This document summarizes a presentation on sperm DNA fragmentation (SDF) and male infertility. It discusses how SDF provides different information than routine semen analysis and is a better prognostic indicator. Elevated SDF is associated with infertility, poor assisted reproductive technology outcomes, and miscarriage. Several methods can assess SDF but differ in their ability to directly or indirectly measure damage. Lifestyle changes like reducing stress and smoking, treating underlying conditions, and using oral antioxidants can help lower SDF. Varicocele repair is also effective at reducing SDF levels in men with the condition.
Sperm DNA Fragmentation (Oxidative stress, DNA damage and apoptosis, Test, Techniques, Relation to other semen parameters, Relationship to leucocytes, Relation to ICSI outcomes, Clinical applications, significance and limitations)
ICSI as it is presently performed is far from an ideal solution because the selection of sperm is based on the judgement of an embryologist, who is looking for the most normal appearing sperm available.
Sperm DNA Fragmentation (Oxidative stress, DNA damage and apoptosis, Test, Techniques, Relation to other semen parameters, Relationship to leucocytes, Relation to ICSI outcomes, Clinical applications, significance and limitations)
ICSI as it is presently performed is far from an ideal solution because the selection of sperm is based on the judgement of an embryologist, who is looking for the most normal appearing sperm available.
Iran march 2011
ABRASCT:
SPERM RETRIEVAL TECHNIQUES FOR THE AZOOSPERMIC MALE
Sandro C. Esteves, MD, PhD
Spermatozoa can be retrieved from either the epididymis or the testis, depending on the type of azoospermia, using different surgical methods such as PESA, MESA, TESA, TESE and micro-TESE.
In obstructive azoospermia (OA), sperm production is normal and gametes can be easily retrieved from the epididymis or the testicle in most cases, irrespective of the technique. PESA or TESA are simple and efficient methods for retrieving epididymal or testicular spermatozoa in men with OA. According to our data on OA, the etiology of the obstruction and the use of fresh or frozen-thawed epididymal/testicular sperm do not seem to affect ICSI outcomes in terms of fertilization, pregnancy, or miscarriage rates.
In cases of nonobstructive azoospermia (NOA), the efficiency of TESA for retrieving spermatozoa is lower than TESE, except in the favorable cases of men with previous successful TESA or testicular histopathology showing hypospermatogenesis. The use of microsurgery during TESE may improve the efficacy of sperm extraction with significantly less tissue removed, which ultimately facilitates sperm processing. Testicular histology results, if available, may be useful to predict the chances to retrieve sperm in men with NOA. Our data demonstrate that micro-TESE performs better than conventional TESE or TESA in cases of maturation arrest and Sertoli cell-only histological patterns, where tubules containing active focus of spermatogenesis can be positively identified using microsurgery. Testicular spermatozoa can be obtained even in the worst case scenario except in the cases of Y chromosome infertility with complete AZFa and/or AZFbmicrodeletions.
In both OA and NOA, sperm retrieval technique itself seems to have no impact on ICSI success rates. The main goal of PESA/TESA/TESE sperm processing is the recovery of a clean sample containing motile sperm. Such specimens are more fragile, and often compromised in motility, as compared to the ones obtained from ejaculates. Laboratory techniques should be carried out with great caution not to jeopardize the sperm fertilizing potential. Surgically-retrieved spermatozoa can be intentionally cryopreserved for future use. Spare left-over specimens that would be discharged after ICSI can also be cryostored. Different strategies can be developed according to each group’s results. If freezing of surgically-retrieved specimens provides results similar to those with the use of fresh sperm, then the use of freezing specimens would be preferable. If not, fresh specimens are preferable.
The reproductive potential of infertile men undergoing ART is related to the type of azoospermia. According to our data, the chances of retrieving spermatozoa (odds ratio [OR] = 43.0; 95% confidence interval [CI]: 10.3-179.5) and of achieving a live birth by ICSI (OR=1.86; 95% CI:l 1.03-2.89) were significantly increased in couples whose male partner had obstructive rather than non-obstructive azoospermia. Children conceived using sperm retrieved from men with OA and NOA should be followed-up because it is still unclear if there is an increased risk of birth defects when ICSI is carried out with non-ejaculated sperm.
References
Esteves SC, Glina S. Recovery of spermatogenesis after microsurgical subinguinal varicocele repair in azoospermic men based on testicular histology. IntBraz J Urol. 2005; 31:541-8.
Verza S Jr, Esteves SC. Sperm defect severity rather than sperm source is associated with lower fertilization rates after intracytoplasmic sperm injection. IntBraz J Urol. 2008,34:49-56.
Esteves SC, Verza S, Prudencio C, Seol B. Sperm retrieval rates (SRR) in nonobstructive azoospermia (NOA) are related to testicular histopathology results but not to the etiology of azoospermia. FertilSteril. 2010; 94(Suppl.):S132.
Esteves SC, Verza S, Prudencio C, Seol B. Success of percutaneous sperm retrieval and i
Iran march 2011
ABRASCT:
SPERM RETRIEVAL TECHNIQUES FOR THE AZOOSPERMIC MALE
Sandro C. Esteves, MD, PhD
Spermatozoa can be retrieved from either the epididymis or the testis, depending on the type of azoospermia, using different surgical methods such as PESA, MESA, TESA, TESE and micro-TESE.
