Diagnosis of male infertility: A need of functional and chromatin evaluation | Actas Urológicas Españolas (English Edition)

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Curti, M. Cánepa, L. Cantú, J.M. Montes" "autores" => array:4 [ 0 => array:4 [ "nombre" => "G." "apellidos" => "Curti" "email" => array:1 [ 0 => "giannicurti@gmail.com" ] "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "¿" "identificador" => "cor0005" ] ] ] 1 => array:2 [ "nombre" => "M." "apellidos" => "Cánepa" ] 2 => array:2 [ "nombre" => "L." "apellidos" => "Cantú" ] 3 => array:2 [ "nombre" => "J.M." "apellidos" => "Montes" ] ] "afiliaciones" => array:1 [ 0 => array:2 [ "entidad" => "Laboratorio Fertilab, Montevideo, Uruguay" "identificador" => "aff0005" ] ] "correspondencia" => array:1 [ 0 => array:3 [ "identificador" => "cor0005" "etiqueta" => "⁎" "correspondencia" => "Corresponding author." ] ] ] ] "titulosAlternativos" => array:1 [ "es" => array:1 [ "titulo" => "Diagnóstico de infertilidad masculina: necesidad de valoraciones funcionales y cromatínicas" ] ] "textoCompleto" => "IntroductionIt is estimated that infertility affects approximately 15% of couples of reproductive age,<a class="elsevierStyleCrossRef" href="#bib0005">1</a> and that in about 50% of the cases, the male factor is involved.<a class="elsevierStyleCrossRef" href="#bib0010">2</a>Semen analysis is now a routine procedure for the determination of the contribution of the male factor in the study of the infertile couple. It is thus an essential tool for clinicians when deciding between algorithms to continue with the diagnosis, or directly between therapeutic alternatives.In search of large-scale standardization of semen analysis, the World Health Organization (WHO) published in 1980 the first edition of its ‘Manual for the examination of human semen and sperm–cervical mucus interaction’.<a class="elsevierStyleCrossRef" href="#bib0015">3</a> Following the progress in this area, the WHO reissued revised and updated versions of the manual in 1987,<a class="elsevierStyleCrossRef" href="#bib0020">4</a> 1992,<a class="elsevierStyleCrossRef" href="#bib0025">5</a> 1999<a class="elsevierStyleCrossRef" href="#bib0030">6</a> and 2010.<a class="elsevierStyleCrossRef" href="#bib0035">7</a> These publications generated controversy on the true value of the recommended characteristics and their reference values.<a class="elsevierStyleCrossRefs" href="#bib0040">8–13</a>The sperm must exhibit a set of unique biological properties to achieve fertilization and normal embryonic development. Given this complexity, it is difficult to diagnose changes in its physiology using the standard semen analysis, this is, the study of the variables recommended in the WHO manuals.<a class="elsevierStyleCrossRef" href="#bib0070">14</a>Other tests, additional to those recommended in the manuals, have been developed in search of a more comprehensive evaluation. Studying both functional and chromatin features of the sperm, the results have been positively correlated with sperm quality and reproductive success, constituting potential diagnostic and prognostic tools<a class="elsevierStyleCrossRefs" href="#bib0075">15–19</a>.The aim of this work was to study the incidence of functional and chromatin alterations in the sperm of a population of patients with marital infertility subdivided according to the normality of the standard semen parameters, according to the latest edition of the WHO manual.<a class="elsevierStyleCrossRef" href="#bib0035">7</a> In turn, we intended to identify and correlate the altered characteristics with greater frequency in the normal subpopulation for standard parameters.Materials and methodsPatients and samplesThis study included 110 patients with marital infertility, whose average age was 38±7 years, selected between March and July 2011. A control group was made up of 6 volunteer fertile donors, aged 25±1 years. After standard sexual abstinence, and following the WHO guidelines,<a class="elsevierStyleCrossRef" href="#bib0035">7</a> the semen samples were collected by masturbation, in sterile containers tested and supplied by the laboratory personnel. All the subjects included in this study signed informed consent forms.The standard semen parameters analyzed were: sample volume, sperm concentration, progressive sperm motility, percentage of normal forms, percentage of immature forms, round cell concentration and concentration of peroxidase-positive cells. We also carried out the following functional tests: sperm survival at 24hours, hypo-osmotic swelling test (HOST), modified stress test (MOST), and the following additional tests: nitroblue tetrazolium test (NBT), aniline blue test (AB), and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay.Standard parameters were studied according to the latest WHO manual<a class="elsevierStyleCrossRef" href="#bib0035">7</a> and they were classified as normal or abnormal taking as a reference the ranges published in the manual. According to the above, the total set of patients was divided into two groups: group A, which included the patients whose samples were classified as normal for all standard parameters; and group B, with the patients whose samples showed alterations in at least one of these parameters.Analysis of standard semenAfter the liquefaction and determination of the volume of the samples, the sperm concentration and motility were analyzed, as well as the round cell concentration, by phase-contrast microscopy (Olympus BX51) using a Makler counting chamber type (Makler counting chamber, Sefi-Medical Instruments, Haifa, Israel). In order to show the peroxidase-positive cells, we used the benzidine–cianosine–H2O2 technique, and the reading of it was carried out by light-field microscopy (Olympus BX51). The sperm morphology was determined by analyzing stained smears according to the Papanicolaou technique described in the last WHO manual.<a class="elsevierStyleCrossRef" href="#bib0035">7</a> Immature sperm forms were counted in the same preparations.Functional tests24-h survivalThe survival rate was determined by the ratio between the percentages of motile sperm counted after and before 24h of incubation at room temperature, of an aliquot of washed semen (half Earle's Balanced Salt Solution [EBSS]+SAH 10%).Hypo-osmotic testThis test was carried out by incubating 100μl of semen for 30min at 37°C, in 1ml of hypo-osmotic solution (d-fructose 0.075M, sodium citrate 0.025M). These conditions cause a passive flow of water through intact and functional membranes into cells, which in turn causes them to swell. This swelling is evidenced by the winding of the sperm tails. Dead sperm or with altered membranes do not swell, and their tails are straight. The result of the test is expressed as the percentage of swollen sperm.Modified stress testThe rate described as a result of this test was calculated as the ratio between the percentages of motile sperm counted before and after 4h of incubation, at 40°C of an aliquot of washed sperm with EBSS+SAH 10%.Additional testsAniline blue testFor this study, smears with fresh semen were performed that were fixed in glutaraldehyde 3% in 0.2M phosphate buffer, and stained with aniline blue solution at 5% in the same buffer. The test result was expressed as the percentage of stained sperm.Nitroblue tetrazolium testIt was diluted 1:1, an aliquot of fresh semen with a NBT solution at 0.1% (Sigma–Aldrich, St. Louis, USA) in phosphate buffered saline (PBS – Dako North America Inc., CA, USA), and incubated for 30min at 37°C. After washing the suspension, smears were performed which were stained with dye according to Wright. Under immersion and 1000× in a light-field microscope, the sperm were classified as NBT-positive as formazan granules were detected therein, and the result of the trial was expressed as the percentage of these sperm.TUNEL trialFor this technique, we used the In Situ Cell Death Detection Kit-Fluorescein (Roche Diagnostics GmbH, Mannheim, Germany) following the manufacturer's specifications. Briefly, aliquots of washed semen were transferred onto multicell slides and fixed in 4% paraformaldehyde in PBS for 30min. After washing in PBS, the preparations were permeabilized in a solution of cold Triton X-100 at 0.1% in sodium citrate at 0.1%. Labeling was subsequently performed in a moist chamber at 37°C with the TUNEL reaction solution. Each experiment included a negative control (omitting the enzyme in the reaction solution) and a positive control (treated with 1mg/ml DNase). As counterstaininig, we used Hoechst 33342 (Molecular Probes, Invitrogen Detection Technologies). The preparations were analyzed under fluorescence microscope (Olympus BX40) and the DNA of the sperm was classified as intact or fragmented (nuclear green fluorescence). The result of the test was expressed as the percentage of sperm with fragmented DNA. The sperm of the negative controls showed no fluorescence, while 100% of the positive controls had their DNA fragmented.Statistical analysisIn order to determine significant differences between groups, we used the one-way ANOVA test. In the cases that showed correlations, we used the Spearman Rank Order Correlation test. Statistical significance was assumed with p values lower than 0.05. The statistic was carried out on the SigmaPlot 11.0 platform (Systat Software Inc.).ResultsFor the full set of patient samples, the average for volume, sperm concentration, progressive motility, and percentage of normal forms was within the reference ranges, while the percentage of immature forms and the round cell concentration was slightly altered. The results of the additional tests were within the reference ranges as well (<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>). Interestingly, only in 3.39% of the samples, all the variables tested were classified as normal. This means that 96.61% of the samples showed at least one of the analyzed parameters altered.