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1.
Zhong Nan Da Xue Xue Bao Yi Xue Ban ; 45(8): 909-915, 2020 Aug 28.
Artigo em Inglês, Chinês | MEDLINE | ID: mdl-33053531

RESUMO

OBJECTIVES: To compare the effects of different preservation methods on biomacromolecules and microbial flora at different time points, and to understand the relationship between different cryopreservation methods, storage time and sample stability. METHODS: Tissue samples were taken from patients who underwent radical gastrectomy in the Third Xiangya Hospital, Central South University. Stool samples were obtained from volunteers. The tissue samples were stored in a refrigerator at -80 ℃ (-80 ℃ refrigerator group) and a liquid nitrogen tank (liquid nitrogen tank group), respectively. According to the preservation method, the stool samples were divided into a -80 ℃ refrigerator group, a liquid nitrogen tank group, a -80 ℃ refrigerator+ethanol absolute group, a liquid nitrogen tank+ethanol absolute group, and a room temperature+ethanol absolute group. Relevant indexes were examined and analyzed at the 0 month (fresh sample), 3th month, 6th month, 9th month and 12th month respectively to evaluate the sample stability with different preservation methods. RESULTS: Compared with the fresh sample, there was no significant changes in the protein concentration and the absorbance value of RNA in the samples in the liquid nitrogen tank group after 12 months of storage (both P>0.05); but the protein concentration of the tissue samples was decreased in the samples of the -80 ℃ refrigerator group after 12 months of storage (P<0.05), while the absorbance value of RNA was increased in the 9th and 12th months (both P>0.05). For the microbial flora of feces samples, at the phylum level, the flora changed significantly with time in the room temperature+ethanol absolute group, and the abundance of bacteroidetes, clostridium and proteobacteria was decreased significantly, while the abundance of firmicutes was increased significantly. At the family level, the abundance of bacteroidaceae, trichospirillaceae and veroniaceae was lower than that in the fresh sample, and the abundance of rumenbacteriaceae was higher than that in the fresh sample. In the 4 cryopreserved groups (-80 ℃ refrigerator group, liquid nitrogen tank group, -80 ℃ refrigerator+ethanol absolute group, liquid nitrogen tank+ethanol absolute group), the abundance of enterobacteriaceae was higher than that in the fresh sample. The analysis of Alpha and Beta diversity indicated that the species diversity, richness and flora structure of samples in the room temperature group were different from those in the fresh sample and other experimental groups. CONCLUSIONS: The protein concentration and RNA purity in the tissue samples preserved by liquid nitrogen tank are relatively stable; the protein concentration and RNA purity of the tissue samples which preserved in the -80 ℃ refrigerator are also relatively stable within half a year; the -80 ℃ refrigerator and liquid nitrogen tank cryopreservation can both maintain the microbial flora stability of the fecal specimen; the room temperature+ethanol absolute is not suitable for long-term preservation of stool samples.


Assuntos
Criopreservação , Nitrogênio , Etanol , Fezes , Humanos
2.
Zhong Nan Da Xue Xue Bao Yi Xue Ban ; 45(8): 994-998, 2020 Aug 28.
Artigo em Inglês, Chinês | MEDLINE | ID: mdl-33053544

RESUMO

The incidence of cervical cancer ranks the first among the gynecological malignant tumors. Given the implementation of widespread cervical cancer screening and the improvement of clinical treatment, the cervical cancer mortality has decreased substantially. However, surgery, radiotherapy, and chemotherapy may impair the ovarian function, resulting in premature ovarian insufficiency and premature climacteric, which has reduced the quality of life in young patients. As a new fertility preservation technology, autologous ovarian tissue cryopreservation and transplantation is developing rapidly and showing great potentiality in preserving ovarian endocrine function of young cervical cancer patients.


