Lecture 6- Female Sex Hormones Flashcards
Learning Outcomes female Reproduction & Pregnancy
*Describe the physiological changes to the endometrium during the menstrual cycle relating these to:
*Endocrine regulation
*Ovarian cycle
*Explain the endocrine control Oogenesis including
*Ovarian and hypothalamo-pituitary hormones
*Feedback variation across the cycle
*Show an understanding of whole body effects of menopause, linking to hormonal effects on other organs
Female Reproductive System components
We’re going to start off looking at the anatomy of Females, just as we’ve Looked at the anatomy of males. So.
You can see here at the top of this diagram
Um, that we have got a fallopian tube.
That’s that thing that looks like it’s got that little Professor Zoidberg from Futurama.
But at the bottom of it. Okay, this is hanging right over the top of the ovary, which is this little white ball underneath.
These are attached to the uterus.
Okay. And this sits right above the bladder.
These three make up our main things that we’re going to look at during this part of the module.
And this is because the ovaries, all the things that not only produce the oocytes which are going to eventually generate a foetus,
but they are also able to produce progesterone and oestrogen, which are the main reproductive hormones in females, but also inhibin and relaxin.
Okay. Now, inhibin should be relatively familiar name to you because we had that in males as well.
But relaxin is a new hormone that we’ll look at only in females okay.
We’ve also got on our list vagina. Obviously that’s the way that things get in and out.
And we’ve also got mammary glands okay.
Mammary glands, for those of you who are not aware are in the breast tissue.
And they are the things that generate milk.
Um, if you are pregnant or have recently had a baby.
*Ovaries produce secondary Oocytes
➢Also produce hormones:
oProgesterone
oOestrogen
oInhibin
oRelaxin
*Fallopian tubes
*Uterus
*Vagina
*Mammary Glands
Oogenesis
Um, okay. So we’re going to start off by thinking a little bit about oogenesis, which is happening after gametogenesis has occurred.
Okay. So gametogenesis is generation of gametes.
oogenesis is generating a follicle.
And it’s also generating the cells that come from that.
So these things are happening concurrently.
We start, just like we did with males with an oogonium.
Much as we had our spermatogonium before. Uh, but in a difference here is that this process of the oogonium becoming a primary oocyte.
This is happening by mitosis, but happens before the birth of this foetus.
Okay. Whereas this process in males is happening constantly.
Okay. So this is why females have a finite number of primary oocytes that will have the opportunity to become mature.
Uh, follicles. So our primary oocyte then undergoes meiosis, and it will generate a first polar body and a secondary oocyte.
We’re going to not talk any more about these first polar bodies, okay?
This is the end of those. And we’re not going to consider them, okay?
Because there is only so much room in one’s head, and there is only so much room in this module to talk about this.
We could have an entire module on it. Okay. So I think we’re just going to say that it generates a first polar body.
And that’s the end. The secondary oocyte is generated.
As ovulation happens.
So as it’s going down the fallopian tube it’s now progressed to a secondary oocyte.
Okay. And that secondary oocyte is ready for fertilisation.
And you can see there’s a lovely spermatozoa stood right here next to our secondary oocyte.
And once those two come together and fertilisation happens you get an ovum that has now two n and it becomes a zygote.
Okay 2n meaning that it has 2 copies of DNA.
Okay. So we are going through a very similar process that happened in males.
So we’re not going to spend ages talking about it.
Okay. But there are some major differences.
Ovaries
there are some major differences between males and females and that also comes in where we see in the ovary.
So remember this is our uterus our fallopian tube with the finger like projections on the end.
And then here is our ovary on the right.
What you’ll see is that we’ve got development as it’s going around in this orientation in a clockwise direction.
They don’t actually move in your ovary.
They stay in one place. But obviously it’s very hard to show you what that will look like.
Otherwise we’ll have lots of slides with things going one on top of the other.
So instead it’s organised in this manner so you can see them all on one sheet.
So what we’ve got to start with is our primordial follicle that has got a primary oocyte inside it.
This is what is generated during development.
Okay. And then the primary follicles are created just shortly after birth.
Okay. And these primary follicles, they sit there much as a great big reserve.
Uh, then for the rest of the female’s life, there are several million of these at birth, and only about 400 of them actually end up being used.
So there’s a relatively large amount of wastage. Okay.