<elsevierMultimedia ident="tbl0005"></elsevierMultimedia>The division of the total set of samples according to the normality of the standard semen parameters resulted in the composition of group A with 53.64% of the samples and group B with 46.36%. Along with this, and taking into account the previous data, it was obtained that for group A, 93.68% of the samples showed altered at least one of the complementary tests.Considering only the altered parameters not included in the recommendations of the last WHO manual,<a class="elsevierStyleCrossRef" href="#bib0035">7</a> or the complementary tests, and analyzing them one by one for each group, we obtained the results presented in <a class="elsevierStyleCrossRef" href="#tbl0010">Table 2</a>. Surprisingly, in group A we found a significant percentage of samples classified as abnormal for some of these parameters. As it might be suspected, these percentages were higher in group B. In the control group, we obtained values included in the reference ranges for all variables.<elsevierMultimedia ident="tbl0010"></elsevierMultimedia>Statistically significant differences were found between groups A and B in all complementary tests and in some standard analysis parameters (volume, concentration, progressive motility, and normal forms). None of the variables showed significant difference when we compared group A to the control group, while the latter differed with statistical significance from group B in the concentration, progressive motility, normal forms, and HOST, AB, NBT and TUNEL tests (<a class="elsevierStyleCrossRef" href="#tbl0015">Table 3</a>).<elsevierMultimedia ident="tbl0015"></elsevierMultimedia>From the correlation analysis between the additional tests, we obtained direct and significant correlations between NBT and AB (r=0.271; p<0.01); NBT and TUNEL (r=0.279; p<0.005); and AB and TUNEL (r=0.258; p<0.01).DiscussionOur results show that by analyzing only the parameters recommended by the WHO in the latest edition of its manual,<a class="elsevierStyleCrossRef" href="#bib0035">7</a> we get a partial view of the actual situation of the studied sample. Even finding these parameters within the reference ranges, and therefore classifying the samples as normal, when performing the other tests, not all these samples were really normal.Several considerations arise from the above observation. First, careful handling of the term ‘normality’ is required, especially taking into account the effect it may have on this particular type of patients consulting for infertility, given the emotional, social, etc. burden to which they are usually subjected. Then, an important role of the manuals and reference ranges is to provide a prognosis to help determine if medical interventions are necessary, if they are, to establish which is the best suited to the patient's conditions and what chances of success exist, or if other actions such as the use of donors or adoption are to be considered.<a class="elsevierStyleCrossRef" href="#bib0065">13</a> Clearly, this becomes difficult if we take into account only the parameters of the standard semen analysis with their reference ranges recommended in the fifth edition of the WHO manual.<a class="elsevierStyleCrossRef" href="#bib0035">7</a>Furthermore, the standard seminal study provides no information on the pathogenesis, that is, the mechanism underlying the observed alteration in a certain parameter.<a class="elsevierStyleCrossRef" href="#bib0065">13</a> Introducing functional tests and the additional tests evaluated in this paper in the seminal routine evaluation means providing valuable information for the diagnosis, prognosis, and etiology of infertility.The fact that a sample may have alterations in the functional tests or in the other sperm parameters evaluated in this paper is documented, even taking standard parameters within the reference ranges.<a class="elsevierStyleCrossRefs" href="#bib0100">20,21</a> In 2002, Saleh et al. described significant levels of sperm DNA damage in normal patients for the standard sperm parameters.<a class="elsevierStyleCrossRef" href="#bib0070">14</a> In line with this, an interesting finding is that published in the papers by Avedaño et al. The authors demonstrated that infertile patients may present fragmentation in the DNA of sperm classified as morphologically normal, and that this condition has a negative impact on the quality of the embryos generated by ICSI.<a class="elsevierStyleCrossRefs" href="#bib0110">22,23</a>Our results are consistent with the previous observations, and they show that the vast majority of the patients whose sperm samples would be classified as normal, according to the WHO criteria for standard semen parameters,<a class="elsevierStyleCrossRef" href="#bib0035">7</a> has at least one of the other tests performed in this paper altered. It is striking that the feature that is more frequently affected in this group of patients is the sperm DNA integrity. If to this is added that its correlation with the sperm fertilizing capacity has been documented for a long time both in vivo and in vitro,<a class="elsevierStyleCrossRefs" href="#bib0120">24,25</a> it is clear that its evaluation should be included in the diagnostic algorithms of male infertility almost routinely. The exception is given by those cases in which from the study of standard semen parameters, a clear diagnosis can be deduced, such as azoospermia.It is now accepted that there are three fundamental mechanisms that give rise to sperm DNA damage: (a) oxidative stress generated by an imbalance between the production of reactive oxygen species by sperm and leukocytes and the seminal plasma antioxidant activity; (b) alterations in the chromatin packaging provided by altered activity of the topoisomerase II enzyme during the replacement of histones with protamines in the sperm maturation process; and (c) ‘abortive apoptosis’, that is, apoptotic events that normally occur during the spermatogenesis go haywire for reasons still unknown, throwing in the ejaculate sperm that have entered the apoptotic pathway.<a class="elsevierStyleCrossRef" href="#bib0130">26</a> While it would be wrong to think that these mechanisms act independently, when what is most likely to take place is a combination of the three of them,<a class="elsevierStyleCrossRef" href="#bib0135">27</a> it is possible to investigate them, especially in cases of patients with high levels of sperm DNA damage.In this paper, we examine two of the mechanisms: The sperm production of reactive oxygen species by the NBT test, and erroneous replacement of histones with protamines through the AB test. Through these trials, we obtain indirect vision of the contribution in the sperm DNA damage of the action of reactive oxygen species and of the errors in the chromatin packaging during sperm maturation. We consider this approach effective, technically simple and inexpensive that can be included in the routine of any laboratory. Moreover, the correlations found between these tests and the percentage of sperm with damaged DNA reinforces the idea that the mechanisms studied indirectly by these two trials are detrimental to the sperm DNA integrity. On the other hand, and because of the correlation we find between both tests, we adhere to the idea that those mechanisms would act in parallel and not individually.Further studies are needed, increasing the number of cases and adding the apoptotic pathway analysis as a third source mechanism of sperm DNA damage, to continue to contribute to the understanding of the etiology of male infertility, and concomitantly to the development of efficient tools for its correct diagnosis.Going beyond the parameters recommended by the WHO,<a class="elsevierStyleCrossRef" href="#bib0035">7</a> deepening in the seminal study by introducing trials evaluating the sperm functionality, as well as its chromatin integrity, results in a more refined diagnosis and a better strategy.Conflict of interestThe authors declare that they have no conflict of interest." "textoCompletoSecciones" => array:1 [ "secciones" => array:10 [ 0 => array:2 [ "identificador" => "xres191966" "titulo" => array:5 [ 0 => "Abstract" 1 => "Objectives" 2 => "Materials and methods" 3 => "Results" 4 => "Conclusions" ] ] 1 => array:2 [ "identificador" => "xpalclavsec179191" "titulo" => "Keywords" ] 2 => array:2 [ "identificador" => "xres191967" "titulo" => array:5 [ 0 => "Resumen" 1 => "Objetivos" 2 => "Materiales y métodos" 3 => "Resultados" 4 => "Conclusiones" ] ] 3 => array:2 [ "identificador" => "xpalclavsec179192" "titulo" => "Palabras clave" ] 4 => array:2 [ "identificador" => "sec0005" "titulo" => "Introduction" ] 5 => array:3 [ "identificador" => "sec0010" "titulo" => "Materials and methods" "secciones" => array:5 [ 0 => array:2 [ "identificador" => "sec0015" "titulo" => "Patients and samples" ] 1 => array:2 [ "identificador" => "sec0020" "titulo" => "Analysis of standard semen" ] 2 => array:3 [ "identificador" => "sec0025" "titulo" => "Functional tests" "secciones" => array:3 [ 0 => array:2 [ "identificador" => "sec0030" "titulo" => "24-h survival" ] 1 => array:2 [ "identificador" => "sec0035" "titulo" => "Hypo-osmotic test" ] 2 => array:2 [ "identificador" => "sec0040" "titulo" => "Modified stress test" ] ] ] 3 => array:3 [ "identificador" => "sec0045" "titulo" => "Additional tests" "secciones" => array:3 [ 0 => array:2 [ "identificador" => "sec0050" "titulo" => "Aniline blue test" ] 1 => array:2 [ "identificador" => "sec0055" "titulo" => "Nitroblue tetrazolium test" ] 2 => array:2 [ "identificador" => "sec0060" "titulo" => "TUNEL trial" ] ] ] 4 => array:2 [ "identificador" => "sec0065" "titulo" => "Statistical analysis" ] ] ] 6 => array:2 [ "identificador" => "sec0070" "titulo" => "Results" ] 7 => array:2 [ "identificador" => "sec0075" "titulo" => "Discussion" ] 8 => array:2 [ "identificador" => "sec0080" "titulo" => "Conflict of interest" ] 9 => array:1 [ "titulo" => "References" ] ] ] "pdfFichero" => "main.pdf" "tienePdf" => true "fechaRecibido" => "2011-11-14" "fechaAceptado" => "2011-11-16" "PalabrasClave" => array:2 [ "en" => array:1 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Keywords" "identificador" => "xpalclavsec179191" "palabras" => array:4 [ 0 => "Sperm DNA fragmentation" 1 => "Terminal deoxynucleotidyl transferase dUTP nick end labeling" 2 => "Nitroblue tetrazolium" 3 => "Sperm function tests" ] ] ] "es" => array:1 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Palabras clave" "identificador" => "xpalclavsec179192" "palabras" => array:4 [ 0 => "Fragmentación del ADN espermático" 1 => "Terminal deoxynucleotidyl transferase dUTP nick end labeling" 2 => "Nitroblue Tetrazolium" 3 => "Pruebas funcionales espermáticos" ] ] ] ] "tieneResumen" => true "resumen" => array:2 [ "en" => array:2 [ "titulo" => "Abstract" "resumen" => "ObjectivesTo evaluate the incidence of functional and chromatin alterations on spermatozoids in patients grouped according to normality of standard semen parameters based on the 5th edition of the World Health Organization (WHO) guidelines. To identify and correlate the most frequently altered characteristics in the normal standard semen parameters sub-population. Materials and methodsA prospective study was performed. It evaluated standard semen parameters (volume, sperm concentration, motility and morphology, round cells, peroxidase-positive cells) according to WHO guidelines, as well as functional tests (24h survival, hypoosmotic swelling test, modified stress test), and additional assays (aniline blue, nitroblue-tetrazolium, TUNEL) in 110 semen samples from patients and 6 from fertile donors (control). Based on standard semen parameters values, patients were divided into two groups (A: all standard parameters normal; B: one altered standard parameter at least). ResultsAt least one of the variables analyzed was altered in 96.61% of the samples. Groups A and B showed statistically significant differences in all the complementary tests. At least one of the complementary tests were altered in 93.68% of the samples in group A, and the most frequently affected variable was sperm DNA fragmentation (16.95%). ConclusionsPerforming a more in-depth seminal study within the routine functional and chromatin assays provides a more precise diagnosis of male infertility. The WHO standards should be considered as a primary approach." ] "es" => array:2 [ "titulo" => "Resumen" "resumen" => "ObjetivosEstudiar la incidencia de alteraciones funcionales y cromatínicas en espermatozoides de pacientes, agrupados según normalidad de los parámetros seminales estándar en función de la quinta edición del manual de la Organización Mundial de la Salud (OMS). Identificar y correlacionar las características alteradas con mayor frecuencia en la subpoblación normal para los parámetros estándar. Materiales y métodosEstudio prospectivo en el que se evaluaron en muestras de semen de 110 pacientes y 6 donantes fértiles (control), los parámetros seminales estándar (volumen, concentración, motilidad, morfología, células redondas, células peroxidasa-positivas) según las directrices de la OMS, así como test complementarios funcionales (sobrevida 24 horas, test hipo-osmótico, test de estrés modificado) y adicionales (azul de anilina, nitroblue-tetrazolium, TUNEL). Según el resultado de los análisis estándar los pacientes se dividieron en dos grupos (A: todos los parámetros estándar normales; B: al menos uno de los parámetros estándar alterado). ResultadosEl 96,61% de las muestras presentó alterada al menos una de las variables analizadas. Los grupos A y B mostraron diferencia estadísticamente significativa en todos los test complementarios. En el grupo A el 93,68% de las muestras presentó al menos un test complementario alterado, y la variable afectada con mayor frecuencia fue la fragmentación del ADN espermático (16,95%). ConclusionesLa profundización en el estudio seminal, introduciendo en la rutina ensayos funcionales y cromatínicos, redunda en un diagnóstico más afinado de la infertilidad del varón. Los parámetros de la OMS deben considerarse como un abordaje primario." ] ] "NotaPie" => array:1 [ 0 => array:2 [ "etiqueta" => "☆" "nota" => "Please cite this article as: Curti G, et al. Diagnóstico de infertilidad masculina: necesidad de valoraciones funcionales y cromatínicas. Actas Urol Esp. 2013;37:100–5." ] ] "multimedia" => array:3 [ 0 => array:7 [ "identificador" => "tbl0005" "etiqueta" => "Table 1" "tipo" => "MULTIMEDIATABLA" "mostrarFloat" => true "mostrarDisplay" => false "tabla" => array:1 [ "tablatextoimagen" => array:1 [ 0 => array:2 [ "tabla" => array:1 [ 0 => """ <table border="0" frame="\n \t\t\t\t\tvoid\n \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t" style="border-bottom: 2px solid black"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t" style="border-bottom: 2px solid black">Value \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t" style="border-bottom: 2px solid black">Reference \t\t\t\t\t\t\n \t\t\t\t</td></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " colspan="3" align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Standard parameters</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Volume (ml) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">3.57±0.16 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">>1.5 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Concentration (mill/ml) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">48.14±3.48 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">>15 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Progressive motility (%) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">55.84±1.84 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">>32 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Normal forms (%) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">7.69±0.45 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">>4 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Immature forms (%) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">3.03±0.93 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t"><2 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Round cells (mill/ml) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">1.00±0.24 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t"><1 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Peroxidase+(mill/ml) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.48±0.20 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t"> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " colspan="3" align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t"></td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " colspan="3" align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Functional tests</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">24h survival (%) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">45.48±2.52 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">>20 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">HOST (%) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">63.06±1.73 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">>60 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">MOST \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.84±0.01 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">>0.50 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " colspan="3" align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t"></td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " colspan="3" align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Additional tests</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Aniline blue (%) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">20.38±0.88 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t"><25 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">NBT (%) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">12.08±0.58 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t"><23 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">TUNEL (%) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">15.39±0.85 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t"><20 \t\t\t\t\t\t\n \t\t\t\t</td></tr></tbody></table> """ ] "imagenFichero" => array:1 [ 0 => "xTab328652.png" ] ] ] ] "descripcion" => array:1 [ "en" => "Seminal parameters evaluated with their corresponding reference ranges. The values are expressed as median±standard error (n=110)." ] ] 1 => array:7 [ "identificador" => "tbl0010" "etiqueta" => "Table 2" "tipo" => "MULTIMEDIATABLA" "mostrarFloat" => true "mostrarDisplay" => false "tabla" => array:1 [ "tablatextoimagen" => array:1 [ 0 => array:2 [ "tabla" => array:1 [ 0 => """ <table border="0" frame="\n \t\t\t\t\tvoid\n \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t" style="border-bottom: 2px solid black"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t" style="border-bottom: 2px solid black">Group A \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t" style="border-bottom: 2px solid black">Group B \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t" style="border-bottom: 2px solid black">Control group \t\t\t\t\t\t\n \t\t\t\t</td></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Immature forms≥2% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">11.86% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">27.45% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0% \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Round cells≥1mill/ml \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">13.25% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">29.41% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0% \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">24h survival≤20% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">13.56% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">41.18% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0% \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">HOST≤60% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">8.47% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">43.14% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0% \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">MOST≤0.50 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">3.92% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0% \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">TUNEL≥20% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">16.95% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">47.06% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0% \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Aniline blue≥25% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">15.25% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">59.90% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0% \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">NBT≥23% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">5.08% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">19.61% \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0% \t\t\t\t\t\t\n \t\t\t\t</td></tr></tbody></table> """ ] "imagenFichero" => array:1 [ 0 => "xTab328653.png" ] ] ] ] "descripcion" => array:1 [ "en" => "Parameters altered depending on the group of patients analyzed. The table shows the parameters not considered by the 2010 WHO manual.