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1.
Medicine (Baltimore) ; 103(8): e37201, 2024 Feb 23.
Artigo em Inglês | MEDLINE | ID: mdl-38394502

RESUMO

BACKGROUND: Testicular cancer (TC) mostly occurs in men aged 14 to 44. Studies have shown that TC seriously damages male fertility, and 6% to 24% of patients with TC were even found to suffer from azoospermia when they are diagnosed. At present, some studies have pointed out that onco-microdissection testicular sperm extraction (mTESE) can extract sperm from tumor testicles. However, there are almost no reports on remedial measures after onco-mTESE failure. Given the valuable opportunity for fertility preservation in patients with TC and azoospermia, it is necessary to provide effective remedial methods for patients with failed onco-mTESE. METHODS: Two young men, who were diagnosed with TC and also found to have azoospermia, tried onco-mTESE while undergoing radical orchiectomy for fertility preservation. However, sperm extraction failed in both patients. Subsequently, the isolated testicular tissue of the patient in case 1 suffered from TC again, and the patient in case 2 was scheduled to receive multiple cycles of gonadotoxic chemotherapy. Because both had a plan to have a birth in the future, we performed remedial mTESE. RESULTS: Sperm was successfully extracted from both patients. The patient recovered well, without complications. The patient couple in case 1 underwent 1 intracytoplasmic sperm injection (ICSI) cycle but did not achieve clinical pregnancy. CONCLUSIONS: There is still an opportunity to extract sperm successfully using onco-mTESE, despite the difficulty of fertility preservation in TC patients with azoospermia. If sperm extraction from the tumor testis fails, implementing remedial mTESE as early as possible would likely preserve the last chance of fertility for these patients.


Assuntos
Azoospermia , Neoplasias Embrionárias de Células Germinativas , Neoplasias Testiculares , Gravidez , Feminino , Humanos , Masculino , Azoospermia/terapia , Azoospermia/complicações , Neoplasias Testiculares/cirurgia , Neoplasias Testiculares/complicações , Microdissecção/métodos , Recuperação Espermática , Sêmen , Espermatozoides/patologia , Estudos Retrospectivos , Testículo/cirurgia , Testículo/patologia
2.
Andrology ; 12(1): 30-44, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37172416

RESUMO

BACKGROUND: There has been no systematic review and meta-analysis to analyze and summarize the predictive factors of successful sperm extraction in salvage microdissection testicular sperm extraction. OBJECTIVES: We aimed to investigate the factors predicting the result of salvage microdissection testicular sperm extraction in patients with non-obstructive azoospermia who failed the initial microdissection testicular sperm extraction or conventional testicular sperm extraction. MATERIALS AND METHODS: We conducted a systematic literature search in PubMed, Web of Science, EMBASE, and the Cochrane Library for literature that described the characteristics of patients with non-obstructive azoospermia who underwent salvage microdissection testicular sperm extraction after failing the initial microdissection testicular sperm extraction or conventional testicular sperm extraction published prior to June 2022. RESULTS: This meta-analysis included four retrospective studies with 332 patients with non-obstructive azoospermia who underwent a failed initial microdissection testicular sperm extraction and three retrospective studies with 177 non-obstructive azoospermia patients who underwent a failed conventional testicular sperm extraction. The results were as follows: among non-obstructive azoospermia patients whose first surgery was microdissection testicular sperm extraction, younger patients (standard mean difference: -0.28, 95% confidence interval [CI]: -0.55 to -0.01) and those with smaller bilateral testicular volume (standard mean difference: -0.55, 95% CI: -0.95 to -0.15), lower levels of follicle-stimulating hormone (standard mean difference: -0.86, 95% CI: -1.18 to -0.54) and luteinizing hormone (standard mean difference: -0.68, 95% CI: -1.16 to -0.19), and whose testicular histological type was hypospermatogenesis (odds ratio: 3.52, 95% CI: 1.30-9.53) were more likely to retrieve spermatozoa successfully, while patients with Sertoli-cell-only syndrome (odds ratio: 0.41, 95% CI: 0.24-0.73) were more likely to fail again in salvage microdissection testicular sperm extraction. Additionally, in patients who underwent salvage microdissection testicular sperm extraction after a failed initial conventional testicular sperm extraction, those with testicular histological type of hypospermatogenesis (odds ratio: 30.35, 95% CI: 8.27-111.34) were more likely to be successful, while those with maturation arrest (odds ratio: 0.39, 95% CI: 0.18-0.83) rarely benefited. CONCLUSION: We found that age, testicular volume, follicle-stimulating hormone, luteinizing hormone, hypospermatogenesis, Sertoli-cell-only syndrome, and maturation arrest were valuable predictors of salvage microdissection testicular sperm extraction, which will assist andrologists in clinical decision-making and minimize unnecessary injury to patients.


