Lucrarile practice de embriologie se vor desfasura pe parcursul procesului de invatamant urmand ca examenul practic sa fie sustinut in sesiunea de examene:

1. Pentru anul I se vor organiza 2 lucrari practice pentru fiecare semestru cu durata de 3 ore iar pentru anul II, 2 lucrari practice cu durata de 2 ore semestrul I) si 1 lucrare practica de 2 ore (semestrul II). Orarul lucrarilor practice se va stabili in prima saptamana a fiecarui semestru iar programarea pe serii si grupe va fi afisata pe site- ul www.embriologie.ro

Fiecare lucrare practica cuprinde 2 parti:

O parte in care se explica aplicabilitatea clinica si practica a notiunilor predate la curs. Inaintea fiecarei lucrari practice studentul are obligatia de a se familiariza cu informatiile cursului !!!!

A doua parte este dedicata microscopului optic si explicarea sectiunilor de microscopie.

Activitatea fiecarui student din timpul semestrului va fi notata si va influenta nota examenului final.

Prezenta - este obligatorie la toate lp-urile. Orice absenta (motivata) trebuie recuperata.

In sesiunea de recuperari organizata in ultimile 2 saptamani ale fiecarui semestru. Absentele nerecuperate nu permit intrarea in examenul practic de embriologie si de aici mai departe in examenul de anatomie. Programarea recuperarilor va fi afisata pe siteul www.embriologie.ro pentru fiecare serie in parte.

Examenul la embriologie se noteaza cu puncte intre 1 si 10. Acestea sunt distribuite astfel:

5 puncte la examenul scris (2 subiecte teoretice fiecare notat cu cate 2,5 puncte)

4 puncte la examenul microscopic (recunoasterea si EXPLICAREA lamei)

1 punct - activitatea din timpul semestrului

Data examenului va fi afisata pe situl www.embriologie.ro. Nota minima de trecere este 5.

Examenul are caracter eliminatoriu (absenta la examen sau o nota mai mica de 5 nu permite trecerea in urmatoarea proba a examenului de anatomie).

De asemenea orice absenta la lucrarile practice din timpul semestrului nerecuperata nu premite accesul in examenul practic. 

Structura anului universitar 2015-2016 -Semestrul II  (14 saptamani)

29.02.2016- 29.04.2016 ( 9 saptamani) – activitate didactica

02.05.2016- 06.05.2016 (1 saptamana) – vacanta 

09.05.2016-10.06.2016 (5 saptamani) – activitate didactica

13.06.2016-08.07.2016 (4 saptamani) – sesiune examene

09.07.2016-31.08.2016 -vacanta de vara

01.09.2016- 25.09.2016 -restante

15.09.2016-25.09.2016 - mariri

Impartirea seriilor pe asistenti (embriologie):

Anul I	Asistent embriologie
Seria 1	Dr. Rusu Ioana
Seria 2	Dr. Negoi Ruxandra
Seria 3	Dr. Ilie Angela
Seria 4	Dr. Negoi Ruxandra
Seria 5	Dr. Rusu Ioana
Seria 6	Dr. Rusu Ioana
Seria 7	Dr. Negoi Ruxandra
Seria 8	Dr. Ilie Angela
ETG seria1	Dr. Tulin Raluca
ETG seria2	Dr. Tulin Raluca

 

Anul II
Seria 1	Dr. Rusu Ioana
Seria 2	Dr. Ilie Angela
Seria 3	Dr. Ilie Angela
Seria 4	Dr. Ilie Angela
Seria 5	Dr. Rusu Ioana
Seria 6	Dr. Tulin Raluca
Seria 7	Dr. Negoi Ruxandra
Seria 8	Dr. Negoi Ruxandra
ETG seria1	Dr. Tulin Raluca
ETG seria2	Dr. Tulin Raluca

Programa lucrarilor practice embriologie – AN I SEM I

Preparate de microscopie optica:

1. Sectiune prin ovar;

2. Sectiune prin testicul;

3. Morula.Blastula;

4. Sectiune transversala embrionara – evidentierea derivatelor foitelor embrionare;

5. Cordon ombilical;

6. Sectiune uter (sobolanca gestanta) – evidentierea placentei si a celorlalte anexe fetale;

7. Sectiune sac vitelin;

 Notiuni teoretice -LP1

1. Factori de reglare a gametogenezei – FSH, LH, Inhibina – celule Leydig, celule Sertoli, celulele tecii foliculare interne, celulele stratului granulos folicular - definitii si implicatii clinice

2. Ciclul ovarian, ciclul uterin – implicatii clinice.

3. Teste de infertilitate masculina – spermograma, test MAR, test Halosperm- definitii, principii, utilitate clinica.

4. Infertilitate feminina

5. Metode contraceptive.

Notiuni teoretice -LP2

6. hCG si testul de sarcina.

