Multiple Anomalies Index Research Articles

Semen Cryopreservation in Adolescents and Young Adults with Hematologic Diseases: from Bed to Benchside.

Purpose: Infertility in adolescents and young adult (AYA) survivors of malignant disease remains a major long-term adverse effect, but semen collection for fertility preservation in fertility centers is not always feasible and makes AYAs uncomfortable. We evaluated the feasibility of collecting sperm samples on the ward versus in fertility centers. Methods: Consecutive hospitalized AYA-aged male patients in the Hematology AYA unit (Saint-Louis Hospital, France) between August 2010 and June 2016 with hematological disease and indication of semen collection (n = 95) were included in this retrospective study. Semen quality was analyzed according to World Health Organization guidelines and was compared according to semen collection place: on the ward (n = 46) or in fertility center (n = 49). Results: The median age was median age 19.1 years (range: 13.7-33.3; interquartile range: 17.1-22.8) and 85 patients successfully collected semen. Sperm collection failure was ∼11% and was comparable between the two modalities as were main sperm quality characteristics (semen volume, sperm concentration, total sperm count, progressive motility and vitality, sperm morphology, and multiple anomalies index). Oligospermia was significantly higher in the samples obtained in fertility center (47.7%) than on the ward (26.8%), p = 0.047. Average frozen straws were comparable, 12.2 ± 6.4 on the ward versus 11.9 ± 6.3 in fertility center. Conclusion: Semen collection on the ward is feasible and would be particularly interesting for AYA male patients without altering semen quality characteristics.

Mild experimental increase in testis and epididymis temperature in men: effects on sperm morphology according to spermatogenesis stages.

A mild increase in testicular and epididymal temperatures in men, bulls and rams (pendulous scrotum) inhibits spermatogenesis and increases the percentage of sperm with an abnormal morphology. However, the stages of spermatogenesis that are most sensitive to a mild increase in testicular temperature in men are unknown. The aim of the present study was to explore the effects of a mild induced increase in testicular and epididymal temperature (i.e., testicular temperature maintained below the core body temperature) on sperm morphology in humans depending on the physiological time of spermatogenesis and epididymal transit. Five healthy volunteers were enrolled in an experimental study in which testicular and epididymal temperatures were increased by maintaining the testes in a supra-scrotal position with a specially designed underwear worn 15±1 h a day for 120 consecutive days. Semen collection was scheduled on specific days depending on spermatogenic stages and epididymal transit. Sperm morphology and the multiple anomalies index (MAI) were analysed before, during and after heating. This mild induced increase in testicular and epididymal temperatures resulted in a significant rise in the percentage of morphologically abnormal spermatozoa on day 34 of heating, which remained elevated throughout the heating period and persisted until 45 days after cessation of heating. The MAI was significantly increased on day 20 throughout the heating period and persisted 45 days after cessation of heating. An increase in the percentage of anomalies in the sperm head, acrosome or tail occurred on days 34 and/or 45 of heating. Abnormal sperm morphology and MAI reverted to control values 73 days after cessation of heating. A mild sustained increase in testicular and epididymal temperature in man leads morphological abnormalities in spermatozoa mainly due to an impairment of spermiogenesis and meiosis.

The association between iron, calcium, and oxidative stress in seminal plasma and sperm quality.

The present study aimed to determine the level of iron and calcium in the seminal plasma of men with different fertility potentials and to examine its relationship with oxidative stress. Seventy-nine sub-fertile patients with asthenoteratozoospermia (AT), n 27; teratoleucozoospermia (TL), n 20; teratozoospermia (Terato), n 32; and 29 healthy donors were included. The ability of spermatozoa to produce reactive oxygen species (ROS) was evaluated by using nitroblue tetrazolium (NBT) staining. The lipid peroxidation end product, malondialdehyde (MDA), and the trace element levels (iron and calcium) were measured spectrophotometrically. Iron and calcium concentrations in seminal plasma of the patient groups were significantly more elevated than the normal group. Nevertheless, both calcium and iron showed strong negative correlations with the total sperm motility and normal sperm morphology, but only iron was positively and significantly associated with multiple anomalies index and seminal leucocyte concentration. On the other hand, the rates of MDA and ROS production in semen were significantly higher in the three abnormal groups than in controls. These two oxidative stress biomarkers were significantly associated with the percentage of atypical forms in semen. However, only semen ROS level was significantly associated with the decreased sperm motility and the sperm leucocytes concentration. Meanwhile, there are positive correlations between seminal iron and calcium content and the studied oxidative stress biomarkers. Oxidative stress and trace element excess are implicated in low sperm quality. Iron and calcium might be the mediators of the effects of oxidative damage and induces lipid peroxidation.