Assuntos
Azoospermia , Oligospermia , Síndrome de Células de Sertoli , Humanos , Masculino , Azoospermia/cirurgia , Azoospermia/patologia , Oligospermia/patologia , Estudos Retrospectivos , Microdissecção/métodos , Recuperação Espermática , Sêmen , Testículo/cirurgia , Testículo/patologia , Espermatozoides/patologia , Hormônio Foliculoestimulante , Hormônio Luteinizante , Hormônio Foliculoestimulante Humano
3.
Reprod Sci ; 31(2): 366-374, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-37749447

RESUMO

Patients with Idiopathic non-obstructive azoospermia (iNOA) can achieve fertility by extracting testicular sperm through microdissection testicular sperm extraction (mTESE). But more than half of iNOA patients still cannot benefit from mTESE. In recent years, some studies had reported that serum hormones may be related to the outcome of sperm retrieval, but few had been verified. We hope to obtain a predictive method that is convenient for clinical application and can help judge the outcome of sperm extraction before implementing mTESE. We performed a retrospective analysis of NOA patients who underwent mTESE in the same andrology center from June 2020 to November 2022. A total of 261 patients with complete data were collected, logistic regression analysis was performed and a predictive model was constructed. Then, from December 2022 to May 2023, one prospective cohort of 48 NOA patients who met the inclusion criteria from the same center was recruited to validate the risk prediction model. We successfully constructed a logistic regression model to predict the outcome of iNOA patients undergoing mTESE and found that higher serum anti-Müllerian hormone (AMH) levels were associated with failure sperm retrieval, resulting in an AMH cut-off of 2.60 ng/ml. The area under the receiver operating curve was 0.811, the sensitivity was 0.870, and the specificity was 0.705. Decision curve analysis demonstrated that the threshold probability was above 4%, and unnecessary mTESE could be reduced using this model. In a prospective cohort at the same center, 85.42% (41/48) of iNOA patients correctly identified the mTESE outcome using this model. A logistic regression model with AMH as an independent predictor can predict mTESE outcomes in iNOA patients. Preoperative selection of mTESE in patients with iNOA using this model had clinical benefit in reducing unnecessary surgery. The model demonstrated good accuracy in a small prospective cohort validation.


Assuntos
Azoospermia , Humanos , Masculino , Azoospermia/diagnóstico , Azoospermia/cirurgia , Estudos Retrospectivos , Microdissecção/métodos , Estudos Prospectivos , Recuperação Espermática , Sêmen , Testículo/cirurgia , Espermatozoides
4.
Andrology ; 12(1): 157-163, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37210678

RESUMO

BACKGROUND: Cryptorchidism is considered to be one of the most common causes of non-obstructive azoospermia. There are several surgical techniques to retrieve sperm in these patients. Microdissection testicular sperm extraction (m-TESE) is a recent sperm retrieval technique which is considered to be a safe, non-blind, and feasible method. OBJECTIVES: This study aimed to investigate sperm retrieval rate (SRR) by the mTESE method in patients who have undergone orchidopexy due to bilateral cryptorchidism. MATERIALS AND METHODS: In this retrospective study, 56 ex-cryptorchid patients, who underwent mTESE due to post orchidopexy azoospermia, were included. Patients with hypogonadotropic hypogonadism, Klinefelter syndrome, azoospermia factors (AZF) microdeletion, or chromosomal translocation were excluded from the study. Data were obtained from medical files. RESULTS: SRR in this study was 46%. Patients were divided into two groups of negative (n = 30) and positive (n = 26) based on the sperm extraction outcomes. There was no statistically significant difference between two groups regarding the mean age at mTESE, mean age at orchidopexy, testicular size, and serum testosterone concentration. However, testicular location, histological patterns, FSH, and LH level showed to have statistically significant relation with sperm retrieval results. But, according to our logistic regression, none of the included variable in the model including FSH, LH, histopathology, and testis location have a significant effect on the presence of the sperm. DISCUSSION: In the present study, SRR was significantly higher in patients with scrotal testis and low level of FSH and LH. CONCLUSIONS: Performing mTESE could be recommended in ex-cryptorchid patients with post orchidopexy NOA. Preoperative testicular biopsy seems to be unnecessary while clinical criteria can perfectly define NOA.


