Manual of Sperm Function Testing in Human Assisted Reproduction Edited by Ashok Agarwal , Ralf Henkel , Ahmad Majzoub Frontmatter More Information

Cambridge University Press 978-1-108-79353-7 — Manual of Sperm Function Testing in Human Assisted Reproduction Edited by Ashok Agarwal , Ralf Henkel , Ahmad Majzoub Frontmatter More Information

Manual of Sperm Function Testing in Human Assisted Reproduction

© in this web service Cambridge University Press www.cambridge.org Cambridge University Press 978-1-108-79353-7 — Manual of Sperm Function Testing in Human Assisted Reproduction Edited by Ashok Agarwal , Ralf Henkel , Ahmad Majzoub Frontmatter More Information

Cambridge Laboratory Manuals in Assisted Reproductive Technology

Titles in the series: Manual of Oocyte Retrieval and Preparation in Human Assisted Reproduction, Rachel Cutting & Mostafa Metwally Manual of Sperm Retrieval and Preparation in Human Assisted Reproduction, Ashok Agarwal, Ahmad Majzoub & Sandro C. Esteves Manual of Intracytoplasmic Sperm Injection in Human Assisted Reproduction: With Other Advanced Micromanipulation Techniques to Edit the Genetic and Cytoplasmic Content of the Oocyte, Gianpiero D. Palermo & Peter Nagy Manual of Embryo Selection in Human Assisted Reproduction, Catherine Racowsky, Jacques Cohen & Nick Macklon Manual of Embryo Culture in Human Assisted Reproduction, Kersti Lundin & Aisling Ahlström Manual of Cryopreservation in Human Assisted Reproduction, Sally Catt, Kiri Beilby & Denny Sakkas

Manual of Sperm Function Testing in Human Assisted Reproduction

Edited by Ashok Agarwal The Cleveland Clinic Foundation, Cleveland, OH, USA Ralf Henkel University of the Western Cape, Bellville, South Africa, Imperial College London, London, UK, and LogixX Pharma Ltd., Theale, Reading, UK Ahmad Majzoub Hamad Medical Corporation, Weill Cornell Medicine – Qatar, Doha, Qatar

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www.cambridge.org Information on this title: www.cambridge.org/9781108793537 DOI: 10.1017/9781108878715 © Cambridge University Press 2021 This publication is in copyright. Subject to statutory exception and to the provisions of relevant collective licensing agreements, no reproduction of any part may take place without the written permission of Cambridge University Press. First published 2021 Printed in the United Kingdom by TJ Books Limited, Padstow Cornwall A catalogue record for this publication is available from the British Library. Library of Congress Cataloging-in-Publication Data Names: Agarwal, Ashok, editor. | Henkel, Ralf (Ralf R.), editor. | Majzoub, Ahmad, editor. Title: Manual of sperm function testing in human assisted reproduction / edited by Ashok Agarwal, Ralf Henkel, Ahmad Majzoub. Description: Cambridge, United Kingdom ; New York, NY : Cambridge University Press, 2020. | Includes bibliographical references and index. Identifiers: LCCN 2020041955 (print) | LCCN 2020041956 (ebook) | ISBN 9781108793537 (paperback) | ISBN 9781108878715 (epub) Subjects: MESH: Spermatozoa–pathology | Infertility, Male–etiology | Semen Analysis–methods | Molecular Diagnostic Techniques–methods | Reproductive Techniques, Assisted Classification: LCC QP255 (print) | LCC QP255 (ebook) | NLM WJ 834 | DDC 571.8/451–dc23 LC record available at https://lccn.loc.gov/2020041955 LC ebook record available at https://lccn.loc.gov/2020041956 ISBN 978-1-108-79353-7 Paperback Cambridge University Press has no responsibility for the persistence or accuracy of URLs for external or third-party internet websites referred to in this publication and does not guarantee that any content on such websites is, or will remain, accurate or appropriate......

Every effort has been made in preparing this book to provide accurate and up-to-date information that is in accord with accepted standards and practice at the time of publication. Although case histories are drawn from actual cases, every effort has been made to disguise the identities of the individuals involved. Nevertheless, the authors, editors, and publishers can make no warranties that the information contained herein is totally free from error, not least because clinical standards are constantly changing through research and regulation. The authors, editors, and publishers therefore disclaim all liability for direct or consequential damages resulting from the use of material contained in this book. Readers are strongly advised to pay careful attention to information provided by the manufacturer of any drugs or equipment that they plan to use.