Assuntos
Preservação da Fertilidade , Neoplasias do Colo do Útero , Criopreservação , Detecção Precoce de Câncer , Feminino , Humanos , Qualidade de Vida
3.
Medicine (Baltimore) ; 99(33): e21660, 2020 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-32872029

RESUMO

This study aimed to investigate the effect of the duration of embryo culture on clinical outcome in vitrified-warmed cycles.This retrospective cohort study enrolled 10,464 infertile patients, with a total of 18,843 vitrified-warmed day 3 embryos from 2012 to 2017 at a single center. The patients were divided into 2 groups: 9470 cycles in the short-term culture group (0.5-8 hours of post-thaw culture) and 994 cycles in the 48 to 72 hours culture group. The independent effect of the following variables on clinical outcomes was determined: duration of post-thaw culture, maternal age, transferred embryos, embryo quality, and endometrial thickness.We found that the pregnancy rate was positively associated with the post-thaw culture time. Ordinary least square regression analyses showed that the duration of post-thaw culture was positively associated with implantation and live birth rates overall. However, the implantation and live birth rates were not significantly associated with the post-thaw culture time in the short-term culture group. Additionally, maternal age and the number of transferred embryos were independent predictors of the implantation and live birth rates. Moreover, the duration of post-thaw culture did not affect live birth weight.These results indicated that the pregnancy rate is positively associated with the duration of post-thaw culture. Therefore, under the condition of not affecting work shifts, properly prolonging the duration of post-thaw culture to improve the outcome of frozen-thawed embryo transfer should be considered.


Assuntos
Técnicas de Cultura Embrionária/métodos , Implantação do Embrião , Transferência Embrionária/métodos , Adulto , Estudos de Casos e Controles , Criopreservação/métodos , Transferência Embrionária/estatística & dados numéricos , Feminino , Humanos , Gravidez , Taxa de Gravidez , Estudos Retrospectivos , Fatores de Tempo , Vitrificação
4.
Anim Sci J ; 91(1): e13451, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32926550

RESUMO

This study assessed the effects of cryoprotectant concentration during equilibration on the efficiency of bovine blastocyst vitrification and the expression of selected developmentally important genes. In vitro produced bovine blastocysts were equilibrated in either 7.5% ethylene glycol (EG) + 7.5% DMSO (Va group) or in 2% EG + 2% DMSO (Vb group) then vitrified on Cryotop® sheets in 16.5% EG + 16.5% DMSO + 0.5M sucrose. After warming, embryos were cultured for 48 hr. Re-expansion, hatching, and the numbers of total and membrane damaged cells were compared among vitrified groups and a control. There was no significant difference between the vitrified groups in survival, cell numbers and the extent of membrane damage. Vitrification increased the number of membrane-damaged cells in both groups, however, in a greater extent in the Vb group. Vitrification increased (p < .05) the expression of the HSP70 gene in Va but not in Vb embryos. The expression of IGF2R, SNRPN, HDAC1, DNMT3B, BAX, OCT4, and IFN-t genes were the same in control and vitrified groups. In conclusion, the concentration of cryoprotectants during equilibration did not affect survival rates; however, normal cell numbers could be maintained only by equilibration in 15% cryoprotectants which was associated with increased HSP70 expression.


Assuntos
Blastocisto , Sobrevivência Celular/efeitos dos fármacos , Criopreservação/métodos , Criopreservação/veterinária , Crioprotetores/farmacologia , Fertilização In Vitro/métodos , Fertilização In Vitro/veterinária , Vitrificação/efeitos dos fármacos , Animais , Bovinos , Células Cultivadas , Relação Dose-Resposta a Droga , Feminino , Expressão Gênica , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP70/metabolismo
5.
Rinsho Ketsueki ; 61(8): 885-887, 2020.
Artigo em Japonês | MEDLINE | ID: mdl-32908051

RESUMO

Dimethyl sulfoxide (DMSO) is used as a cryoprotectant for peripheral blood stem cells (PBSC) preservation. Dimethyl sulfide (DMS) is a metabolite of DMSO secreted through patients' breath after PBSC infusion. It possesses malodor causing an unpleasant environment. We evaluated the efficacy of a photocatalyst environment purifier, which has the potential to lyse toxic substances, in reducing DMS malodor. High DMS concentration in the air after PBSC infusion rapidly decreased after operating the device. Our results suggest that photocatalytic reaction has the potential to reduce the DMS odor associated with PBSC infusion.