In obstructive azoospermia (OA), sperm production is normal and gametes can be easily retrieved from the epididymis or the testicle in most cases, irrespective of the technique. PESA or TESA are simple and efficient methods for retrieving epididymal or testicular spermatozoa in men with OA. According to our data on OA, the etiology of the obstruction and the use of fresh or frozen-thawed epididymal/testicular sperm do not seem to affect ICSI outcomes in terms of fertilization, pregnancy, or miscarriage rates.
In cases of nonobstructive azoospermia (NOA), the efficiency of TESA for retrieving spermatozoa is lower than TESE, except in the favorable cases of men with previous successful TESA or testicular histopathology showing hypospermatogenesis. The use of microsurgery during TESE may improve the efficacy of sperm extraction with significantly less tissue removed, which ultimately facilitates sperm processing. Testicular histology results, if available, may be useful to predict the chances to retrieve sperm in men with NOA. Our data demonstrate that micro-TESE performs better than conventional TESE or TESA in cases of maturation arrest and Sertoli cell-only histological patterns, where tubules containing active focus of spermatogenesis can be positively identified using microsurgery. Testicular spermatozoa can be obtained even in the worst case scenario except in the cases of Y chromosome infertility with complete AZFa and/or AZFbmicrodeletions.
In both OA and NOA, sperm retrieval technique itself seems to have no impact on ICSI success rates. The main goal of PESA/TESA/TESE sperm processing is the recovery of a clean sample containing motile sperm. Such specimens are more fragile, and often compromised in motility, as compared to the ones obtained from ejaculates. Laboratory techniques should be carried out with great caution not to jeopardize the sperm fertilizing potential. Surgically-retrieved spermatozoa can be intentionally cryopreserved for future use. Spare left-over specimens that would be discharged after ICSI can also be cryostored. Different strategies can be developed according to each group’s results. If freezing of surgically-retrieved specimens provides results similar to those with the use of fresh sperm, then the use of freezing specimens would be preferable. If not, fresh specimens are preferable.
The reproductive potential of infertile men undergoing ART is related to the type of azoospermia. According to our data, the chances of retrieving spermatozoa (odds ratio [OR] = 43.0; 95% confidence interval [CI]: 10.3-179.5) and of achieving a live birth by ICSI (OR=1.86; 95% CI:l 1.03-2.89) were significantly increased in couples whose male partner had obstructive rather than non-obstructive azoospermia. Children conceived using sperm retrieved from men with OA and NOA should be followed-up because it is still unclear if there is an increased risk of birth defects when ICSI is carried out with non-ejaculated sperm.
References
Esteves SC, Glina S. Recovery of spermatogenesis after microsurgical subinguinal varicocele repair in azoospermic men based on testicular histology. IntBraz J Urol. 2005; 31:541-8.
Verza S Jr, Esteves SC. Sperm defect severity rather than sperm source is associated with lower fertilization rates after intracytoplasmic sperm injection. IntBraz J Urol. 2008,34:49-56.
Esteves SC, Verza S, Prudencio C, Seol B. Sperm retrieval rates (SRR) in nonobstructive azoospermia (NOA) are related to testicular histopathology results but not to the etiology of azoospermia. FertilSteril. 2010; 94(Suppl.):S132.
Esteves SC, Verza S, Prudencio C, Seol B. Success of percutaneous sperm retrieval and i
Role of sperm index in embryo quality what to do - 17th iranian congressSandro Esteves
17th International Congress of the Iranian Association for Fertility and Reproductive Medicine
Tehran– March 2011
Abstract
ROLE OF SPERM INDEXES IN EMBRYO QUALITY: WHAT TO DO?
Sandro C. Esteves, MD, PhD
Spermatozoa are highly specializedcells with the purpose of not onlydelivering competent paternal DNA to the oocyte but also to provide a robust epigenetic contribution to embryogenesis. The identification of sperm fertility markers and the ability to selecthealthy spermatozoa for ART have a dual objective of choosing the best treatment strategy and optimizing ART outcomes. Currently, sperm indexes determination in the clinical setting is generally based on cell morphology and DNA content. Both sperm morphology and DNA integrity results, obtained from raw semen samples, have been shown to be of prognostic value for unassisted and assisted conception and useful in the selection of the best assisted conception modality.
These assays, however,provide an assessment of the distribution of cells in a given ejaculatethat may not be representative of the sperm population used in the ART treatment cycle. In fact, severe teratozoospermia,using Kruger’s strict criteria on pre-ART semen analysis, does notcorrelate to fertilization and embryo formation (including blastocyst development) in ICSI cycles. Nonetheless, if a more holistic approach to sperm morphology is taken, two prognostic groups can still be identified in cases of severeteratozoospermia (<4% normal) because certain morphology patterns and sperm abnormalities are known to affect ICSI outcomes. The first group includes mostly genetically determined sperm pattern defects, such asglobozoospermia, short tail syndrome and small-headed spermatozoa (in most cases combined with very small acrosomes). All of these types represent untreatable conditions that have been associated with abnormal sperm function andpoor ART outcomes. The second group includes unspecifiedor non-genetically determined sperm defects or patternscaused by environmental factors, medication, infection and related infertility conditions, including varicocele. Treatment of these conditions has been shown to optimize sperm morphology indexes with a positive impact on ART outcomes. Although the technician microscopically selects morphologically normal individual sperm during ICSI, form normalcy does not necessarily imply normal DNA content. As such, sperm DNA testing has been advocated to be an independent and reliable marker of fertility potential since sperm chromatin andDNA integrity is essential to ensure that the fertilizing sperm cansupport normal embryonic development of the zygote.At present, conflicting reports exist on the role of sperm DNA fragmentation index for embryo development, and it is apparent that DNA fragmentation does not significantly impair zygote and cleaving embryo morphology because major activation of the embryonic genome only beginafter the 4-cell stage. These observations do no underscore the importance of finding ways to increase sperm DNA integrity, since it has been suggested that DNA fragmentation is associated with late paternal effects that may lead to early miscarriages or diseases in the offspring. The etiology of sperm DNA damage is multi-factorial and may be due to primary (ageing, cryptorchidism, genetic defects, idiopathic) and or secondary (drugs, environmental, tobacco smoking, genital tract inflammation, infection,testicular hyperthermia and varicoceles) factors. Specific or non-specific treatments, including antioxidant supplements, are generally associated with reduced levels of sperm DNA damage and/or improved fertility potential.
