Reproduksi Semester II.

(1)

PENGANTAR

FISIOLOGI REPRODUKSI

Kuliah 1

Rahmatina B. Herman

Bagian Fisiologi

(2)

Reproduction

Reproduction is process to maintain

continuation of species by which

- new individuals of a species are produced - genetic material is passed from generation

to generation

Cell division in a multicellular organism is

necessary for growth and it involves passing of genetic material from parent cells to daughter cells

(3)

The Reproductive System

does not contribute to homeostasis

is not essential for survival of an individual

But still plays a i porta t i a perso ’s life,

e.g. the manner:

- in which people relate as sexual beings contributes in significant ways to

psychosocial behavior

- how people view themselves

(4)

The Reprodu tive “yste …..

Reproductive function also has a profound effect on society:

- universal organization of societies into

family units provide a stable environment that is conducive for perpetuating our

species

- on other hand, population explosion and its resultant drain on dwindling resources

have led to worldwide concern with means by which reproduction can be limited

(5)

The Reprodu tive “yste …..

Reproductive capability depends on intricate

relationship among hypothalamus, anterior

pituitary, reproductive organs, and target cells

of sex hormones

These relationship employ many of regulatory

mechanisms used by other body systems for

maintaining homeostasis, such as

(6)

The Reprodu tive “yste …..

Sexual behavior and attitudes are deeply influenced by emotional factors and socio-cultural mores of the society in which the individual lives

However, Reproductive Physiology will

concentrate on basic sexual and reproductive functions that are under nervous and hormonal control, and will not examine physiological and social ramifications of sexual behavior

(7)

The Reprodu tive “yste …..

The organ of male and female may be grouped by function

Testes and ovaries (gonads), function in production of gametes: sperm and ova Gonads also secrete hormones

The ducts of reproductive systems transport, receive, and store gametes

Accessory sex glands produce materials that support gametes

(8)

The Reproductive System

…..

In females, the breasts are also considered accessory reproductive organs

The externally visible portions of reproductive system are known as external genitalia

The production of gametes and fluid, and their discharge into ducts classify the gonads as

exocrine glands

Whereas the production of hormones classify the gonads as endocrine glands

(9)

Secondary Sexual Characteristic

Secondary sexual characteristic are many external

characteristics not directly involved in

reproduction

That distinguish male and female

Development and maintenance governed by

testosterone in males and estrogen in females Progesterone has no influence on secondary

sexual characteristic

Axillary and pubic hair growth is not secondary sexual characteristic

(10)

Secondary Sexual Characteristic

..…

In some species, secondary sexual

characteristic are great importance in courting

a d ati g ehavior e.g. to attra t fe ale’s

attention)

In humans, attraction the opposite sex not only influenced by secondary sexual characteristic but also strongly affected by the complexities of human society and cultural behavior

(11)

Overview of Functions and Organs of Male Reproductive System

The essential reproductive functions of male are:

1. Production of sperm (spermatogenesis) by testes (in skin-covered sac: scrotum)

2. Delivery of sperm to female – semen by - male reproductive tract: epididymis, vas deferens, ejaculatory duct

- urethra (in penis)

3. Male accessory sex glands: providing bulk of semen: seminal vesicle, prostate,

(12)

(13)

Overview of Functions and Organs

of Female Reproductive System

Female’s role i reprodu tio is ore o pli ated:

1. Production of ova (oogenesis) by ovaries

2. Reception of sperm: vagina-cervix

3. Reception of sperm and ovum to a common site for union (fertilization or conception): Fallopian tube

4. Maintenance of the developing fetus until it can survive in outside world (gestation or pregnancy), including formation of placenta (organ exchange between mother and fetus): uterus

5. Giving birth to the baby (parturition)

6. Nourishing the infant after birth by milk production (lactation): mammae

(14)

Overview of Functions and Organs

of Fe ale Reprodu tive “yste …..

Product of fertilization: embryo

During first 2 months of intrauterine

development when tissue differentiation is taking place

Developing living being is recognizable as human: fetus

- no further tissue differentiation

(15)

Overview of Functions and Organs

of Fe ale Reprodu tive “yste …..

