The document discusses semen analysis and spermatogenesis. It describes the two steps of spermatogenesis as spermatocytogenesis and spermiogenesis. The seminiferous tubules of the testis are identified as the site of sperm formation. Semen is defined as the fluid ejaculated during orgasm that contains sperm and secretions from various glands. Normal values for semen parameters according to WHO are provided. The steps of semen analysis and sample collection are outlined in detail.
Sperm Function Tests are the keystones of evaluating functional condition of sperms. The fertility potential of a sperm will be decided not only with the number & motility but with the functional competence which is of utmost importance.
Semen is a “ thick, viscous, creamy, slightly yellowish or grayish” substance made up of spermatozoa — commonly known as sperm — and a fluid called seminal plasma, secret from the male reproductive organs.
The function of seminal plasma are:
To provide motility to sperm
To provide nutrition to spermatozoa
Sperm Function Tests are the keystones of evaluating functional condition of sperms. The fertility potential of a sperm will be decided not only with the number & motility but with the functional competence which is of utmost importance.
Semen is a “ thick, viscous, creamy, slightly yellowish or grayish” substance made up of spermatozoa — commonly known as sperm — and a fluid called seminal plasma, secret from the male reproductive organs.
The function of seminal plasma are:
To provide motility to sperm
To provide nutrition to spermatozoa
Intrauterine insemination (IUI) is a fertility treatment that involves placing sperm inside a woman’s uterus to facilitate fertilisation. The goal of IUI is to increase the number of sperm that reach the fallopian tubes and subsequently increase the chance of fertilisation.
TThe effect of interval from semen collection to IUI, does it matterNeelam Ohri
IUI is widely practiced modality for infertility treatment. Despite all efforts success rate has not reached beyond 16% pregnancy rate/cycle. The variation in success rate is very high as there are many variables for success like different age groups, different causes of infertility, different treatment protocols such as the nature of induction of ovulation, the capacity of processing the semen in the laboratory and insemination techniques involved in different studies.
sperm assessment- traditional and novel approaches.pptxDeepekaTS
The latest WHO recommendations,2010 are based on semen parameters from approximately 2000 fertile men, from eight countries and three continents, whose partners achieved pregnancy within 12 months of unprotected sexual intercourse.
Pitfalls- huge shift in the lower reference values, one sided criteria.
Reference limits shouldn’t be over-interpreted
Interpret along with clinical history and physical examination.
Explore natural remedies for syphilis treatment in Singapore. Discover alternative therapies, herbal remedies, and lifestyle changes that may complement conventional treatments. Learn about holistic approaches to managing syphilis symptoms and supporting overall health.
Report Back from SGO 2024: What’s the Latest in Cervical Cancer?bkling
Are you curious about what’s new in cervical cancer research or unsure what the findings mean? Join Dr. Emily Ko, a gynecologic oncologist at Penn Medicine, to learn about the latest updates from the Society of Gynecologic Oncology (SGO) 2024 Annual Meeting on Women’s Cancer. Dr. Ko will discuss what the research presented at the conference means for you and answer your questions about the new developments.
Title: Sense of Smell
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 primary categories of smells and the concept of odor blindness.
Explain the structure and location of the olfactory membrane and mucosa, including the types and roles of cells involved in olfaction.
Describe the pathway and mechanisms of olfactory signal transmission from the olfactory receptors to the brain.
Illustrate the biochemical cascade triggered by odorant binding to olfactory receptors, including the role of G-proteins and second messengers in generating an action potential.
Identify different types of olfactory disorders such as anosmia, hyposmia, hyperosmia, and dysosmia, including their potential causes.
Key Topics:
Olfactory Genes:
3% of the human genome accounts for olfactory genes.
400 genes for odorant receptors.
Olfactory Membrane:
Located in the superior part of the nasal cavity.
Medially: Folds downward along the superior septum.
Laterally: Folds over the superior turbinate and upper surface of the middle turbinate.
Total surface area: 5-10 square centimeters.
Olfactory Mucosa:
Olfactory Cells: Bipolar nerve cells derived from the CNS (100 million), with 4-25 olfactory cilia per cell.
Sustentacular Cells: Produce mucus and maintain ionic and molecular environment.
Basal Cells: Replace worn-out olfactory cells with an average lifespan of 1-2 months.
Bowman’s Gland: Secretes mucus.
Stimulation of Olfactory Cells:
Odorant dissolves in mucus and attaches to receptors on olfactory cilia.