7. Celule germinale primordiale, implicatii clinice – tumori germinale.

8. Anomalii placentare – mola hidatiforma; coriocarcinom.

9. Teste uzuale pentru determinarea malformatiilor fetale – dublu test, triplu test, amniocenteza, biopsia de vilozitati coriale

 

 

Programa lucrarilor practice embriologie – AN I SEM II

Preparate microscopie optica:

1. Diverticul laringo-traheal

2. Muguri pulmonari

3. Cord si plamani fetali - sectiune transversala, frontala, sagitala

4. Arcuri branhiale - sectiune transversala; frontala

5. Cavitate bucala primitiva

6. Cavitate bucala definitiva

7. Timus fetal

 

Notiuni teoretice LP1

1. Anomalii faciale - implicatii clinice

2. Anomalii tiroidiene - implicatii clinice

Notiuni teoretice LP2

1.Circulatia fetala. Modificari circulatorii postnatale.

2. Hematopoeza embrionara. Eritropoeza si sinteza de hemoglobina.

3. Anomalii cardiace - clasificare, mecanisme embriologice, implicatii clinice

4. Notiuni de dinamica respiratorie i.u. si postnatal

5. Anomalii respiratorii frecvente - implicatii clinice

 

Programa lucrarilor practice embriologie – AN II SEM I

Preparate microscopie optica:

1.Sectiune sagitala fetala -cavitati seroase, diaphragm;

2.Stomac -pozitie intermediara;

3.Sectiune transversala embrionara -structura hepatica primitiva;

4.Sectiune transversala fetala - structura hepatica definitiva;

5.Evolutia mugurilor pancreatici;

6.Hernie fiziologica

7.Pronefros, Mezonefros, Metanefros;

8.Creasta genitala, gonada primitiva;

Aplicatii clinice ale notiunilor de embriologie prezentate la curs LP 1 

Duct hepatic accesor. Atrezie biliara extrahepatica.
Hernia fiziologica.Omfalocel congenital. Gastroschizis.Anomalii de pozitie si structura intestin subtire.
Diverticuli intestin subtire ; intestin gros; Maladia Hirschprung; Anomalii ano-rectale.

Aplicatii clinice ale notiunilor de embriologie prezentate la curs LP 2

Anomalii renale de pozitie, numerice, de forma.Rinichi polichistic.

Testicul ectopic. Hernie inghinala congenitala. Hidrocel. Criptorhidism. Sindrom adrenogenital.

Anomalii peniene: bifid, dublu, micropenis, agenezie. Epispadias. Hipospadias - implicatii clinice.

Implicatii clinice si utilitatea MIF (Mullerian Inhibiting Factor) in dezvoltarea cailor genitale la ambele sexe.

Vestigii embrionare. Anomalii uter, trompe uterine, vagin - implicatii clinice

 

Programa lucrarilor practice embriologie – AN II SEM II

Notiuni teoretice LP1

1. Formare glob ocular
2. Anomalii oculare - implicatii clinice 3. Formare si evolutie a urechii interne. 4. Anomalii otice - implicatii clinice

Preparate microscopie optica LP1

1. Cupa optica  si Glob ocular fetal

3. Otocisst 

­

Grupa/assist SERIA 4/ AN1	Data	ora
Dr Bulescu/ 31,32	21.11/05.12	10-13
Dr Cristea/33,34,36	24.11/8.12	17-20
Dr Stroica/35,38,40	24.11/08.12	8-11
Dr Zamfir/37,39	28.11/12.12	15-18

 

Grupa/assist SERIA 3/ AN1	Data	ora
Dr Breazu/21,22,23	23.11/07.12	13-16
Dr Ursut/ 24,25	21.11/05.12	16-19
Dr Oprea/ 26,27	24.11/8.12	14-17
Dr Pantu/ 28,29,30	23.11/07.12	8-11

 

Grupa/assist SERIA 8/ AN1	Data	ora
Dr Ungureanu/ 71,72,73	22.11/06.12	17-20
Dr Breazu/ 74,75	21.11/05.12	13-16
Dr Trancau/ 76,77	24.11/08.12	11-14
Dr Proca/Dr Diaconescu 78,79,80	20.11/04.12	14-17

 

Grupa/assist SERIA 2/ AN2	Data	ora
Dr Chirculescu/ 11,12	22.11/5.12	12-14
Dr Stanescu/ 13,14	23.11/07.12	10-12
Dr Marinescu/ 15,16	21.11/05.12	18-20
Dr Dumitru/Dr Bistran/ 17,18,19	23.11/6.12	8-10

 

Grupa/assist SERIA 6/ AN2	Data	ora
Dr Stanciulescu/48,49,50	20.11/04.12	18-20
Dr Tulin/51,52,53	20.11/04.12	12-14
Dr Cristescu/54,55	22.11/06.12	14-16
Dr Mihalea/ 56,57	22.11/06.12	16-18

First year – Seria 1, Practical Lab 2

Monday, 13^th November, 9:00 – 12:00 a.m. – Groups 3, 5, 7

Monday, 17^th November, 9:00 – 12:00 a.m. – Groups 2, 8

Monday, 11^th December, 9:00 – 12:00 a.m. – Groups 1, 4, 6

 

First year – Seria 2, Practical Lab 2

Tuesday, 14^th November, 5:30 – 8:00 p.m. – Groups 9, 10, 11

Tuesday, 28^th November, 5:30 – 8:00 p.m. – Groups 14, 15, 16

Tuesday, 12^th December, 5:30 – 8:00 p.m. – Groups 12, 13

 