Variability in the morphologic assessment of human sperm: use of the strict criteria recommended by the World Health Organization in 2010

To determine the variability in the recognition of normal sperm and various sperm defects using the strict criteria recommended by the World Health Organization (5th edition, 2010). Sperm morphologic assessment by three experienced evaluators. Image processing laboratory and reproduction research institute. Semen donors from a sperm bank. The morphology of 5,296 sperm was evaluated using statistical analyses of variability. The proportion and coefficients of variation (CVs) of normal sperm, defects of specific parts, and the categories of defects were measured. The degree of agreement between any two of the three evaluators was calculated. The multiple anomalies index, teratozoospermia index, sperm deformity index, and the CVs were also measured. The CVs of normal sperm, multiple anomalies index, teratozoospermia index, and sperm deformity index were 4.80%, 4.14%, 5.75%, and 6.81%, respectively. A broader range (4.80%-132.97%) of CVs was observed for the recognition of various defects. The coefficients of the degree of agreement concerning specific morphologic parts of sperm varied (0.387-0.607), with lower relative values for the head and mid-piece than for the tail and cytoplasm. The sperm head is more difficult to evaluate than the other parts using the criteria recommended by the World Health Organization in 2010. The degree of agreement concerning specific parts and various defects varied in broad ranges. A stricter definition for each defect is needed.

Effect of the serum testosterone/estradiol ratio alteration on sperm defect and fertility

Objective To study the effect of the serum testosterone/estradiol ratio (T/E2) alteration on sperm defect and fertility. Methods The testosterone, estradiol, FSH, LH, PRL,sperm parameters and sperm morphology of 90 men were analyzed and the T/E2 and multiple anomalies index (MAI) were calculated. The patients were divided into 3 groups; T/E2≤10 (Ⅰ), T/E2＞10 (Ⅱ), and T/E2＞20 (Ⅲ). Results The sperm concentration and motility among the 3 groups were not significantly different (P＞0. 05). The percentages of the sperm whose head, neck and tail were abnormal declined gradually with the increase of the T/E2 in serum. The percentage of sperm head defeet of group Ⅰ was significantly higher than that of group Ⅲ (t=2. 482, P=0. 016) and that of sperm neck defect of groups Ⅰ and Ⅱ were significantly higher than that of group Ⅲ (t=4.113, 2. 050, P=0.000, 0. 046, respectively). The percentage of sperm tail defect among 3 groups was significantly different (t=2. 722, 3. 996, 3. 110, P=0. 008, 0. 000, 0. 003, respectively). The SDI of group Ⅱ was significantly higher than that of group Ⅰ (t= -2. 293, P= 0. 025). But the TZI increased gradually with the increase of the serum T/E2 and the TZI of groups Ⅱ and Ⅲ were significantly higher than that of group Ⅰ (t=2. 285, 2. 727, P=0. 025, 0. 009, respectively). The percentage of the men in group Ⅰ whose partners became pregnant was 29. 5% and those of groups Ⅱ and Ⅲ were 50% and 42.9%, respectively. Although the percentage among three groups was not different statistically (x2 = 3. 285, 0. 854, 0. 199, P= 0. 070, 0. 355, 0. 655, respectively), the relative risks of groups Ⅱ and Ⅲ were 2.4 and 1.8 times of that of group Ⅰ. There were 25, 15, 7 cases of idiopathic infertility among the 3 groups, respectively. The relative risk of I group was 1.5 and 1.3 times of that of Ⅱ and Ⅲ groups. The correlation analysis showed that the T/E2 in serum had significantly negative correlation with the percent of the sperm head or neck or tail defects (r= -0. 209, -0. 316 and -0. 335,respectively and P= 0. 048, 0. 002 and 0. 001, respectively). Conclusions The decrease of T/E2 in serum was correlative with the decrease of fertility probability, but it did not alter the sperm density and the sperm motility. It showed that the level of the T/E2 in serum was important for spermatogenesis and sperm fertilizing capability. Key words: Testosterone; Estradiol; Testosterone/estradiol ratio in serum; Sperm defect ; Fertility

