Sperm origins and concentration do not impact the clinical outcomes in intracytoplasmic sperm injection cycles / 亚洲男科学杂志(英文版)

In the present study, we evaluated the impact of sperm origins and concentration on the clinical outcomes of intracytoplasmic sperm injection (ICSI) cycles. A total of 1201 ICSI cycles were retrospectively analyzed for male azoospermia or oligozoospermia between January 2015 and December 2015 in the Peking University Third Hospital. Patients were divided into three groups (Group 1 vs Group 2/3; surgically extracted sperm vs ejaculated sperms): Group 1 included 343 ICSI cycles and Group 2 analyzed 388 cycles on semen with sperm concentration <5 × 106 ml-1 (severe oligozoospermia group). Group 3 included 470 cycles with sperm concentration between 5 × 106 ml-1 and 15 × 106 ml-1 (mild oligozoospermia group). Fertilization rates, clinical pregnancy rates, and live birth rates were analyzed and compared among groups of different semen origins and concentrations on the oocyte retrieval day. Group 2 showed a lower fertilization rate than Group 3 (62.9% ± 21.6% vs 66.8% ± 22.1%,P< 0.05). There were no statistically significant differences in clinical pregnancy rate per transfer (51.3%, 46.7%, and 50.0%, respectively), live birth rate per transfer (44.4%, 40.9%, and 41.4%, respectively), accumulative live birth rate (58.3%, 51.0%, and 52.1%, respectively), twin birth rate (18.4%, 10.6%, and 12.6%, respectively), and birth defects rate (0, 0.3%, and 0.2%, respectively) among three groups. The results of this study indicated that sperm origins and concentration do not impact the clinical outcomes in ICSI cycles.

Update in the evaluation of the azoospermic male

Approximately 1% of all men in the general population suffer from azoospermia, and azoospermic men constitute approximately 10 to 15% of all infertile men. Thus, this group of patients represents a significant population in the field of male infertility. A thorough medical history, physical examination and hormonal profile are essential in the evaluation of azoospermic males. Imaging studies, a genetic workup and a testicular biopsy (with cryopreservation) may augment the workup and evaluation. Men with nonobstructive azoospermia should be offered genetic counseling before their spermatozoa are used for assisted reproductive techniques. This article provides a contemporary review of the evaluation of the azoospermic male.

The importance of semen analysis in the context of azoospermia

Azoospermia is a descriptive term referring to ejaculates that lack spermatozoa without implying a specific underlying cause. The traditional definition of azoospermia is ambiguous, which has ramifications on the diagnostic criteria. This issue is further compounded by the apparent overlap between the definitions of oligospermia and azoospermia. The reliable diagnosis of the absence of spermatozoa in a semen sample is an important criterion not only for diagnosing male infertility but also for ascertaining the success of a vasectomy and for determining the efficacy of hormonal contraception. There appears to be different levels of rigor in diagnosing azoospermia in different clinical situations, which highlights the conflict between scientific research and clinical practice in defining azoospermia.

Alteraciones del espermiograma en pacientes asistidos en la Unidad de Biología de la Reproducción de la Universidad de Chile / Alterations in the semen analysis of patients attending the reproductive biology unit at the University of Chile

En Chile, 10 a 15 por ciento de las parejas son consideradas como infértiles y el factor masculino es responsable en un 50 por ciento de los casos. El espermiograma, es un examen fundamental para el diagnóstico inicial de parejas infértiles. Objetivo: Determinar cambios en cuatro parámetros del espermiograma de mayor valor diagnóstico, según edad, estableciendo el parámetro alterado de mayor frecuencia. Métodos: Se realizo un estudio descriptivo retrospectivo de una muestra de 100 pacientes atendidos por problemas de fertilidad entre los años 2004 y 2009, clasificándolos en cuatro grupos etarios. Resultados: Al evaluar la concentración espermática, el 33 por ciento presenta: 5 baja concentración. El 86 por ciento de los pacientes presento astenozoospermia. El 81 por ciento de los pacientes presento anormalidad en la morfología espermática. La viabilidad espermática fue anormal en el 8 por ciento de los pacientes, siendo significativamente más alto en el grupo etario de mayor edad. Conclusiones: Los parámetros estudiados muestran un alto porcentaje de anormalidad en la población en estudio. Al comparar entre grupos, el grupo de mayor edad (sobre los 47 &los) presenta un aumento significativo del- porcentaje de alteraciones en morfología, motilidad y viabilidad respecto a los otros grupos etarios, estableciéndose la edad como un factor negativo en la calidad espermática. La movilidad corresponde al parámetro mas frecuentemente alterado seguido por la morfología espermática a medida que el varón consultante envejece.