Taken in conjunction, it is apparent that there is no unique sperm factor able to predict embryo development, but several candidate biomarkers are involved in this complex process.As a result, a wide variety of techniques have been proposed, including externalization of phosphotidylserine (magnetic-activated cell sorting),cell
Optimizing male infertility treatment in ART- Dr Parul Katiyar, Max Hospitals...Dr Parul Katiyar
Male factor itself is responsible for infertility in approx 30-40% couples and contributes to infertility in another approx 20%. In many men having normal sperm parameters on semenogrom,
sperms do not function in a manner necessary for fertility and can still cause infertility. This often goes undetected unless specifically sought for. This presentation presents a clinical approach to male factor infertility, specifically related to functional aspects of male fertility.
Dr. Michael Dyck - Impact of the Boar on Herd Fertility John Blue
Impact of the Boar on Herd Fertility - Dr. Michael Dyck, University of Alberta, from the 2012 Allen D. Leman Swine Conference, September 15-18, St. Paul, Minnesota, USA.
Analysis of an Alternative Method for Determining Sperm DNA Integrity and Com...Genea Limited
Presented by: Linda Morrison
Co-authors: Uli Schmidt, Nico Foley, Rebecca Dorey, Steven McArthur, Genea, Sydney
Presented at the World Congress on Human Reproduction 2011, Melbourne
Aula Indice de Fragmentação de DNA espermático: como avaliar e tratar. Conrad...Conrado Alvarenga
Aula Indice de Fragmentação de DNA espermático: como avaliar e tratar. Conrado Alvarenga M.D. - XI Congresso Paulista de Medicina Reprodutiva, 2017 - mesa de andrologia.
Curso de imersão em medicina reprodutiva Arte Academy - Aula no Arte Academy ...Conrado Alvarenga
Curso de imersão em medicina reprodutiva Arte Academy - Aula no Arte Academy primeira edição, ministrada no Tivolli em SP, no dia 15 de setembro de 2018 - Conrado Alvarenga
Sperm DNA Fragmentation : Role in natural and assisted conception: Recent adv...Shivani Sachdev
Male factor infertility is responsible for >40% of couples presenting for ART. Conventional SA continues to be the only routine test to diagnose this condition. Current SA is based on 5th edition of the WHO manual (2010) . All normal values shifted to lower centile compared to previous edition. Less men now classified as infertile (Murray et al 2012). Sperm DNA damage - used to denote abnormal genetic material which in turn may lead to male sub fertility/ IVF failure / miscarriage.
DNA Fragmentation Index of Sperm - Expressed as DFI- or percentage of the number of cells with defects in protamination of DNA structure in the evaluated sperm cells
The integrity of paternal genome is of paramount importance in the initiation of viable pregnancy. The fragmented DNA is incompatible with normal embryonic development
Sperm chromatin anomalies are often present in men with abnormal semen analyses 8% of men with normal semen parameters have abnormal sperm DNA integrity
Zini A.Biologic variability of sperm DNA denaturation in infertile men. Urology 2001
We discuss the various tests used and recent concepts and techniques and what are the newer treatment options
Novel concepts in male factor infertility: clinical and laboratory perspectivesSandro Esteves
Presentation Objectives:
1. Update on the WHO reference values for semen parameters, and understand the role of sperm DNA fragmentation testing to decision-making strategies;
2. Learn how to counsel azoospermic men seeking fertility, and the role of gonadotropin therapy in this infertility condition;
3. Understand the benefits of microsurgery to both sperm retrieval and varicocele treatment;
4. Appraise the role of medical and surgical interventions to infertile men undergoing ART.
Air quality: is it that important? And if so, how to measure and control it?Sandro Esteves
Quality and Risk Management in the IVF Laboratory; Redlara Brasil, Belo Horizonte, 14-15 September 2016
Content:
1.Air quality: is it that important?