To my father Professor RC Agarwal (late) for instilling the virtues of honesty, dedication, and hard work. To my wonderful wife, Meenu, sons, Rishi and Neil-Yogi, for their unconditional love and support. To Prof. Kevin Loughlin (Harvard Medical School), Prof. Anthony Thomas (late) (Cleveland Clinic), and Prof. Edmund Sabanegh (Cleveland Clinic) for their friendship, guidance, and support and for making an indelible positive impression on my life. To my associates at work, large number of researchers and students, and most importantly the patients who placed their trust in our work. –Ashok Agarwal

To my late parents Waldemar and Helga Henkel for instilling the values of hard work, perseverance, and dedication. To my academic teachers, Professor Christoph Kirchner (Philipps University, Marburg, Germany) and Professor Wolf-Bernhard Schill (Justus Liebig University, Giessen, Germany) for guiding me in my academic career and continuous support. To my wife, Adv. Sharon Henkel for her unconditional love and support, without which I would not be able to do this work. –Ralf Henkel

I would like to dedicate this book to my wife, Zeinab, and our two lovely daughters, Sarah and Tala, for their love and support; to my mentors, Dr. Edmund Sabanegh and Prof. Ashok Agarwal (Cleveland Clinic), for their guidance and support and for all the opportunities they made possible. –Ahmad Majzoub

Contents

List of Contributors ix Short Biography xii Foreword xiii Denny Sakkas Preface xv

Introduction 1 10 Capacitation and Acrosome Reaction: R. John Aitken Histochemical Techniques to Determine Acrosome Reaction 81 1 Standard Semen Examination Analysis: Brett Nixon, Shenae L. Café, Elizabeth G. Manual Semen Analysis 6 Bromfield, Geoffry N. De Iuliis, Matthew D. Dun Lars Björndahl 11 Zona Binding: Competitive Sperm- 2 Standard Semen Analysis: Computer-Assisted Binding Assay 93 Semen Analysis 11 De Yi Liu, Ying Zhong, Zi-Na Wen Liana Maree 12 Zona Binding: Hemizona Assay 100 3 Standard Semen Analysis: Home Shubhadeep Roychoudhury, Daniel Franken Sperm Testing 23 Manesh Kumar Panner Selvam, Christopher 13 Oolemma Binding: Sperm Douglas, Ashok Agarwal Penetration Assay 106 Anup Shah, Kathleen Hwang 4 Standard Semen Analysis: Leukocytospermia 31 14 Oxidative Stress Testing: Direct Tests 111 Renata Finelli, Ralf Henkel Renata Finelli, Manesh Kumar Panner Selvam, Ashok Agarwal 5 Standard Semen Analysis: Morphology 39 Roelof Menkveld 15 Oxidative Stress Testing: Indirect Tests 123 Rakesh Sharma, Kathy Robert, Ashok Agarwal 6 Sperm Vitality: Eosin-Nigrosin Dye Exclusion 47 16 Chromatin Condensation: Aniline Rakesh Sharma, Ashok Agarwal Blue Stain 142 Jesse JP, Vidhu Dhawan, Rima Dada 7 Sperm Vitality: Hypo-Osmotic Swelling Test 52 Alexandre Rouen, Rajasingam S Jeyendran 17 Chromatin Condensation: Chromomycin A3 (CMA3) Stain 151 8 Determination of Mitochondrial Membrane Sara Marchiani, Lara Tamburrino, Francesca Potential by Flow Cytometry in Human Benini, Sandra Pellegrini, Elisabetta Baldi Sperm Cells 58 Nicolas Germain, Nathalie Jouy, Carole 18 Sperm Chromatin Structure: Toluidine Marchetti, Philippe Marchetti Blue Staining 156 Mohammed Nasr-Esfahani, Marziyeh Tavalaee 9 Capacitation and Acrosome Reaction: Fluorescence Techniques to Determine 19 DNA Damage: TdT-Mediated dUTP Acrosome Reaction 72 Nick-End-Labelling Assay 163 Raúl Sánchez, Fabiola Zambrano, Rakesh Sharma, Concetta Iovine, vii Pamela Uribe Ashok Agarwal