In Chile, 10 to 15 percent of the couples are considered as infertile. Since the male factor is responsible of 50 percent of the cases, spermogram is an essential test for initial diagnosis of the infertile couple. Objective: To analyze the frequency of change in four spermogram parameters -according to age- to determine their diagnostic value. Method: A descriptive retrospective study of spermogram data from 100 patients -subdivided in four age groups- analyzed in our Unit for fertility problems between 2004 and 2009 was performed. Results: In sperm count, 33 percenr showed an abnormally low concentration. An 86 percent of the patients has astenozoospermia. 81 percent of the patients showed abnormal sperm morphology. Sperm viability was subnormal in 8 percent of the patients, being significantly higher in the oldest group. Conclusions: The seminal parameters analyzed revealed a high percentage of anomalies in the studied population. The oldest group had significant percentages of anomalies in sperm motility, morphology and viability, thus corroborating that age is a negative factor that affects semen quality. Sperm motility was the most frequently altered parameter followed by sperm morphology in the population under study.

Role of testicular fine-needle aspiration cytology in infertile men with clinically obstructive azoospermia.

BACKGROUND: Azoospermia due to obstruction of the vaso-epididymal junction is one of the few surgically correctable causes of male infertility. In patients where all clinical and laboratory parameters suggest a vaso-epididymal junction block amenable to surgery, failure to find normal spermatogenesis on fine-needle aspiration cytology (FNAC) of the testis may necessitate a change in treatment modality to the more expensive intracytoplasmic sperm injection. We evaluated the validity of FNAC findings in predicting failure of surgical exploration when clinical parameters suggest otherwise. METHODS: Infertile, azoospermic men in whom the semen volume and fructose content, testis size, follicle-stimulating hormone level were normal and the vas deferens was palpable with no evident cause for obstruction, underwent FNAC of the testis to confirm the presence of normal spermatogenesis before surgical exploration. Men with hypospermatogenesis or maturation arrest on FNAC and a normal karyotype with absence of Y chromosome microdeletion were offered assisted reproduction or surgical exploration to identify a reconstructable obstruction. Men who chose surgery were included in the study and the findings on exploration were compared with the FNAC reports. RESULTS: Of the 10 men who satisfied the inclusion criteria, 6 had hypospermatogenesis and in 4 FNAC showed maturation arrest. On surgical exploration, none had sperm in the epididymis. A biopsy of the testis taken at the time of exploration confirmed the FNAC findings. CONCLUSION: Clinical parameters are insufficient for diagnosing obstructive azoospermia. FNAC can accurately evaluate the testicular pathology and predict whether or not surgical exploration should be undertaken.

Search and identification of spermatozoa and spermatids in the ejaculate of non-obstructive azoospermic patients

OBJECTIVE: To search and to identify spermatozoa and spermatids, present in the ejaculate of non-obstructive azoospermic patients. MATERIALS AND METHODS: 27 patients, aged between 18 and 48 years, with initial diagnosis compatible with non-obstructive azoospermia, underwent up to 3 seminal samples, with assessment of macroscopic and microscopic parameters differentiated for each sample. In the first sample, 5 æL of semen were analyzed in a Horwell chamber in order to assess the presence or absence of spermatozoa. The procedure was repeated with 2 other aliquots. In the absence of spermatozoa, the entire sample was transferred to a conic tube and following centrifugation the sediment was freshly analyzed. The second seminal sample was collected only when no spermatozoa were found in the first sample and the research was performed in the same way. In cases where spermatozoa were not seen, the sample was centrifuged and the obtained sediment was stained by the panoptic method and observed under common light microscopy (1250X). The third seminal sample was collected only in cases when patients had not shown spermatozoa in the first and second seminal samples. RESULTS: 4/27 (14.8 percent) patients presented spermatozoa in the first seminal sample and 6/23 (26.1 percent), in the second seminal sample. No spermatozoa were seen in the third sample, however, 11/17 (64.7 percent) presented spermatids. CONCLUSION: In clinical situations where the initial diagnosis is non-obstructive azoospermia, one single routine seminal analysis is not enough to confirm this diagnosis and the analysis of the centrifuged sediment can have relevant clinical consequences. Among patients considered non-obstructive azoospermic, when duly assessed, 37 percent presented spermatozoa and 64.7 percent, spermatids.

