroides Gonadais/farmacologia , Hemólise , Progesterona/farmacologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Preservação de Sangue/métodos , Cálcio/metabolismo , Criopreservação/métodos , Eritrócitos/citologia , Eritrócitos/efeitos dos fármacos , Feminino , Hormônios Esteroides Gonadais/administração & dosagem , Hemólise/efeitos dos fármacos , Humanos , Menopausa , Pessoa de Meia-Idade , Progesterona/administração & dosagem , Adulto Jovem
16.
Vox Sang ; 114(7): 711-720, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31373012

RESUMO

BACKGROUND AND OBJECTIVES: There is a growing concern for shortage in future blood supply, caused by a predicted decrease in eligible blood donors and simultaneous increase in recipients. Cryopreservation of split red blood cell units could increase stock supply by reducing waste of rare blood. This would be particularly useful in paediatric care where very small volumes often are transfused. The aim of this study was to develop a cryopreservation protocol for split units using the closed-system automated cell processor ACP215, as such protocols are currently missing. MATERIALS AND METHODS: Using a pool-and-split design, red blood cell units (N = 8) were glycerolized and frozen, either as standard volume reference units, or further divided into three smaller split units each. After thawing/deglycerolization, the supernatant of the smaller splits was reduced by additional centrifugation, and new SAGM was added to 60% haematocrit. The units were analysed for storage lesion effects up to ten days post-thawing. RESULTS: Haemolysis and extracellular potassium ion levels were lower in the split units than in the whole units from day three onwards. The metabolic parameters pH, ATP, glucose and lactate were also lower, though likely caused by lower additive solution pH rather than storage. CONCLUSION: Split units of red blood cells can be successfully cryopreserved using the ACP215. In addition to favourably low haemolysis and potassium, they also have higher haematocrit than corresponding whole units and enable involvement of fewer donors at repeated transfusions. These characteristics are all desirable features in paediatric care.


Assuntos
Preservação de Sangue/métodos , Criopreservação/métodos , Eritrócitos/metabolismo , Antígenos de Grupos Sanguíneos , Criança , Eritrócitos/efeitos dos fármacos , Congelamento/efeitos adversos , Glucose/metabolismo , Glicerol/farmacologia , Hemólise , Humanos , Ácido Láctico/metabolismo , Potássio/metabolismo
17.
Acta Clin Croat ; 58(1): 147-156, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31363337

RESUMO

Although breast cancer (BC) occurs more often in older women, it is the most commonly diagnosed malignancy in women of childbearing age. Owing to the overall advancement of modern medicine and the growing global trend of delaying childbirth until later age, we find ever more younger women diagnosed and treated for BC who have not yet completed their family. Therefore, fertility preservation has emerged as a very important quality of life issue for young BC survivors. This paper reviews currently available options for fertility preservation in young women with early-stage BC and highlights the importance of a multidisciplinary approach to fertility preservation as a very important quality of life issue for young BC survivors. Pregnancy after BC treatment is considered not to be associated with an increased risk of BC recurrence; therefore, it should not be discouraged for those women who want to achieve pregnancy after oncologic treatment. Currently, it is recommended to delay pregnancy for at least 2 years after BC diagnosis, when the risk of recurrence is highest. However, BC patients of reproductive age should be informed about the potential negative effects of oncologic therapy on fertility, as well as on the fertility preservation options available, and if interested in fertility preservation, they should be promptly referred to a reproductive specialist. Early referral to a reproductive specialist is an important factor that increases the likelihood of successful fertility preservation. Embryo and mature oocyte cryopreservation are currently the only established fertility preservation methods but they require ovarian stimulation (OS), which delays initiation of chemotherapy for at least 2 weeks. Controlled OS does not seem to increase the risk of BC recurrence. Other fertility preservation methods (ovarian tissue cryopreservation, cryopreservation of immature oocytes and ovarian suppression with gonadotropin-releasing hormone agonists) do not require OS but are still considered to be experimental techniques for fertility preservation.


Assuntos
Criopreservação/métodos , Preservação da Fertilidade/métodos , Recuperação de Oócitos/métodos , Indução da Ovulação/métodos , Adulto , Neoplasias da Mama/terapia , Feminino , Humanos , Infertilidade Feminina/prevenção & controle , Adulto Jovem
18.
Plast Reconstr Surg ; 144(3): 679-683, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31461026

RESUMO

BACKGROUND: Although cryopreservation has been widely used in clinical practice, it remains limited to small or thin bloodless tissues with a simple structure and function. In August of 2002, the authors performed the first successful transplantation of a cryopreserved vascularized rat hind limb. Subsequently, the authors extended this cryopreservation technique to the preservation of human fingers. METHODS: In December of 2002 and December of 2003, the authors performed two in situ implantations of cryopreserved amputated fingers for two patients by means of the "two-step" and programmed cryopreservation methods. In case 1, computed tomographic angiography was performed on the affected hand 6 months after surgery. In case 2, pieces of skin were obtained from the thawed amputated finger for pathologic examination before replantation. RESULTS: One finger was cryopreserved for 81 days and the other for 5 days. Both fingers were replanted successfully. Computed tomographic angiography revealed a patent radial proper digital artery in case 1. The pathologic results of case 2 showed satisfactory skin cell morphology. After 15- and 14-year follow-up assessments of the two patients, the replanted fingers achieved satisfactory appearance and function. CONCLUSIONS: The authors confirmed the effectiveness of deep cryopreservation for the long-term preservation of human fingers. The current application scope of these cryopreservation techniques includes small limbs with minimal amounts of muscle tissue. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, V.


Assuntos
Amputação Traumática/cirurgia , Criopreservação/métodos , Traumatismos dos Dedos/cirurgia , Reimplante/métodos , Adulto , Humanos , Masculino , Resultado do Tratamento , Adulto Jovem
19.
Biopreserv Biobank ; 17(5): 378-386, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31464512

RESUMO

Sugar kelp (Saccharina latissima) is an economically important species, and natural populations provide diverse and productive habitats as well as important ecosystem services. For seaweed aquaculture to be successful in newly emerging industry in Europe and other Western countries, it will have to develop sustainable production management strategies. A key feature in this process is the capacity to conserve genetic diversity for breeding programs aimed at developing seed stock for onward cultivation, as well as in the management of wild populations, as potentially interesting genetic resources are predicted to disappear due to climate change. In this study, the cryopreservation of male and female gametophytes (haploid life stage) of S. latissima by different combinations of two-step cooling methods and cryoprotectants was explored. We report here that cryopreservation constitutes an attractive option for the long-term preservation of S. latissima gametophytes, with viable cells in all treatment combinations. The highest viabilities for both male and female gametophytes were found using controlled-rate cooling methods combined with dimethyl sulfoxide 10% (v/v). Morphological normal sporophytes were observed to develop from cryopreserved vegetative gametophytic cells, independent of treatment. This indicates that cryopreservation is a useful preservation method for male and female S. latissima gametophytes.


Assuntos
Criopreservação/métodos , Células Germinativas Vegetais/citologia , Feófitas/citologia , Aquicultura , Proliferação de Células , Sobrevivência Celular , Dimetil Sulfóxido/química , Alga Marinha/crescimento & desenvolvimento
20.
PLoS One ; 14(7): e0220055, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31329628

RESUMO

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 [1] 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.


Assuntos
Criopreservação/métodos , Células-Tronco Mesenquimais/citologia , Apoptose , Sobrevivência Celular , Células Cultivadas , Humanos , Células-Tronco Mesenquimais/metabolismo , Tempo
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