Inter-individual variability in the morphological assessment of human sperm: effect of the level of experience and the use of standard methods.

The assessment of human sperm morphology is not devoid of methodological problems, making comparison among laboratories difficult. The main goal of this study was to assess how the experience and compliance to the recommended procedures can modulate it. Sixty-two technicians and biologists of varying degrees of andrological experience classified the same 100 sperm in a blind study from images captured on a video microscope and projected onto a screen. David's morphology classification which distinguishes normal and abnormal sperm and calculates a Multiple Anomalies Index (MAI) was used. These sperm were also classified by two trained and experienced biologists using videotaped images to provide reference values. The inter-participant coefficient of variation was 40% for normal sperm, lower for five defects and, 12% for MAI. Experienced participants had the closest results to the reference values. Moreover, participants not routinely using the recommended methodologies, regardless of their experience level, consistently showed marked differences compared with the reference values. The present data confirm the wide variability in the assessment of normal sperm and show a lower variability for several anomalies and MAI. They underscore the role that experience plays and the importance of following the recommended methodologies for relevant and comparable results.

Time to pregnancy and semen parameters: a cross-sectional study among fertile couples from four European cities.

In fertile populations, little is known about the association between semen parameters and time to pregnancy (TTP). Pregnant women from Copenhagen, Edinburgh, Paris and Turku who conceived without medical intervention were asked for their TTP (942 couples), and their partners provided a semen sample. The proportion of morphologically normal sperm and the multiple anomalies index (MAI, ratio of the total number of anomalies to the number of abnormal sperm) were centrally estimated. We estimated rate ratios for the occurrence of a pregnancy by a discrete survival model, adjusted for sexual activity and female factors affecting fecundity. Increasing sperm concentration influenced TTP up to 55 x 10(6)/ml. The proportion of morphologically normal sperm influenced TTP up to 39% according to David's criteria, and this association held among the subjects with a sperm concentration >55 x 10(6)/ml. For strict criteria, the threshold value was 19% normal sperm. An increase of 0.5 in MAI was associated with an adjusted rate ratio for the occurrence of a pregnancy of 0.68 (95% confidence interval: 0.54-0.85). These results highlight the importance of sperm morphology parameters and indicate that the effect of proportion of normal sperm on TTP may be independent of sperm concentration.

Familial sperm polyploidy induced by genetic spermatogenesis failure: Case report

We report a case of oligoasthenoteratozoospermia in a 40 year-old patient with a familial history that revealed multiple cases of infertility and perinatal deaths. The patient's semen sample contained 2x10(6) spermatozoa/ml, with an overall progressively motile population of <5%. Cytological analysis revealed a teratozoospermia with 100% of abnormal macrocephalic sperm heads and an irregular acrosomal cap in 38% of cells. Moreover, 72% of spermatozoa carried multiple flagella (2-5). The midpiece was elongated and/or enlarged with cytoplasmic droplets in 15% of cells. The multiple anomalies index (MAI) was 3.3 (normal value = 1.6), reflecting the high incidence of spermatozoal morphological abnormalities in this patient. Ultrastructural analysis revealed the presence of 2 or 3 vacuolated nuclei per sperm head. The acrosome was abnormal and the chromatin, partially packaged, appeared rough. In some cases, a large amount of cytoplasm containing vacuoles was observed around the nucleus and the acrosome. The mitochondrial helix was disorganized. Chromosome analysis performed on blood cells revealed a normal karyotype. Three-colour fluorescence in-situ hybridization (FISH) analysis of 1148 spermatozoa showed 21.6% to be diploid, 62.4% triploid, 13.3% quadriploid and 2.7% hyperploid (<4n). In conclusion, we suggest that this case could result from a genetically induced spermiation failure, the origin of which is discussed.