Now, once a female reaches, um, puberty, they will start to generate.
Or allow several primary follicles every month that will compete with each other to be the strongest one,
okay, and only one follicle will then develop into a secondary follicle.
It’s really important that you listen to the language here.
An oocyte is the egg if you like.
I hate using that word because it’s very non-scientific.
But the follicle is the oocyte.
And the cells that are inside the little pocket that it sits in, and the cells that are all around the outside of the little pocket that it sits in.
Okay, that is a follicle. Try not to get those mixed up because that can cause confusion.
Okay. So this secondary follicle has still got a primary oocyte.
sat inside it.
But what you’ll notice is, is that this secondary follicle that’s now been selected is the strongest one and looks the best,
has now got much more fluid inside the follicle.
And those of you who are really looking carefully, we’ll see that there’s lots of yellow cells around the inside of that follicle.
And we’ll talk about those in a minute. So that follicle continues to grow both in size but also in that fluid that’s inside.
And so does the oocyte okay.
And as it gets larger it then matures into a graafian follicle or mature follicle as
it’s sometimes called just before ovulation is about to happen and ovulation happens.
It’s the release of this now secondary oocyte as it’s about to enter the, um, fallopian tube.
Okay. So that release, that ovulation okay, marks the end of the follicular phase of the ovarian cycle.
Okay. Not really surprising because you’ve been calling everything a follicle up till now.
So that’s why it’s called a follicular phase. Ovulation is always the point at which the oocyte is released.
And this is usually just referred to as ovulation. But then after that process has happened, it’s considered to be the luteal phase.
And the reason that that’s considered to be the luteal phase is because this little kind of orangy, yellowy structure, called the
corpus luteum, and the corpus luteum is formed from the remnants of that follicle.
So you can see as the ovulation happens, there’s going to be a little bit of bloods because it’s literally just burst out of.
The follicle. Okay. So much like when you scratch yourself, you get a little clot, you get a little scab on there.
The same kind of thing happens on the ovary. Okay?
Somethings just burst out of there. It’s almost like bursting a spot.
Okay. You’re going to have a little scab on there afterwards, okay.
While healing happens underneath. And part of that healing process is the corpus luteum shrivelling up.
And you can see this little blood clot in there.
And as it degenerates into a smaller one and then a smaller one again, just like your scab would on your hand, it then becomes the corpus albicans.
Okay. Now this structure’s really important during pregnancy.
It’s really important to know what those cells of the follicle do.
So we’ll shift our focus now to focus on what those cells are.
So you can see that this is our oocyte in the middle.
And unfortunately the colour coding is not the same which is really annoying.
But this pink stuff kind of pinky purple stuff here in the follicle the same as this yellow stuff that’s in the zoomed in image of the follicle
Okay. And these cells are the granulosa cells.
Okay. And the granulosa cells. They’re the cells that generate oestrogen.
Okay.
They’re the ones that make everything grow and they maintain the oocyte, making sure that it’s going to be looked after until, uh, ovulation happens.
The cells that are around the outside. They’re outside the membrane of the follicle.
I’ll call the theca cells. Okay. So if you think about when you’re looking in males, you’ve your seminiferous tubule and you had cells that were outside and you had cells that were inside, it’s very similar.
Okay. Very similar situation to what was going on there.
So our theca cells aren’t able to be part of the follicle itself,
but they secrete hormones that will then move into the follicle and affect both the oocyte and
The granulosa cells are insidece.
those follicles look like.
They look like big sort of velvety pockets. Okay.
And this is on the surface of the ovary. This happens in one place.
And then the next month or whatever, there will be,
some other place and that scar or that kind of corpus albicans then becomes almost like a scab,
or a scar that then stays on the ovary until the end of that person’s life.
Okay. soif you happened to look at an autopsy, for example, you would see evidence of this process happening
Female Sex Hormones
So now we’re going to talk about what hormones control which part of the cycle.
And again we go back to this beautiful diagram of the hypothalamus and the pituitary.
Okay. And previously when we talked about males we talked about the gonadotropin cells that would generate FSH and LH.
And we go back to those. So it’s exactly the same set of hormones that are involved in the ovarian cycle.
Ovarian Hormones
so.