2. How to control?
3. How to measure?
Public lecture - Stem Cell and Male InfertilitySandro Esteves
Reproductive Andrology Workshop III
17-21 January 2016 - Kuwait City - KUWAIT
Organized by: Al Jahra Reproductive Medicine Unit - Ministry of Health
Public Lecture - Stem Cell and Male Infertility
Clinical management of men with nonobstructive azoospermia - Role of IVF Labo...Sandro Esteves
Reproductive Andrology Workshop III
17-21 January 2016 - Kuwait City - KUWAIT
Organized by: Al Jahra Reproductive Medicine Unit - Ministry of Health
Lecture 5: Role of IVF Laboratory in Nonobstructive Azoospermia
Clinical management of men with nonobstructive azoospermia - Sperm Retrieval ...Sandro Esteves
Reproductive Andrology Workshop III
17-21 January 2016 - Kuwait City - KUWAIT
Organized by: Al Jahra Reproductive Medicine Unit - Ministry of Health
Lecture 4: Sperm Retrieval Methods in Nonobstructive Azoospermia
Clinical management of men with nonobstructive azoospermia - Steps Before Spe...Sandro Esteves
Reproductive Andrology Workshop III
17-21 January 2016 - Kuwait City - KUWAIT
Organized by: Al Jahra Reproductive Medicine Unit - Ministry of Health
Lecture 3: Steps Before Sperm Retrieval in Nonobstructive Azoospermia
Couples presenting to the infertility clinic- Do they really have infertility...Sujoy Dasgupta
Dr Sujoy Dasgupta presented the study on "Couples presenting to the infertility clinic- Do they really have infertility? – The unexplored stories of non-consummation" in the 13th Congress of the Asia Pacific Initiative on Reproduction (ASPIRE 2024) at Manila on 24 May, 2024.
ARTIFICIAL INTELLIGENCE IN HEALTHCARE.pdfAnujkumaranit
Artificial intelligence (AI) refers to the simulation of human intelligence processes by machines, especially computer systems. It encompasses tasks such as learning, reasoning, problem-solving, perception, and language understanding. AI technologies are revolutionizing various fields, from healthcare to finance, by enabling machines to perform tasks that typically require human intelligence.
These lecture slides, by Dr Sidra Arshad, offer a quick overview of physiological basis of a normal electrocardiogram.
Learning objectives:
1. Define an electrocardiogram (ECG) and electrocardiography
2. Describe how dipoles generated by the heart produce the waveforms of the ECG
3. Describe the components of a normal electrocardiogram of a typical bipolar leads (limb II)
4. Differentiate between intervals and segments
5. Enlist some common indications for obtaining an ECG
Study Resources:
1. Chapter 11, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 9, Human Physiology - From Cells to Systems, Lauralee Sherwood, 9th edition
3. Chapter 29, Ganong’s Review of Medical Physiology, 26th edition
4. Electrocardiogram, StatPearls - https://www.ncbi.nlm.nih.gov/books/NBK549803/
5. ECG in Medical Practice by ABM Abdullah, 4th edition
6. ECG Basics, http://www.nataliescasebook.com/tag/e-c-g-basics
micro teaching on communication m.sc nursing.pdfAnurag Sharma
Microteaching is a unique model of practice teaching. It is a viable instrument for the. desired change in the teaching behavior or the behavior potential which, in specified types of real. classroom situations, tends to facilitate the achievement of specified types of objectives.
Ethanol (CH3CH2OH), or beverage alcohol, is a two-carbon alcohol
that is rapidly distributed in the body and brain. Ethanol alters many
neurochemical systems and has rewarding and addictive properties. It
is the oldest recreational drug and likely contributes to more morbidity,
mortality, and public health costs than all illicit drugs combined. The
5th edition of the Diagnostic and Statistical Manual of Mental Disorders
(DSM-5) integrates alcohol abuse and alcohol dependence into a single
disorder called alcohol use disorder (AUD), with mild, moderate,
and severe subclassifications (American Psychiatric Association, 2013).
In the DSM-5, all types of substance abuse and dependence have been
combined into a single substance use disorder (SUD) on a continuum
from mild to severe. A diagnosis of AUD requires that at least two of
the 11 DSM-5 behaviors be present within a 12-month period (mild
AUD: 2–3 criteria; moderate AUD: 4–5 criteria; severe AUD: 6–11 criteria).
The four main behavioral effects of AUD are impaired control over
drinking, negative social consequences, risky use, and altered physiological
effects (tolerance, withdrawal). This chapter presents an overview
of the prevalence and harmful consequences of AUD in the U.S.,
the systemic nature of the disease, neurocircuitry and stages of AUD,
comorbidities, fetal alcohol spectrum disorders, genetic risk factors, and
pharmacotherapies for AUD.
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These simplified slides by Dr. Sidra Arshad present an overview of the non-respiratory functions of the respiratory tract.
Learning objectives:
1. Enlist the non-respiratory functions of the respiratory tract
2. Briefly explain how these functions are carried out
3. Discuss the significance of dead space
4. Differentiate between minute ventilation and alveolar ventilation
5. Describe the cough and sneeze reflexes
Study Resources:
1. Chapter 39, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 34, Ganong’s Review of Medical Physiology, 26th edition
3. Chapter 17, Human Physiology by Lauralee Sherwood, 9th edition
4. Non-respiratory functions of the lungs https://academic.oup.com/bjaed/article/13/3/98/278874
- Video recording of this lecture in English language: https://youtu.be/lK81BzxMqdo
- Video recording of this lecture in Arabic language: https://youtu.be/Ve4P0COk9OI
- Link to download the book free: https://nephrotube.blogspot.com/p/nephrotube-nephrology-books.html
- Link to NephroTube website: www.NephroTube.com
- Link to NephroTube social media accounts: https://nephrotube.blogspot.com/p/join-nephrotube-on-social-media.html
MANAGEMENT OF ATRIOVENTRICULAR CONDUCTION BLOCK.pdfJim Jacob Roy
Cardiac conduction defects can occur due to various causes.