Assuntos
Azoospermia , Criptorquidismo , Síndrome de Klinefelter , Humanos , Masculino , Orquidopexia , Estudos Retrospectivos , Microdissecção/métodos , Sêmen , Testículo/cirurgia , Testículo/patologia , Espermatozoides/patologia , Criptorquidismo/cirurgia , Criptorquidismo/patologia , Recuperação Espermática , Hormônio Foliculoestimulante
5.
Int Urol Nephrol ; 55(9): 2177-2182, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37330932

RESUMO

OBJECTIVE: To compare sperm retrieval rates between unilateral and bilateral microdissection testicular sperm extraction (MD-TESE) procedures in patients with non-obstructive azoospermia and to contribute to the literature by comparing them with literature data. METHODS: This prospective study included 84 males with primary infertility who had azoospermic NOA, who had been married for at least one year, and whose female partners did not have a history of infertility. The study was conducted between January 2019 and January 2020. MD-TESE was applied bilaterally to 48% (n:41) (Group 1) and unilaterally to 52% (n:43) (Group 2) of the patients, and sperm retrieval rates were compared. RESULTS: There was no statistically significant difference between Group 1 patients and Group 2 patients in terms of sperm availability (61%, 56.5%, p=0.495, respectively). In addition, while no complications were observed in unilateral MD-TESEs, 3 complications were observed in bilateral MD-TESEs. CONCLUSIONS: In our study, it was determined that there was no significant difference between the groups in terms of sperm availability in patients with NOA. Considering the operative time and complication rates of bilateral MD-TESE in patients diagnosed with NOA and the possible MD-TESE procedures that may be performed later, we believe that unilateral MD TESE is a more preferable procedure for the patient and surgeon in this patient group.


Assuntos
Azoospermia , Humanos , Masculino , Feminino , Azoospermia/cirurgia , Estudos Prospectivos , Microdissecção/métodos , Estudos Retrospectivos , Sêmen , Testículo/cirurgia , Espermatozoides , Recuperação Espermática
6.
Urology ; 176: 82-86, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-37054923

RESUMO

OBJECTIVE: To assess the incidence of discordant testicular pathology in men undergoing bilateral microdissection testicular sperm extraction (mTESE) and association with sperm retrieval rate. METHODS: We performed a retrospective single-institutional review of all patients undergoing mTESE from 2007 to 2021 and aggregated clinical history, physical exam, semen analysis, and operative findings. Specimens with discordant pathology were re-reviewed by an experienced genitourinary.ßpathologist and categorized in a standardized fashion. Data were analyzed using SPSS. RESULTS: One hundred fourteen non-obstructive azoospermia.ßmen undergoing 132 mTESEs were identified within the study period. Eighty-five percent (112/132) of cases had pathology specimens available, and within this specific cohort the success rate was 41.9% (47/112). A total of 206 pathological reports resulted including 52.4% Sertoli cell only, 4.9% Leydig cell hyperplasia, 8.7% fibrosis, 16.5% maturation arrest, and 17.5% hypospermatogenesis. Twelve percent of testicles had more than 1 pathologic diagnosis. Sixty-six men had synchronous bilateral testicular pathology, and 11/66 (16.6%) had at least partially discordant pathology on initial review. Focused re-review by a genitourinary pathologist confirmed exclusively discordant pathology in 7/66 (10.6%) cases, with a sperm retrieval rate of 57% (4/7). The sperm retrieval rate.ßin men with discordant pathology was not significantly different from those with concordant pathology. CONCLUSION: Over 1 in 10 men undergoing mTESE may have discordant pathology between testicles, though this may not affect sperm retrieval rate.ßat the time of procedure. Clinicians should consider submitting bilateral testicular specimens for pathology to (1) clarify their outcomes data, and (2) assist with clinical decision-making and surgical planning if a repeat mTESE is indicated.


Assuntos
Azoospermia , Testículo , Humanos , Masculino , Testículo/cirurgia , Testículo/patologia , Microdissecção/métodos , Estudos Retrospectivos , Recuperação Espermática , Sêmen , Espermatozoides/patologia , Azoospermia/cirurgia
7.
Methods Mol Biol ; 2642: 365-373, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36944888

RESUMO

Plants display a great diversity of particular cell types that obviously perform functions and regulations that are essential for successful growth and development, whether under optimal or adverse conditions. The functions performed by each of these particular cell types must be associated with specific transcriptomic, proteomic, and metabolic profiles that cannot be disentangled by analyzing whole plant organs and tissues. Laser microdissection is a technique for the collection of specific cell types in plant organs and tissues comprising heterogeneous cell populations. It has been successfully used for physiological and molecular studies. Laser microdissection can be applied to any plant species as long as it is possible to reliably identify the cell types of interest. Here, we describe step by step, using citrus as a model plant, a fast, simple, easy to perform, and experimentally validated protocol to collect cells from the abscission zone, a specific tissue that is difficult to access and whose activity is important in the response of plants to adverse environmental conditions.