First Year – Seria 3, Practical Lab 2

Monday, 13^rd November, 5:00 – 8:00 p.m. – Groups 17,18,19

Monday, 27^th November, 5:00 – 8:00 p.m. – Groups 20, 21

Monday, 11^th December, 5:00 – 8:00 p.m. – Groups 22, 23

 

Lp 2 S7, anI

6.12 ora 13-16 gr. 63, 63( dr. E. Ungureanu)

13.12 ora13-16 gr. 67, 68( dr. O. Trancau)

10.01.ora 13-16 gr. 65, 66( dr. I. Giuvarasteanu) si gr. 69( dr. I. Proca)

17.01. Ora 13-16 gr. 61, 62( dr. C. Stanescu)

         

                                       Dr. Angela Ilie

/kcfinder/upload/files/subjects.FB.pdf

EMBRYOLOGY EXAM SUBJECTS- FIRST YEAR, SECOND SEMESTER

1. Neurocranium in mammals
2. Desmocranium
3. Chondrocranium
4. Chondrocranium-chordal segment
5. Chondrocranium-prechordal segment
6. Development of the hypophysis
7. Branchial region-general features
8. First pharyngeal arch
9. Second pharyngeal arch
10. Third pharyngeal arch
11. Fourth and sixth pharyngeal arches
12. Pharyngeal clefts
13. Pharyngeal pouches
14. Development of the tongue
15. Development of the face
16. Development of the palate
17. Abnormalities in development of the face
18. Development of the larynx
19. Formation and evolution of the lung buds
20. Pseudoglandular period in lung development
21. Canalicular period in lung development
22. Terminal saccular period in lung development
23. Alveolar period in lung development
24. Abnormalities in the development of the respiratory system
25. Formation and evolution of the primitive heart tube
26. Subdivisions of heart tube and their evolution
27. Septum formation in the common atrium and in the atrioventricular canal
28. Septum formation in the common ventricle and in the conus cordis (bulbus cordis)
29. Abnormalities in the development of the cardiovascular system
30. Formation of the aortic arches
31. Evolution of the third aortic arch
32. Evolution of the fourth aortic arch
33. Evolution of the sixth aortic arch
34. Abnormalities in the evolution of the arterial arches

Subjects for practical embryology exam, first semester, RALUCA TULIN, MD, PhD

1.Spermatogenesis and blood testes barrier

2.Spermiogenesis. Sperm characteristics

3.Oogenesis and arrested prophase

4.Follicular cycle

5.Ovulation. Corpus lutheum. Corpus albicans

6.Endometrial (uterine) cycle

7.Trial of gametes in the female genital tract

8.Insemniation. Capacitation. Acrosome reaction

9.Fertilization. Superfecundation. Superfetation.

10.Cleavege. Implantation. Decidual reaction.

11.Second week - the evolution of the embryonic disck

12.Second week - the evolution of trophoblast

13.Third week – development of the intraembryonic mesoderm and formation of the notochord

14.Third week - evolution of trophoblast

15.Formarea and evolution of neural plate

16.Embryonic layers- main derivatives

17.Somites – formation and development

18. Formation	of blood and	circulatory system. Formation of
intraembryonic	and extraembryonic cavity.

19.Fetal period - general features

20.Amniotic cavity –development, evolution, role, characteristics of amniotic fluid

21.Fetal annexes: umbilical cord, yolk sac, allantois

22.Placenta- formation, external configuration

23.Placenta - structure

24.Placenta - functions

25.Development of the limbs. Malformations.

26. Primordial germ cells- clinical implications (germinal tumors)

27. Regulation of spermatogenesis.

28. Determination of ovulation

29. Male infertility tests

30. MAR (Mixed Antiglobulin Reaction) TEST and Halosperm

31. Female contraception

32. Human Chorionic Gonadotropin and pregnancy test

33. Choriocarcinoma

34. Usual tests for fetal malformations

35. Assisted reproduction technology (ART)

 

 

 

 

26. Primordial germ cells- clinical implications (germinal tumors)

 

Primordial germ cells (PGC) appear among endoderm in the dorsal part of the yolk sac. They migrate along dorsal mesentery of the hindgut (posterior part of the primitive gut) into the gonadal ridge (located on the posterior wall of celomic cavity) and induce formation of the gonad (ovary or testis). The gonad development is related with the type of the chromosomes of the PGC:

• PGC  that carry 46,XX – stimulate development of the ovary
• PGC  that carry 46,XY – stimulate development of the testis

PGC transform into sperm cells or oocytes.

Germ cell tumors (germinal tumors) are malignant (cancerous) or nonmalignant (benign, noncancerous) tumors that are comprised mostly of germ cells. Germ cells are the cells that develop in the embryo (fetus, or unborn baby) and become the cells that make up the reproductive system in males and females. These germ cells follow a midline path through the body after development and descend into the pelvis as ovarian cells or into the scrotal sac as testicular cells. Most ovarian tumors and testicular tumors are of germ cell origin. The ovaries and testes are called gonads.

 

Tumor sites outside the gonad are called extragonadal sites. The tumors also occur along the midline path and can be found in the head, chest, abdomen, pelvis, and sacrococcygeal (lower back) area.