Involves a cascade effect through G-proteins and second messengers, leading to depolarization and action potential generation in the olfactory nerve.
Quality of a Good Odorant:
Small (3-20 Carbon atoms), volatile, water-soluble, and lipid-soluble.
Facilitated by odorant-binding proteins in mucus.
Membrane Potential and Action Potential:
Resting membrane potential: -55mV.
Action potential frequency in the olfactory nerve increases with odorant strength.
Adaptation Towards the Sense of Smell:
Rapid adaptation within the first second, with further slow adaptation.
Psychological adaptation greater than receptor adaptation, involving feedback inhibition from the central nervous system.
Primary Sensations of Smell:
Camphoraceous, Musky, Floral, Pepperminty, Ethereal, Pungent, Putrid.
Odor Detection Threshold:
Examples: Hydrogen sulfide (0.0005 ppm), Methyl-mercaptan (0.002 ppm).
Some toxic substances are odorless at lethal concentrations.
Characteristics of Smell:
Odor blindness for single substances due to lack of appropriate receptor protein.
Behavioral and emotional influences of smell.
Transmission of Olfactory Signals:
From olfactory cells to glomeruli in the olfactory bulb, involving lateral inhibition.
Primitive, less old, and new olfactory systems with different path
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.
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
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.
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.
Tom Selleck Health: A Comprehensive Look at the Iconic Actor’s Wellness Journeygreendigital
Tom Selleck, an enduring figure in Hollywood. has captivated audiences for decades with his rugged charm, iconic moustache. and memorable roles in television and film. From his breakout role as Thomas Magnum in Magnum P.I. to his current portrayal of Frank Reagan in Blue Bloods. Selleck's career has spanned over 50 years. But beyond his professional achievements. fans have often been curious about Tom Selleck Health. especially as he has aged in the public eye.
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Introduction
Many have been interested in Tom Selleck health. not only because of his enduring presence on screen but also because of the challenges. and lifestyle choices he has faced and made over the years. This article delves into the various aspects of Tom Selleck health. exploring his fitness regimen, diet, mental health. and the challenges he has encountered as he ages. We'll look at how he maintains his well-being. the health issues he has faced, and his approach to ageing .
Early Life and Career
Childhood and Athletic Beginnings
Tom Selleck was born on January 29, 1945, in Detroit, Michigan, and grew up in Sherman Oaks, California. From an early age, he was involved in sports, particularly basketball. which played a significant role in his physical development. His athletic pursuits continued into college. where he attended the University of Southern California (USC) on a basketball scholarship. This early involvement in sports laid a strong foundation for his physical health and disciplined lifestyle.
Transition to Acting
Selleck's transition from an athlete to an actor came with its physical demands. His first significant role in "Magnum P.I." required him to perform various stunts and maintain a fit appearance. This role, which he played from 1980 to 1988. necessitated a rigorous fitness routine to meet the show's demands. setting the stage for his long-term commitment to health and wellness.
Fitness Regimen
Workout Routine
Tom Selleck health and fitness regimen has evolved. adapting to his changing roles and age. During his "Magnum, P.I." days. Selleck's workouts were intense and focused on building and maintaining muscle mass. His routine included weightlifting, cardiovascular exercises. and specific training for the stunts he performed on the show.
Selleck adjusted his fitness routine as he aged to suit his body's needs. Today, his workouts focus on maintaining flexibility, strength, and cardiovascular health. He incorporates low-impact exercises such as swimming, walking, and light weightlifting. This balanced approach helps him stay fit without putting undue strain on his joints and muscles.
Importance of Flexibility and Mobility
In recent years, Selleck has emphasized the importance of flexibility and mobility in his fitness regimen. Understanding the natural decline in muscle mass and joint flexibility with age. he includes stretching and yoga in his routine. These practices help prevent injuries, improve posture, and maintain mobilit
The hemodynamic and autonomic determinants of elevated blood pressure in obes...
Semen Analysis
1. Dr. Furquan Alam
PG Resident,
Department of Biochemistry,
SGRRIMHS & SMI Hospital, Dehradun.
SEMEN ANALYSIS
2. SPERMATOGENESIS
The sperm formation involves two steps :
1) Spermatocytogenesis
In the first step spermtogenic cells form rounded cells called
spermatids.
2) Spermiogenesis
In the second step spermatids differentiate into specialized
cells known as sperms.