Assuntos
Filtros de Ar , Células-Tronco de Sangue Periférico , Criopreservação , Humanos , Sulfetos
6.
Blood Adv ; 4(17): 4147-4150, 2020 09 08.
Artigo em Inglês | MEDLINE | ID: mdl-32886750

RESUMO

Donor registries and transplantation societies recommend cryopreservation of unrelated donor hemopoietic progenitor cell (HPC) products before the recipient commences conditioning therapy to mitigate the donor and travel risks associated with the COVID-19 pandemic. However, little is known regarding the postthaw quality of such allogeneic products or the effect of precryopreservation storage and processing on these characteristics. We investigated the postthaw CD34+ cell recovery and viability of 305 allogeneic HPC products cryopreserved at 9 laboratories across Australia. Median postthaw CD34+ cell recovery was 76% and ranged from 6% to 122%. Longer transit time before cryopreservation, white cell count (WCC) during storage, and complex product manipulation before cryopreservation were independently associated with inferior postthaw CD34+ cell recovery. Longer precryopreservation transit time and WCC were also associated with inferior postthaw CD34+ cell viability. We conclude that although postthaw CD34+ cell recovery and viability of cryopreserved allogeneic HPC is generally acceptable, there is a significant risk of poor postthaw product quality, associated with prolonged storage time, higher WCC, and complex product manipulation precryopreservation. Awareness of expected postthaw recovery and practices that influence it will assist collection, processing, and transplant centers in optimizing outcomes for transplant recipients.


Assuntos
Antígenos CD34/análise , Criopreservação , Células-Tronco Hematopoéticas/citologia , Sobrevivência Celular , Infecções por Coronavirus/epidemiologia , Transplante de Células-Tronco Hematopoéticas , Humanos , Pandemias , Pneumonia Viral/epidemiologia , Transplante Homólogo
7.
Crit Rev Oncol Hematol ; 154: 103092, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32896752

RESUMO

Fertility preservation in women with cervical cancer is a demanding but evolving issue. Some remarkable achievements have been reached, in particular the improvement of primary and secondary prevention and the broadening of the indications for conservative surgery up to FIGO 2018 stage IB2. Natural pregnancy rate and the rate of obstetrics complications following conservative approach is satisfactory even if not optimal. On the other hand, the use of classic strategies for fertility preservation such as oocytes or ovarian cortex freezing is extremely limited, being the uterus compromised by treatment in a high proportion of cases. In fact, the availability of uterine surrogacy can play a role in the counseling and the decision-making process. The recent advent of uterus transplantation is fascinating but, at present, cannot be viewed as a realistic solution.


Assuntos
Preservação da Fertilidade , Neoplasias do Colo do Útero , Aconselhamento , Criopreservação , Feminino , Humanos , Oócitos , Ovário , Gravidez
8.
Anim Sci J ; 91(1): e13453, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32926526

RESUMO

This study was performed to evaluate reproductive performance after non-surgical embryo transfer (Ns-ET) of 10-15 porcine expanded blastocysts (ExBs) that had been vitrified and warmed (V/W) using the micro volume air cooling (MVAC) method. The effect of asynchrony between the donor and recipient estrous cycle was investigated. Ns-ET was conducted in recipients whose estrous cycle was asynchronous to that of donors by a delay of 2, 1, or 0 days. In the 2-day and 1-day groups, the similar farrowing rates (27.3% and 25.0%) and survival rates to term (13.9% and 15.7%) were obtained after Ns-ET of V/W ExBs. None of the recipients in 0-day group farrowed. Artificial insemination (AI) prior to Ns-ET was then evaluated. Ten-15 V/W ExBs were transferred non-surgically to 12 recipients whose estrous cycles were asynchronous to that of donors by a 2-day delay. All of the recipients produced piglets, and all (100.0%) delivered piglets were derived from the transferred V/W ExBs. The survival rate of V/W ExBs to term was 25.2%. These results demonstrate that Ns-ET of V/W ExBs using MVAC can facilitate piglet production, even if 10-15 embryos are transferred. Moreover, piglets were obtained stably when AI was performed prior to Ns-ET.