Germ cell tumors are rare. Germ cell tumors account for about 2 -4 % of all cancers in children and adolescents younger than age 20.  Germ cell tumors can spread (metastasize) to other parts of the body. The most common sites for metastasis are the lungs, liver, lymph nodes, and central nervous system. Rarely, germ cell tumors can spread to the bone, bone marrow, and other organs.

 Symptoms vary depending on the size and location of the tumor:

• A tumor, swelling, or mass that can be felt or seen
• Elevation of alpha-fetoprotein (AFP) and  beta-human chorionic gonadotropin (ß-HCG)
• Constipation, incontinence, and leg weakness if the tumor is in the sacrum compressing structures
• Abdominal pain or abnormal shape, or irregularity in, testicular size
• Shortness of breath or wheezing if tumors in the chest are pressing on the lungs

 The most common types of germ cell tumors include:

1. Teratomas. Teratomas contain cells from the three germ layers: ectoderm, mesoderm, and endoderm. Teratomas can be malignant or benign, depending on the maturity and other types of cells that may be involved. Teratomas are the most common germ cell tumor found in the ovaries. Sacrococcygeal (tail bone, or distal end of spinal column) teratomas are the most common germ cell tumors found in childhood. Because these sacrococcygeal tumors are often visible from the outside of the body, diagnosis is made early and treatment and/or surgery are initiated early, making the prognosis for this type of germ cell tumor very favorable.
2. Germinomas. Germinomas are malignant germ cell tumors. Germinomas are also termed dysgerminoma when located in the ovaries; and seminoma when located in the testes. Among children, germinoma, or dysgerminoma, occurs most frequently in the ovary of a prepubescent or adolescent female. Dysgerminoma is the most common malignant ovarian germ cell tumor seen in children and adolescents.
3. Endodermal sinus tumor or yolk sac tumors.  These tumors are most commonly found in the ovary, testes, and sacrococcygeal areas (tail bone, or distal end of spinal column). When found in the ovaries and testes, they're often very aggressive, malignant, and can spread rapidly through the lymphatic system and other organs in the body. Most yolk sac tumors will require surgery and chemotherapy, regardless of stage or presence of metastasis, because of the aggressive nature and recurrence of the disease.
4. Choriocarcinoma. Choriocarcinoma is a very rare, but often malignant germ cell tumor that arises from the cells in the chorion layer of the placenta. These cells may form a tumor in the placental cells during pregnancy and spread (metastasize) to the infant and mother. When the tumor develops during pregnancy, it's called gestational choriocarcinoma. If a nonpregnant young child develops choriocarcinoma from the chorion cells that originated from the placenta that are still in the body, the term used is nongestational choriocarcinoma.
5. Embryonal carcinoma. Embryonal carcinoma cells are malignant cells that occur most often in the testes. These types of cells have the ability to rapidly spread to other parts of the body.

 

 

27. Regulation of spermatogenesis.

I. Hormonal factors:
1. FSH stimulate Sertoli cell growth and stimulate testes growth
stimulate late stage of spermatogenesis
2. LH  stimulate testosterone secretion from Leydig cell
3. Testosterone stimulate development of germinal epithelia and is needed for complete maturation of spermatozoa

4. Inhibin produced by Seroli cells, Inhibits FSH

II. Temperature factors. Optimum temperature for spermatogenesis is 35. The  factors maintainig temperature at 35:
a-testes outside abdominal cavity
b-scrotum has no fat
c-blood vessels arranged in counter current manner to incrase surface area and increase heat loss

III. Diet:  vitamins  and proteins are needed  as they increase DNA division
IV. Hypoxia,bacterial toxin - decrease spermatogenesis
V. Atomic radiation - large dose of xray damage germinal epithelia
VI. Alcohol,pesticides -  lead to abnormal sperm production
VII. Prolactin hormone (PRL) inhibit testosterone so inhibit spermatogenesis

 

Anti-Müllerian hormone also known as AMH.

 It inhibits the development of the Müllerian ducts (paramesonephric ducts – responsable for the development of uterine tubes, uterus and vagina in female) in the male embryo. It has also been called Müllerian inhibiting factor (MIF), Müllerian-inhibiting hormone (MIH), Müllerian-inhibiting substance (MIS), and Anti-paramesonephric hormone(APH). In adult testes (MEN), AMH is expressed in the Sertoli cells and in adult ovaries (FEMALES) in granulosa cells surrounding the oocytes.

In men - AMH measurements have also become widely used in the evaluation of testicular presence and function in infants with intersex conditions, ambiguous genitalia, and cryptorchidism. A measurable value of AMH in a boy with bilateral cryptorchidism is predictive of undescended testes, while an undetectable value is highly suggestive of anorchia or ovaries. In humans, androgens both induce spermatogenesis and repress AMH.  AMH is undetectable  or very low in female infants.

 

                                       

28. Determination of ovulation:

Every cycle, Follicle Stimulating Hormone (FSH) promotes the development of around 5-12 follicles, with the most dominant follicle being released at ovulation. The growing follicles produce oestrogen. As a result of high oestrogen levels in your blood, Luteinizing Hormone (LH) triggers ovulation.