The sperm deformity and the sperm multiple anomalies indexes in patients who underwent unilateral orchectomy and preventive radiotherapy

Two new indices, the sperm deformity and the sperm multiple anomalies index, were assessed in 16 male patients who underwent unilateral orchectomy owing to seminoma and in preventive radiotherapy of paraortic and of the iliac lymph glands homolateral to the testis excised. The aim was to assess the value of these two morphological sperm parameters in distinguishing the semen between the above mentioned group of men from the semen of men with other causes of infertility. It was found that: (1) the values of the sperm deformity index and those of the sperm multiple anomalies index were not significantly different between men who underwent unilateral orchectomy and preventive radiotherapy and the men with other causes of subfertility and (2) the values of the above indices were significantly higher in the semen specimens of men who underwent unilateral orchectomy and preventive radiotherapy and in the semen specimens of men with other causes of subfertility than in the semen specimens of fertile men. It is concluded that testicular response, as far as sperm morphology is concerned, is the same irrespective of the cause exerting a negative effect on spermatogenesis.

Flow cytometric light scattering analysis, acrosome reaction, reactive oxygen species production and leukocyte contamination of semen preparation in prediction of fertilization rate in vitro.

The effects on fertilization of the morphology of spermatozoa, acrosome reaction, reactive oxygen species (ROS) production, leukocyte contamination and the light scattering in flow cytometry of semen preparation were investigated in 73 couples undergoing their first in-vitro fertilization treatment. All men had normal concentrations of spermatozoa with sufficient motility (> or = 50%) and yield (> or = 6 x 10(6)) in the semen preparation on the day of oocyte retrieval. The light scattering properties of all cells present in the semen preparation, as assessed with flow cytometry, were correlated with fertilization. The proportion of metaphase I oocytes was also correlated with the fertilization rate of all collected oocytes and of metaphase II oocytes. ROS production, leukocyte contamination, spontaneous or calcium ionophore-stimulated acrosome reaction, percentage of normal morphology, and multiple anomalies index had no independent contribution.

Predicting the fertilizing potential of human sperm suspensions in vitro: importance of sperm morphology and leukocyte contamination

To determine the relationships between sperm function tests and fertilization of human oocytes in vitro. Analysis of infertile patients undergoing IVF therapy. Diagnostic Andrology Laboratory and Assisted Conception Service. Forty-one couples who underwent IVF-ET therapy were studied. None. The ability of human spermatozoa to achieve fertilization in vitro was examined in relation to numerous criteria of semen quality, including the conventional semen profile, the computer-aided assessment of sperm movement, ionophore-induced acrosome reaction, acridine orange staining, sperm morphology, and chemiluminescent signals induced by phorbol ester and N-formyl-methionyl-leucyl-phenylalanine (FMLP). Significant correlations were observed between fertilization rates and several attributes of the sperm preparations, including elements of sperm function (acrosome reaction), movement (percentage motile, hyperactivation, the amplitude of lateral sperm head displacement), morphology (normal morphology, midpiece defects, multiple anomalies index), nuclear normality (acridine orange staining), and reactive oxygen species generation (chemiluminescence induced by phorbol ester and FMLP). In a stepwise multiple regression analysis, an accurate prediction of fertilization rates was obtained using a multiple regression equation incorporating six variables of which sperm morphology and FMLP-induced chemiluminescence were the most informative. A set of criteria have been identified that accurately predict the fertilizing potential of human sperm suspensions in vitro and that place particular emphasis on sperm morphology and the degree of leukocyte contamination.