Contents

20 DNA Damage: Sperm Chromatin 24 Clinical Value of Sperm Function Tests 234 Structure Assay 192 Ahmad Majzoub Don Evenson 25 Future Developments: Sperm 21 DNA Damage: COMET Assay 202 Proteomics 245 Luke Simon, Benjamin R. Emery, Douglas Manesh Panner, Selvam Kumar, Saradha T. Carrell Baskaran, Ashok Agarwal 22 DNA Damage: Halo Sperm Test 213 Conclusion 256 Jaime Gosálvez, José Luís Fernández Ahmad Majzoub, Ralf Henkel, Ashok Agarwal 23 DNA Damage: Fluorescent In-Situ Hybridization 228 Sezgin Gunes Index 259

viii

Contributors

Ashok Agarwal, Ph.D. Shenae L. Cafe, B Biotech (Hons) Professor of Urology at Case Western Reserve Priority Research Centre for Reproductive Science, University; Director of Andrology Center; Director, School of Environmental and Life Sciences, American Center for Reproductive Medicine, The University of Newcastle, Callaghan, NSW 2308, Cleveland Clinic Foundation, Cleveland, Ohio, USA Australia, and Hunter Medical Research Institute, Pregnancy and Reproduction Program, R. John Aitken FRSE, FRSN, FAHMS, FAA New Lambton Heights, NSW 2305, Australia. Distinguished Laureate Professor of Biological Sciences, Priority Research Centre for Reproductive Douglas T. Carrell Science, University of Newcastle, Callaghan NSW Department of Surgery (Urology), University of Utah 2308, Australia School of Medicine, Salt Lake City, UT, USA, and Department of Human Genetics, University of Utah Elisabetta Baldi, Ph.D. School of Medicine, Salt Lake City, UT, USA Associate Professor of Clinical Pathology, Department of Clinical and Experimental Medicine, Rima Dada, M.D., Ph.D. University of Florence, Viale Pieraccini 6, 50139 Laboratory for Molecular Reproduction and Genetics, Florence, Italy Department of Anatomy, AIIMS, N Delhi Saradha Baskaran, Ph.D. Christopher Douglas, BS. American Center for Reproductive Medicine, American Center for Reproductive Medicine, Cleveland Clinic, Cleveland, OH, USA Cleveland Clinic, Cleveland, OH, USA, and Texas College of Osteopathic Medicine, Fort Worth, Francesca Benini TX, USA Senior Embryologist, Centro Procreazione Assistita Demetra (Florence, Italy) Matthew D. Dun, B Biotech (Hons), Ph.D. Hunter Medical Research Institute, Pregnancy and Lars Björndahl, M.D. Ph.D. Reproduction Program, New Lambton Heights, NSW Licensed Swedish Physician, Specialist in Clinical 2305, Australia; Cancer Signaling Research Group, Chemistry, Senior Consultant Physician, Andrology School of Biomedical Sciences and Pharmacy, Faculty Laboratory Director, ANOVA – Karolinska of Health and Medicine, University of Newcastle, University Hospital. Callaghan, NSW 2308, Australia; and Priority Research Centre for Cancer Research Innovation and Elizabeth G. Bromfield, B Biotech (Hons), Ph.D. Translation, Hunter Medical Research Institute, Priority Research Centre for Reproductive Science, Lambton, NSW 2305, Australia. School of Environmental and Life Sciences, The University of Newcastle, Callaghan, NSW 2308, Benjamin R. Emery Australia, and Hunter Medical Research Institute, Department of Surgery (Urology), University Pregnancy and Reproduction Program, New of Utah School of Medicine, Salt Lake City, Lambton Heights, NSW 2305, Australia UT, USA ix