Ovulation occurs around 12-24 hours after the LH surge, when the mature follicle bursts through the ovarian wall. If sperm is waiting for the egg, or intercourse happends at this point the pregnancy can occor. For 90% of women, cycle length can vary from 23-35 days, with ovulation occurring mid-cycle. Other factors (e.g. stress) can result in ovulation being delayed, until as late as the third or fourth week.  Not every woman has a 28 day cycle, nor ovulates on day 14. A normal, healthy menstrual cycle should be about 26-32 days in length.

 

1. Ovulation Pain or Mittelschmerz

The most uncomfortable ovulation symptom for some women is ovulation pain or mittelschmerz – a German word meaning mid (mittel) pain (schmerz). For these women, ovulation causes a sudden, constant pain in their lower abdomen. It’s important to understand that painful ovulation is not normal. A mild sensation is normal, but pain is not. It could be a sign you have ovarian cysts, adhesions from previous abdominal surgery, or other heath issues.

 

2. Basal Body Temperature - taking  temperature every morning upon waking (close to the same time every day). After ovulation, we notice the temperature normally rises, and stays that way until  next period. If woman becomes pregnant, the temperature stays higher. This is how some women know when to expect their period, noting a drop in temperature around the time their period is due.

 

3. Cervical Mucus  is one of the more reliable ovulation symptoms. The mucus changes in response to being at fertile or infertile stages of the cycle. It’s also a good indicator of when fertility has returned after having a baby. Cervical mucus changes with fluctuations in hormones (e.g. oestrogen). Following a period, mucus will typically be dry before becoming sticky, then creamy, then watery, before its most fertile state – clear, slippery and stretchy. Highly fertile mucus looks like raw egg white. This best aids the sperm on its passage to the egg, and provides an alkaline protection from the vagina’s acidic environment. As the woman get older, she has fewer days of egg-white cervical mucus (EWCM). For example, a woman in her 20s might have up to five days of EWCM, whereas women in their late 30s might have one or two days at most.

 

4. There are some ovulation prediction tests on the market. They can help detect changes in the body that signal ovulation is near. An ovulation prediction kit works like a pregnancy test, except it measures your levels of LH, which indicate that you will be ovulating in the next 12-24 hours.

 

 

29. Male infertility tests

Semen analysis - check the sperm count, motility and morphology

This is a very simple and important test and should be done early in the evaluation process. Sometimes the test should be done 2, or even 3 times to get an accurate reflection of the numbers and their variation over time.

The most important parameters in a semen analysis are:

1. Concentration (often called "count") - how many sperm are in each ml of semen?
2. Motility - what percent of them are swimming forward?
3. Morphology - what percent of them are normally shaped?

Cutoff values for normal vary somewhat, depending on the lab and the interpreter.

• The World Health Organization's 5th edition of "normal semen analysis" values are shown below

 

Semen Analysis Parameter	Normal Values
Volume	1.5 ml or more
pH	> or equal to 7.2
Sperm concentration	15,000,000/ml or more
Total motility	40% or more
Progressive motility	32% or more
Morphology	4% or more normal forms (Strict criteria)
Vitality	58% or more live
White blood cells	Less than 1,000,000/ml

 

If a severe male factor defect is found, the amount of testing on the female is often reduced. Treatments, such as inseminations or in vitro fertilization can then be started more directly.

                                     
Human sperm stained for morphology assessment - part of the male fertility testing workup

Morphology: The evaluation of sperm size, shape and appearance characteristics should be assesed by carefully observing a stained sperm sample under the microscope. The addition of colored "dyes" (stains) to the sperm allow the observer to distinguish important normal landmarks (characteristics) as well as abnormal findings. 

Abnormal heads : Many different sperm head abnormalities may be seen. Large heads (macrocephalic), small heads (microcephalic) and an absence of identifiable head are all seen in evaluations. Tapering sperm heads, pyriform heads (teardrop shape) and duplicate or double heads have been seen. Overall (gross) abnormalities in appearance may be termed "amorphous" changes.

Abnormal tails : Coiling and bending of the tail are sometimes seen. Broken tails of less than half normal length should be categorized abnormal. Double, triple and quadruple tails are seen and are abnormal. Cytoplasmic droplets along the tail may indicate an immature sperm.

 

Definitions of "abnormal" counts:

Polyzoospermia: Excessively high sperm concentration
Oligozoospermia: Sperm count less than 15 million/ml
Hypospermia: Semen volume < 1.5 ml
Hyperspermia: Semen volume > 5.5 ml
Aspermia: No semen volume
Pyospermia: Leukocytes (germ fighter cells) present in semen
Hematospermia: Red blood cells present in semen
Asthenozoospermia: Sperm motility < 40%
Teratozoospermia: > 4% of sperm seen are of abnormal form
Necrozoospermia: Nonviable ("dead") sperm
 

30. MAR (Mixed Antiglobulin Reaction) TEST and Halosperm

MAR test  (Mixed Antiglobulin Reaction)
The direct MAR test detects the presence of antibodies bound to spermatozoa. A solution of spermatozoa is mixed with a solution of latex beads bound to monoclonal anti- human IgA or to monoclonal anti- human IgG. Antisperm antibodies The presence of antibodies bound to membrane antigens on spermatozoa is characteristic of immunologically derived infertility. This occurs in approximately 8% of infertile males. The limitations of the procedure. The direct MAR test can only be carried out on mobile spermatozoa. Samples containing a low concentration of or weakly motile spermatozoa may result in false negatives. 
When anti-sperm antibodies are present, mobile spermatozoa will bind to the latex beads in an agglutination reaction. The resulting complexes will be as large as the magnitude of the immunological reaction. 
Reference values: 
• < 10% - negative; 
• 10 – 39% - borderline (suspicion of immunologically derived infertility); 
• > 40% - pozitive (high probability of immunologically derived infertility).