Atrioventricular conduction blocks ( AV blocks ) are classified into 3 types.
This document describes the acute management of AV block.
Pulmonary Thromboembolism - etilogy, types, medical- Surgical and nursing man...VarunMahajani
Disruption of blood supply to lung alveoli due to blockage of one or more pulmonary blood vessels is called as Pulmonary thromboembolism. In this presentation we will discuss its causes, types and its management in depth.
Lung Cancer: Artificial Intelligence, Synergetics, Complex System Analysis, S...Oleg Kshivets
RESULTS: Overall life span (LS) was 2252.1±1742.5 days and cumulative 5-year survival (5YS) reached 73.2%, 10 years – 64.8%, 20 years – 42.5%. 513 LCP lived more than 5 years (LS=3124.6±1525.6 days), 148 LCP – more than 10 years (LS=5054.4±1504.1 days).199 LCP died because of LC (LS=562.7±374.5 days). 5YS of LCP after bi/lobectomies was significantly superior in comparison with LCP after pneumonectomies (78.1% vs.63.7%, P=0.00001 by log-rank test). AT significantly improved 5YS (66.3% vs. 34.8%) (P=0.00000 by log-rank test) only for LCP with N1-2. Cox modeling displayed that 5YS of LCP significantly depended on: phase transition (PT) early-invasive LC in terms of synergetics, PT N0—N12, cell ratio factors (ratio between cancer cells- CC and blood cells subpopulations), G1-3, histology, glucose, AT, blood cell circuit, prothrombin index, heparin tolerance, recalcification time (P=0.000-0.038). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and PT early-invasive LC (rank=1), PT N0—N12 (rank=2), thrombocytes/CC (3), erythrocytes/CC (4), eosinophils/CC (5), healthy cells/CC (6), lymphocytes/CC (7), segmented neutrophils/CC (8), stick neutrophils/CC (9), monocytes/CC (10); leucocytes/CC (11). Correct prediction of 5YS was 100% by neural networks computing (area under ROC curve=1.0; error=0.0).
CONCLUSIONS: 5YS of LCP after radical procedures significantly depended on: 1) PT early-invasive cancer; 2) PT N0--N12; 3) cell ratio factors; 4) blood cell circuit; 5) biochemical factors; 6) hemostasis system; 7) AT; 8) LC characteristics; 9) LC cell dynamics; 10) surgery type: lobectomy/pneumonectomy; 11) anthropometric data. Optimal diagnosis and treatment strategies for LC are: 1) screening and early detection of LC; 2) availability of experienced thoracic surgeons because of complexity of radical procedures; 3) aggressive en block surgery and adequate lymph node dissection for completeness; 4) precise prediction; 5) adjuvant chemoimmunoradiotherapy for LCP with unfavorable prognosis.
Title: Sense of Taste
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the structure and function of taste buds.
Describe the relationship between the taste threshold and taste index of common substances.
Explain the chemical basis and signal transduction of taste perception for each type of primary taste sensation.
Recognize different abnormalities of taste perception and their causes.
Key Topics:
Significance of Taste Sensation:
Differentiation between pleasant and harmful food
Influence on behavior
Selection of food based on metabolic needs
Receptors of Taste:
Taste buds on the tongue
Influence of sense of smell, texture of food, and pain stimulation (e.g., by pepper)
Primary and Secondary Taste Sensations:
Primary taste sensations: Sweet, Sour, Salty, Bitter, Umami
Chemical basis and signal transduction mechanisms for each taste
Taste Threshold and Index:
Taste threshold values for Sweet (sucrose), Salty (NaCl), Sour (HCl), and Bitter (Quinine)
Taste index relationship: Inversely proportional to taste threshold
Taste Blindness:
Inability to taste certain substances, particularly thiourea compounds
Example: Phenylthiocarbamide
Structure and Function of Taste Buds:
Composition: Epithelial cells, Sustentacular/Supporting cells, Taste cells, Basal cells
Features: Taste pores, Taste hairs/microvilli, and Taste nerve fibers
Location of Taste Buds:
Found in papillae of the tongue (Fungiform, Circumvallate, Foliate)
Also present on the palate, tonsillar pillars, epiglottis, and proximal esophagus
Mechanism of Taste Stimulation:
Interaction of taste substances with receptors on microvilli
Signal transduction pathways for Umami, Sweet, Bitter, Sour, and Salty tastes
Taste Sensitivity and Adaptation:
Decrease in sensitivity with age
Rapid adaptation of taste sensation
Role of Saliva in Taste:
Dissolution of tastants to reach receptors
Washing away the stimulus
Taste Preferences and Aversions:
Mechanisms behind taste preference and aversion
Influence of receptors and neural pathways
Impact of Sensory Nerve Damage:
Degeneration of taste buds if the sensory nerve fiber is cut
Abnormalities of Taste Detection:
Conditions: Ageusia, Hypogeusia, Dysgeusia (parageusia)
Causes: Nerve damage, neurological disorders, infections, poor oral hygiene, adverse drug effects, deficiencies, aging, tobacco use, altered neurotransmitter levels
Neurotransmitters and Taste Threshold:
Effects of serotonin (5-HT) and norepinephrine (NE) on taste sensitivity
Supertasters:
25% of the population with heightened sensitivity to taste, especially bitterness
Increased number of fungiform papillae
1. Delhi & Chennai, India 2014
Sperm DNA Fragmentation
in Male Infertility
Sandro C. Esteves, MD, PhD
Director, ANDROFERT
Campinas, Brazil
2. Contents
Role of SDF on fertility
SDF and ART outcome
Methods for SDF assessment
Management Strategies
Esteves, 2
ANDROFERT, Referral Center for Male Reproduction
4. Why semen analysis is not enough
Esteves, 4
ANDROFERT, Referral Center for Male Reproduction
5. Sperm Function
Deliver intact DNA into
the oocyte
Intact chromatin is critical
for the production of a
viable pregnancy
DNA is a manual of
instructions
Esteves, 5
ANDROFERT, Referral Center for Male Reproduction
7. What are the lesions associated with
Sperm DNA Fragmentation?