List of Contributors

Donald P Evenson, Ph.D., HCLD and Reproduction, Imperial College London, President and Director, SCSA Diagnostics, Inc., London, UK Brookings, SD; Emeritus Distinguished Professor, Kathleen Hwang, M.D. South Dakota State University, Brookings, SD; Adjunct Professor, Sanford Medical School, Dept Associate Professor of Urology, Department of OB/GYN, University of South Dakota, Sioux Falls, Urology, University of Pittsburgh Medical Center, SD; former faculty, Sloan Kettering Research Pittsburgh, Pennsylvania Institute, NY, USA Nathalie JOUY, Ph.D. José Luís Fernández University of Lille, CNRS, Inserm, CHU Lille, Institut Unidad de Genética, Instituto de Investigación Pasteur de Lille, US 41 - UMS 2014 - PLBS, F-59000 Biomédica de A Coruña (INIBIC), Complexo Lille, France Hospitalario Universitario de A Coruña (CHUAC), De Yi Liu, Ph.D. Sergas, Universidade de A Coruña (UDC), Spain, and Chengdu Xinan Gynecology Hospital, Chengdu, Laboratorio de Genética, Centro Oncológico de China and Department of Obstetrics & Gynecology, Galicia, A Coruña, Spain. University of Melbourne, Australia Renata Finelli Concetta Iovine American Centre for Reproductive Medicine, American Center for Reproductive Medicine, Cleveland Clinic, Cleveland, Ohio, USA Cleveland Clinic, Cleveland, OH, USA Daniel R Franken, Ph.D. Geoffry N. De Iuliis, BSc (Hons), Ph.D. Emeritus Professor, Department of Obstetrics and Priority Research Centre for Reproductive Science, Gynecology, Tygerberg Hospital, Stellenbosch School of Environmental and Life Sciences, University, Tygerberg, South Africa; Emeritus The University of Newcastle, Callaghan, NSW 2308, Professor, Department of Obstetrics and Gynecology, Australia, and Hunter Medical Research Institute, University of the Free State, Bloemfontein, South Pregnancy and Reproduction Program, New Africa; Former Adjunct Professor, Eastern Virginia Lambton Heights, NSW 2305, Australia. Medical School, Norfolk, VA; and Consultant, SARhealth Line Chennai, India Jesse JP, M.D. Laboratory for Molecular Reproduction and Genetics, Nicolas GERMAIN, M.D. Department of Anatomy, AIIMS, N Delhi University of Lille, CNRS, Inserm, CHU Lille, UMR9020-U1277 - CANTHER - Cancer Rajasingam S. Jeyendran, B.V.Sc., M.S., Ph.D., Heterogeneity Plasticity and Resistance to Therapies, HCLD, ALD F-59000 Lille, France Androlab Inc., Chicago, IL 60611, USA

Jaime Gosálvez Ahmad Majzoub Unidad de Genética, Facultad de Biología, Department of Urology, Hamad Medical Corporation Universidad Autónoma de Madrid, Madrid, Spain. Doha, Qatar, and Weill Cornell Medicine – Qatar, Doha, Qatar Sezgin GunesOndokuz Mayis University Medical Faculty Department of Medical Carole MARCHETTI, M.D., Ph.D. Biology, 55139 Atakum, Samsun, Turkey Cerballiance 44 avenue Max Dormoy, F-59000 Lille, France Ralf Henkel American Center for Reproductive Medicine, Philippe MARCHETTI, M.D., Ph.D. Cleveland Clinic, Department of Urology, Cleveland, University of Lille, CNRS, Inserm, CHU Lille, Ohio, USA; Department of Medical Bioscience, UMR9020-U1277 - CANTHER - Cancer x University of the Western Cape, Bellville, South Heterogeneity Plasticity and Resistance to Therapies, Africa; and Department of Metabolism, Digestion F-59000 Lille, France

List of Contributors

Sara Marchiani, Ph.D. Sandra Pellegrini, M.D. Scholarship at Sodc di Medicina della Sessualità e gynecologist, Centro Procreazione Assistita Demetra Andrologia afferente al DAI Materno Infantile, Azienda (Florence, Italy) Ospedaliero Universitaria Careggi, Florence, Italy Anup A. Shah, M.D. Liana Maree Ph.D. Department of Urology, University of Pittsburgh Department of Medical Biosciences, University of the Medical Center, Pittsburgh, Pennsylvania Western Cape, Bellville, South Africa Rakesh Sharma Roelof Menkveld, Ph.D. American Center for Reproductive Medicine, Department of Obstetrics and Gynaecology, Faculty Cleveland Clinic, Cleveland, USA of Medicine and Health Sciences, Stellenbosch University, Tygerberg 7505, South Africa Luke Simon Department of Surgery (Urology), University Mohammad Hossein Nasr-Esfahani, Ph.D of Utah School of Medicine, Salt Lake City, Department of Animal Biotechnology, Reproductive UT, USA Biomedicine Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran and Isfahan Tamburrino Lara, Ph.D. Fertility and Infertility Center, Isfahan, Iran Research Fellowship at Department of Clinical and Experimental Medicine, University of Florence, Viale Brett Nixon, BSc (Hons), Ph.D. Pieraccini 6, 50139 Florence, Italy Priority Research Centre for Reproductive Science, School of Environmental and Life Sciences, The Marziyeh Tavalaee, Ph.D University of Newcastle, Callaghan, NSW 2308, Department of Animal Biotechnology, Australia, and Hunter Medical Research Institute, Reproductive Biomedicine Research Center, Pregnancy and Reproduction Program, New Royan Institute for Biotechnology, ACECR, Lambton Heights, NSW 2305, Australia. Isfahan, Iran. Kathy A. Robert Pamela Uribe, Ph.D. American Center for Reproductive Medicine, Center of Excellence in Translational Medicine- Cleveland Clinic, Cleveland, Ohio, USA Scientific and Technological Bioresource Nucleus (CEMT – BIOREN), Department of Internal Alexandre Rouen, M.D., Ph.D. Medicine, Faculty of Medicine, Universidad de La Medical Genetics Department, Trousseau Hospital, Frontera, Temuco, Chile Paris, France Vidhu Dhawan, M.D. Shubhadeep Roychoudhury, Ph.D. Laboratory for Molecular Reproduction and Genetics, Assistant Professor, Department of Life Science and Department of Anatomy, AIIMS, N Delhi Bioinformatics, Assam University, Silchar, India, and Visiting Professor, Department of Morphology, Zi-Na Wen, M.Sc. Physiology and Animal Genetics, Mendel University Chengdu Xinan Gynecology Hospital, Chengdu, in Brno, Czech Republic. China Raúl Sánchez, M.D. Fabiola Zambrano, Ph.D. Center of Excellence in Translational Medicine- Center of Excellence in Translational Medicine- Scientific and Technological Bioresource Nucleus Scientific and Technological Bioresource Nucleus (CEMT – BIOREN), Department of Preclinical (CEMT – BIOREN), Department of Preclinical Sciences, Faculty of Medicine, University of La Sciences, Faculty of Medicine, University of La Frontera, Temuco, Chile. Frontera, Temuco, Chile.