Assuntos
Blastocisto , Sobrevivência Celular , Criopreservação/métodos , Criopreservação/veterinária , Técnicas de Cultura Embrionária/veterinária , Transferência Embrionária/métodos , Transferência Embrionária/veterinária , Inseminação Artificial/métodos , Inseminação Artificial/veterinária , Suínos/fisiologia , Vitrificação , Animais , Ciclo Estral , Feminino , Reprodução
9.
BMJ ; 370: m2519, 2020 08 05.
Artigo em Inglês | MEDLINE | ID: mdl-32759285

RESUMO

OBJECTIVE: To compare the ongoing pregnancy rate between a freeze-all strategy and a fresh transfer strategy in assisted reproductive technology treatment. DESIGN: Multicentre, randomised controlled superiority trial. SETTING: Outpatient fertility clinics at eight public hospitals in Denmark, Sweden, and Spain. PARTICIPANTS: 460 women aged 18-39 years with regular menstrual cycles starting their first, second, or third treatment cycle of in vitro fertilisation or intracytoplasmic sperm injection. INTERVENTIONS: Women were randomised at baseline on cycle day 2 or 3 to one of two treatment groups: the freeze-all group (elective freezing of all embryos) who received gonadotropin releasing hormone agonist triggering and single frozen-thawed blastocyst transfer in a subsequent modified natural cycle; or the fresh transfer group who received human chorionic gonadotropin triggering and single blastocyst transfer in the fresh cycle. Women in the fresh transfer group with more than 18 follicles larger than 11 mm on the day of triggering had elective freezing of all embryos and postponement of transfer as a safety measure. MAIN OUTCOME MEASURES: The primary outcome was the ongoing pregnancy rate defined as a detectable fetal heart beat after eight weeks of gestation. Secondary outcomes were live birth rate, positive human chorionic gonadotropin rate, time to pregnancy, and pregnancy related, obstetric, and neonatal complications. The primary analysis was performed according to the intention-to-treat principle. RESULTS: Ongoing pregnancy rate did not differ significantly between the freeze-all and fresh transfer groups (27.8% (62/223) v 29.6% (68/230); risk ratio 0.98, 95% confidence interval 0.87 to 1.10, P=0.76). Additionally, no significant difference was found in the live birth rate (27.4% (61/223) for the freeze-all group and 28.7% (66/230) for the fresh transfer group; risk ratio 0.98, 95% confidence interval 0.87 to 1.10, P=0.83). No significant differences between groups were observed for positive human chorionic gonadotropin rate or pregnancy loss, and none of the women had severe ovarian hyperstimulation syndrome; only one hospital admission related to this condition occurred in the fresh transfer group. The risks of pregnancy related, obstetric, and neonatal complications did not differ between the two groups except for a higher mean birth weight after frozen blastocyst transfer and an increased risk of prematurity after fresh blastocyst transfer. Time to pregnancy was longer in the freeze-all group. CONCLUSIONS: In women with regular menstrual cycles, a freeze-all strategy with gonadotropin releasing hormone agonist triggering for final oocyte maturation did not result in higher ongoing pregnancy and live birth rates than a fresh transfer strategy. The findings warrant caution in the indiscriminate application of a freeze-all strategy when no apparent risk of ovarian hyperstimulation syndrome is present. TRIAL REGISTRATION: Clinicaltrials.gov NCT02746562.