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DR. FURQUAN ALAM 2
3. Site Of Sperm Formation
Seminiferous tubules of Testis
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DR. FURQUAN ALAM 3
6. Seminal Fluid
Semen is body fluid that is ejaculated at the time of
orgasm, contain sperm & secretion of seminal vesicle,
prostate, Cowper's gland & urethral gland.
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DR. FURQUAN ALAM 6
7. Source Volume Characteristics
Urethral and bulbourethral
glands
0.1-0.2cc Viscous, clear
Testes, epididymides, vas
deferentia
0.1-0.2cc Sperm present
Prostate 0.5-1.0cc Acidic,watery
Seminal vesicles 1.0-3.0cc Gelatinous, fructose positive
Complete ejaculate 1.5-5.0cc Liquefies in 20-25min
Semen
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DR. FURQUAN ALAM 7
8. Semen is viscous, neutral or slightly alkaline & whitish
opaque.
60 % semen volume is derived from seminal vesicle which is
also a major source of high FRUCTOSE content of semen.
Seminal vesicle secretion also provide the substrate for the
coagulation of the semen following ejaculation.
About 20 % of the volume of semen is contributed by the
prostate gland.
It is milky in appearance & also rich in proteolytic enzymes are responsible
for the liquefaction of semen.
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DR. FURQUAN ALAM 8
9. About 10 -15 % of semen volume is also contributed by
epididymis, vasdeferens, cowper’s gland & uretheral gland.
Less than 5 % of semen volume is contributed by Spermatozoa.
The process of ejaculation result in the mixing of these distinct
fraction of semen.
These enter the urethra individually in the rapid succession.
The third & final fraction consist of mucoid secretion resulting from
emptying of seminal vesicle.
The semen specimen collected for routine examination should
contain all above mention fraction.
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DR. FURQUAN ALAM 9
11. Normal Values From WHO Manuals, Editions 2- 4 And Lower
Reference Limits From New 5th WHO Manual (2010)
Semen parameter
WHO edition and year
2nd - 1987 3rd - 1992 4th - 1999 5th - 2010
Volume (ml) 2.0 2.0 2.0 1.5
Sperm concentration (106/ml) 20 20 20 15
Total sperm count (106) 40 40 40 39
Motility (% progressive) 50 50 50 32
Vitality (% live) 50 75 75 58
Morphology (%
normal)
50 30 (15) 4
3/13/2016 8:00:38 PM DR. FURQUAN ALAM
11
12. Steps: Semen Analysis
In the first 5 minutes:
Placing the specimen container on the bench or in an
incubator (37 °C) for liquefaction.
Between 30 and 60 minutes:
Assessing liquefaction and appearance of the semen.
Measuring semen volume.
Measuring semen pH (if required).
Preparing a wet preparation for assessing microscopic
appearance, sperm motility and the dilution required for
assessing sperm number.
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DR. FURQUAN ALAM 12
13. Steps Of Semen Analysis
Between 30 and 60 minutes:
Assessing sperm vitality (if the percentage of motile cells is
low).
Making semen smears for assessing sperm morphology.
Making semen dilutions for assessing sperm concentration.
Assessing sperm number.
Performing the mixed antiglobulin reaction (MAR) test (if
required).
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14. Steps
Between 30 and 60 minutes:
Assessing peroxidase-positive cells (if round cells are present).
Preparing spermatozoa for the immunobead test (if required).
Centrifuging semen (if biochemical markers are to be assayed).
Within 3 hours:
Sending samples to the microbiology laboratory (if required).
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15. Steps
After 4 hours:
Fixing, staining and assessing smears for sperm
morphology.
Later on the same day (or on a subsequent day if
samples are frozen):
Assaying accessory gland markers (if required).
Performing the indirect immunobead test (if
required).
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DR. FURQUAN ALAM 15
16. Sample Collection:
Preparation:
The sample should be collected in a private room near
the laboratory, in order to limit the exposure of the
semen to fluctuations in temperature and to control the
time between collection and analysis.
The sample should be collected after a minimum of 2
days and a maximum of 7 days of sexual abstinence. If
additional samples are required, the number of days of
sexual abstinence should be as constant as possible at
each visit.
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DR. FURQUAN ALAM 16
17. Sample Collection:
Preparation:
The man should be given clear written and spoken instructions
concerning the collection of the semen sample. These should
emphasize that the semen sample needs to be complete and
that the man should report any loss of any fraction of the
sample.