<a class="elsevierStyleCrossRef" href="#bib0035">7</a> The values correspond to the percentage of samples of the total in each group." ] ] 2 => array:7 [ "identificador" => "tbl0015" "etiqueta" => "Table 3" "tipo" => "MULTIMEDIATABLA" "mostrarFloat" => true "mostrarDisplay" => false "tabla" => array:2 [ "leyenda" => "The p values in bold show statistical significance." "tablatextoimagen" => array:1 [ 0 => array:2 [ "tabla" => array:1 [ 0 => """ <table border="0" frame="\n \t\t\t\t\tvoid\n \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="" valign="\n \t\t\t\t\ttop\n \t\t\t\t" style="border-bottom: 2px solid black"> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t" style="border-bottom: 2px solid black">Group A \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t" style="border-bottom: 2px solid black">Group B \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t" style="border-bottom: 2px solid black">Control group \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t" style="border-bottom: 2px solid black">A vs. B \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t" style="border-bottom: 2px solid black">A vs. control \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t" style="border-bottom: 2px solid black">B vs. control \t\t\t\t\t\t\n \t\t\t\t</td></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " colspan="7" align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Standard parameters</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Volume (ml) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">3.94±0.22 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">3.14±0.21 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">3.73±1.05 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.012 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.769 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.403 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Concentration (×106ml−1) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">64.25±4.37 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">29.51±4.27 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">73.01±13.03 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p<0.001 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.549 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.004 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Proggresive motility (%) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">64.93±1.03 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">45.33±3.21 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">63.33±3.77 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p<0.001 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.835 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.021 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Normal forms (%) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">10.71±0.50 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">4.19±0.40 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">11.33±1.38 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p<0.001 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.724 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p<0.001 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Immature forms (%) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">2.06±1.13 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">4.16±1.51 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.23±0.11 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.254 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.658 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.344 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Round cells (×106ml−1) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.69±0.19 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">1.37±0.46 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.42±0.22 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.150 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.794 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.369 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Peroxidase+(×106ml−1) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.17±0.03 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.83±0.43 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.24±0.19 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.096 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.944 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.594 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " colspan="7" align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t"></td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " colspan="7" align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Functional tests</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">24h survival (%) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">54.39±3.16 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">33.52±3.38 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">53.67±11.00 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p<0.001 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.945 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.061 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">HOST (%) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">67.56±0.67 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">57.65±2.24 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">73.17±4.91 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p<0.001 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.265 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.002 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">MOST \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.87±0.01 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.80±0.02 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">0.87±0.04 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.001 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.994 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.137 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " colspan="7" align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t"></td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " colspan="7" align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Additional tests</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Aniline blue (%) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">17.14±1.00 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">24.20±1.32 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">15.50±3.45 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p<0.001 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.666 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.023 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">NBT (%) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">10.86±0.68 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">13.57±0.96 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">8.33±1.20 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.019 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.315 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.040 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">TUNEL (%) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">12.71±1.00 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">18.74±1.45 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="char" valign="\n \t\t\t\t\ttop\n \t\t\t\t">10.00±1.88 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p<0.001 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.455 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">p=0.018 \t\t\t\t\t\t\n \t\t\t\t</td></tr></tbody></table> """ ] "imagenFichero" => array:1 [ 0 => "xTab328654.png" ] ] ] ] "descripcion" => array:1 [ "en" => "Seminal parameters studied in groups A, B, and control, and comparisons between groups. The values of the variables are expressed as median±standard error. The comparisons were performed using the one-way ANOVA test; the p values are shown." ] ] ] "bibliografia" => array:2 [ "titulo" => "References" "seccion" => array:1 [ 0 => array:2 [ "identificador" => "bibs0005" "bibliografiaReferencia" => array:27 [ 0 => array:3 [ "identificador" => "bib0005" "etiqueta" => "1" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "How we are born" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:1 [ 0 => "S.K. Dey" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1172/JCI42380" "Revista" => array:6 [ "tituloSerie" => "J Clin Invest" "fecha" => "2010" "volumen" => "120" "paginaInicial" => "952" "paginaFinal" => "955" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/20364092" "web" => "Medline" ] ] ] ] ] ] ] ] 1 => array:3 [ "identificador" => "bib0010" "etiqueta" => "2" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Male infertility: role of genetic background" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:6 [ 0 => "A. Ferlin" 1 => "F. Raicu" 2 => "V. Gatta" 3 => "D. Zuccarello" 4 => "G. Palka" 5 => "C. Foresta" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "Reprod BioMed Online" "fecha" => "2007" "volumen" => "14" "paginaInicial" => "734" "paginaFinal" => "745" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/17579990" "web" => "Medline" ] ] ] ] ] ] ] ] 2 => array:3 [ "identificador" => "bib0015" "etiqueta" => "3" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "WHO laboratory manual for the examination of human semen and sperm–cervical mucus interaction" "autores" => array:1 [ 0 => array:2 [ "colaboracion" => "World Health Organization" "etal" => false ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Libro" => array:4 [ "edicion" => "1st ed." 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Diagnosis of male infertility: A need of functional and chromatin evaluation

Diagnóstico de infertilidad masculina: necesidad de valoraciones funcionales y cromatínicas

G. Curti

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giannicurti@gmail.com

Corresponding author.