Female reproductive tract consists of:

Ovaries

Oviduct s (Fallopian tubes)

- pick up ova on ovulation and serve as fertilization site

Uterus, thick-walled hollow: responsible for

- maintaining fetus during development , and - expelling it at the end of pregnancy

Cervical canal

- small opening of cervix

- pathway for sperm to uterus then to oviduct - passageway for delivery of baby from uterus

Cervix

(16)

Overview of Functions and Organs

of Fe ale Reprodu tive “yste ……

Vagina

- expandable tube

- connects uterus to external environment

Vaginal opening

- located in perineal region

- between urethral opening and anal opening

Hymen

- thin mucus membrane partially covering vaginal opening

• Labia minora and labia majora

- skin folds surrounding vaginal and urethral openings

• Clitoris

(17)

(18)

(19)

(20)

Sex Determination and Differentiation

Reproductive cells each contain a half set of chromosomes

Gametogenesis is accomplished by meiosis

The sex of and individual is determined by combination of sex chromosomes

Sexual differentiation along male or female lines depends on the presence/ absence of masculinizing determinant

(21)

(22)

(23)

Parents with diploid (46 chr) somatic cells

Mother

Father

Meiotic division of germ cells

Haploid Ovum Haploid Sperm

Fertilization

Diploid fertilized Ovum

Mitosis

(24)

Ovum with X sex chromosome Fertilized by

Sperm with Y sc Sperm with X sc

Embryo with XY sc Genetic sex Embryo with XX sc

Sex-determining region of Y chr (SRY) stimulates Production of H-Y antigen

In plasma membrane of undifferentiated gonad

H-Y antigen directs differentiation of gonads into testes

No Y chr, so no SRY and no H-Y antigen

With no H-Y antigen, undifferentiated gonads

develop into ovaries

Gonadal sex

(25)

Testosterone

Promotes development of undifferentiated external genitalia along male lines

(e.g. penis, scrotum)

Testes secrete hormone and factor

Phenotype sex Mullerian-inhibiting factor Dihydrotestosterone (DHT) Converted to Degeneration of Mullerian ducts Transforms Wolfian ducts into male reproductive tract

(e.g. epididymis, ductus deferens, ejaculatory duct,

(26)

(27)

(28)

Absence of testosterone

Undifferentiated external genitalia along female lines

(e.g. clitoris. labia)

Ovaries does not secrete hormone and factor

Phenotype sex

Absence of Mullerian- inhibiting factor Degeneration of

Wolfian ducts

Mullerian ducts develop Into female reproductive tract (e.g. oviducts, uterus)

(29)

Errors in Sexual Differentiation

Genetic sex and phenotype sex are usually

compatible

Occasionally, discrepancies occur

between genetic and anatomic sexes

(30)

Errors i “e ual Differe tiatio …..

1. If testes in a genetic male fail to properly

differentiate and secrete hormones, the result is the development of an apparent anatomic female in a genetic male, who, of course will be sterile.

Similarly, genetic males whose target cells lack receptors for testosterone are feminized, even though their testes secrete testosterone

(31)

Errors i “e ual Differe tiatio …..

2. Testosterone acts on Wolfian ducts to convert them into a male reproductive tract;

If testosterone derivative dihydrotestosterone (DHT) that responsible for masculinization of external genitalia because of genetic deficiency of the enzyme which converts testosterone

into DHT, results in a genetic male with testes and a male reproductive tract but with female external genitalia

(32)

Errors i “e ual Differe tiatio …..

3. Adrenal gland normally secretes a weak androgen, dehydroepiandrosterone in

insufficient quantities to masculinize females. If, pathologically excessive secretion of this hormone in a genetically female fetus during critical developmental stages imposes

differentiation of reproductive tract and genitalia along males lines

(33)

Errors i “e ual Differe tiatio …..

Sometimes, the discrepancies between genetic sex and apparent sex are not recognized until puberty, when discovery produces

psychologically traumatic gender identity crisis For instance: a masculinized genetic female

with ovaries, but with male type external

genitalia may be reared as a boy until puberty. When breast enlargement and lack of beard growth signal an apparent problem

(34)

Errors i “e ual Differe tiatio …….