The following information should be recorded on the report
form: the man’s name, birth date and personal code number,
the period of abstinence, the date and time of collection, the
completeness of the sample, any difficulties in producing the
sample, and the interval between collection and the start of the
semen analysis.
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DR. FURQUAN ALAM 17
18. Sample Collection for Diagnostic/ Research purpose:
The sample should be obtained by masturbation and ejaculated into
a clean, wide-mouthed container made of glass or plastic, from a
batch that has been confirmed to be non-toxic for spermatozoa.
The specimen container should be kept at ambient temperature,
between 20 °C and 37 °C, to avoid large changes in temperature
that may affect the spermatozoa after they are ejaculated into it. It
must be labelled with the man’s name and identification number, and
the date and time of collection.
The specimen container is placed on the bench or in an incubator
(37 °C) while the semen liquefies.
Note in the report if the sample is incomplete, especially if the first,
sperm-rich fraction may be missing. If the sample is incomplete, a
second sample should be collected, again after an abstinence period
of 2–7 days.
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19. Sterile collection of semen for assisted reproduction:
This is performed as for diagnostic collection but the
specimen containers, pipette tips and pipettes for
mixing must be sterile.
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20. Sterile collection of semen for microbiological analysis:
In this situation, microbiological contamination from non-semen sources
(e.g. commensal organisms from the skin) must be avoided. The
specimen containers, pipette tips and pipettes for mixing must be sterile.
The man should:
Pass urine.
Wash hands and penis with soap, to reduce the risk of contamination of
the specimen with commensal organisms from the skin.
Rinse away the soap.
Dry hands and penis with a fresh disposable towel.
Ejaculate into a sterile container.
Note: The time between collection of the semen sample and the start of
the investigation by the microbiological laboratory should not exceed 3
hours.
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21. Collection of semen at home:
A sample may be collected at home in exceptional circumstances, such as a
demonstrated inability to produce a sample by masturbation in the clinic or the
lack of adequate facilities near the laboratory.
The man should be given clear written and spoken instructions concerning the
collection and transport of the semen sample. These should emphasize that the
semen sample needs to be complete, i.e. all the ejaculate is collected, including
the first, sperm-rich portion, and that the man should report any loss of any
fraction of the sample. It should be noted in the report if the sample is incomplete.
The man should be given a pre-weighed container, labelled with his name and
identification number.
The man should record the time of semen production and deliver the sample to
the laboratory within 1 hour of collection.
During transport to the lab., the sample should be kept between 20 °C - 37 °C.
The report should note that the sample was collected at home or another
location outside the laboratory.
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DR. FURQUAN ALAM 21
22. Collection of semen by condom:
A sample may be collected in a condom during sexual intercourse only in
exceptional circumstances, such as a demonstrated inability to produce a sample
by masturbation.
Only special non-toxic condoms designed for semen collection should be used;
such condoms are available commercially.
The man should be given information from the manufacturer on how to use the
condom, close it, and send or transport it to the laboratory.
The man should record the time of semen production and deliver the sample to
the laboratory within 1 hour of collection.
During transport to the lab., the sample should be kept between 20 °C and 37 °C.
The report should note that the sample was collected by means of a special
condom during sexual intercourse at home or another location outside the
laboratory.
Note: Ordinary latex condoms must not be used for semen collection because they
contain agents that interfere with the motility of spermatozoa (Jones et al., 1986).
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DR. FURQUAN ALAM 22
23. Safe handling of specimens:
Semen samples may contain dangerous infectious agents
(e.g. human immunodeficiency virus (HIV), hepatitis
viruses or herpes simplex virus) and should therefore be
handled as a biohazard. If the sample is to be processed
for bioassay, intra-uterine insemination (IUI), in-vitro
fertilization (IVF) or intracytoplasmic sperm injection (ICSI),
or if semen culture is to be performed, sterile materials and
techniques must be used. Safety guidelines as outlined in
Appendix 2 should be strictly followed; good laboratory
practice is fundamental to laboratory safety (WHO, 2004).
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DR. FURQUAN ALAM 23
24. Precautions
Specimen should not be collected in ordinary condoms
since the powder or lubricant applied to the condom
may be spermicidal.
The container in which the semen sample is collected
should be free from detergent.
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25. Storage
The semen specimen should be examined immediately
after collection.
It should be kept at room temperature.
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26. Indications For Analysis:
To determine the fertility of the man.