, M. Cánepa, L. Cantú, J.M. Montes

Laboratorio Fertilab, Montevideo, Uruguay

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the World Health Organization &#40;WHO&#41; published in 1980 the first edition of its &#8216;Manual for the examination of human semen and sperm&#8211;cervical mucus interaction&#8217;&#46;<a class="elsevierStyleCrossRef" href="#bib0015"><span class="elsevierStyleSup">3</span></a> Following the progress in this area&#44; the WHO reissued revised and updated versions of the manual in 1987&#44;<a class="elsevierStyleCrossRef" href="#bib0020"><span class="elsevierStyleSup">4</span></a> 1992&#44;<a class="elsevierStyleCrossRef" href="#bib0025"><span class="elsevierStyleSup">5</span></a> 1999<a class="elsevierStyleCrossRef" href="#bib0030"><span class="elsevierStyleSup">6</span></a> and 2010&#46;<a class="elsevierStyleCrossRef" href="#bib0035"><span class="elsevierStyleSup">7</span></a> These publications generated controversy on the true value of the recommended characteristics and their reference values&#46;<a class="elsevierStyleCrossRefs" href="#bib0040"><span class="elsevierStyleSup">8&#8211;13</span></a></p><p id="par0020" class="elsevierStylePara elsevierViewall">The sperm must exhibit a set of unique biological properties to achieve fertilization and normal embryonic development&#46; Given this complexity&#44; it is difficult to diagnose changes in its physiology using the standard semen analysis&#44; this is&#44; the study of the variables recommended in the WHO manuals&#46;<a class="elsevierStyleCrossRef" href="#bib0070"><span class="elsevierStyleSup">14</span></a></p><p id="par0025" class="elsevierStylePara elsevierViewall">Other tests&#44; additional to those recommended in the manuals&#44; have been developed in search of a more comprehensive evaluation&#46; Studying both functional and chromatin features of the sperm&#44; the results have been positively correlated with sperm quality and reproductive success&#44; constituting potential diagnostic and prognostic tools<a class="elsevierStyleCrossRefs" href="#bib0075"><span class="elsevierStyleSup">15&#8211;19</span></a>&#46;</p><p id="par0030" class="elsevierStylePara elsevierViewall">The aim of this work was to study the incidence of functional and chromatin alterations in the sperm of a population of patients with marital infertility subdivided according to the normality of the standard semen parameters&#44; 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After standard sexual abstinence&#44; and following the WHO guidelines&#44;<a class="elsevierStyleCrossRef" href="#bib0035"><span class="elsevierStyleSup">7</span></a> the semen samples were collected by masturbation&#44; in sterile containers tested and supplied by the laboratory personnel&#46; All the subjects included in this study signed informed consent forms&#46;</p><p id="par0040" class="elsevierStylePara elsevierViewall">The standard semen parameters analyzed were&#58; sample volume&#44; sperm concentration&#44; progressive sperm motility&#44; percentage of normal forms&#44; percentage of immature forms&#44; round cell concentration and concentration of peroxidase-positive cells&#46; We also carried out the following functional tests&#58; sperm survival at 24<span class="elsevierStyleHsp" style=""></span>hours&#44; hypo-osmotic swelling test &#40;HOST&#41;&#44; modified stress test &#40;MOST&#41;&#44; and the following additional tests&#58; nitroblue tetrazolium test &#40;NBT&#41;&#44; aniline blue test &#40;AB&#41;&#44; and terminal deoxynucleotidyl transferase dUTP nick end labeling &#40;TUNEL&#41; assay&#46;</p><p id="par0045" class="elsevierStylePara elsevierViewall">Standard parameters were studied according to the latest WHO manual<a class="elsevierStyleCrossRef" href="#bib0035"><span class="elsevierStyleSup">7</span></a> and they were classified as normal or abnormal taking as a reference the ranges published in the manual&#46; According to the above&#44; the total set of patients was divided into two groups&#58; group A&#44; which included the patients whose samples were classified as normal for all standard parameters&#59; and group B&#44; with the patients whose samples showed alterations in at least one of these parameters&#46;</p></span><span id="sec0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0080">Analysis of standard semen</span><p id="par0050" class="elsevierStylePara elsevierViewall">After the liquefaction and determination of the volume of the samples&#44; 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of an aliquot of washed semen &#40;half Earle&#39;s Balanced Salt Solution &#91;EBSS&#93;<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>SAH 10&#37;&#41;&#46;</p></span><span id="sec0035" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0095">Hypo-osmotic test</span><p id="par0060" class="elsevierStylePara elsevierViewall">This test was carried out by incubating 100<span class="elsevierStyleHsp" style=""></span>&#956;l of semen for 30<span class="elsevierStyleHsp" style=""></span>min at 37<span class="elsevierStyleHsp" style=""></span>&#176;C&#44; in 1<span class="elsevierStyleHsp" style=""></span>ml of hypo-osmotic solution &#40;<span class="elsevierStyleSmallCaps">d</span>-fructose 0&#46;075<span class="elsevierStyleHsp" style=""></span>M&#44; sodium citrate 0&#46;025<span class="elsevierStyleHsp" style=""></span>M&#41;&#46; These conditions cause a passive flow of water through intact and functional membranes into cells&#44; which in turn causes them to swell&#46; This swelling is evidenced by the winding of the sperm tails&#46; Dead sperm or with altered membranes do not swell&#44; and their tails are straight&#46; The result of the test is expressed as the percentage of swollen sperm&#46;</p></span><span id="sec0040" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0100">Modified stress test</span><p id="par0065" class="elsevierStylePara elsevierViewall">The rate described as a result of this test was calculated as the ratio between the percentages of motile sperm counted before and after 4<span class="elsevierStyleHsp" style=""></span>h of incubation&#44; at 40<span class="elsevierStyleHsp" style=""></span>&#176;C of an aliquot of washed sperm with EBSS<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>SAH 10&#37;&#46;</p></span></span><span id="sec0045" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0105">Additional tests</span><span id="sec0050" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0110">Aniline blue test</span><p id="par0070" class="elsevierStylePara elsevierViewall">For this study&#44; 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Under immersion and 1000&#215; in a light-field microscope&#44; the sperm were classified as NBT-positive as formazan granules were detected therein&#44; and the result of the trial was expressed as the percentage of these sperm&#46;</p></span><span id="sec0060" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0120">TUNEL trial</span><p id="par0080" class="elsevierStylePara elsevierViewall">For this technique&#44; we used the In Situ Cell Death Detection Kit-Fluorescein &#40;Roche Diagnostics GmbH&#44; Mannheim&#44; Germany&#41; following the manufacturer&#39;s specifications&#46; Briefly&#44; aliquots of washed semen were transferred onto multicell slides and fixed in 4&#37; paraformaldehyde in PBS for 30<span class="elsevierStyleHsp" style=""></span>min&#46; After washing in PBS&#44; the preparations were permeabilized in a solution of cold Triton X-100 at 0&#46;1&#37; in sodium citrate at 0&#46;1&#37;&#46; Labeling was subsequently performed in a moist chamber at 37<span class="elsevierStyleHsp" style=""></span>&#176;C with the TUNEL reaction solution&#46; Each experiment included a negative control &#40;omitting the enzyme in the reaction solution&#41; and a positive control &#40;treated with 1<span class="elsevierStyleHsp" style=""></span>mg&#47;ml DNase&#41;&#46; As counterstaininig&#44; we used Hoechst 33342 &#40;Molecular Probes&#44; Invitrogen Detection Technologies&#41;&#46; The preparations were analyzed under fluorescence microscope &#40;Olympus BX40&#41; and the DNA of the sperm was classified as intact or fragmented &#40;nuclear green fluorescence&#41;&#46; The result of the test was expressed as the percentage of sperm with fragmented DNA&#46; The sperm of the negative controls showed no fluorescence&#44; while 100&#37; of the positive controls had their DNA fragmented&#46;</p></span></span><span id="sec0065" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0125">Statistical analysis</span><p id="par0085" class="elsevierStylePara elsevierViewall">In order to determine significant differences between groups&#44; we used the one-way ANOVA test&#46; In the cases that showed correlations&#44; we used the Spearman Rank Order Correlation test&#46; Statistical significance was assumed with p values lower than 0&#46;05&#46; The statistic was carried out on the SigmaPlot 11&#46;0 platform &#40;Systat Software Inc&#46;&#41;&#46;</p></span></span><span id="sec0070" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0130">Results</span><p id="par0090" class="elsevierStylePara elsevierViewall">For