Less dramatic cases of inappropriate sex differentiation often appear as sterility problems

Therefore, important to diagnose any

problems in sexual differentiation in infancy. It can be reinforced, if necessary, with surgical

and hormonal treatment, so that psychosexual development can proceed as normally as

(35)

(36)

Tugas

1. Hubungan sistem limbik (limbic system)

dengan pengaturan fungsi seks

2. Hubungan kelenjar pineal (pineal body)

(37)

DASAR-DASAR

BIOMOLEKULER

REPRODUKSI WANITA

Rahmatina B. Herman

Bagian Fisiologi

Fakultas Kedokteran Universitas Andalas

(38)

Sex Determination and Differentiation

Reproductive cells/gamete each contain a half set of chromosomes (haploid)

Gametogenesis is accomplished by meiosis

The sex of and individual is determined by

combination of sex chromosomes

Sexual differentiation along male or female lines depends on the presence/absence of

(39)

(40)

(41)

Absence of testosterone

Undifferentiated external genitalia along female lines

(e.g. clitoris. labia)

Ovaries does not secrete hormone and factor

Phenotype sex

Absence of Mullerian- inhibiting factor

Degeneration of Wolfian ducts

Mullerian ducts develop Into female reproductive tract (e.g. oviducts, uterus)

(42)

Summary of 4 possible defects produced by maternal nondisjunction of sex chromosomes at the time of meiosis

The YO combination is believed to be lethal, the fetus dies in utero

/Ovarian agenesis/Turner syndrome

(43)

Female Reproductive System

Reproductive system of women shows regular cyclic

changes that may be regarded as periodic preparation for fertilization and pregnancy

In humans and primate, the cycle is a menstrual cycle and its conspicuous feature is periodic vaginal bleeding that

occurs with the shedding of uterine mucosa (menstruation)

In other mammal: the sexual cycle is called estrous cycle, no episodic vaginal bleeding occurs, but the underlying endocrine events are essentially similar

- in some species: ovulation occurs spontaneously - in other species: ovulation is induced by copulation

(44)

Ovaries

Primary female reproductive organs Perform dual function:

- producing ova (oogenesis)

- secreting female sex hormones:

estrogen and progesterone which act together to: > promote fertilization of ovum

> prepare female reproductive system for pregnancy

Containing various levels of follicle development

(45)

Histology of the ovary. The arrows indicate the sequence of developmental stages that occur as part of ovarian cycle

(46)

Oogenesis

Undergo numerous mitotic divisions

± 7 month after conception, fetal oogonia cease dividing

From this point on, no new germ cells are generated

Still in the fetus, all oogonia develop into

(47)

Primary Oocyte

Begin a first meiotic division by replicating their DNA However, they do not complete the division in the fetus

Accordingly, all the eggs present at birth are primary

oocytes containing 46 chromosomes, each with two

sister chromatids

Cells are said to be in a state meiotic arrest

State meiotic arrest continues until puberty and the onset of renewed activity in ovaries

Only primary oocytes destined for ovulation will ever

complete the first meiotic division, for it occurs just before the egg is ovulated

(48)

Pri ary Oo yte…..

Each daughter cells receives 23 chromosomes, each with 2 chromatids

One of the two daughter cells, secondary oocytes retains virtually all cytoplasm (other is first polar body)

Thus, the primary oocytes:

- Already as large as the egg will be

- Passes on to be secondary oocyte half of its

chromosomes but almost all of its nutrient-rich cytoplasm

(49)

Secondary Oocyte

The second meiotic division occurs in a

fallopian tube after ovulation, but only if the secondary oocyte is fertilized (penetrated by a sperm)

Daughter cells each receive 23 chromosomes, each with a single chromatid

One of the two daughter cells, termed an ovum retains nearly all cytoplasm (other is second

(50)

Final Result of Oogenesis

Net result of oogenesis is

that

each primary oocyte

can produce only one

(51)

(52)

(53)

Summary of Oogenesis

Birth Puberty Oogonia Chromosomes Per cell Chromatids Per cell

46 2

2 46

23 1

(54)

Summary of

Ooge esis…..

Oogonia: mitotic divisions until ± 7 month after

conception

Mitosis of oogo iu → primary oocyte

Meiosis of primary oocyte, but do not complete

(beginning of the 1st eioti divisio → eioti arrest

Primary oocyte at birth containing 46 chromosomes 1st meiotic division is completed just before ovulation

→ secondary oocyte

2nd meiotic division occurs in a fallopian tube after

ovulation, but only if the secondary oocyte is fertilized (penetrated by a sperm)

(55)

Comparison of Spermatogenesis and Oogenesis

Spermatogenesis

Three major stages:

1. Mitotic proliferation

2. Meiosis

3. Packaging/ spermiogenesis: physically

reshaping/ remodeling

± 64 days, from spermatogonium to mature sperm

Up to several hundred million sperm may reach maturity daily

(56)

(57)