After a male has undergone vasectomy to check the
completeness of the procedure (retrograde ejaculation).
In medico-legal situations such as disputes about the
paternity of a child.
After reversal of vasectomy to confirm the success of the
procedure.
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27. History To Be Noted:
Name
Date and time of collection
Length of abstinence
Interval between collection and analysis
History of fever
Drug intake
Alcohol abuse
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30. NOTE:
Prolonged abstinence will lead to increased volume, but
reduced motility.
The patient should evacuate his bladder before specimen
collection. If retrograde ejaculation is suspected, a post-
ejaculate urine sample is collected and examined for presence
of sperms.
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31. Assessment Of Semen:(according To WHO)
Standard tests:
1. Volume
2. pH
3. Sperm concentration
4. Total sperm count
5. Motility
6. Morphology
7. Vitality
8. White blood cells
9. Immunobead test
10.MAR test
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32. Optional tests:
Alpha galactosidase (neutral): 20mU or more.
Zinc (total): 2.4 micromol or more
Citric acid(total): 52 micromol or more
Acid phosphatase: 200U or more
Fructose: 13 micromol or more
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33. Volume:
Measured by aspirating into a pipette or by using a syringe(non-toxic 1,2 or
5ml): Normal: 1.5ml or more.
Low volume: <1.5ml
B/l ejaculatory duct obstruction
B/l congenital vasal aplasia
Inadequate erection & improper mood at collection
Incomplete collection
High volume: >10ml: Dilutional Oligozoospermia
Aspermia:
Absence of ejaculate
Retrograde ejaculation
Anejaculation
B/l ejaculatory duct obstruction
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DR. FURQUAN ALAM 33
34. Colour:
Homogenous grey opalescent appearance (Normal).
After prolonged abstinence, slightly yellow.
Deep yellow- Pyospermia.
Rust colour - Small bleedings in seminal vesicles.
Red or brown indicates presence of blood.
Trauma to the genital tract.
Inflammation.
Tumor of the genital tract.
Increased turbidity indicates inflammatory process in some part
of the reproductive tract.
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35. Viscosity:
Freshly ejaculated semen is highly viscous due to
substrate produced by seminal vesicles. The
coagulum liquefies spontaneously to form a
translucent, viscous fluid in a three stage process.
Action of a prostatic clotting enzyme.
Liquefaction is initiated by enzymes of prostatic
origin.
Protein fragments are further degraded into free
amino acids and ammonia.
Failure to liquefy indicates inadequate prostatic
secretion. To liquefy, add bromeline, plasmin or
chymotrypsin.
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DR. FURQUAN ALAM 35
36. Normal liquefaction time: 20-60min
Viscosity of liquefied semen can be
estimated by: Gentle aspiration into a 5ml pipette, then
allowing the semen to drop by gravity. Observe the length of the
thread.
Normal semen leaves as small discrete drops.
Increased viscosity is associated with poor
invasion of cervical mucus in post-coital studies as well as
decreased ability to fertilize ovum.
Absence of viscosity points to reduced cell content.
The semen from males with b/l congenital absence of vas
deferentia & seminal vesicles fails to coagulate due to absence of
substrate.
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DR. FURQUAN ALAM 36
37. pH:
Measured by using a pH meter or pH paper.
Normal: 7.2-8
Semen is the strongest buffer in the body.
Seminal vesicle & vas deference secretions alkaline
Prostatic secretion acidic(due to citric acid, proteolytic enzymes,
acid phosphatase)
Motility is reduced in acidic medium.
pH<7 is associated with largely prostatic secretions due to
congenital aplasia of vas & seminal vesicles and when contaminated
with urine.
pH>8 is associated with acute infection of prostate, seminal vesicles
or epididymis.
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38. Sperm Concentration:
WBC Micropipette Semen-0.5mark
Diluting Fluid- 11 mark
Charge in counting chamber.
Count the number in four corner squares.
Sperm/ml = Nx10x20x1000 = Nx50,000
4
If it is very viscid, add mucolytic agent in 1:1 dilution
and multiply by 2.
If count is high(>100m), then use higher dilution.
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DR. FURQUAN ALAM 38
39. Composition of diluting fluid:
1. Sodium bicarbonate- 5g. Counteracts mucus and allows even
dilution of this viscous fluid.
2. Phenol-1ml. Kills sperms and stops their movement. Also acts
as preservative.