Assuntos
Peso ao Nascer , Blastocisto , Criopreservação , Fertilização In Vitro/métodos , Transferência de Embrião Único/métodos , Aborto Espontâneo/epidemiologia , Adulto , Gonadotropina Coriônica/sangue , Feminino , Humanos , Nascimento Vivo , Ciclo Menstrual , Complicações do Trabalho de Parto/epidemiologia , Gravidez , Taxa de Gravidez , Nascimento Prematuro/epidemiologia , Fatores de Tempo
10.
Adv Exp Med Biol ; 1252: 181-187, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32816280

RESUMO

Fertility preservation includes all the necessary steps that have to be taken in order to implement a woman's aim for preserving and enhancing her future chances of having her own biological offspring.There are two available choices for patients to maintain their fertility.The first category includes all the preventive methods following the principle of reduction of gonadotoxicity, in terms of using fewer toxic drugs to the ovaries and similarly milder radiation regimens. It also includes the administration of agents that would keep the ovaries suppressed, to minimize the harmful effects of systemic therapy on the oocytes.The second includes necessary artificial reproductive technology procedures for collecting and freezing either gametes (oocytes) or zygotes (embryos). This can be accomplished with immature oocyte pickup followed by in vitro maturation (IVM). Alternatively, mature oocytes can be collected after minimal ovarian stimulation with very small doses or no medication at all, or through a natural cycle.Beyond that, promising is the modern option of ovarian tissue cryopreservation for future transplantation, while the option of oocyte donation is actually an option for childbearing, not for fertility.


Assuntos
Neoplasias da Mama , Aconselhamento , Preservação da Fertilidade , Fertilidade , Criança , Criopreservação , Feminino , Humanos , Oócitos
11.
PLoS One ; 15(8): e0237766, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32822364

RESUMO

Semen contains epithelial cells that can be cultured in vitro. For somatic cell nuclear transfer applications, it is essential to know whether clone(s) produced from semen-derived epithelial cells (SedECs) are healthy and reproductively competent. In this study, the semen and fertility profile of a cloned bull (C1) that was produced from a SedEC were compared with its donor (D1) and with two cloned bulls (C2, C3) that were produced from commonly used skin-derived fibroblast cells (SkdFCs). We observed variations in some fresh semen parameters (ejaculated volume and mass motility), frozen-thawed sperm parameters (plasma membrane integrity, and computer-assisted semen analysis (CASA) indices), but values are within the normal expected range. There was no difference in sperm concentration of ejaculated semen and frozen-thawed semen parameters which include sperm motility, percentage of live and normal morphology sperm, and distance traveled through oestrus mucus. Following in vitro fertilization (IVF) experiments, zygotes from C1 had higher (P < 0.05) cleavage rates (81%) than C2, C3, and D1 (71%, 67%, and 75%, respectively); however, blastocyst development per cleaved embryo and quality of produced blastocysts did not differ. The conception rate of C1 was 46% (7/15) and C2 was 50% (8/15) following artificial insemination with frozen-thawed semen. Established pregnancies resulted in births of 7 and 6 progenies sired by C1 and C2, respectively, and all calves show no signs of phenotypical abnormalities. These results showed that semen from a cloned bull derived from SedECs is equivalent to semen from its donor bull and bulls cloned from SkdFCs.


Assuntos
Búfalos/fisiologia , Clonagem de Organismos/veterinária , Células Epiteliais/citologia , Sêmen/citologia , Animais , Criopreservação/veterinária , Feminino , Fertilidade , Fertilização , Fertilização In Vitro/veterinária , Inseminação Artificial/veterinária , Masculino , Análise do Sêmen/veterinária , Motilidade Espermática
12.
Sci Total Environ ; 744: 140785, 2020 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-32707413