Assuntos
Microdissecção , Proteômica , Microdissecção/métodos , Plantas/genética , Perfilação da Expressão Gênica , Lasers
8.
Lab Invest ; 103(7): 100133, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-36990152

RESUMO

Cell-specific microRNA (miRNA) expression estimates are important in characterizing the localization of miRNA signaling within tissues. Much of these data are obtained from cultured cells, a process known to significantly alter miRNA expression levels. Thus, our knowledge of in vivo cell miRNA expression estimates is poor. We previously demonstrated expression microdissection-miRNA-sequencing (xMD-miRNA-seq) to acquire in vivo estimates, directly from formalin-fixed tissues, albeit with a limited yield. In this study, we optimized each step of the xMD process, including tissue retrieval, tissue transfer, film preparation, and RNA isolation, to increase RNA yields and ultimately show strong enrichment for in vivo miRNA expression by qPCR array. These method improvements, such as the development of a noncrosslinked ethylene vinyl acetate membrane, resulted in a 23- to 45-fold increase in miRNA yield, depending on the cell type. By qPCR, miR-200a increased by 14-fold in xMD-derived small intestine epithelial cells, with a concurrent 336-fold reduction in miR-143 relative to the matched nondissected duodenal tissue. xMD is now an optimized method to obtain robust in vivo miRNA expression estimates from cells. xMD will allow formalin-fixed tissues from surgical pathology archives to make theragnostic biomarker discoveries.


Assuntos
MicroRNAs , MicroRNAs/genética , MicroRNAs/metabolismo , Microdissecção/métodos , Células Epiteliais/metabolismo , Formaldeído , Perfilação da Expressão Gênica
9.
Nat Methods ; 20(4): 590-599, 2023 04.
Artigo em Inglês | MEDLINE | ID: mdl-36928074

RESUMO

Our understanding of nerve regeneration can be enhanced by delineating its underlying molecular activities at single-neuron resolution in model organisms such as Caenorhabditis elegans. Existing cell isolation techniques cannot isolate neurons with specific regeneration phenotypes from C. elegans. We present femtosecond laser microdissection (fs-LM), a single-cell isolation method that dissects specific cells directly from living tissue by leveraging the micrometer-scale precision of fs-laser ablation. We show that fs-LM facilitates sensitive and specific gene expression profiling by single-cell RNA sequencing (scRNA-seq), while mitigating the stress-related transcriptional artifacts induced by tissue dissociation. scRNA-seq of fs-LM isolated regenerating neurons revealed transcriptional programs that are correlated with either successful or failed regeneration in wild-type and dlk-1 (0) animals, respectively. This method also allowed studying heterogeneity displayed by the same type of neuron and found gene modules with expression patterns correlated with axon regrowth rate. Our results establish fs-LM as a spatially resolved single-cell isolation method for phenotype-to-genotype mapping.


Assuntos
Proteínas de Caenorhabditis elegans , Caenorhabditis elegans , Animais , Caenorhabditis elegans/genética , Microdissecção/métodos , Neurônios/fisiologia , Lasers , Análise de Sequência de RNA , MAP Quinase Quinase Quinases , Proteínas de Caenorhabditis elegans/genética
10.
Investig Clin Urol ; 64(1): 41-50, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36629064