Azoospermie: ICSI avec prelevement synchrone versus asynchrone du sperme

We review the improvement made in the management of infertile couples with obstructive and non obstructive azoospermia thanks to ICSI. We present the difficulties encountered in predicting the presence or the absence of spermatozoa in non obstructive azoospermia according to clinical and complementary investigations. Testicular biopsy is the only effective test. We recommend the practice of non synchronic testicular biopsy and sperm freezing to avoid unnecessary ovarian hyperstimulation

Comparison of the outcome of intracytoplasmic sperm injection using fresh and frozen-thawed epididymal spermatozoa obtained by percutaneous epididymal sperm aspiration.

The study was conducted to compare the outcome of intracytoplasmic sperm injection (ICSI) between fresh and frozen-thawed epididymal spermatozoa retrieved by percutaneous epididymal sperm aspiration (PESA) from obstructive azoospermic men. Overall, 53 PESA procedures were performed in 42 obstructive azoospermic men, followed by ICSI procedures with either fresh (n = 40) or frozen-thawed (n = 13) epididymal spermatozoa. Comparing all ICSI cycles with fresh and frozen-thawed epididymal spermatozoa, the fertilization rates (77.4 vs 86.8%) and the cleavage rate (91.3 vs 95.1%) were not statistically different. A total of 64 embryo transfers were performed: 48 embryo transfers after the use of fresh epididymal spermatozoa and 16 embryo transfers after the use of frozen-thawed spermatozoa. The overall pregnancy rate per embryo transfer was also similar between both groups (40.8 vs 40%). The implantation rate per embryo (18.2 vs 12.7%), clinical pregnancy per embryo transfer (36.7 vs 33.3%) and delivery/ongoing pregnancy rate (36.7 vs 33.3%) were not statistically different. In conclusion, there were no significant differences of the outcome of intracytoplasmic sperm injection using fresh and frozen-thawed epididymal spermatozoa obtained by PESA.

Investigation of sex chromosome abnormalities in teratozoospermia of infertile men using fluorescence in situ hybridization.

Fluorescence in situ hybridization is a fast and efficient method of investigating chromosomal abnormalities in human spermatozoa. In this study, we have established the frequency of sex chromosome disomy (XX, XY and YY) in teratozoospermia (98-100% abnormal morphology) of infertile men compared with normospermic men using double-probe FISH procedures. A total of 40,000 sperms were scored in each group of semen for chromosome X and Y with overall hybridization efficiency of 97.00 per cent and 98.02 per cent in infertile men and normal men, respectively. It was found that the frequency of disomy XX, XY and YY was significantly higher in infertile men compared with normal men (P < 0.05). It is concluded that the infertile men showed increased frequencies of sex chromosomal abnormalities.

Microdeleçöes do cromossoma Y em homens azoospérmicos e oligoospérmicos severos / Y-Chromosome microdeletions in azoospermic and severel oligozoospermic men

O fator masculino é responsável por aproximadamente metade dos casos de infertilidade do casal. Em mais de 60 por cento dos casos a origem da funçäo testicular diminuída näo é conhecida, podendo haver muitas anomalias genéticas näo identificadas. Microdeleçöes do braço longo do cromossoma Y säo associadas com falência da espermatogênese e tem sido usado para definir três regiöes do Yq (AZFa, AZFb e AZFc) que estäo correntemente deletados em homens inférteis. Em torno de 10 a 15 por cento dos azoospérmicos e 5 a 10 por cento dos homens severemente oligospérmicos tem microdeleçöes do Yq. Técnicas de reproduçäo assistida, principalmente Injeçäo Intra Citoplasmática de Esperma em associaçäo com retirada de esperma testicular, representam uma eficiente terapia para estes pacientes. Os autores fazem uma revisäo atual das microdeleçöes do cromossoma Y e suas conseqüências na fertilidade masculina.