3. Distilled water-100ml.
A man should not be termed oligozoospermic until atleast 3
samples are evaluated at an interval of 3wks and 3 months.
Azoospermia is a condition where the semen sample has no
spermatozoa in a fresh sample or in a centrifuged resuspended
sample.
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DR. FURQUAN ALAM 39
41. Motility:
Routinely used technique:
Place a drop of liquefied semen on a glass slide.
Cover with cover slip and rim its edges with Vaseline.
Examine under 40X with reduced illumination.
Count the number actively motile sperms out of 200.
Calculate the percentage.
For accuracy:
Cover slip- 22mm x 22mm
Semen- 10µl , depth of 20µm.
Phase contrast microscopy – 400x/600x at 37 °C.
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DR. FURQUAN ALAM 41
43. Viability:
If motility is < 40% a viability should be performed.
Supravital staining by eosin Y with nigrosin. Dead cells take up the
stain. 100 sperms are counted. Live/Dead sperm ratio calculated.
Hypo osmotic saline test (HOS):
Sperms are added to hypo osmotic saline and incubated at 37deg C.
Swelling of the sperm tail is examined under phase contrast
microscope. Sperms which have active membrane swell after
30mins.
Many immotile live sperms direct towards immotile cilia syndrome.
Do electron microscopy.
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DR. FURQUAN ALAM 43
44. Normal Motility: > 32%
If less than this, asthenozoospermia.
Causes:
Cold
Radiation
Spermicides, Pesticides
Prolonged heat exposures
Prolonged abstinence
Autoimmunity
Progressive loss of motility by 5%/hr after 3hrs.
After liquefaction or
within 60mins after
ejaculation.
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DR. FURQUAN ALAM 44
45. Sperm Morphology:
Thin smears similar to blood smears are made feathering
technique.
Before staining, mucoid material is removed by gentle washing with
semen dilution fluid. Then wash gently with buffered distilled water.
Several staining techniques are used
1. Pap is the best.
2. Haematoxylene technique.
3. Giemsa.
4. Leishman/basic fuchsin (0.25% acqeous).
5. Crystal violet.
6. Diff-Quick stain
On basic fuchsin: Sperm head caps: light blue.
Nuclear post: dark blue.
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DR. FURQUAN ALAM 45
46. Sperm morphology: Ideal spermatozoon
Menkveld et al. 1991, WHO 1999, ESHRE 2002
Head oval shaped regular contour
Length: 4-5.5 micron
Width 2.5-3.5 micron
Darker posterior region
Base of head should be broad
Single tail symmetrically attached
BORDERLINE FORMS =
ABNORMAL
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DR. FURQUAN ALAM 46
47. Computer Aided Sperm Analysis:
Automation seeks to establish standard methods of
analysis and set acceptable levels of accuracy in
measuring various parameters to promote
interlaboratory comparisons.
Advantages:
Subjective errors avoided
Motility can be assessed quantitatively
Capable of handling many samples without undergoing
fatigue
Morphological defects can be made out
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DR. FURQUAN ALAM 47
48. Agglutination Of Sperms:
Motile sperms stick to each other in various
orientations like- head to head, midpiece to midpiece,
tail to tail or in combinations depending on the
specificity of antisperm antibodies. Agglutination
points to immunological cause of infertility.
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DR. FURQUAN ALAM 48
50. Antisperm Antibodies:
Can occur in the:
a)Serum of male or female
b)Seminal plasma
c)Spermatozoa
Effects:
a)Lowered progressive motility.
b)Decreased ability to penetrate cervical mucus.
c)Decreased ability to penetrate egg.
Antibodies are found to react with:
a)The front part of acrosome.
b)Post-nuclear cap
c)Tail piece
d)Equatorial part of acrosome
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DR. FURQUAN ALAM 50
51. Antisperm antibodies are found in following
conditions:
Testicular disease
Autoimmune azoospermatogenesis
Following vasectomy
Repeated infections
Obstruction of ducts
Cryptorchidism
Varicocele
Testicular biopsy
Trauma
Torsion
Genetic predisposition
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DR. FURQUAN ALAM 51
52. Techniques of detecting antibodies:
Agglutination
Immobilization
Precipitation
Complement fixation
Passive hemagglutination
Cytotoxicity
For screening,
Mixed antiglobulin reaction (MAR) test
Immunobead method
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DR. FURQUAN ALAM 52
53. Other Cells:
Round cells :
1) Germinal cells (single or double highly condensed nucleus with
abundant cytoplasm).