Follicle

From the time of birth, there are many primordial follicles, each containing 1 primary oocyte

Progression of some primordial follicles to preantral and early antral stages occurs

- throughout infancy and childhood, and

- then during the entire menstrual cycle

Therefore, although most of follicles in ovaries are

still primordial, there are also always present a

relatively constant few number of preantral and early antral follicles

(58)

Menstrual Cycle

At the start of each menstrual cycle, 10-25 the

follicles begin to develop into larger follicles

In humans, usually one of the larger follicles in one ovary starts to grow rapidly on ± the 6thday, becomes

the dominant follicle

The dominant follicle continues to develop, and others (in both ovaries) regress and become a

degenerative process called atresia (an example of

programmed cell death, or apoptosis)

(59)

Ovulation

Mature follicle (Graafian follicle): ± 1,5 cm in

dia eter, that it alloo s out o ovary’s surfa e

Ovulation occurs when the thin walls of follicle and ovary at site where they are joined rupture because of enzymatic digestion

Secondary oocyte surrounded by its tightly

adhering zona pellucida and granulosa cells, as well as cumulus, is carried out of ovary and onto ovarian surface by antral fluid

(60)

Ovulatio …..

Occasionally, 2 or more follicles reach

maturity and more than 1 egg may be

ovulated

This is the most common of cause of

multiple births

In such cases, siblings are fraternal, not

identical, because the eggs carry different

sets of genes

(61)

Follicular steroid hormones (progesterone)

Proteolytic enzymes (collagenase)

Follicular hyperemia and

Prostaglandin secretion

Weakened follicle wall Plasma transudation into follicle

Degeneration

of stigma Follicle swelling Follicle rupture

Evagination of ovum

(62)

(63)

Indicators of Ovulation

A surge in LH secretion triggers ovulation

- Ovulation normally occurs ± 9 h after the peak of LH surge - The ovum lives for ± 72 h after ovulation, but it is

fertilizable for a much shorter time

Research shows:

> Intercourse on the day of ovulation: pregnancy 36% > Intercourse on days after ovulation: pregnancy 0

> Intercourse 1-2 d before ovulation: pregnancy 36%

> A few pregnancies resulted from intercourse 3-5 d before ovulation (8% on day 5 before ovulation)

- Thus, some sperms can survive in the female genital tract and fertilize the ovum for up to 120 h before ovulation, but the most fertile period is clearly 48 h before ovulation

(64)

I di ators of Ovulatio …..

A change (usually rise) in basal body temperature

caused by secretion of progesterone, since progesterone is thermogenic

- The rise starts 1-2 d after ovulation

- Obtaining an accurate temperature chart should use a digital thermometer and take oral/rectal temperatures in the morning before getting out of bed

- Temperature change at the time of ovulation is probably caused by the increase in progesterone secretion

(65)

Basal body temperature and plasma LH and FSH concentrations (mean ± SE) during the normal human menstrual cycle

(66)

Formation of Corpus Luteum

After mature follicle discharges its antral fluid and egg, it collapses around antrum and undergoes rapid transformation

Granulosa cells enlarge greatly, and entire glandlike structure formed, known as corpus luteum (CL)

CL secretes estrogen, progesterone, inhibin

If the discharged egg (now in a fallopian tube) is not fertilized, CL reaches its maximum development

within ± 10 days.

CL then rapidly degenerates by apoptosis

It leads to menstruation and beginning of a new menstrual cycle

(67)

Granulosa Cell

Primordial follicles surrounded by a single layer of

granulosa cells

Granulosa cells secrete:

- estrogen ,

- small amounts of progesterone just before

ovulation

- peptide hormone inhibin

During childhood granulosa cells secrete: - nourishment for ova

(68)

Gra ulosa Cell…..

Further development from primordial follicle stage is characterized by

- an increase in size of oocyte

- a proliferation of granulosa cells into multiple layers

- separation of oocyte from inner granulosa cells by a

thick of material: zona pellucida

Granulosa cells produce one or more factors that act on primary oocytes to maintain them in meiotic arrest

(69)

Gra ulosa Cell…..