Localization of antibodies on spermatozoa and sperm movement characteristics are good predictors of in vitro fertilization success in cases of male autoimmune infertility

To define sperm factors related to in vitro fertilization (IVF) failure in cases of antisperm autoimmunity. A detailed analysis of sperm morphology, movement characteristics, acrosomal function, and antibody binding was performed on the sperm population selected on a discontinuous two-layer Percoll gradient and used for IVF. The results were compared retrospectively between fertilizing (n = 13) and nonfertilizing (n = 11) sperm populations. Twenty-one infertile couples undergoing 24 cycles of IVF treatment because of antisperm autoimmunity were included in this study. Fertilizing and nonfertilizing sperm populations were not different with respect to the percentage of motility, the normal morphology, the multiple anomalies index, the acrosome abnormalities, and the spontaneous or induced acrosome reaction. If the proportion of spermatozoa coated with either immunoglobulin (Ig)A or IgG antibodies was similar in the two groups, their localization was often different: antibodies were mainly on the sperm heads in the cases of fertilization failure. There were significant differences between fertilizing and nonfertilizing sperm samples in several movement parameters. Among them, the amplitude of lateral head displacement (ALH) was the most significantly correlated with fertilization success. Finally, spermatozoa were poorly bound to the zona pellucida (ZP) when fertilization failed, whereas high numbers of spermatozoa were attached to the ZP when fertilization occurred. Fertilization failure in patients with antisperm antibodies may be the result of several factors in which the impact of the antibodies on the membrane function play a critical role. Movement parameters, particularly the ALH and the localization of antibodies on migrated spermatozoa could predict the IVF failure or success more accurately than the proportion of antibody-coated spermatozoa in the inseminated populations. The fertilization failure was associated with an incapacity of spermatozoa to bind to ZP.

Male factors and the likelihood of pregnancy in infertile couples. I. Study of sperm characteristics.

A prospective study of 394 infertile men was conducted over 3 years following a primary semen analysis. The cumulative pregnancy rate was 43 and 64% after 1 and 3 years, respectively. The pregnancy rate was significantly higher in the secondary infertile group. The study of various sperm factors and the occurrence of pregnancy showed that they were not of equal significance in predicting male fertility potential. The percentage of pregnancies decreased significantly only when the sperm concentration was less than 5 x 10(6)/ml. The pregnancy rate increased significantly with the percentage of motile sperm. The percentage of sperm with normal morphology was also found to be significantly higher when a pregnancy occurred than when the couple remained infertile (43.6% vs 37.7%). In a detailed morphological analysis of the sperm, six abnormalities (microcephaly, double head, amorphous head, cytoplasmic droplet, bent tail and coiled tail) were found to be significantly more frequent when a pregnancy did not occur. The most predictive value was given by the Multiple Anomalies Index (MAI), which is the mean number of abnormalities observed per abnormal sperm. The pregnancy rate was significantly lower after both 1 and 3 years when the MAI was greater than 1.6. Multivariate analysis showed that the best prognostic indicator of fertility was given by the percentage of motile sperm and the MAI, particularly in patients with primary infertility.

Male factors and the likelihood of pregnancy in infertile couples. II. Study of clinical characteristics--practical consequences.

A prospective study of 394 infertile men was conducted over 3 years following a primary semen analysis. Simple comparisons between the groups of men whose partners conceived and those that did not, showed that the mean duration of infertility in the primary infertile group was significantly shorter when a pregnancy was observed, while age differed significantly between the two populations in the secondary infertile group. Multivariate analyses, taking these variables and sperm characteristics into account, showed that prognostic variables for the occurrence of pregnancy were the duration of infertility, sperm motility and the multiple anomalies index in the primary infertile group, and age of the male partner and percentage of normal sperm in the secondary infertile group. Tables for estimating the probability of observing a pregnancy as a function of these criteria are presented. In this study no positive influence of treatment on the occurrence of a pregnancy was found during the 3 years following the first semen analysis.