Defects in DNA structure:
Single-strand DNA break (ss-DB)
Double-strand DNA break (ds-DB)
Base deletion or modification
Inter or intra-strand cross linkage
damaged base
single-strand
break mis-match
double-strand
break
intra-strand
crosslink
inter-strand
crosslink
Esteves et al 2013; Alvarez and Gosálbez 2011; Ward 2011
Esteves, 7
ANDROFERT, Referral Center for Male Reproduction
8. What are the biological
mechanisms of SDF?
Protamination Failure
Replacement of histone to protamines during
spermiogenesis
Oxidative Stress
Epididymis transit
Post-ejaculation: leukocytes, immature sperm,
abnormal levels seminal plasma antioxidants
Apoptosis
During sperm maturation (testis & epididymis)
Fernández et al. 2009; Alvarez and Sakkas 2010; Agarwal et al. 2013
Esteves, 8
ANDROFERT, Referral Center for Male Reproduction
9. What are the external factors
leading to increased SDF?
Environmental factors
Phtalate exposure, radiation,
temperature
Diseases
Varicocele, GTI, fever
Life-style
Obesity, smoking
DNA Damage
Aging
Kort et al. 2006; Rubes et al 2007; Viloria et al 2007; Esteves & Agarwal 2011
Esteves, 9
ANDROFERT, Referral Center for Male Reproduction
10. SDF and Male Infertility Etiologies
Gosálbez et al. 2013
Esteves, 10
ANDROFERT, Referral Center for Male Reproduction
11. Frequency of elevated SDF in men
with normal semen analysis
52
143
Normal semen analysis results (WHO)
Elevated SDF (36.4%)
Esteves, 11
ANDROFERT, Referral Center for Male Reproduction
12. SDF and Infertility: Why bother?
Esteves, 12
ANDROFERT, Referral Center for Male Reproduction
13. IUI Outcome and SDF
Live Birth Rates with
Intrauterine Insemination
19%
OR = 0.07
[95% CI: 0.01-0.48]
1.5%
Normal
Esteves, 13
Bungum et al. Hum Reprod 2007
Elevated
ANDROFERT, Referral Center for Male Reproduction
14. IVF Outcome and SDF
Pregnancy by Method in
Cases of Elevated Sperm DNA
Fragmentation
42%
Meta-analysis of 16 studies and
2,969 couples
Increased miscarriage in couples
undergoing IVF/ICSI with high
sperm DNA damage
26%
Risk ratio (RR) = 2.16
95% CI: 1.54-3.03; p<0.00001
IVF
ICSI
Robinson et al. Hum Reprod 2012
Bungum et al. Hum Reprod 2007
Esteves, 14
ANDROFERT, Referral Center for Male Reproduction
15. SDF and reproductive success
Points to consider (1)
Oocyte repair capability and severity
of damage
Repair likely to occur at pronuclei stage (prior syngamy)
Low levels breaks can be repaired (especially ss-DBs)
Repair ability decrease with female age
Menezo et al 2007; Genescá et al. 1992; Obe et al. 2002
Esteves, 15
ANDROFERT, Referral Center for Male Reproduction
16. SDF and reproductive success
Points to consider (2)
Site of damage
Coding DNA (exons)
represent ~3% of genome
Gosálbez et al. 2013; Dada et al. 2012
Esteves, 16
ANDROFERT, Referral Center for Male Reproduction
17. SDF and Male Infertility
Key Messages (1)
SDF
gives
different
informa4on
than
rou4ne
semen
analysis,
and
of
be;er
prognos4c
value
SDF
is
mainly
oxida4ve-‐stress
mediated
during
sperm
transit
through
the
epididymis
Elevated
SDF
associated
with
infer4lity,
poor
ART
outcome
and
miscarriage
Reproduc4ve
outcome
related
to
oocyte
repair
capacity
as
well
as
severity
and
site
of
DNA
damage
Esteves,
17
ANDROFERT,
Referral
Center
for
Male
Reproduc4on
18. What are the methods for SDF
assessment?
Direct
Incorporation of probes at the site of damage
e.g. TUNEL, ISNT
Indirect
Susceptibility of DBs to denature in a acid solution
e.g. Sperm chromatin structure assay (SCSA), sperm
chromatin dispersion test (SCD), Comet assay
Chromatin compaction
Incorporation of probes to nuclear proteins
e.g. Aniline blue, toluidine blue
Gosálbez et al 2013; Esteves & Agarwal 2011; Esteves et al. 2013
Esteves, 18
ANDROFERT, Referral Center for Male Reproduction
19. Comparison Between SDF Methods
Fertility and Sterility 2014; 101(1):58-63.