RESUMO

PURPOSE: This study evaluated the predictors of sperm retrieval (SR) in non-mosaic Klinefelter syndrome (KS) patients undergoing microsurgical testicular sperm extraction (mTESE). The cutoff values of the predictors of SR and overall pregnancy rates after intracytoplasmic sperm injection (ICSI) were analyzed for the positive SR (PSR) cases. MATERIALS AND METHODS: The study was a dual-center retrospective study. Overall 118 patients with KS underwent mTESE between January 2011 and July 2021. Clinicopathological factors including comorbidities, endocrine profiles, and testicular volumes were analyzed. RESULTS: A total of 58 patients showed PSR (49.2%) and 60 patients (50.8%) had negative SR (NSR). The mean overall age of the patients was 32.5 years. The NSR patients had a significantly greater prevalence of obesity, diabetes mellitus, and cerebrovascular disease. The PSR group had a significantly higher left testis mean volume (p=0.039). The differences between the two study groups regarding follicular-stimulating hormone, luteinizing hormone, and testosterone variations at 1 and 3 months after mTESE were insignificant. Preoperative mean neutrophil-to-lymphocyte ratio was significantly greater in the NSR group (p=0.011), but the platelet-to-lymphocyte ratio showed no significant difference between the two study groups. A live child birth was achieved in 53.4% of the PSR patients. Multivariate logistic analysis showed that total testicular volume >3.93 mL, left testis volume >1.79 mL, and neutrophil-to-lymphocyte ratio ≤1.82 were significantly associated with PSR. CONCLUSIONS: mTESE-ICSI is a feasible method for KS patients to have a child, and total testicular volume, left testis volume, and neutrophil-to-lymphocyte ratio might be predictors of successful SR.


Assuntos
Síndrome de Klinefelter , Testículo , Adulto , Feminino , Humanos , Masculino , Gravidez , Síndrome de Klinefelter/complicações , Síndrome de Klinefelter/patologia , Microdissecção/métodos , Estudos Retrospectivos , Sêmen , Injeções de Esperma Intracitoplásmicas , Recuperação Espermática , Espermatozoides , Testículo/patologia
11.
Pathobiology ; 90(3): 199-208, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-35952628

RESUMO

Histopathology has historically been the critical technique for the diagnosis and treatment of human disease. Today, genomics, transcriptomics, and proteomics from specific cells, rather than bulk tissue, have become key to understanding underlying disease mechanisms and rendering useful diagnostic information. Extraction of desired analytes, i.e., nucleic acids or proteins, from easily accessible formalin-fixed paraffin-embedded tissues allows for clinically relevant activities, such as sequencing biomarker mutations or typing amyloidogenic proteins. Genetic profiling has become routine for cancers as varied as non-small cell lung cancer and prostatic carcinoma. The five main tissue dissection techniques that have been developed thus far include: bulk scraping, manual macrodissection, manual microdissection, laser-capture microdissection, and expression microdissection. In this review, we discuss the importance of tissue dissection in clinical practice and research, the basic methods, applications, as well as some advantages and disadvantages for each modality.


Assuntos
Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Humanos , Prognóstico , Dissecação/métodos , Microdissecção/métodos , Fixação de Tecidos/métodos , Inclusão em Parafina/métodos
12.
J Chin Med Assoc ; 86(3): 300-305, 2023 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-36346204

RESUMO

BACKGROUND: Microdissection testicular sperm extraction (mTESE) is the mainstay of sperm retrieval in men with nonobstructive azoospermia (NOA). Some experts believe that a resting period of ≥6 months is required between procedures to ensure better sperm retrieval rates and fewer complications. However, no scientific studies have investigated how long patients should wait before arranging a second mTESE procedure. This retrospective study aimed to evaluate whether good success rates are still achieved when a repeat mTESE procedure is performed within 6 months. METHODS: Total 146 patients with NOA who underwent mTESE twice from the same testis between May 2012 and September 2019 were retrospectively collected. These patients were categorized into three groups according to the time interval between the two mTESE surgeries, with 44, 60, and 42 patients undergoing a repeat mTESE after <3 months (group I), between 3 and 6 months (group II), and >6 months (group III) after the first procedure, respectively. RESULTS: No significant differences were observed between groups in terms of patient characteristics and preoperative hormone profiles. Overall sperm retrieval rates did not differ among the three groups (93.2%, 90.0%, and 88.1% in groups I, II, and III, respectively [ p = 0.719]), nor did fertility outcomes, including rates of fertilization, biochemical pregnancy, clinical pregnancy, and cumulative live births. CONCLUSION: Sperm retrieval rates for repeat mTESE procedures were consistently high, even when the second procedure was performed within 6 months of the first. Repeat mTESE within 6 months is not a limitation for patients with NOA, if required clinically.


Assuntos
Azoospermia , Testículo , Gravidez , Feminino , Humanos , Masculino , Azoospermia/cirurgia , Estudos Retrospectivos , Microdissecção/métodos , Estudos de Viabilidade , Sêmen , Espermatozoides
13.
Semin Reprod Med ; 41(6): 267-272, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-38262439

RESUMO

Nonobstructive azoospermia (NOA) is among the most common causes of male infertility. For men with NOA seeking fertility treatment, microdissection testicular sperm extraction (microTESE) is the best option for retrieving sperm, which can be used with in vitro fertilization-intracytoplasmic sperm injection to achieve pregnancy in their partner. With the aid of the operating microscope, microTESE allows for thorough evaluation of the testis tissue and selection of seminiferous tubules that appear most capable of sperm production. Rates of success with microTESE vary depending on the underlying cause of NOA and the center at which the procedure is performed. Not all patients are candidates for microTESE, and those who are candidates should be counseled on the likelihood of sperm retrieval and the potential for changes in postoperative testis function.