Inner layer of granulosa cells remains closely associated with oocyte by means of

cytoplasmic processes that traverse zona

pellucida and form gap junctions with oocyte

Nutrients and chemical messengers are passed to oocyte through gap junctions

Granulosa cells produce one or more factors that act on primary oocytes to maintain them in meiotic arrest

(70)

Theca Formation

As follicle grows by mitosis of granulosa cells, connective tissue cells surrounding granulosa cells differentiate and form layers known as

theca

Shortly after theca formation,

- Primary oocyte reaches full size (115 m in diameter)

- Antrum (fluid-filled space) begins to for in the midst of granulosa cells as result of fluid they secrete

(71)

Illustration of an ovary shows sequential development of follicle, the formation of corpus luteum and follicular atresia

(72)

Process of Atresia

Atresia is not limited to just antral follicles, follicles

can undergo atresia at any stage

This process is already occurring in utero so that the 2-4 million follicles and eggs are present at

birth represent only a small fraction of those

present at earlier time in the fetus

Atresia then continues all through pubertal life so

that only 200,000-400,000 follicles remain when

active reproductive life begins.

(73)

Pro ess of Atresia…..

Therefore, 99,99 % of ovarian

follicles present at birth will

undergo atresia

(74)

Sites of Synthesis of Ovarian Hormone

Estrogen is synthesized and released into blood:

- during follicular phase mainly by granulosa cells

- after ovulation, by CL

Progesterone is synthesized and released into

blood:

- in very small amounts by granulosa and theca cells just before ovulation

- major source is CL (after ovulation)

Inhibin is synthesized and released into blood: - by granulosa cells

(75)

Interactions between theca and granulosa cells in estradiol synthesis and secretion

(76)

(77)

Ovarian Cycle

1. The follicular phase:

- ovarian follicle growth

- ovulation

2. The luteal phase:

(78)

Uterine Cycle

1. Proliferative phase:

- estrogen  estrogen phase - before ovulation

2. Secretory phase:

- progesterone  progestational phase - after ovulation

3. Menstruation

(79)

Relative concentrations of anterior pituitary gland hormones (FSH – LH) and ovarian Hormones (estrogen – progesterone) during a normal female sexual cycle. Note the relationship of the hormones to the ovarian and uterine cycles

(80)

(81)

Cyclical Changes in Cervix

The mucosa of cervix does not undergo cyclical desquamation

There are regular changes in cervical mucus:

- Estrogen makes the mucus thinner, watery, and more

alkali e → pro otes the survival a d tra sport of sper s

- At the time of ovulation the mucus is: > thinnest and fern-like pattern on slide

> its elasti ity i reases → a drop a e stret hed i to a

long (8 - ≥ , a d thi thread

- Progesterone makes the mucus thick, tenacious, and cellular

- After ovulation and during pregnancy: thick, no pattern

(82)

Microscopic of patterns formed of cervical mucus on dried smeared slide. Progesterone makes the mucus thick and cellular.

In anovulatory, no progesterone is present to inhibit fern-like pattern Estrogen:

fern-like pattern

Estrogen,

no progesterone: Fern-like pattern Progesterone:

(83)

Cyclical Changes in Vagina

Under influence of estrogen:

- the vaginal epithelium becomes cornified that can be identified in the vaginal smear

Under influence of progesterone: - secretion of thick mucus

- the vaginal epithelium proliferates and becomes infiltrated with leukocytes

The cyclical changes in vaginal smear in rats are relatively marked; in humans and other species are similar, but not so clear cut

(84)

Cyclical Changes in Breast

Although lactation normally does not occur until the end of pregnancy, cyclical changes take place in the breasts during the menstrual cycle

Estrogens cause proliferation of mammary ducts Progesterone causes: growth of lobules and alveoli

The breast swelling, tenderness, and pain experienced by many women during the 10 day preceding

e struatio ← due to diste tio of the du ts,

hyperemia, and edema of the breast interstitial tissue All the changes regress along with symptoms, during menstruation

(85)

Normal Menstruation

Menstrual blood is predominantly arterial, only 25% of the blood being of venous origin

Containing tissue debris, prostaglandins, and relatively large amount of fibrinolysin from endometrial tissue

Fi ri olysi lyses lots → o lots i e strual lood

Usual duration is 3-5 d, but 1-8 d can occur normally The average amount of blood lost is 30 ml (range

normally from light spotting – 80 ml)

The amount of blood affected by various factors,

including the thickness of endometrium, medication, and diseases that affect clotting mechanism

(86)

Anovulatory Cycles

Anovulatory cycles are common for the first 12-18 months after menarche and before the onset of menopause

Whe ovulatio does ot o ur → o CL → effe ts of

progesterone on endometrium are absent

Estrogens continue to cause growth, and proliferative endometrium becomes thick enough to break down and begins to slough