the full set of patient samples&#44; the average for volume&#44; sperm concentration&#44; progressive motility&#44; and percentage of normal forms was within the reference ranges&#44; while the percentage of immature forms and the round cell concentration was slightly altered&#46; The results of the additional tests were within the reference ranges as well &#40;<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#41;&#46; Interestingly&#44; only in 3&#46;39&#37; of the samples&#44; all the variables tested were classified as normal&#46; This means that 96&#46;61&#37; of the samples showed at least one of the analyzed parameters altered&#46;</p><elsevierMultimedia ident="tbl0005"></elsevierMultimedia><p id="par0095" class="elsevierStylePara elsevierViewall">The division of the total set of samples according to the normality of the standard semen parameters resulted in the composition of group A with 53&#46;64&#37; of the samples and group B with 46&#46;36&#37;&#46; Along with this&#44; and taking into account the previous data&#44; it was obtained that for group A&#44; 93&#46;68&#37; of the samples showed altered at least one of the complementary tests&#46;</p><p id="par0100" class="elsevierStylePara elsevierViewall">Considering only the altered parameters not included in the recommendations of the last WHO manual&#44;<a class="elsevierStyleCrossRef" href="#bib0035"><span class="elsevierStyleSup">7</span></a> or the complementary tests&#44; and analyzing them one by one for each group&#44; we obtained the results presented in <a class="elsevierStyleCrossRef" href="#tbl0010">Table 2</a>&#46; Surprisingly&#44; in group A we found a significant percentage of samples classified as abnormal for some of these parameters&#46; As it might be suspected&#44; these percentages were higher in group B&#46; In the control group&#44; we obtained values included in the reference ranges for all variables&#46;</p><elsevierMultimedia ident="tbl0010"></elsevierMultimedia><p id="par0105" class="elsevierStylePara elsevierViewall">Statistically significant differences were found between groups A and B in all complementary tests and in some standard analysis parameters &#40;volume&#44; concentration&#44; progressive motility&#44; and normal forms&#41;&#46; None of the variables showed significant difference when we compared group A to the control group&#44; while the latter differed with statistical significance from group B in the concentration&#44; progressive motility&#44; normal forms&#44; and HOST&#44; AB&#44; NBT and TUNEL tests &#40;<a class="elsevierStyleCrossRef" href="#tbl0015">Table 3</a>&#41;&#46;</p><elsevierMultimedia ident="tbl0015"></elsevierMultimedia><p id="par0110" class="elsevierStylePara elsevierViewall">From the correlation analysis between the additional tests&#44; we obtained direct and significant correlations between NBT and AB &#40;<span class="elsevierStyleItalic">r</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;271&#59; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;01&#41;&#59; NBT and TUNEL &#40;<span class="elsevierStyleItalic">r</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;279&#59; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;005&#41;&#59; and AB and TUNEL &#40;<span class="elsevierStyleItalic">r</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;258&#59; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;01&#41;&#46;</p></span><span id="sec0075" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0135">Discussion</span><p id="par0115" class="elsevierStylePara elsevierViewall">Our results show that by analyzing only the parameters recommended by the WHO in the latest edition of its manual&#44;<a class="elsevierStyleCrossRef" href="#bib0035"><span class="elsevierStyleSup">7</span></a> we get a partial view of the actual situation of the studied sample&#46; Even finding these parameters within the reference ranges&#44; and therefore classifying the samples as normal&#44; when performing the other tests&#44; not all these samples were really normal&#46;</p><p id="par0120" class="elsevierStylePara elsevierViewall">Several considerations arise from the above observation&#46; First&#44; careful handling of the term &#8216;normality&#8217; is required&#44; especially taking into account the effect it may have on this particular type of patients consulting for infertility&#44; given the emotional&#44; social&#44; etc&#46; burden to which they are usually subjected&#46; Then&#44; an important role of the manuals and reference ranges is to provide a prognosis to help determine if medical interventions are necessary&#44; if they are&#44; to establish which is the best suited to the patient&#39;s conditions and what chances of success exist&#44; or if other actions such as the use of donors or adoption are to be considered&#46;<a class="elsevierStyleCrossRef" href="#bib0065"><span class="elsevierStyleSup">13</span></a> Clearly&#44; this becomes difficult if we take into account only the parameters of the standard semen analysis with their reference ranges recommended in the fifth edition of the WHO manual&#46;<a class="elsevierStyleCrossRef" href="#bib0035"><span class="elsevierStyleSup">7</span></a></p><p id="par0125" class="elsevierStylePara elsevierViewall">Furthermore&#44; the standard seminal study provides no information on the pathogenesis&#44; that is&#44; the mechanism underlying the observed alteration in a certain parameter&#46;<a class="elsevierStyleCrossRef" href="#bib0065"><span class="elsevierStyleSup">13</span></a> Introducing functional tests and the additional tests evaluated in this paper in the seminal routine evaluation means providing valuable information for the diagnosis&#44; prognosis&#44; and etiology of infertility&#46;</p><p id="par0130" class="elsevierStylePara elsevierViewall">The fact that a sample may have alterations in the functional tests or in the other sperm parameters evaluated in this paper is documented&#44; even taking standard parameters within the reference ranges&#46;<a class="elsevierStyleCrossRefs" href="#bib0100"><span class="elsevierStyleSup">20&#44;21</span></a> In 2002&#44; Saleh et al&#46; described significant levels of sperm DNA damage in normal patients for the standard sperm parameters&#46;<a class="elsevierStyleCrossRef" href="#bib0070"><span class="elsevierStyleSup">14</span></a> In line with this&#44; an interesting finding is that published in the papers by Aveda&#241;o et al&#46; The authors demonstrated that infertile patients may present fragmentation in the DNA of sperm classified as morphologically normal&#44; and that this condition has a negative impact on the quality of the embryos generated by ICSI&#46;<a class="elsevierStyleCrossRefs" href="#bib0110"><span class="elsevierStyleSup">22&#44;23</span></a></p><p id="par0135" class="elsevierStylePara elsevierViewall">Our results are consistent with the previous observations&#44; and they show that the vast majority of the patients whose sperm samples would be classified as normal&#44; according to the WHO criteria for standard semen parameters&#44;<a class="elsevierStyleCrossRef" href="#bib0035"><span class="elsevierStyleSup">7</span></a> has at least one of the other tests performed in this paper altered&#46; It is striking that the feature that is more frequently affected in this group of patients is the sperm DNA integrity&#46; If to this is added that its correlation with the sperm fertilizing capacity has been documented for a long time both in vivo and in vitro&#44;<a class="elsevierStyleCrossRefs" href="#bib0120"><span class="elsevierStyleSup">24&#44;25</span></a> it is clear that its evaluation should be included in the diagnostic algorithms of male infertility almost routinely&#46; The exception is given by those cases in which from the study of standard semen parameters&#44; a clear diagnosis can be deduced&#44; such as azoospermia&#46;</p><p id="par0140" class="elsevierStylePara elsevierViewall">It is now accepted that there are three fundamental mechanisms that give rise to sperm DNA damage&#58; &#40;a&#41; oxidative stress generated by an imbalance between the production of reactive oxygen species by sperm and leukocytes and the seminal plasma antioxidant activity&#59; &#40;b&#41; alterations in the chromatin packaging provided by altered activity of the topoisomerase II enzyme during the replacement of histones with protamines in the sperm maturation process&#59; and &#40;c&#41; &#8216;abortive apoptosis&#8217;&#44; that is&#44; apoptotic events that normally occur during the spermatogenesis go haywire for reasons still unknown&#44; throwing in the ejaculate sperm that have entered the apoptotic pathway&#46;<a class="elsevierStyleCrossRef" href="#bib0130"><span class="elsevierStyleSup">26</span></a> While it would be wrong to think that these mechanisms act independently&#44; when what is most likely to take place is a combination of the three of them&#44;<a class="elsevierStyleCrossRef" href="#bib0135"><span class="elsevierStyleSup">27</span></a> it is possible to investigate them&#44; especially in cases of patients with high levels of sperm DNA damage&#46;</p><p id="par0145" class="elsevierStylePara