Esteves, 19
ANDROFERT, Referral Center for Male Reproduction
21. Sperm Chromatin Dispersion (SCD)
Susceptibility of DNA to denaturation
with formation of single-strand (ss)
DNA from pre-existing single or
double strand breaks;
Combination of DNA denaturation used
in SCSA and protein depletion used in
the comet assay;
Difference in the pattern of forming a
loop (halo) around lysed and acid
treated nuclear membrane carcass
reflects the overall chromatin structure.
Fernández et al. 2003, 2005 Gosálvez et al. 2006
Esteves, 21
ANDROFERT, Referral Center for Male Reproduction
22. Correlation between SCD and TUNEL
SCD more sensitive than
TUNEL.
Important to distinguish
between the methods as they
differently evaluate SDF.
20.6
SCD
TUNEL
11.5
Fertil Steril 2014; 101(1):58-63
% SDF
Esteves, 22
ANDROFERT, Referral Center for Male Reproduction
23. Diagnostic accuracy of SCD in men with
unexplained infertility
Despite poorly correlated, SCD
may discriminate men with
normal and abnormal sperm DNA
damage with ~70% accuracy
when compared with TUNEL.
Fertil Steril 2014; 101(1):58-63
Esteves, 23
ANDROFERT, Referral Center for Male Reproduction
24. Which is the best method for SDF?
Laborintensive
TUNEL
SCSA
Comet
SCD
Esteves, 24
Expensive
equipment
Analysis
Subjectivity
Validation
& Standardization
++++
++
++++
+
+++
++++
+++
+
++
+
+++
++
++
++++
+
+++
ANDROFERT, Referral Center for Male Reproduction
25. SDF and Male Infertility
Key Messages (2)
Several
methods
available
to
assess
SDF
Methods
differen4ally
assess
SDF
and
cannot
determine
nature
or
e4ology
of
damage
Best
method
yet
to
be
determined
SCD
is
a
quick
and
easy
assay
to
assess
SDF
Esteves,
25
ANDROFERT,
Referral
Center
for
Male
Reproduc4on
26. What can we do to reduce SDF?
Antioxidants and life-style
changes
Treatment of underlying condition
Avoid iatrogenic SDF
Wong et al., 2000; Wong et al. 2002; Comhaire and Mahmoud, 2003; Agarwal and Said, 2004;
Bansal and Bilaspuri, 2010; Gosálbez et al. 2009, 2011; Esteves et al. 2011; Sánchez-Martín et al 2013
Esteves, 26
ANDROFERT, Referral Center for Male Reproduction
27. Oral Antioxidants
Outcome
No.
studies
Effect
size
(OR;
95%
CI)
Live
birth
3
4.85
[1.92,
12.24]
Pregnancy
rate
15
4.18
[2.65,
6.59]
DNA
fragmenta4on
1
-‐13.80
[-‐17.50,
-‐10.10]
6-‐16
No
effect
Miscarriage,
sperm
count,
sperm
mo4lity
Showell MG et al. Cochrane Database Syst Rev 2011
Esteves, 27
ANDROFERT, Referral Center for Male Reproduction
28. Oral Antioxidants
Beneficial
No effect
Kodama 1997
Dawson, 1992
Kessopoulou, 1995
Vezina, 1996
Vicari, 2001; 2002
Lenzi, 2003; 2004
Cavallini, 2004
Comhaire, 2005
Grecco 2005
Menezo 2007
Tremellen 2007
Giovenco, 1987
Moilanen, 1993
Iwanier, 1995
Rolf, 1999
Sigman, 2006
Piomboni 2008
Gil Villa 2009
Esteves, 28
l Short-term use
appear to be safe
Detrimental
long-term use and high
doses;
increased mortality in
cancer populationbased studies.
l Caution against
indiscriminate use of
high dosages for
long periods
Heinonen, 1994
Lonn, 2005
Bjelakovic, 2007
ANDROFERT, Referral Center for Male Reproduction
29. Oral Antioxidants
How I prescribe
Vitamin C 500mg; Vitamin E 400 mg
Folic acid 2 mg, Zinc 25 mg
Selenium 26 mcg
Minimum 2 months
From initiation of sperm production to ejaculation
Old concept ~80 days
New concept ~60 days
Misell LM et al. J Urol. 2006
Esteves & Agarwal. Novel concepts in male infertility. Int Braz J Urol 2011
Esteves, 29
ANDROFERT, Referral Center for Male Reproduction
30. Decrease No. Leukocytes in Semen
% DNA Damage (SCSA) 39%
34%
25%
granulocyte
Normal
Abnormal
macrophage
lymphocyte
Abnl &
Leukocytospermia
Endtz
test
Henkel R et al, AJA 2007; Alvarez et al. Fertil Steril 2002
Esteves, 30
ANDROFERT, Referral Center for Male Reproduction
31. Treatment of Subclinical GTI and
Associated Inflammatory Changes
Antibiotics
• Yanushpolsky et al, 1995; Erel et al., 1997
• Branigan et al., 1995
Cicloxigenase-2
Inhibitors
Antihistamines
• Oliva & Mutigner, 2006
Antioxidants
Esteves, 31
• Lackner et al., 2006
• Gambera et al., 2007
• Tremellen et al., 2007
• Piombini et al., 2008
ANDROFERT, Referral Center for Male Reproduction
32. Subclinical Male Genital Tract Infection
Anti-bacterial
properties (Zinc)
Azitromycin 1.0g single dose (couple)+ frequent ejaculation
(every 2-3 days) + Antioxidants
42% leukocytospermia resolution (N=278)
Esteves, 32
ANDROFERT, Referral Center for Male Reproduction
33. Varicocele Surgery
Twelve studies comparing SDF
in pts. with and without
varicocele:
SDF higher in varicocele
Mean difference = 9.9%
(95% CI: 9.2-10.5; p<0.0001)
Miyaoka & Esteves. Adv Urol 2012
Agarwal, Esteves, Hamada. Nature Urol Rev 2013;
Wang YJ et al. Reprod Biomed Online. 2012;25:307-14.