The time it takes for bleeding usually < 28 d from the last menstrual period

(87)

(88)

FISIOLOGI ANDROGEN

Kuliah 2

Rahmatina B. Herman Bagian Fisiologi

(89)

Testes

Perform dual function of producing sperm and secreting testosterone

± 80% of testicular mass consists of highly coiled

se i iferous tu ules → sper atoge esis

Leydig cells/ interstitial cells

- in connective tissue between seminiferous tubules - are endocrine cells that produce testosterone

- Less easily destroyed (by x-ray, excessive heat)

Thus, portion of testes that produce sperm and

portion that secrete testosterone are distinct structurally and functionally

(90)

(91)

Androgen

Steroid hormones

Derived from cholesterol precursor molecule Masculinizing effects

Produced by: - Testes

- Adrenal glands

- Ovary

Consists of: - Testosterone

- Dihydrotestosterone

(92)

Testosterone

Once produced:

- Some of testosterone is secreted into

circulation, where it is transported to its target sites of action

- A substantial portion goes into the lumen of seminiferous tubules, where it plays an

(93)

(94)

…..Testostero e

In circulation, testosterone primarily bound to plasma protein:

- Loosely bound with plasma albumin

- More tightly bound with a beta globulin: sex hormone-binding globulin

Become fixed to the tissues (converted into

dihydrotestosterone) especially target organs: - prostate gland

(95)

…..Testostero e

The testosterone that dose not become fixed to the tissues is rapidly converted (mainly by the

liver) into:

- androsterone

- dehydroepiandrosterone



conjugated as glucoronide or sulfate



excreted into - gut in the liver bile - urine through the kidney

(96)

Functions of Testosterone

Responsible for the distinguishing characteristics

of the masculine body

Most of testostero e’s a tio s ulti ately fu tio

to ensure delivery of sperm to the female The effects can be group into 5 categories:

1. Effects on reproductive system before birth/ during fetal life

2. Effects on sex-specific tissues after birth

3. Other reproduction-related effects

4. Effects on secondary sexual characteristics 5. Non reproductive actions

(97)

Effects of Testosterone

1. Before birth/ During fetal life

- Secreted by genital ridges/ fetal testes

- Stimulated by placental chorionic gonadotropin/

human chorionic gonadotropin (HCG)

- Responsible for:

> Development of male characteristics:

Masculinizes reproductive tract and external

genitalia

(98)

….Effe ts of Testostero e

2. After birth

- Until puberty:

> Testosterone secretion ceases

> Reproductive system remain small and nonfunctional

- At Puberty:

> Leydig cells start secreting testosterone once again, stimulated by pituitary gonadotropin

hormone through the brain-testicular axis

> Testosterone is responsible for growth and

maturation of entire male reproductive system

(99)

….Effe ts of Testostero e

(100)

….Effe ts of Testostero e

2. After birth

...

- At Puberty:

> Promotes growth and maturation of

reproductive system: testes , penis, scrotum enlarge 8 X before the age of 20 years

> Essential for producing sperm (spermatogenesis) > Maintains reproductive tract throughout

adulthood

Once initiated at puberty, testosterone secretion and spermatogenesis occur continuously

(1)

Accuracy and precision of the assay for NE with amitriptyline as internal

standard (n=4)

Conc

of

NE

Intra-assay variation

(Intra-day variation)

Inter-assay variation

(Inter-day variation)

Peak ratio

Mean



SD

CV

(%)

Peak ratio

Mean



SD

CV

(%)

1 run

2 run

3 run

4 run

1 run

2 run

3 run

4 run

8000 4000 2000 1000 500 100 (LLOQ) 1.67 0.90 0.41 0.22 0.121 0.024 1.62 0.86 0.42 0.20 0.118 0.028 1.70 0.82 0.44 0.19 0.129 0.026 1.65 0.88 0.40 0.20 0.124 0.021

1.66  0.03 0.87  0.03 0.42  0.02 0.20  0.01 0.123  0.005 0.025  0.003

1.81 3.45 4.76 5.00 4.07 12.00 1.67 0.90 0.41 0.22 0.121 0.024 1.59 0.82 0.40 0.20 0.117 0.026 1.68 0.86 0.43 0.19 0.119 0.027 1.72 0.83 0.44 0.21 0.127 0.020

1.660.05 0.850.04 0.420.02 0.210.01 0.1210.004 0.024  0.03