Síndrome de Young: infecções respiratórias de repetição e azoospermia / Young's syndrome: repeated respiratory tract infections and azoospermia

INTRODUÇÃO: A síndrome de Young é uma variante da discinesia ciliar primária, caracterizada pela ocorrência de infecções respiratórias de repetição e obstrução congênita do epidídimo. APRESENTAÇÃO DO CASO: Os autores apresentam um caso de rinossinusite e pneumonias de repetição em um paciente de 28 anos do sexo masculino. Dosagem de sódio e cloro no suor e pesquisa de imunodeficiências celulares e humorais resultaram negativas. O espermograma revelou azoospermia, embora a espermatogênese estivesse mantida, conforme achado na biópsia de testículo. DISCUSSÃO: O diagnóstico foi de síndrome de Young, sendo este o primeiro caso relatado no Brasil. CONCLUSÃO: Os autores alertam para a importância desse diagnóstico, dadas suas implicações para aconselhamento genético, além do diagnóstico diferencial a ser feito com a fibrose cística.

Análise crítica da utilidade do teste de clonidina - homônio de crescimento (GH) no diagnóstico do homen infértil / Critical analysis of reliabillity of clonidine test - growth hormone (GH) used in diagnosis of infertile male

O presente trabalho visa analisar a confiabilidade do teste de clonidina - GH (TC-GH) no diagnóstico de uma insuficiência relativa de GH em uma população de homens inférteis. Vinte e sete pacientes foram classificados em 3 grupos de acordo com a dose de clonidina. Os grupos I (clonidina 0,150mg) e II (clonidina 0,200mg) tinham 9 e 6 pacientes oligozoospérmicos, respectivamente. O grupo III (clonidina 0,400mg) tinha 5 pacientes oligozoospérmicos (subgrupo IIIa) e 7 normozoospérmicos (subgrupo IIIb). Trinta e um TC-GH foram realizados e vinte e nove deles foram analisados; cinco (17,2 por cento) resultaram normais (1 no grupo I, 1 no grupo III e 3 no grupo IIIb). As respostas do GH após 60, 90 e 120 minutos do estímulo não mostraram diferença significativa nos grupos I, II e III nem entre os subgrupos IIIa e IIIb. Conclui-se que o TC-GH não é de utilidade no diagnóstico de uma relativa insuficiência de GH no homem infértil, independente da dose de clonidina. A razão é a incapacidade da clonidina produzir um estímulo de GH confiável no homem adulto.

Extraçäo de espermatozóide testicular (TESE) em azoospermia não obstrutiva: aspectos urológico e anatômico / Testicular sperm extraction (TESE) for nonobstructive azoospermia: urological and anatomical issues

A abordagem do paciente com diagnóstico de azoospermia näo obstrutiva se modificou com o passar dos anos, tendo em vista o advento da técnica de injeçäo intracitoplasmática de espermatozóides (ICSI). Com a extraçäo de espermatozóides do testículo (TESE) o estudo de infertilidade masculina näo se restringe a anatomia estrutural do tecido, mas acrescenta-se o apoio do laboratório de espermatologia. Na quantificaçäo espermática se valoriza o tamanho do fragmento, a presença de espermatozóide e células precursoras. Outro ponto importante é o estudo do comportamento citogenético do cromossomo Y. No presente artigo, a técnica de TESE é descrita e sua importância destacada com novas perspectivas no estudo da infertilidade do homem.