Esteves, 33
ANDROFERT, Referral Center for Male Reproduction
34. Effect of Varicocele Surgery on SDF
Meta-analysis of seven studies
evaluating the effect of
varicocele repair on SDF
SDF decreased after repair
Mean difference = 3.4%
(95% CI: -4.1 to -2.6; p<0.0001)
Wang YJ et al.
Reprod Biomed Online. 2012;25:307-14.
Esteves, 34
ANDROFERT, Referral Center for Male Reproduction
36. TESA-ICSI and SDF
Sperm
% TUNEL +
% CPR
Ejaculated
23.6
6
Testicular
4.8
44
<0.001
<0.05
P value
Greco et al. Hum Reprod 2005
Esteves, 36
ANDROFERT, Referral Center for Male Reproduction
37. Difference in SDF between
Testicle and Ejaculate
DNA damage in
Testicular
Spermatozoa
three-fold lower
compared with
Ejaculated
Spermatozoa*
*Absolute differences between two specimens ranging from -3.3% to -56.3%.
Moskovtsev et al. Fertil Steril 2010
Esteves, 37
ANDROFERT, Referral Center for Male Reproduction
39. TESA-ICSI: ANDROFERT
• 93 patients enrolled; Mean age: 37.5 yo.
• SDF (Halosperm®): 39.0% ± 15.4% [range: 21%-88%]
• 53 pts. with live birth data (Sept. 2013)
80
60
40
70.5
61.5
53.6
54.8
51
p=0.21
p=0.62
20
p=0.50
25
11.7
CPR (%)
Miscarriage (%)
33.3
p=0.70
48.4
25
p=0.17
0
2PN (%)
TQE (%)
Ejaculate
Esteves, 39
LBR (%)
TESA
ANDROFERT, Referral Center for Male Reproduction
40. What can we do to decrease SDF?
Ø Oral antioxidants
Ø Life-style modifications, including quit
smoking and weight loss
Ø Identify and treat underlying condition
(GTI and varicocele)
Ø Consider TESA-ICSI
Esteves, 40
ANDROFERT, Referral Center for Male Reproduction
41. Dynamic Nature of SDF
Iatrogenic damage
Esteves, 41
ANDROFERT, Referral Center for Male Reproduction
42. Abstinence Period and Sperm
Processing
Serial ejaculation every 24h for 4 days:
25% reduction SDF
Density centrifugation post-3h
ejaculation: 44% reduction
Gosálbez et al. Fertil Steril 2011
One-day abstinence: Reduction in ~90% pts.
Esteves, 42
Pons et al. 2013
ANDROFERT, Referral Center for Male Reproduction
43. Annexin-V microbeads and
MACS columns
Microbeads conjugated with
Annexin-V
Esteves, 43
ANDROFERT, Referral Center for Male Reproduction
44. Sperm Selection
Annexin-V columns
Parameters
Pre-MACS
Annexin V neg.
Annexin V
pos.
Count
1 x 106/ml
600.000/ml
400.000/ml
Viability
49%
64%
0
Progressive
motility
30%
43%
0
SDF
34%
10%
70%
Cleaved
caspase-3
8%
7%
20%
Ø Annexin-V negative fraction: IVF, ICSI, IUI
Ø Annexin-V positive fraction: discard
Esteves, 44
Rawe et al., RBM Online 20:320, 2010
ANDROFERT, Referral Center for Male Reproduction
45. Laboratory handling of ejaculated
and testicular specimens
Esteves SC & Varghese A, J Hum Reprod Sci 2012
Esteves, 45
ANDROFERT, Referral Center for Male Reproduction
46.
47. What we can do to avoid iatrogenic SDF
Ø Short abstinence periods (1 day) and serial ejaculation
Ø Instruct patients to deliver specimens 1- 2h before
ART is to be performed
Ø Process specimens as soon as possible
Ø Keep specimens at room T using appropriate culture
media
Ø Incubation time after processing should not exceed 4h
Ø Thaw cryopreserved specimens just before performing
ART
Esteves, 47
ANDROFERT, Referral Center for Male Reproduction
48. Take-home Messages
SDF provides information that is different
and of better prognostic value than
semen analysis.
SDF mainly occur during sperm transit through
the epididymis, and it is mediated by ROS.
Oocyte can repair ssDNA but not dsDNA
damage.
Sperm chromatin dispersion (SCD) test is
the simplest method to assess SDF in
routine clinical practice.
Esteves, 48
ANDROFERT, Referral Center for Male Reproduction
49. Take-home Messages
SDF has a negative effect on reproductive
potential, both in vivo and in vitro.
Strategies to reduce SDF includes antioxidant
therapy, treatment of subclinical GTI,
varicocele repair, and TESA-ICSI.
Avoid iatrogenic damage: short abstinence
periods, laboratory sperm selection and
proper sperm handling.
Esteves, 49
ANDROFERT, Referral Center for Male Reproduction