Assuntos
Azoospermia , Testículo , Gravidez , Feminino , Humanos , Masculino , Testículo/cirurgia , Microdissecção/métodos , Estudos Retrospectivos , Sêmen , Azoospermia/cirurgia , Espermatozoides
14.
Eur Rev Med Pharmacol Sci ; 26(19): 7176-7181, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-36263526

RESUMO

OBJECTIVE: Azoospermia is a cause of infertility in a subgroup of infertile men. Sperm retrieval techniques including testicular sperm aspiration (TESA) and microscopic testicular sperm extraction (mTESE) are widely used. In this study, we have reviewed our findings regarding mTESE performed following a negative TESA outcome. PATIENTS AND METHODS: This is a retrospective chart review study that included 41 infertile patients who underwent mTESE after a negative TESA outcome. Charts were reviewed for demographic data, type of infertility, and type of azoospermia. Hormone level analysis was done for follicle-stimulating hormone, luteinizing hormone, and testosterone. Testicular volume was estimated by ultrasound. RESULTS: The study included 41 patients who underwent mTESE following a negative TESA outcome. Most patients had primary infertility (n = 32; 78%). Of the 41 patients, four had a previous history of either TESE or orchidopexy, and two had a history of varicocelectomy before the recent percutaneous TESA procedure. There was no significant association between sperm retrieval and the different surgical procedures that had been performed. Of the 41 patients, 27 had positive sperm retrieval by mTESE with a success rate of 65.9%. CONCLUSIONS: The positive sperm retrieval rate of mTESE performed following a negative TESA outcome was reasonable (65.9%). No significant correlations were identified with all variables studied.


Assuntos
Azoospermia , Recuperação Espermática , Humanos , Masculino , Microdissecção/métodos , Azoospermia/cirurgia , Estudos Retrospectivos , Sêmen , Testículo/diagnóstico por imagem , Testículo/cirurgia , Hormônio Foliculoestimulante , Testosterona , Hormônio Luteinizante
15.
STAR Protoc ; 3(3): 101600, 2022 09 16.
Artigo em Inglês | MEDLINE | ID: mdl-36042886

RESUMO

Cell response variability is a starting point in cancer drug resistance that has been difficult to analyze because the tolerant cell states are short lived. Here, we present fate-seq, an approach to isolate single cells in their transient states of drug sensitivity or tolerance before profiling. The drug response is predicted in live cells, which are laser-captured by microdissection before any drug-induced change can alter their states. This framework enables the identification of the cell-state signatures causing differential cell decisions upon treatment. For complete details on the use and execution of this protocol, please refer to Meyer et al. (2020).


Assuntos
Diagnóstico por Imagem , Microdissecção , Lasers , Microdissecção/métodos
16.
Eur Urol ; 82(4): 377-384, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-35599183

RESUMO

BACKGROUND: Surgical extraction of testicular spermatozoa is needed in men with nonobstructive azoospermia (NOA) who wish to become biological fathers. Based on available uncontrolled studies with unspecific patient selection, microdissection testicular sperm extraction (mTESE), having a sperm retrieval rate (SRR) of 50%, is considered the most efficient sperm retrieval procedure. However, no randomized clinical trials for comparison of different sperm retrieval procedures exist. Testicular sperm aspiration (TESA) is simple and commonly used, and we hypothesized that this technique using multiple needle passes would give similar SRRs to mTESE. OBJECTIVE: To compare mTESE and multiple needle-pass TESA in men with NOA. DESIGN, SETTING, AND PARTICIPANTS: A randomized clinical trial was performed between June 2017 and April 2021, with inclusion of 100 men with NOA from four centers in Denmark and Sweden. All participants received treatment at the same institution. INTERVENTION: Participants were randomized to mTESE (n = 49) or multiple needle-pass TESA (n = 51). Patients with failed multiple needle-pass TESA proceeded directly to salvage mTESE. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: The primary outcome was SRR. Secondary outcomes included complications and changes in reproductive hormones after surgery. RESULTS AND LIMITATIONS: Spermatozoa were retrieved in 21/49 (43%) men after mTESE and in 11/51 (22%) men after multiple needle-pass TESA (rate difference -0.21; 95% confidence interval -0.39 to -0.03; p = 0.02). The combined SRR for multiple needle-pass TESA + salvage mTESE was 15/51 (29%). No complications occurred after multiple needle-pass TESA only, while 5/89 (6%) men having mTESE experienced a complication requiring surgical intervention. Overall, no statistically significant differences in reproductive hormones were observed between groups after 6 mo. Limitations include the low number of patients in secondary outcome data. CONCLUSIONS: In direct comparison, SRR was higher in mTESE than in multiple needle-pass TESA. PATIENT SUMMARY: Men with azoospermia need surgical extraction of spermatozoa to become biological fathers. In this randomized trial, we compared two surgeries (microdissection testicular sperm extraction [mTESE] and testicular sperm aspiration [TESA]) and found that mTESE gives a higher sperm retrieval rate than multiple needle-pass TESA.