Manesh Kumar Panner Selvam, Ph.D. Ying Zhong, M.D. xi American Center for Reproductive Medicine, Chengdu Xinan Gynecology Hospital, Chengdu, Cleveland Clinic, Cleveland, OH, USA China

Short Biography

Dr. Ashok Agarwal is the Head of Andrology Center Digestion and Reproduction at Imperial College and Director of Research at the American Center for London, London, UK. He is also Honorary Professor Reproductive Medicine. He holds these positions at at the Universidad Peruana Cayetano Heredia, Lima, The Cleveland Clinic Foundation, where he is Peru, and Editor-in-Chief of Andrologia. Ralf has Professor of Surgery (Urology) at the Lerner College published more than 165 original and review articles of Medicine of Case Western Reserve University. as well as 46 book chapters and supervised more than Ashok was trained in Male Infertility and Andrology 70 Hons/M.Sc./M.D./Ph.D. postgraduate students’ at the Brigham and Women’s Hospital and Harvard theses. For his research, Ralf received 16 awards. Ralf Medical School and has over 28 years of experience in has received 26 research grants for numerous research directing busy male infertility diagnostic and fertility projects to investigate the impact of oxidative stress preservation services. He is an editor of over on sperm functions, DNA fragmentation and 40 medical text books on male infertility, assisted fertilization, as well as the effects of Herbal Medicine reproductive technology (ART), fertility preservation, on male reproductive functions including prostate DNA damage, and antioxidants. cancer and benign prostatic hyperplasia. Ralf Henkel, B.Ed., Ph.D., Habil., studied Biology and Dr. Ahmad Majzoub is a Consultant at the Chemistry at the University of Marburg, Germany, department of Urology and the program director of and obtained his Ph.D. in 1990. Ralf continued his the andrology and male infertility fellowship at post-doctoral training and obtained his Habilitation Hamad Medical Corporation, Doha, Qatar. He is an at the University of Giessen, School of Medicine in Arab Board Certified Urologist and has undergone 1998. From 1998 to 2004, he was an Assistant, Clinical and Research Fellowship in Andrology at Associate and Extraordinary Professor, at the Justus- the world-famous Glickman Urological and Liebig University of Giessen, Germany, before he Kidney Institute and the American Center for accepted a Full Professorship at the Department of Reproductive Medicine at Cleveland Clinic Urology at the University of Jena, Germany. From Foundation, Cleveland, United States. He is actively 2005 to 2020, he was Professor and Head of involved in the field of Medical Research with over Department at the Department of Medical Bioscience 150 research publications at peer reviewed journals at the University of the Western Cape, Bellville, South and several book chapters mainly focusing on Africa. Currently, he is Chief Scientific Advisor at Andrology and men’s health. He also co-edited LogixX Pharma, Reading, UK, and holds a Visiting several books and special issues on various aspects Reader Position in the Department of Metabolism, of male infertility.