elsevierViewall">In this paper&#44; we examine two of the mechanisms&#58; The sperm production of reactive oxygen species by the NBT test&#44; and erroneous replacement of histones with protamines through the AB test&#46; Through these trials&#44; we obtain indirect vision of the contribution in the sperm DNA damage of the action of reactive oxygen species and of the errors in the chromatin packaging during sperm maturation&#46; We consider this approach effective&#44; technically simple and inexpensive that can be included in the routine of any laboratory&#46; Moreover&#44; the correlations found between these tests and the percentage of sperm with damaged DNA reinforces the idea that the mechanisms studied indirectly by these two trials are detrimental to the sperm DNA integrity&#46; On the other hand&#44; and because of the correlation we find between both tests&#44; we adhere to the idea that those mechanisms would act in parallel and not individually&#46;</p><p id="par0150" class="elsevierStylePara elsevierViewall">Further studies are needed&#44; increasing the number of cases and adding the apoptotic pathway analysis as a third source mechanism of sperm DNA damage&#44; to continue to contribute to the understanding of the etiology of male infertility&#44; and concomitantly to the development of efficient tools for its correct diagnosis&#46;</p><p id="par0155" class="elsevierStylePara elsevierViewall">Going beyond the parameters recommended by the WHO&#44;<a class="elsevierStyleCrossRef" href="#bib0035"><span class="elsevierStyleSup">7</span></a> deepening in the seminal study by introducing trials evaluating the sperm functionality&#44; as well as its chromatin integrity&#44; results in a more refined diagnosis and a better strategy&#46;</p></span><span id="sec0080" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0140">Conflict of interest</span><p id="par0165" class="elsevierStylePara elsevierViewall">The authors declare that they have no conflict of interest&#46;</p></span></span>"
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          "identificador" => "xres191966"
          "titulo" => array:5 [
            0 => "Abstract"
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            2 => "Materials and methods"
            3 => "Results"
            4 => "Conclusions"
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          "titulo" => "Keywords"
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          "titulo" => array:5 [
            0 => "Resumen"
            1 => "Objetivos"
            2 => "Materiales y m&#233;todos"
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          "identificador" => "xpalclavsec179192"
          "titulo" => "Palabras clave"
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        4 => array:2 [
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          "titulo" => "Introduction"
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          "identificador" => "sec0010"
          "titulo" => "Materials and methods"
          "secciones" => array:5 [
            0 => array:2 [
              "identificador" => "sec0015"
              "titulo" => "Patients and samples"
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            1 => array:2 [
              "identificador" => "sec0020"
              "titulo" => "Analysis of standard semen"
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            2 => array:3 [
              "identificador" => "sec0025"
              "titulo" => "Functional tests"
              "secciones" => array:3 [
                0 => array:2 [
                  "identificador" => "sec0030"
                  "titulo" => "24-h survival"
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                1 => array:2 [
                  "identificador" => "sec0035"
                  "titulo" => "Hypo-osmotic test"
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                2 => array:2 [
                  "identificador" => "sec0040"
                  "titulo" => "Modified stress test"
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            3 => array:3 [
              "identificador" => "sec0045"
              "titulo" => "Additional tests"
              "secciones" => array:3 [
                0 => array:2 [
                  "identificador" => "sec0050"
                  "titulo" => "Aniline blue test"
                ]
                1 => array:2 [
                  "identificador" => "sec0055"
                  "titulo" => "Nitroblue tetrazolium test"
                ]
                2 => array:2 [
                  "identificador" => "sec0060"
                  "titulo" => "TUNEL trial"
                ]
              ]
            ]
            4 => array:2 [
              "identificador" => "sec0065"
              "titulo" => "Statistical analysis"
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        6 => array:2 [
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          "titulo" => "Results"
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          "titulo" => "Discussion"
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        8 => array:2 [
          "identificador" => "sec0080"
          "titulo" => "Conflict of interest"
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          "titulo" => "References"
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    "pdfFichero" => "main.pdf"
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    "fechaRecibido" => "2011-11-14"
    "fechaAceptado" => "2011-11-16"
    "PalabrasClave" => array:2 [
      "en" => array:1 [
        0 => array:4 [
          "clase" => "keyword"
          "titulo" => "Keywords"
          "identificador" => "xpalclavsec179191"
          "palabras" => array:4 [
            0 => "Sperm DNA fragmentation"
            1 => "Terminal deoxynucleotidyl transferase dUTP nick end labeling"
            2 => "Nitroblue tetrazolium"
            3 => "Sperm function tests"
          ]
        ]
      ]
      "es" => array:1 [
        0 => array:4 [
          "clase" => "keyword"
          "titulo" => "Palabras clave"
          "identificador" => "xpalclavsec179192"
          "palabras" => array:4 [
            0 => "Fragmentaci&#243;n del ADN esperm&#225;tico"
            1 => "Terminal deoxynucleotidyl transferase dUTP nick end labeling"
            2 => "Nitroblue Tetrazolium"
            3 => "Pruebas funcionales esperm&#225;ticos"
          ]
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      ]
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        "titulo" => "Abstract"
        "resumen" => "<span class="elsevierStyleSectionTitle" id="sect0010">Objectives</span><p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">To evaluate the incidence of functional and chromatin alterations on spermatozoids in patients grouped according to normality of standard semen parameters based on the 5th edition of the World Health Organization &#40;WHO&#41; guidelines&#46; To identify and correlate the most frequently altered characteristics in the normal standard semen parameters sub-population&#46;</p> <span class="elsevierStyleSectionTitle" id="sect0015">Materials and methods</span><p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">A prospective study was performed&#46; It evaluated standard semen parameters &#40;volume&#44; sperm concentration&#44; motility and morphology&#44; round cells&#44; peroxidase-positive cells&#41; according to WHO guidelines&#44; as well as functional tests &#40;24<span class="elsevierStyleHsp" style=""></span>h survival&#44; hypoosmotic swelling test&#44; modified stress test&#41;&#44; and additional assays &#40;aniline blue&#44; nitroblue-tetrazolium&#44; TUNEL&#41; in 110 semen samples from patients and 6 from fertile donors &#40;control&#41;&#46; Based on standard semen parameters values&#44; patients were divided into two groups &#40;A&#58; all standard parameters normal&#59; B&#58; one altered standard parameter at least&#41;&#46;</p> <span class="elsevierStyleSectionTitle" id="sect0020">Results</span><p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">At least one of the variables analyzed was altered in 96&#46;61&#37; of the samples&#46; Groups A and B showed statistically significant differences in all the complementary tests&#46; At least one of the complementary tests were altered in 93&#46;68&#37; of the samples in group A&#44; and the most frequently affected variable was sperm DNA fragmentation &#40;16&#46;95&#37;&#41;&#46;</p> <span class="elsevierStyleSectionTitle" id="sect0025">Conclusions</span><p id="spar0020" class="elsevierStyleSimplePara elsevierViewall">Performing a more in-depth seminal study within the routine functional and chromatin assays provides a more precise diagnosis of male infertility&#46; The WHO standards should be considered as a primary approach&#46;</p>"
      ]
      "es" => array:2 [
        "titulo" => "Resumen"