Assuntos
Azoospermia , Recuperação Espermática , Azoospermia/complicações , Azoospermia/cirurgia , Feminino , Hormônios , Humanos , Masculino , Microdissecção/métodos , Estudos Retrospectivos , Sêmen , Espermatozoides , Testículo/cirurgia
17.
J Assist Reprod Genet ; 39(6): 1297-1303, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35469372

RESUMO

PURPOSE: To define the risk of hypogonadism following microdissection testicular sperm extraction in cases of non-obstructive azoospermia. While sperm retrieval by open testicular sperm extraction can be associated with an increased risk of hypogonadism, there is limited data addressing which procedures and which patients harbor the greatest risk. METHODS: We report on a community-acquired, nested, case-cohort of non-obstructive azoospermic patients referred to one clinic after failed bilateral microdissection testicular sperm extraction. Patients were health-matched (1:2) to surgically naïve controls and divided into 2 cohorts based on risk factors for hypogonadism. Among microdissection patients, we compared total testosterone and gonadotropin levels before and > 6 months after surgery. Biochemical hypogonadism was defined as a total serum testosterone level ≤ 300 ng/dL. Hormone levels were compared to risk-matched controls. Comparative statistics were used to assess hormone levels within and between cohorts. RESULTS: There were no significant differences in baseline testosterone levels between microdissection patients (n = 26) and risk-matched controls (n = 52). At a mean of 26 months (range 6.2-112.8) post-procedure, mean testosterone levels decreased significantly (73 ng/dL or 16%; CI - 27, - 166; p < 0.01, paired t-test). Among microdissection patients with baseline testosterone > 300 ng/dL, 8/22 (36%) experienced hypogonadism post-procedure. There was a corresponding increase in follicle stimulating hormone (p = 0.05) and a trending increase in luteinizing hormones (p = 0.10). CONCLUSION: A durable decrease in testosterone levels occurs after failed microdissection testicular sperm extraction regardless of baseline risk of hypogonadism. In addition, a significant proportion of eugonadal patients will become hypogonadal after failed testicular microdissection procedures.


Assuntos
Azoospermia , Hipogonadismo , Azoospermia/genética , Azoospermia/cirurgia , Estudos de Coortes , Humanos , Masculino , Microdissecção/efeitos adversos , Microdissecção/métodos , Estudos Retrospectivos , Recuperação Espermática , Espermatozoides , Testículo/cirurgia , Testosterona
18.
Reprod Biol Endocrinol ; 20(1): 61, 2022 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-35365173

RESUMO

BACKGROUND: Microdissection testicular sperm extraction (micro-TESE) in combination with ICSI can make paternity possible for non-obstructive azoospermia (NOA) patients. Testicular sperm can be successfully retrieved in nearly half of NOA patients. Nevertheless, not many convincing protocols are established to improve sperm retrieval rate (SRR). The goal of this study was to evaluate whether gonadotropins therapy before micro-TESE could improve sperm retrieval rate and affect the ICSI outcomes in non-obstructive azoospermia patients with hypergonadotropic hypogonadism. METHODS: This retrospective cohort study included a total of 569 non-obstructive azoospermia men who underwent micro-TESE with or without 3-month of preoperative hCG / hCG plus highly purified urinary FSH (uFSH) between January 2016 and December 2019. The primary outcome was the sperm retrieval rate of micro-TESE. RESULTS: Sperm was found in 27 patients among 395 NOA men who accepted preoperative gonadotropins treatment (6.8%, 27/395) in post-treatment semen analysis for ICSI. One hundred forty nine out of 542 patients could successfully obtain enough sperm for ICSI through the micro-TESE (overall SRR = 27.5%). There was a statistically significant difference in the SRR between the preoperative gonadotropins treatment and non-gonadotropins treatment groups (31.2%, 115/368 vs. 19.5%, 34/174, P = 0.006). In the multivariable analysis with IPTW according to the propensity score, there was a significant association between preoperative gonadotropins treatment and the SRR (OR, 1.59; 95% CI: 1.02-2.52; P = 0.042). No differences in the clinical pregnancy rate, live birth delivery rate, or miscarriage rate were observed between the two groups. CONCLUSION: Preoperative gonadotropins therapy seems to have a role in improving SRR in NOA patients with hypergonadotropic hypogonadism. We found that gonadotropins therapy had no effect on ICSI clinical outcomes and live birth.