You can see at the top of this slide where we start to think about our ovarian hormones that we’ve got in very tiny writing and I apologise.
and Cholesterol. Okay. And just like we had in males, cholesterol can be used to generate progesterone
And it can also generate oestrogen of which these three, estradiol, estrone and estriol, are considered to be oestrogens.
Okay. Some of the same enzymes are involved in the generation of progesterone and testosterone, as they are in aldosterone and cortisol.
Okay. But we’re not going to go into those steps that were involved.
Instead, we’re going to point out that aromatase is the is the enzyme that converts
testosterone and androgens into estrone and beta estradiol (E2) and estriol,
which is E3. And of these, beta estradiol is the most potent.
So from now on, when we say oestrogen, we mean beta estradiol.
Okay, the others we’re not going to worry too much about because they are a lot less impactful.
Now, the reason why I talked to you about what cells were in The follicle is because.
Much like we had in the males our theca cells that are on the outside of the follicle are able to generate progesterone and androgens.
And as I mentioned earlier, they will do this to support the growth of the secondary oocyte in the granulosa cells that are inside the follicle.
They do this in response to luteinizing hormone LH binding to its receptor.
That’s on the outside of the theca cells.
Okay, now they do this only if low density lipoprotein is present.
Why does it need low density lipoprotein? Those of you who’ve been on biochemistry.
Because it’s full of cholesterol. Absolutely. Okay, so it can’t start this process.
If we don’t have cholesterol. Okay.
So in the presence of both good cholesterol and of luteinising hormone, we can generate progesterone and androgens.
And these then are able to move into the granulosa cells that are inside the
follicle and in the presence again of cholesterol and luteinising hormone.
Not only can these cells generate progesterone, but the FSH is needed to bind to the FSA receptor so that we can generate aromatase or activate aromatase.
And if that is the case, then we can generate oestrogen.
Okay, so in the presence of FSH, we can generate oestrogen in the absence of FSH, but in the presence of LH We can generate progesterone and androgens.
Okay. And those progesterone and androgens are really needed.
to generate the oestrogens. So we need to have all of these things coming in together.
And it all needs to be synchronised. And so we need something to synchronise it which will come on into a minute.
Ovarian Hormones summary
So in terms of those ovarian hormones, if they’re generated from cholesterol, they have that ring structure, which means they are steroid hormones.
Okay. So mainly synthesised from cholesterol or if there’s no cholesterol.
And you could make it in your liver uh, from acetyl-CoA.
And as those of you who are doing biochemistry will know that these act through nuclear hormone receptors,
or you might have come across them in this module as intracellular receptors.
Uh, with Professor Johnson. So these nuclear hormone receptors.
Different receptors to what we had in males.
Androgen receptor is responsible. Responsive.
Testosterone. Oestrogen receptor binds to oestrogen.
Okay. So oestrogens only secreted by the ovaries in non-pregnant females.
Okay. And as I mentioned before, beta estradiol is the most potent.
And behind that, by quite a lot something like 17 times less potent is estrone
And estriol is about 100 times less potent than estrone.
Okay. In terms of progestins of which progesterone is one.
It is the most important in females. Okay. We need this because we need it as a precursor to generate oestrogen,
but also because it has its own role in maintaining, the growth and development of the endometrium.
So if somebody is going to get pregnant, it’s really important.
progesterone can get produced by both sets of cells,
partially because it needs the luteinizing hormone and the uh receptor and the, um, low density lipoprotein receptor.
Okay. So.
In non-pregnant females.
So most of us in this room at least, um, if you’re female, then, um, this is produced by the corpus luteum.
Okay. It’s not produced in the follicular phase.
Okay. So early on it’s not really produced.
There might be a very small amount of it, but not very much of it is happening okay.
It’s mostly produced by. The corpus luteum.
Of course there is some leakage.
❖Steroid hormones (synthesised from cholesterol mainly or Acetyl-CoA)
❖Act through nuclear (hormone) receptors.
OESTROGENS:
*Only secreted by ovaries in non-pregnant females
*Potency: β-Estradiol> estrone> estriol
PROGESTINS:
*Progesterone is the most important
*Only secreted by Corpus Luteum in non-pregnant females
Female Sex Hormone breakdown
So if we look then I’m going to go back to this.
If we look then at the sex hormones and what order they occur in our hypothalamus at the top makes gonadotropin releasing hormone,
just as we had in the males.
And though that leads to the production of luteinizing hormone and FSA from the pituitary, from those gonadotroph cells, just as happened in males.