Les obstructions congenitales ou acquises de l'epididyme humain: Consequences et diagnostic etiologique

In this study, concerning 86 patients submitted to surgical exploration for sterility, has been achieved with a double objective to determines the precise histological nature of the lesions of the sperm ducts mainly of the epididymis and determine the etiological diagnosis of the epididymal obstruction. We've showen that whatever the level of the acquired obstruction of the epididymis, it will lead to the formation of an inflammatory home and to a situation combining the lessening of light, stase and massive arrival of spermiophage of destroying almost completely the spermatozoa. The simultaneous consideration of the 2 values of the <-glucosidase and the plasma FSH permit of reducing the mistakes in interpreting the origin of azoospermia. However we have produced evidence of diagnostic traps corresponding to a fibrose of rete testis where all the biological parameters are normal. W've showen that, there is a correlation between the presence of epididymal pathology and a drop in epididymal markers which can be found in severe oligospermia [which can be epididymal in origin and not testicular], also when there is non abnormalities in the spermogramm. This last situation can occur in " invisible" abnormalities of sperm maturation in the epididymis

Pulmonary alveolar microlithiasis: diagnosed by sputum examination and transbronchial biopsy.

A case of pulmonary alveolar microlithiasis presenting with uncharacteristic radiological features who was diagnosed on the basis of sputum examination and confirmed by transbronchial biopsy, is being reported. Azoospermia resulting in primary infertility and increased lung volumes by helium dilution test were other noteworthy features of the case.

Caracterización de fracciones mayoritarias del plasma seminal humano: estudio comparativo entre individuos normospérmicos, azoospérmicos, necrozoospérmicos, oligozoospérmicos y vasectomizados / Caracterization of major human seminal plasma fractions: comparative study with normospermic, azoospermic, necrozoospermic, oligozoospermic and vasectomised donors

El plasma seminal humano (PSH) de individuos normospérmicos y con diferentes patologías fue analizado por cromatografía de afinidad en Concanavalina-A Sepharose. Se obtuvieron dos fracciones mayoritarias, F1 y F2, en proporciones variables y con diferencias en la composición de monosacáridos y aminoácidos de acuerdo con el tipo de patología. Galactosa es el principal miembro, juntamenete con fucosa y proporciones variables de manosa, N-acetilglucosamina y N-acetilgalactosamina. Se encontraron diferencias en la estructura primaria de los glicanos (enlaces N- y O-glicosídicos) presentes en las fracciones de los individuos con diferentes patologías. En F1 todas las cadenas de oligosacáridos son del tipo O-glicosídica correspondiendo a glicoproteínas tipo mucina; en F2 se encontró una relación molar O-/N- es 5,3/1. La fracción de F1 de individuos azoospérmicos y oligozoospérmicos son del tipo O-glicosídico mientras que en vasectomizados la relación molar O-/N- es 1,7/1. En todos los casos, F2 presenta cadenas de oligosacáridos O- y N-glicosídicas en diferentes proporciones, con la excepción del plasma seminal de vasectomizado que pertenece al tipo O-glicosídico. Los resultados obtenidos sugieren que la composición de las fracciones F1 y F2 de donantes con diferentes patologías está caracterizada por diferencias importantes en el contenido de carbohidratos y pépticos

A comparison of Makler counting chamber and improved Neubauer hemocytometer in sperm concentration measurement.

Evaluation of male fertility is based predominantly on results from semen analysis and determination of the sperm concentration is one of the main parameters of the analysis. To assess the accuracy of sperm concentration measurements by Makler counting chamber, manual sperm counting of 55 semen samples were made using a Makler counting chamber, and compared with concentration values measured using an improved Neubauer hemocytometer according to the World Health Organization guideline (standard procedure). Results were classified according to the standard procedure before statistical analyses. Both values correlated well. Sperm concentration obtained with Makler counting chamber was not statistically different from those determined by improved Neubauer hemocytometer in semen samples with concentrations over 40 x 10(6)/ml. But using Makler counting chambers caused a shift concentrations, which were overestimated significantly (p less than 0.0001) in semen samples with concentrations less than 40 x 10(6)/ml. Overall, Makler chamber counts were 11.2 per cent higher. Although less complicated than the improved Neubauer hemocytometer method, measurement of sperm concentration by Makler counting chamber is an inaccurate method, especially in semen samples with concentrations less than 40 x 10(6).

LDH-x in azoospermia: a new diagnostic alternative to vasography and testicular biopsy.