The presence of antisperm antibodies can interfere with sperm function and zona binding and the acrosome reaction. The presence of sperm antibodies reacting with antigen(s) on the spermatozoa is considered as typical and specific for immunological infertility. These antibodies are found in approximately 8% of infertile mem Antisperm antibodies belong to different immunological classes, but only those of the IgG and the IgA class are clinically relevant. Antisperm antibodies when bound to complement C3 of the IgA class, which mainly have agglutinating properties rarely occur without antibodies of the IgG class , but their meaning for male infertility may be more important. Indeed, patients combining sperm antibodies of the IgA class with IgG antibodies, or presenting IgA antibodies alone have very little chance of impregnating their partner through natural ways. Hence, detection of antibodies of the IgA class is of the utmost importance both for diagnosis and prognosis.

The bulk of the IgA class antisperm antibodies are secreted by the accessory sex glands. They are present on the spermatozoa and sometimes in seminal plasma, but usually are absent in serum. Therefore, testing for antisperm antibodies of the IgA class on serum is not recommended . It may be considered to search for sperm antibodies of the IgA class in seminal plasma, in cases with low sperm concentration or motility, although the possible clinical meaning of these antibodies is questionable. These spermatozoa are mixed with the beads which are coated with antihuman antiIgA. The formation of mixed agglutinates of motile spermatozoa with beads indicates the presence of IgA antisperm antibodies on the spermatozoa.

 

HALOSPERM

is a diagnostic test based on the sperm chromatin dispersion technique of spermatozoa nuclei. Spermatozoa DNA fragmentation (SDF) 
– Factors which may influence spermatozoa DNA fragmentaion: some medication with toxic effects, fever, smoking, drugs, infectious diseases, age, varicocele, long-term abstinence. 
Underlying basis involves differential DNA decondensation which results in a characteristic halo around spermatozoal head with intact DNA and no halo around spermatozoa with fragmented
Target population 
• Infertile males with varicocele have a high percentage of spermatozoa with fragmented DNA. The Halosperm test measures the degree of severity of this condition 
• In infections (especially with Chlamydia trachomatis and Mycoplasma), Halosperm test allows doctors to monitor the effectiveness of antibiotic treatment and helps them pick healthiest sample for assisted reproduction procedures; 
Interpretation– reference values. 
• SDF<30% à within normal range; 
• SDF>30% à high rate of occurrence of fragmented DNA

                                                                             

 

31.Female contraception
A. HORMONAL 
1. Birth control pill (E2+P or only P) –INHIBIT OVULATION, cervical mucous becomes impermeable, inhibits proliferation of uterine lining and thus prevents ovum implantation 
2. Hormone patch (5/5cm) –applied 3weeks/month with a 1 week break (E2+P)
3. Vaginal ring – inserted 3weeks/month (by user) (E2+P)
4. Injection every 3 months (high concentration of P) –parenteral administration, acts on cervical mucous and ovulation 
5. Implant (high concentration of P) – inserted by doctor, in the arm- kept for 3 years 
6. Intrauterine device- inserted by doctor, kept for 5 years 

 

B. NONHORMONAL
1. Intrauterine device 
2. Spermicides 
3. Condoms 
4. Diafragms 

 

NATURAL
1. Body temperature- rises by 0.3-0.5 degrees after ovulation and remains elevated until the menstrual phase 
2. Monitor change in cervical mucosa 

PERMANENT 
1. Tubal ligation/vasectomy 

C. EMERGENCY 
1. Pill with high concentration of P (Postinor)
2. Pill with high concentration of E+P 
3. IUD inserted after 5 days max.

32. Human Chorionic Gonadotropin and pregnancy test
Secretion
Hcg is a glycoprotein hormone produced by the developing placenta (syncitiotrophoblast) shortly after fertilization (5th-6th day ) after the embryo attaches to the uterine lining 
Function
Stimulate progesterone and estrogen secretion
Prevent menstruation; Promote endometrium growth

Pregnancy home test
1. It is a colloidal gold/antibody complex based rapid chromatographic immunoassay for the detection of hCG in urine 
2. hCG can be detected in urine as early as 10 days after conception/ blood 7 days
3. hCG concentrations peak at about 9-12 weeks into pregnancy
4. The amount of hCG will vary greatly with gestational age and between individuals 
5. It detects the presences of hCG in urine specimen at the sensitivity of low concentration

 