RESUMO

Simple cryopreservation of anaerobic ammonium-oxidation (anammox) consortia has become a promising preservation technology for the fast start-up of the anammox process. Here, we use genome-resolved metagenomics and metatranscriptomics to understand of the microbial interaction in a simple and effective resuscitation process for long-term cryopreserved anammox consortia by sequential addition of anammox SBR supernatant. Performance results showed that sequential addition of anammox supernatant significantly reduced the resuscitation time of the granule-based anammox process from 40 to 20 days. Genome-centric metagenomics were used to recover 19 high-quality draft genomes of anammox and heterotrophic bacteria. Comparative metatranscriptomic analysis was conducted to examine the gene expression of Candidatus Kuenenia stuttgartiensis, the dominant anammox bacterium, and heterotrophic bacteria to better understand their potential interactions. Proteobacteria-affiliated bacteria found in the supernatant were highly active in producing the secondary metabolites molybdopterin cofactor and folate which are needed for growth of the auxotrophic anammox bacteria. In addition, the significantly higher expression levels of hzsA and CO2-fixtion genes in the Candidatus Kuenenia genome indicated the anammox bacteria were likely more active and growing faster after sequential anammox supernatant addition during the resuscitation process. The resuscitation treatment pulse assays confirmed that sequential addition of supernatant was an effective way for the rapid resuscitation of anammox consortia. Our findings offer the first evidence of cross-feeding during the rapid resuscitation of cryopreserved anammox consortia.


Assuntos
Reatores Biológicos , Nitrogênio , Anaerobiose , Criopreservação , Metagenômica , Oxirredução
14.
PLoS One ; 15(6): e0234481, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32589634

RESUMO

BACKGROUND: Is freeze-all strategy effective in terms of cumulative live birth rates (CLBRs) in all patients? METHODS: This retrospective single-center study analyzed the CLBRs of 2523 patients undergoing fresh or electively frozen blastocyst transfer cycles. In 1047, cycles, the fresh embryo transfer (ET) strategy was applied for the 1st ET, whereas electively frozen ET (e-FET) was performed in 1476 cycles. Female age ≤ 37 and blastocysts frozen via vitrification were included. The patients in each arm were further stratified into four subgroups according to the number of oocytes retrieved as follows: Group A: 1-5, group B: 6-10, group C: 11-15 and group D: 16-25 oocytes retrieved. The primary endpoint was the CLBR. The secondary endpoints were the ovarian hyperstimulation syndrome (OHSS) rate and the live birth rates (LBRs) following fresh ETs and e-FETs for the first transfers. RESULT(S): The CLBR was similar between the fresh ET and e-FET arms in group A (35/76 (46.1%) vs 29/67 (43.3%), p = 0.74) and group B (165/275 (60%) vs 216/324 (66.7%), p = 0.091), whereas significantly higher rates were detected in favor of the e-FET arm within group C (328/460 (71.3%) vs 201/348 (57.8%), p<0.001) and group D (227/348 (65.2%), vs 446/625 (71.5%), p<0.001). The OHSS rate was also found to be higher in the fresh ET arm among group C (12/348 (3.4%) vs 0/460 (0%), p<0.001) and group D (38/348 (10.9%) vs 3/625 (0.5%), p<0.001) patients than e-FET arm. Perinatal and obstetrical outcomes were nonsignificantly different between fresh and e-FET arms. However, the birth weights were significantly lower for fresh ET, 3064 versus 3201 g for singletons (p<0.001). CONCLUSION: Compared with a fresh-transfer strategy, the e-FET strategy resulted in a higher CLBR among patients with >10 oocytes retrieved during stimulated cycles.