Activities of different isozymes of lactate dehydrogenase were studied in semen samples from fertile, and different groups of infertile men. No isozyme except the sperm specific LDH-x or C4 showed any difference in activity with changes in sperm density. Activity of LDH-x exhibited a trend positive to changes in sperm density and became absent in obstructive and vasectomized azoospermia. Among other types of azoospermia, the activity became very high in maturation arrest but very low in patients with testicular biopsy showing germ cell aplasia, such as in SCOS and ghostlike tubules. Diagnostic usefulness of the estimation of LDH-x activity in semen is discussed.

Ultra-estrutura ciliar à microscopia eletrônica de transmissäo / Ciliary ultrastructure at transmission electron microscopy

A síndrome da discinesia ciliar é uma das hipóteses diagnósticas que devem ser consideradas em indivíduos com sintomatologia crônica pulmonar e de vias aéreas superiores. Nesta doença, os cilios do epitélio respiratório säo imóveis ou entäo apresentam um batimento descoordenado. Qualquer uma dessas situaçöes acaba por prejudicar o transporte mucociliar, um dos mais importantes mecanismos de defesa do aparelho respiratório. Acredita-se que anomalias estruturais ciliares possam levar a funcionamento inadequado dos cílios. Este trabalho teve como principais objetivos a identificaçäo dos elementos da ultra-estrutura ciliar normal à microscopia eletrônica de transmissäo e também a análise desta ultraestrutura em pacientes com diagnóstico clínico de síndrome de discinesia ciliar. Participaram do estudo oito pacientes e quatro controles normais. Quatro dos pacientes eram também portadores da síndrome de Kartagener. A comparaçäo dos resultados obtidos em controles e casos permitiu o diagnóstico de anomalias ultra-estruturais em quatro pacientes. Os defeitos identificados foram: deficiência de braços externos de dineína em dois pacientes, e deficiência de braços internos de dineína e alteraçäo de filamentos radiais em outros dois pacientes. Três pacientes com diagnóstico clínico de discinesia ciliar dmonstraram ter ultra-estrutura ciliar normal. Deste grupo, fazia parte um indivíduo com síndrome de Kartagener. O oitavo paciente apresentou também ultra-estrutura ciliar normal, fato que somado à presença de azoospermia obstrutiva leva ao diagnóstico de síndrome de Young. Alguns achados ultra-estruturais deste trabalho discordam das informaçöes da literatura sobre o assunto. Este é o caso, por exemplo, do pé basal, estrutura que a maioria dos autores apresenta como única; no entanto, foi bastante freqüente neste material a observaçäo de pés basais duplos e até triplos, tanto em controles como em casos

La valoración de la biopsia testicular del varón estéril utilizando métodos cualitativos y cuantitativos / Acsessment of testicle byopsy in the sterile male using qualitative and qualitative methods

Con el propósito de establcer relaciones cuantitativas entre los diferentes componentes del túbulo seminífero que correlaciona con estados patológicos, se analizaron 12 biopsias testiculares de pacientes oligozoospérmicos y azoospérmicos. A cada paciente se le midieron los niveles séricos de LH, FSH y testorerona por lo menos en una ocasión y se les practicó espermocultivo. Las biopsias testiculares se procesaron de acuerdo con las técnicas de rutina para microscopía electrónica. Los cortes semifinos se valoraron cualitativamente de acuerdo con la arquitectura tubular y la composición celular intratubular. Para la valoración cuantitativa se emplearon 2 métodos: 1) el método del "score count" del Johnsen y 2) un estudio morfométrico determinando el diámetro tubular, las áreas tubular total, de pared tubular, membrana basal y epitelio germinativo, para lo cual se empleó un equipo analizador de imágenes IBAS 2000. Se encontraron espermatozoides en cinco biopsias de ocho pacientes reportados como azoospérmicos y en dos pacientes oligozoospérmicos no se encontraron espermatozoides en los túbulos analizados. Utilizando el método del "score count" se encontró un rango de 1.00 (correspondiendo a dos pacientes con esclerosis tubular) a 7.41 (paciente con hipoespermatogénesis). El diámetro tubular varió de 140m (con un score de 1.93) a 207 m (con un score de 7.41). El área tubular varió de 24094 m2 a 70220 m2. Los métodos cuantitativos permiten un conocimiento más profundo de los diferentes eventos morfológicos que concurren en el proceso de la espermatogénesis en pacientes con patología, sin embargo, se requiere de más investigación en este sentido.