Okay. And the difference then is that FSH.
Stimulates the growth of the follicle. That you can see here.
So in the ovary are follicles going from small to medium to large.
Whereas LH is going to literally just control.
The timing here okay.
It is important in an eye opening oestrogen to be produced.
But it is essential for ovulation timing.
It does that all on its own, so LH stimulates ovulation.
It also stimulates the corpus luteum to continue producing some of the hormones that are needed if fertilisation occurs.
So. Where we have our corpus luteum over here.
It will produce progesterone relaxin inhibin and oestrogens.
Because remember it’s still got those theca cells and it’s still got the granulosa cells although it’s shrinking.
They’re still alive. Okay. At this point
So they are still able to produce. Progesterone and oestrogen.
Okay. Progesterone mostly stimulates the endometrium.
It stimulates the growth of the endometrium.
And we’ll touch on that in a minute. Okay. And it also has the ability to cause negative feedback which is what this dotted line going back up is
That will reduce the amount of luteinizing hormone that’s being released from the anterior pituitary.
And it will have a negative feedback on.
The gonadotropin releasing hormone being released from the hypothalamus.
Not only does this structure produce progesterone, but as we said before,
this can produce oestrogen, although it is much lower than what is produced from the follicle.
Okay, so in terms of oestrogen, mostly its job is to support growth of the endometrium so that it’s ready After ovulation has happened to enable a fertilised ovum to implant into the endometrium. So from the corpus luteum, that’s what its job is.
Okay. It’s got a slightly different job during the follicular phase, which we’ll come on to in a minute.
However, we’ve also got inhibin here. Remember inhibin inhibits the release of FSH.
So it’s also got a negative feedback happening.
This new hormone here relaxin inhibits contractions.
Okay. So during that phase of when someone is pregnant it makes sure.
That the uterine muscle is smooth. There’s no contraction going on until right at the end when it’s needed.
And then there’s a large release of relaxin and it slowly builds up.
It also makes sure, that there is a relaxation, hence its name of tendons that hold together the pelvic bones.
Because if you’ve ever looked at a skeleton, the pelvis is actually several bones that are held together with tendons,
and those stretch and lengthen during pregnancy with the release of this relaxin.
Okay, and that’s the reason why if you know anyone who’s been pregnant, they’re was told, don’t do heavy exercise,
uh, while you’re pregnant, don’t do heavy exercise for at least six weeks after you have the baby.
And actually, the relaxin can go on and affect your body for about a year afterwards,
because there’s a probability that you could really quite injure yourself,
because those tendons are a lot more stretchy everywhere in your body, uh, than they would have been before.
So if we look at FSH now, then in terms of FSH, it stimulates that initial growth of the ovarian follicle,
which we knew already that granulosa cells multiply.
But then also it will induce the release of oestrogens.
And of course the Oestrogens will then increase that in response as well.
So we’re getting lots of generation of growth going on in that follicle.
And the oestrogen will also. Stimulate a negative feedback back to the hypothalamus.
So this hypothalamus is getting controlled by lots of different things.
Okay. It’s slightly different to what is seen in the males.
Inhibin, as we said before, also feeds back along with oestrogen to control the release of FSH.
And so during the ovulatory phase, most of the oestrogen is generated from those ovarian follicles.
Not very much else other than inhibin is released from here.
Because FSH has a slightly different role here, it’s kept relatively low until closer to ovulation.
Female Sex Hormones phases during the month
Inhibin, as we said before, also feeds back along with oestrogen to control the release of FSH.
And so during the ovulatory phase, most of the oestrogen is generated from those ovarian follicles.
Not very much else other than inhibin is released from here.
Because FSH has a slightly different role here, it’s kept relatively low until closer to ovulation.
So you can see that here on this graph.
Usually we say that the length of the menstrual cycle or ovarian cycle for a woman would be 28 days.
So four weeks. And if you take a contraceptive pill that will be the case.
However, uh, it can be shorter than this or it can be longer than this.
Okay. So from now on, when I say a day number, it relates to a standard cycle.
It is not the same for everybody. Everybody is different.
Okay. So normally we’d expect follicle stimulating hormone and luteinizing hormone to be
relatively low along with oestrogen and progesterone until about today 8 to 10.