Assuntos
Criopreservação , Transferência Embrionária/métodos , Fertilização In Vitro/métodos , Nascimento Vivo/epidemiologia , Recuperação de Oócitos/estatística & dados numéricos , Manejo de Espécimes/métodos , Adulto , Feminino , Humanos , Síndrome de Hiperestimulação Ovariana/epidemiologia , Estudos Retrospectivos
15.
PLoS One ; 15(6): e0234339, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32516324

RESUMO

Hypothermic storage of boar semen provides the possibility to omit antibiotics from semen extenders so long as sperm quality is maintained and bacterial growth prevented. The objective of this study was to determine an optimal cooling-rate frame for boar semen preserved at 5°C in an antibiotic-free extender. Semen from eight boars extended in AndroStar® Premium was cooled from 30°C to 5°C using seven different cooling rates, ranging initially from 0.01 to 0.36°C min-1 and reaching 5°C between 2 h and 24 h after dilution. Sperm motility, membrane integrity, membrane fluidity, mitochondrial membrane potential and the response to the capacitation stimulus bicarbonate remained at a high level for 144 h at 5°C when the semen was initially cooled in a cooling-rate frame ranging from 0.01 to 0.09°C min­1 in the temperature zone from 30 to 25°C, followed by 0.02 to 0.06°C min-1 to 10°C and 0.01 to 0.02°C min­1 to the final storage temperature. A cooling rate of 0.07°C min-1 in the temperature zone from 30 to 10°C led to a reduced response to bicarbonate (P < 0.01) and fast cooling to 5°C within 1 h with a cooling rate of 0.31°C min-1 resulted in lower values (P > 0.05) of all sperm parameters. In a further experiment, slow cooling with a holding time of 6 h at 22°C induced after 6 h storage a temporary increase in Escherichia coli of 0.5 × 103 to 2.4 × 103 CFU mL-1 in the sperm-free inoculated extender. Overall, the load of mesophilic bacteria in the stored semen was below 6 × 103 CFU mL-1, a level that is not regarded as critical for sperm quality. In conclusion, appropriate cooling protocols were established for the antibiotic-free storage of boar semen at 5°C, allowing the application of hypothermic preservation in research and in artificial insemination.


Assuntos
Criopreservação/métodos , Preservação do Sêmen/métodos , Manejo de Espécimes/métodos , Animais , Secreções Corporais/efeitos dos fármacos , Líquidos Corporais/efeitos dos fármacos , Crioprotetores/farmacologia , Masculino , Sêmen/efeitos dos fármacos , Sêmen/metabolismo , Motilidade Espermática/efeitos dos fármacos , Espermatozoides/fisiologia , Sus scrofa/metabolismo , Suínos , Temperatura
16.
Nat Commun ; 11(1): 2976, 2020 06 12.
Artigo em Inglês | MEDLINE | ID: mdl-32532991

RESUMO

Pre-clinical heart transplantation studies have shown that ex vivo non-ischemic heart preservation (NIHP) can be safely used for 24 h. Here we perform a prospective, open-label, non-randomized phase II study comparing NIHP to static cold preservation (SCS), the current standard for adult heart transplantation. All adult recipients on waiting lists for heart transplantation were included in the study, unless they met any exclusion criteria. The same standard acceptance criteria for donor hearts were used in both study arms. NIHP was scheduled in advance based on availability of device and trained team members. The primary endpoint was a composite of survival free of severe primary graft dysfunction, free of ECMO use within 7 days, and free of acute cellular rejection ≥2R within 180 days. Secondary endpoints were I/R-tissue injury, immediate graft function, and adverse events. Of the 31 eligible patients, six were assigned to NIHP and 25 to SCS. The median preservation time was 223 min (IQR, 202-263) for NIHP and 194 min (IQR, 164-223) for SCS. Over the first six months, all of the patients assigned to NIHP achieved event-free survival, compared with 18 of those assigned to SCS (Kaplan-Meier estimate of event free survival 72.0% [95% CI 50.0-86.0%]). CK-MB assessed 6 ± 2 h after ending perfusion was 76 (IQR, 50-101) ng/mL for NIHP compared with 138 (IQR, 72-198) ng/mL for SCS. Four deaths within six months after transplantation and three cardiac-related adverse events were reported in the SCS group compared with no deaths or cardiac-related adverse events in the NIHP group. This first-in-human study shows the feasibility and safety of NIHP for clinical use in heart transplantation. ClinicalTrial.gov, number NCT03150147.