xii

Foreword

Infertility is ranked as the fifth highest serious global new classes of sperm function tests and improve disability (among populations under the age of 60) by older ones. the World Health Organization. Human Assisted In this excellent publication, many of the stalwarts Reproduction Treatments have enabled many infertile of sperm research and other up and coming research- couples to discard the tag of infertility. Nowhere has ers have provided their impactful insights into the the impact of Assisted Reproduction been more intricacies of preparation and analysis of semen. The apparent than in the treatment of male infertility. book, entitled a Manual of Sperm Function Testing in Male infertility is known to solely affect one third of Human Assisted Reproduction, outlines the experi- all infertile couples and in another 10–20 percent of ences of many leaders in the field. It provides chapters the couples seeking medical assistance it is paired with on semen analysis, different assay techniques and a female factor resulting in the fact that male factor finally the clinical utility of the old and new sperm infertility is found as a reason of infertility in about function tests. The book finishes with a look into the half of all infertile couples. future of sperm diagnostics using techniques adapted Sadly, sperm biology has taken a back seat in from genomics, transcriptomics, proteomics and meta- Assisted Reproduction and unfortunately, the male bolomics. The editors, Professor Ashok Agarwal, is somewhat overlooked as a major clinical con- Professor Ralf Henkel and Dr Ahmad Majzoub, are tributor to a couple’s chances of achieving a live all highly respected members in the field and bring birth. Much of this arose because of the advent of together their foresight into how sperm function tests intra-cytoplasmic sperm injection (ICSI) in the should and will be used in the future. early 1990s. This also led to a loss of interest in It is my hope that this book will be a resource not developing sperm function tests. Although ICSI has only to clinicians and scientists already practicing in proven a savior for many men, it is now becoming the field of Assisted Reproduction, but will encourage evident that there is more than meets the eye about young researchers and clinicians to embrace research sperm. It is now clear that paternal factors may be in the area of sperm function so that future patients responsible for miscarriages and repeated failures can benefit from their discoveries. after Assisted Reproduction. It is also clear that the sperm epigenome can impact present and future Denny Sakkas Ph.D. pregnancies and offspring. Finally, the knowledge Chief Scientific Officer, Boston IVF that sperm quality declines with age has refocused Associate Professor our attention on the male. The tide has therefore Department of OBGYN swung and researchers are now looking to develop Yale University of Medicine

xiii

Preface

Infertility is a global health concern as it affects over contributed 26 state-of-the-art chapters for this book. 190 million couples at reproductive age of which half We have provided detailed information on the role of is due to a male factor. According to the literature, functional sperm parameters in physiology and path- about 50 percent of the global incidence of male ology with essential up-to-date diagnostic and clinical infertility is due to known causes, while the causes information to improve andrological diagnosis and for the remaining percent are unknown. Since male treatment. The most common diagnostic tests to infertility seriously affects not only the physical well- determine sperm function are described in a clinically being of individual patients, but also the psychological relevant manner. health of couples desiring their own children, it is the The combination of basic and clinical information responsibility of clinicians and scientists to identify makes our book one of its kind and highly suited for the causes of male infertility and possible treatment fertility practitioners, andrologists, urologists, med- options. This includes an assessment of the function- ical doctors, reproductive professionals and research ality of the male germ cells in order to predict the students. We are grateful to our contributors, who are chances of a man to father a child. The primary aim distinguished leaders in the field with extensive of a standard semen analysis according to the WHO experience from around the world. guidelines is to evaluate the basic semen parameters, This book would not have been possible without the which over the years have been found to be insuffi- excellent support of Cambridge University Press. We cient in predicting the male fertilizing potential. The are thankful to Nick Dunton, Senior Acquisition Editor increased knowledge about the sperm functional par- and Camille Lee-Own, Editorial Project Manager for ameters such as capacitation, acrosome reaction, zona excellent management of this project. The editors are binding or chromatin condensation has increased our grateful to their families for their love and support. We understanding of the physiology and pathology of genuinely hope that this volume will support and enrich fertilization, and possible treatment options. This your clinical practice in clinical andrology. has also resulted in the development of several advanced diagnostic methods and treatments for male Cleveland, OH, USA Ashok Agarwal, Ph.D., infertility in cases of impaired sperm function. HCLD (ABB), EMB (ACE) Our book provides a detailed description of the Bellville, South Africa Ralf Henkel, BEd, Ph.D., currently known sperm function properties including and London, UK Habil. their physiological and pathological background. Renowned experts from 14 countries have generously Doha, Qatar Ahmad Majzoub, M.D.