Assuntos
Azoospermia , Azoospermia/cirurgia , Estudos de Coortes , Feminino , Gonadotropinas/uso terapêutico , Humanos , Masculino , Microdissecção/métodos , Gravidez , Estudos Retrospectivos , Espermatozoides
19.
Andrologia ; 54(6): e14419, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35266170

RESUMO

The aim of this study is to evaluate the value of the haematologic inflammatory parameters in predicting sperm retrieval rates during microdissection testicular sperm extraction (micro-TESE).159 patients diagnosed with non-obstructive azoospermia were included in the study. After excluding the patients that do not fit the inclusion criteria, age, smoking status, body-mass index, serum luteinizing hormone, follicle-stimulating hormone, total testosterone levels and neutrophil, lymphocyte and platelet counts were recorded. Neutrophil-lymphocyte ratio, platelet-lymphocyte ratio and systemic immune-inflammation index were calculated. The primary outcome was defined as the presence of spermatozoa during the procedure and the association between the candidate predictors and primary endpoint were evaluated by logistic regression analysis. Then, a baseline model from age, smoking, body-mass index and hormonal levels was built. Ratios and indexes were included, respectively, and were compared by multivariate analyses. Each of all three parameters was an independent predictor of obtaining spermatozoa during micro-TESE procedure (all p < 0.001). Even though all three parameters were significant, neutrophil-lymphocyte ratio and systemic immune-inflammation index stood out as more powerful than platelet-lymphocyte ratio (p < 0.08, p < 0.08 respectively). Additionally, adding these parameters individually to the baseline model significantly empowered the predictive value (all p < 0.001). Using haematologic inflammatory parameters in the prediction of sperm presence during microdissection testicular sperm extraction may be helpful when consulting the patient with having a better foresight of the procedural outcomes.


Assuntos
Azoospermia , Microdissecção , Humanos , Inflamação , Linfócitos , Masculino , Microdissecção/métodos , Neutrófilos , Estudos Retrospectivos , Recuperação Espermática , Espermatozoides , Testículo/cirurgia
20.
Andrologia ; 54(6): e14401, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35243681

RESUMO

Testicular sperm retrieval (TSR) techniques are valuable in the context of severe idiopathic male factor infertility; however, there are few studies in the literature examining the long-term impact of TSR on testicular function. The objective was to determine whether testicular sperm aspiration (TESA) or microdissection testicular sperm extraction (micro-TESE) worsens the pre-existing spermatogenesis deficiency in men with either cryptozoospermia or severe oligozoospermia. The study population consisted of 145 men with either cryptozoospermia or severe oligozoospermia that underwent TESA or micro-TESE and had long-term post-operative semen analyses (SA). Patients with SA prior to and following TSR were included (n = 24). Amongst them, 16 men underwent TESA and 8 underwent micro-TESE. The follow-up SA was obtained at a mean of 3.0 ± 2.0 years following TSR (range: 0.3-8.3 years) amongst all participants. The post-operative semen parameters in the TESA group were similar to the pre-intervention parameters (p > 0.1). Similarly, the micro-TESE cohort did not demonstrate significant alterations in semen parameters post-intervention (p > 0.05). None of the men in the study became azoospermic following the TSR. Our study indicates TESA or micro-TESE do not appear to worsen the pre-existing spermatogenesis deficiencies in cryptozoospermic and oligozoospermic men over a long-term period. Larger studies are required to corroborate these findings.


Assuntos
Azoospermia , Infertilidade Masculina , Oligospermia , Azoospermia/etiologia , Azoospermia/cirurgia , Humanos , Masculino , Microdissecção/métodos , Oligospermia/etiologia , Estudos Retrospectivos , Recuperação Espermática , Espermatogênese , Testículo/cirurgia
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