Assuntos
Criopreservação/métodos , Transplante de Coração/métodos , Ensaios Clínicos Controlados não Aleatórios como Assunto , Preservação de Órgãos/métodos , Adulto , Idoso , Feminino , Rejeição de Enxerto , Transplante de Coração/instrumentação , Humanos , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Avaliação de Resultados em Cuidados de Saúde/métodos , Avaliação de Resultados em Cuidados de Saúde/estatística & dados numéricos , Perfusão , Estudos Prospectivos , Fatores de Tempo , Doadores de Tecidos , Listas de Espera
17.
Crit Rev Oncol Hematol ; 151: 102981, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32485429

RESUMO

The topic of fertility preservation in patients with a lymphoproliferative disease offers new aspects of debate, due to the introduction of novel chemotherapeutic regimens and small molecules in the clinical landscape. Cancer related infertility is mostly dependent on gonadotoxic treatments and fertile female patients are today addressed to the oocyte cryopreservation or to ovarian cortex fragment cryopreservation. These methods present advantages and disadvantages, which will be discussed in the present review, together with the options for male patients. The recent discovery of functional ovarian stem cells (OCSs) in woman ovarian cortex, opens new avenues offering a innovative procedure for fertility preservation through as model of regenerative medicine. Here, we review the gonadotoxic potential of "classical" chemotherapeutic treatments as well as of "novel" targeted therapies actually employed for lymphoproliferative neoplasms in young patients and revisit both the today available and future chances to preserve and restore fertility after the cancer healing.


Assuntos
Antineoplásicos/efeitos adversos , Preservação da Fertilidade , Fertilidade/efeitos dos fármacos , Transtornos Linfoproliferativos/dietoterapia , Neoplasias/tratamento farmacológico , Ovário/efeitos dos fármacos , Insuficiência Ovariana Primária/induzido quimicamente , Criopreservação , Feminino , Humanos , Transtornos Linfoproliferativos/patologia , Masculino , Oócitos
19.
J Vis Exp ; (159)2020 05 19.
Artigo em Inglês | MEDLINE | ID: mdl-32510499

RESUMO

The central nervous system (CNS) is comprised of the brain and spinal cord and is enveloped by the meninges, membranous layers serving as a barrier between the periphery and the CNS. The CNS is an immunologically specialized site, and in steady state conditions, immune privilege is most evident in the CNS parenchyma. In contrast, the meninges harbor a diverse array of resident cells, including innate and adaptive immune cells. During inflammatory conditions triggered by CNS injury, autoimmunity, infection, or even neurodegeneration, peripherally derived immune cells may enter the parenchyma and take up residence within the meninges. These cells are thought to perform both beneficial and detrimental actions during CNS disease pathogenesis. Despite this knowledge, the meninges are often overlooked when analyzing the CNS compartment, because conventional CNS tissue extraction methods omit the meningeal layers. This protocol presents two distinct methods for the rapid isolation of murine CNS tissues (i.e., brain, spinal cord, and meninges) that are suitable for downstream analysis via single-cell techniques, immunohistochemistry, and in situ hybridization methods. The described methods provide a comprehensive analysis of CNS tissues, ideal for assessing the phenotype, function, and localization of cells occupying the CNS compartment under homeostatic conditions and during disease pathogenesis.


Assuntos
Sistema Nervoso Central/citologia , Sistema Nervoso Central/imunologia , Meninges/citologia , Meninges/imunologia , Animais , Encéfalo/citologia , Encéfalo/imunologia , Agregação Celular , Criopreservação , Doenças Desmielinizantes/patologia , Doenças Desmielinizantes/virologia , Feminino , Antígenos Comuns de Leucócito/metabolismo , Camundongos , Inclusão em Parafina , Medula Espinal/citologia , Medula Espinal/imunologia , Theilovirus/fisiologia , Fixação de Tecidos
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