33. Choriocarcinoma is a highly malignant germ cell tumour which usually follows an abnormal pregnancy with a hydatidiform mole. It may also occur after a spontaneous abortion, and rarely, may follow a normal pregnancy. The tumour metastasizes early, by means of vascular invasion and blood spread.
Molar pregnancy is an abnormal form of pregnancy in which a non-viable fertilized egg implants in the uterus and will fail to come to term. These villi grow in clusters that resemble grapes. A molar pregnancy can develop when fertilized egg had not contained an original maternal nucleus. The products of conception may or may not contain fetal tissue. A complete mole is caused by a single (incidence is about 90%) or two (incidence is about 10%) sperm combining with an egg which has lost its DNA (the sperm then reduplicates forming a "complete" 46 chromosome set) The genotype is typically 46,XX (diploid) due to subsequent mitosis of the fertilizing sperm, but can also be 46,XY (diploid). 46,YY (diploid) is not observed. A partial mole occurs when an egg is fertilized by two sperm or by one sperm which reduplicates itself yielding the genotypes of 69,XXY (triploid) or 92,XXXY (tetraploid). Complete hydatidiform moles have a higher risk of developing into choriocarcinoma — a malignant tumor of trophoblast cells — than do partial moles. Hydatidiform moles should be treated by evacuating the uterus by uterine suction or by surgical curettage as soon as possible after diagnosis, in order to avoid the risks ofchoriocarcinoma. Patients are followed up until their serum human chorionic gonadotrophin (hCG) level has fallen to an undetectable level. Invasive or metastatic moles (cancer) may require chemotherapy and often respond well to methotrexate. The chances of having another molar pregnancy are approximately 1%.

 

34. Usual tests for fetal malformations:

Types of prenatal tests:

1. Screening tests determine if the baby has an increased risk of having a particular problem such as Down syndrome or a neural tube defect. They are not diagnostic and an increased risk result does not mean the baby will definitely be affected.

• Prenatal screening tests include:

> Ultrasound

> Early pregnancy (first trimester) screening: nuchal translucency ultrasound together with testing of the mother’s blood

> Second trimester screening: testing the mother’s blood (maternal serum screening)

2. Diagnostic tests determine if the baby has, or will develop after birth, a genetic condition. Sampling procedures to obtain cells for chromosome analysis or specific genetic tests are invasive. • Prenatal diagnostic tests include:

> Ultrasound

> Chorionic villus sampling (CVS)

> Amniocentesis

Prenatal diagnostic tests should not be considered routine, but rather offered as a choice to women.

1. Combined first trimester test (double test)

This test is designed to identify women at increased risk of having a baby with Down syndrome, but it can sometimes also identify other problems. The test has three parts.

The first is a blood test at 10 to 12 weeks of pregnancy.  The second is an ultrasound, called a nuchal translucency or NT test, at 11 to13 weeks. The third part is the woman’s age, which is also taken into account.

These three pieces of information are combined to calculate the risk that the baby has Down syndrome. Couples with an increased risk will be offered genetic counselling to consider their choices; the choice of whether or not to have a diagnostic test – either chorionic villus sampling or amniocentesis – to check the baby’s chromosomes.

Nuchal translucency is used to estimate if a baby is at an increased risk of having a chromosomal abnormality. It uses ultrasound to see and measure a fluid filled sac at the back of the unborn baby's neck during early pregnancy. The nuchal translucency test, which is part of the first trimester screening test, can sometimes be done on its own, without the blood test. This ultrasound is carried out between 11 and 13 weeks of pregnancy and is reasonably accurate, but not as accurate as the combined first trimester screening test.

 

• It is recommended that the NT screening test be done in conjunction with a maternal blood test. Two proteins present in the maternal blood are measured:

1.  PAPP-A (pregnancy associated plasma protein) and

2. free ß-subunit of human chorionic gonadotrophin (free ß-hCG).

Levels of these proteins vary, but tend to be different in women who are carrying fetuses with Down syndrome or trisomy 18.

Increased free ß-hCG with decreased PAPP-A is suggestive of Down syndrome, while decreased levels of both analytes is suggestive of trisomy 18.

• By having the blood test in combination with the NT screening test, around 85-90% of babies who have Down syndrome and occasionally other problems will be picked up, compared to 70% or less using NT on its own.

• Approximately 5% of combined first trimester screening tests give an increased risk result. This figure varies depending on maternal age. Women with an increased risk result should be offered a diagnostic test. The majority of increased risk results are not due to Down syndrome, and most of these babies will be healthy.

• Results are provided as risks for Down syndrome and the other chromosomal trisomy 18, at the time of screening  Note then that this is not a risk of delivering an affected fetus. Approximately 30% of babies with Down syndrome do not survive to term.

The factors that need to be entered into the risk calculation algorithm should be noted on the request form including:

• LMP (last menstrual period) & EDD (expected day of delivery) and weight

• Maternal age and previous child with a chromosomal abnormality

• Date and location of ultrasound scan

• Any other information requested on the form, eg ethnicity, IVF details

 

2. Second trimester maternal serum screening

This blood test is best done between 15 and 17 weeks of pregnancy, but it can be carried out between 14 and 20 weeks. The second trimester screening test is suitable for women who did not have either the first trimester screening test or the nuchal translucency test. It can tell parents whether the baby is at increased risk of Down syndrome (and/or some other chromosomal alterations) or a neural tube defect, which is a problem in the development of the spinal cord and/or brain. The second trimester test is not as accurate as the first trimester screening test.

• The optimal time to have this test performed is between 15 and 17 weeks, but it can be performed until 20 weeks.