        "resumen" => "<span class="elsevierStyleSectionTitle" id="sect0035">Objetivos</span><p id="spar0025" class="elsevierStyleSimplePara elsevierViewall">Estudiar la incidencia de alteraciones funcionales y cromat&#237;nicas en espermatozoides de pacientes&#44; agrupados seg&#250;n normalidad de los par&#225;metros seminales est&#225;ndar en funci&#243;n de la quinta edici&#243;n del manual de la Organizaci&#243;n Mundial de la Salud &#40;OMS&#41;&#46; Identificar y correlacionar las caracter&#237;sticas alteradas con mayor frecuencia en la subpoblaci&#243;n normal para los par&#225;metros est&#225;ndar&#46;</p> <span class="elsevierStyleSectionTitle" id="sect0040">Materiales y m&#233;todos</span><p id="spar0030" class="elsevierStyleSimplePara elsevierViewall">Estudio prospectivo en el que se evaluaron en muestras de semen de 110 pacientes y 6 donantes f&#233;rtiles &#40;control&#41;&#44; los par&#225;metros seminales est&#225;ndar &#40;volumen&#44; concentraci&#243;n&#44; motilidad&#44; morfolog&#237;a&#44; c&#233;lulas redondas&#44; c&#233;lulas peroxidasa-positivas&#41; seg&#250;n las directrices de la OMS&#44; as&#237; como test complementarios funcionales &#40;sobrevida 24 horas&#44; test hipo-osm&#243;tico&#44; test de estr&#233;s modificado&#41; y adicionales &#40;azul de anilina&#44; nitroblue-tetrazolium&#44; TUNEL&#41;&#46; Seg&#250;n el resultado de los an&#225;lisis est&#225;ndar los pacientes se dividieron en dos grupos &#40;A&#58; todos los par&#225;metros est&#225;ndar normales&#59; B&#58; al menos uno de los par&#225;metros est&#225;ndar alterado&#41;&#46;</p> <span class="elsevierStyleSectionTitle" id="sect0045">Resultados</span><p id="spar0035" class="elsevierStyleSimplePara elsevierViewall">El 96&#44;61&#37; de las muestras present&#243; alterada al menos una de las variables analizadas&#46; Los grupos A y B mostraron diferencia estad&#237;sticamente significativa en todos los test complementarios&#46; En el grupo A el 93&#44;68&#37; de las muestras present&#243; al menos un test complementario alterado&#44; y la variable afectada con mayor frecuencia fue la fragmentaci&#243;n del ADN esperm&#225;tico &#40;16&#44;95&#37;&#41;&#46;</p> <span class="elsevierStyleSectionTitle" id="sect0050">Conclusiones</span><p id="spar0040" class="elsevierStyleSimplePara elsevierViewall">La profundizaci&#243;n en el estudio seminal&#44; introduciendo en la rutina ensayos funcionales y cromat&#237;nicos&#44; redunda en un diagn&#243;stico m&#225;s afinado de la infertilidad del var&#243;n&#46; Los par&#225;metros de la OMS deben considerarse como un abordaje primario&#46;</p>"
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        "nota" => "<p class="elsevierStyleNotepara" id="npar0005">Please cite this article as&#58; Curti G&#44; et al&#46; Diagn&#243;stico de infertilidad masculina&#58; necesidad de valoraciones funcionales y cromat&#237;nicas&#46; Actas Urol Esp&#46; 2013&#59;37&#58;100&#8211;5&#46;</p>"
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                  \t\t\t\t">55&#46;84<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>1&#46;84&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Normal forms &#40;&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Immature forms &#40;&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Round cells &#40;mill&#47;ml&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Peroxidase<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>&#40;mill&#47;ml&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttop\n
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                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>24<span class="elsevierStyleHsp" style=""></span>h survival &#40;&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>HOST &#40;&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttop\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;84<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;01&nbsp;\t\t\t\t\t\t\n
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                  """
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Proggresive motility &#40;&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttop\n
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                  \t\t\t\t\ttable-entry\n
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                  \t\t\t\t\ttop\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
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                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
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                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
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                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
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                  \t\t\t\t\ttop\n
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                  \t\t\t\t\ttable-entry\n
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                  \t\t\t\t\ttable-entry\n
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                  \t\t\t\t\ttop\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
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                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
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                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;096&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;944&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
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                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " colspan="7" align="left" valign="\n
                  \t\t\t\t\ttop\n
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                  \t\t\t\t  " colspan="7" align="left" valign="\n
                  \t\t\t\t\ttop\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
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                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">33&#46;52<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>3&#46;38&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
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                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
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                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
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                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic"><span class="elsevierStyleBold">p</span></span><span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleBold">&#61;</span><span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleBold">0&#46;001</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;994&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
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                  \t\t\t\t  " colspan="7" align="left" valign="\n
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                  \t\t\t\t"><span class="elsevierStyleVsp" style="height:0.5px"></span></td></tr><tr title="table-row"><td class="td" title="\n
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                  \t\t\t\t  " colspan="7" align="left" valign="\n
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                  \t\t\t\t"><span class="elsevierStyleItalic">Additional tests</span></td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Aniline blue &#40;&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t\ttable-entry\n
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                  \t\t\t\t\ttop\n
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                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t"><span class="elsevierStyleItalic"><span class="elsevierStyleBold">p</span></span><span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleBold">&#61;</span><span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleBold">0&#46;019</span>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t"><span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;315&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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Article information

ISSN: 21735786
Original language: English

DOI:10.1016/j.acuroe.2011.11.012

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Year/Month	Html	Pdf	Total

2023 March	1	2	3
2018 February	3	1	4
2018 January	15	0	15
2017 December	7	2	9
2017 November	11	0	11
2017 October	32	4	36
2017 September	28	1	29
2017 August	22	2	24
2017 July	24	2	26
2017 June	18	2	20
2017 May	23	3	26
2017 April	11	1	12
2017 March	19	12	31
2017 February	21	3	24
2017 January	18	0	18
2016 December	16	3	19
2016 November	14	2	16
2016 October	42	8	50
2016 September	25	5	30
2016 August	21	4	25
2016 July	19	2	21
2016 June	10	5	15
2016 May	14	15	29
2016 April	13	9	22
2016 March	17	13	30
2016 February	15	15	30
2016 January	23	17	40
2015 December	15	10	25
2015 November	18	8	26
2015 October	19	13	32
2015 September	20	7	27
2015 August	11	4	15
2015 July	11	4	15
2015 June	8	1	9
2015 May	9	3	12
2015 April	28	7	35
2015 March	21	6	27
2015 February	11	3	14
2014 July	1	0	1
2014 June	1	0	1
2014 May	2	1	3
2014 January	3	0	3
2013 December	6	2	8
2013 November	9	2	11
2013 October	17	2	19
2013 September	7	3	10
2013 July	2	2	4

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