2) Leucocytes < 1 million/ml or 1-2/hpf. Increased no. in infection of
reproductive tract.
RBCs : normally absent.
Present in
1. TB of seminal vesicles
2. rupture of blood vessels
3. Infection of prostate
4.Vit. C deficiency
Epithelial cells from urogenital tract.
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DR. FURQUAN ALAM 53
55. Determination of fructose:
Procedure:
Pipette 5 ml of resorcinol reagent in a test tube. [Resorcinol reagent:
resorcinol-50mg, Conc.HCl-33ml, distilled water-67ml]
Add 0.5 ml of semen.
Mix and place in a boiling waterbath for 5min or heat.
Observations:
Red colored ppt. in 30 seconds.
In quantitative assays, this is compared with a known fructose standard at
490nm.
Normal level of fructose: 150-300mg/dl.
Reduced levels:
Seminal vesicle dysfunction.
High sperm count
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DR. FURQUAN ALAM 55
56. Microbiological Assays:
Indications:
1) Accessory gland infection.
2) High number of leucocytes in semen(>1million/ml)
Precautions should be taken to avoid contamination.
Culture should be done to detect both aerobic & anaerobic
organisms.
If >1000CFUs/ml, then do antibiotic sensitivity tests.
E. Coli can cause sperm agglutination and immobilization. This
is mediated by mannose and mannose binding cell surface
structures present on both cell types.
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DR. FURQUAN ALAM 56
57. Sperm Function Tests:
Factors responsible for defective sperm function:
Peroxidase damage by excessive generation of reactive oxygen
species.
High activity levels of creatine phosphokinase and a low ratio of
muscle( CK-M) to the combined activities of muscle and brain isoforms
( CK-MB ) abnormal activities of mid piece.
The tests include:
a. Sperm penetration assay (SPA)
b. Hemizona assay
c. Acrosin assay
d. Hypo osmotic saline test
e. Cervical mucous penetration test
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58. a. Sperm penetration assay:
Uses zona denuded golden hamster eggs as hosts for
the penetration of human sperms. This measures
Sperm capacitation
Sperm oocytes fusion
Sperm incorporation in oocytes
Decondensation of sperm chromatin
b. Hemizona assay:
Utilizes unfertilized oocytes
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DR. FURQUAN ALAM 58
59. c. Acrosin assay:
Measures acrosin, a trypsin like serine protienase
specific to sperm acrosome. It is responsible for sperm
penetration of the zona pellucida, after its release is
triggered by the binding of sperm to zona.
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DR. FURQUAN ALAM 59
60. D. Hypo-osmotic Swelling Test:
For successful union test of the spermatozoa with female
gametes, integrity of sperm membrane is very essential.
Sperm capacitation, acrosomal reaction & penetration of
egg is also dependent upon membrane integrity of
spermatozoa.
Therefore assessment of membrane function is a useful
indicator of fertilizing ability of spermatozoa.
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DR. FURQUAN ALAM 60
61. E. Cervical mucous penetration test/ Post coital test/ Sims-
Huhner test:
Aims :
To study the quality of cervical mucous.
To know the ability of spermatozoa to penetrate the cervical mucous and
maintain activity.
After 8-10 hrs of coitus during the ovulatory phase, the endocervical
mucous is collected in a Luer syringe. The volume, colour and viscosity
(Spinbarkeit ) of the mucous noted. A drop of mucous is placed on a slide
and examined for the presence of sperms.
Atleast 10 motile sperms should be present per hpf.
The material should be examined for leucocytes, erythrocytes and
trichomanads.
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62. Examination For The Presence Of Sperms In
Medicolegal Cases:
Obtaining the sample:
1. From vagina: direct aspiration or saline lavage.
2. From clothing or other fabrics: preliminary scan with UV light
green fluorescence 1sq cm of stained fabric soak in 1-2ml of
physiological saline for 1hr. Then fluid is subjected to tests.
Tests:
1)Examination for sperms:
Direct smears from vagina
Smears from aspirate
Washings from fabric after centrifugation Stain with H & E.
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64. 2) Determination of acid phosphatase:
More sensitive 2500 king armstrong units
3) Blood group substances
4) Florence test: screening method
Depends on presence of cholines
5) Precipitin test: Using specific antiserum by capillary tube
reaction.
6) Determination of sperm specific LDH isoenzyme.
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DR. FURQUAN ALAM 64