• Second trimester maternal serum screening uses a blood test in conjunction with the maternal age, gestational age and maternal weight to calculate a risk figure for Down syndrome. This screening test may also detect pregnancies with an increased risk for trisomy 18 and neural tube defects.

• Maternal blood contains hormones and proteins produced by the fetus and placenta, including  1.alphafetoprotein (AFP),

2.unconjugated estriol (μE3),

3. free ß-subunit of human chorionic gonadotrophin (free ß-hCG),

4. and inhibin A.

• Levels tend to be altered in pregnancies affected by Down syndrome, trisomy 18 or neural tube defects. In Down syndrome, the levels of AFP and μE3 tend to be reduced, and free ß-hCG and inhibin A increased. In neural tube defects AFP may be increased and, in trisomy 18, levels of all these substances are decreased.

The quadruple test measures four analytes (AFP, μE3, free ß-hCG and inhibin A), whilst the triple test measures three analytes. Detection rates are improved when four analytes are used.

Using the quadruple test with ultrasound dating: > 75% of fetuses with Down syndrome are detected by second trimester maternal serum screening, and approximately 5% of tests give an increased risk result. Women aged 40 and over have higher detection rates.

 

3. Chorionic villus sampling (CVS)

This procedure is usually performed from 11 weeks, routinely between 11 and 13 weeks gestation 2. It should not be performed prior to 10 weeks gestation due to the risk of limb defects.

A sample of chorionic villus (pre-placental tissue) is removed by a fine needle, either transabdominally, or less frequently transvaginally, under ultrasound guidance.

The tissue is used for chromosome analysis, and in some specific situations, may be used for diagnosis (DNA or biochemical analysis) of a genetic condition where there is a family history.

It is a procedure that can be performed earlier in pregnancy than amniocentesis, and has the benefits of an early scan that may detect anencephaly. The miscarriage rate, in experienced hands, is estimated at ~1% above the background risk.

 

4. Amniocentesis

The procedure is performed from 15 weeks routinely, to approximately 19 to 20 week gestation.  A sample of amniotic fluid is removed from around the fetus by a fine needle, under ultrasound guidance. Testing is performed on amniotic fluid cells, most of which originate from the fetus, compared with CVS, where placental cells are tested.

Discomfort is usually minimal, though a very small number of women experience pain as the needle passes through the peritoneum.  The miscarriage rate is lower than that for CVS and estimated to be around 0.5% above the background risk in experienced hands.

Amniocentesis may not detect mosaicism, because of the limited number of cells counted in a routine test.  As amniocentesis is a second trimester test, the pregnancy is more advanced when results become available, compared with CVS.

 NB: amniocentesis is not the preferred screening test for neural tube defects. An ultrasound at 18 to 20 weeks gestation is more accurate.

 

35. Assisted reproduction technology (ART)

1. In vitro fertilization (IVF):  Eggs are combined with sperm in a laboratory. After fertilization, the resulting embryos develop for three to five days before being transferred to the uterus.
2. Artificial insemination – Intrauterine Insemination -IUI (introduction of semen directly into uterine cavity). Simple, inexpensive procedure that can be done monthly; May follow natural ovarian and uterine cycle; May require ovarian stimulation 
3. Intracytoplasmic sperm injection (ICSI): Approximately two-thirds of all IVF cycles done in the United States use ICSI as the fertilization method. In this procedure, a single sperm is injected directly into your egg, rather than placing many sperm next to the egg, as in IVF. After fertilization, the resulting embryos develop for three to five days before being transferred to the uterus.
4. GIFT is short for Gamete Intrafallopian Transfer, and it was once a very common and popular form of ART. However, it is not used that much anymore. GIFT is where the woman’s eggs and man’s sperm are combined in a lab. The eggs are then inserted into the fallopian tubes. In GIFT treatments, the fertilization takes place inside a woman’s body and not inside a lab. GIFT is a good option for couples who want a more natural ART treatment, that allows the fertilization to take place inside the woman’s body naturally.
5. ZIFT is short for Zygote Intrafallopian Transfer, and it is much like GIFT. The eggs are mixed with the sperm in a lab, and then placed back inside the woman’s fallopian tubes. The difference with ZIFT is that the eggs are not inserted back into the woman’s body until the eggs have already been fertilized. As with GIFT, this process was once very popular, but has now become less common with the increased success of IVF.
6. Donor egg or embryo: If one of the partners in not able to conceive with her own gamets, he/she can have IVF treatment using eggs donated by another woman or sperm donated by another man. The donor egg is combined with  partner's sperm, and the resulting embryo is transferred to mother uterus. This procedure can also be done with a donated embryo or donated sperm.
7. Surrogacy (gestational carrier): Surrogacy is the process in which another woman carries a baby that is made from either your egg and your husband’s sperm, or donor egg and sperm. The surrogate is simply the carrier for the baby during pregnancy.
   
   • 1790 – birth of first baby after artificial insemination using husband’s sperm in U.K. 
   • 1890 – birth of first baby after artificial insemination with sperm from donor 
   • 1978 – Louise Brown – first baby conceived in a test tube p
   • 1984- first birth of frozen embryos 
   • 1992 – first birth after ICSI
   • 1996 – first IVF baby in Romania