Hey now, at that point,
luteinizing hormone starts to rise quite sharply because what it’s trying to do here is it’s trying to ensure that ovulation is going to happen.
Remember I said to you, that’s what it’s really important for. Luteinizing hormone has to spike really high for ovulation to be induced.
If there’s no peak then luteinizing hormone will not have caused ovulation and there will be no release of the secondary oocyte.
Okay. About day 6 to 8.
Oestrogen starts to rise as this is starting to move up slightly.
This makes sense because remember we need our luteinizing hormone and a little bit of FSH if we’re going to generate oestrogen.
And of course it’s now driving a much greater urgency of growth at this
point because you’ve got now one selected strongest follicle that needs to expand in size.
Luteinizing hormone peaks and FSH.
has a small peak just around the same time.
Okay. But then instead of it dropping quite rapidly after, which happens to LH here, this stays relatively high for until about day 14 or 15.
So ovulation is happening about day 12 to 14.
as this peaks
After the oocyte has been released the oocyte takes about 2 to 4 days to move down the fallopian tube, and if it’s going to be fertilised, it will need to happen within that time.
Okay. Meanwhile, quietly in the background, progesterone has been slowly increasing as it comes towards, um, ovulation and just after,
because it’s trying to make sure that the endometrium is ready and raring to go for this, uh, potentially fertilised ovum to arrive.
And this progesterone continues to rise until about day 22.
And if there’s no fertilised ovum that is embedded in the endometrium, then this will start to drop off.
and at day 24 or so, is when menstruation starts to happen.
Menstrual cycle
Okay. So.
How does this relate to what’s going on here?
Here is our ovarian cycle. We’ve talked about what happens in the ovary.
And I’ve talked a little bit about what happens in the endometrium.
But I hadn’t talked about any particular structures in detail until now.
So during this part of the ovulatory phase where oestrogens are being produced.
You can see that the endometrium near the end of the menstrual phase along With the release of oestrogen the cells start to get much larger.
And then as progesterone starts to rise about day ten onwards,
you can see that there’s an increase in the amount of blood supply that’s provided during that time.
The reason for this is that obviously, if a fertilised ovum comes along and embeds in the endometrium, it needs the supply of blood.
It needs a supply of nutrients. And these are all going to come from this well prepared endometrium.
It’s like the hormones have been prepping this really lovely nice comfy bed for this fertilised ovum.
Come along and lay in it
Okay. But if that process doesn’t happen and something that’s fertilised doesn’t come along by the time it gets to day 24 and 25,
it goes, okay, nobody’s coming and starts packing away.
Okay? And when I say starts packing away, it means it’s got to reduce that thickness.
And so it’s actually cells that are shrivelling up and dying off.
And we actually see those that are coming out as a period okay.
So when we say menstruation is happening and it looks like blood, but these are just old dying cells that are coming out.
And that continues, as we said, back in to about day 1 to 4 probably.
Of that beginning of the cycle.
Secondary Female Sex Characteristics
In terms of secondary sex characteristics, which we talked about males about deep voice, lots of muscle, wide shoulders, all of those kinds of things.
Uh, in females, these are that we get the lay down of more adipose tissue.
So, uh, breasts develop and you can see here that all that yellow stuff in here is, um, fat that’s deposited, which happens post puberty.
The mammary glands that are in here that you can see that look like little purple grapes in this diagram.
These are only doing something if the person is pregnant.
So they start to expand and they start to fill with milk okay.
They are not otherwise doing anything. They are there, but they are not, uh, fully developed until somebody, um, undergoes pregnancy.
And it’s the oestrogens that we, um, experience.
The levels of oestrogens go up, um, that induce this growth in the breast tissue, which is why sometimes people report that,
for example, they have sensitive, um, breasts around the time of, um, their menstrual cycle.
Um, and that might possibly be related to the high levels of oestrogen that are seen during the cycle.
Of course, after puberty, oestrogen doesn’t just have an effect on breasts or cause your thumb to get larger,
because that generally happens as you, um, go through puberty.
But it also causes the growth of the uterus and the fallopian tubes and, uh, the vagina and the vulva,
for example, readying it so that it is ready for supporting an embryo if it was required.
Because if you think about evolutionary speaking, people didn’t live as long as we do now.
So millions of years ago, people may have had a baby as early as maybe 12 to 15 years old.
So there’s lots of development in terms of, um, deposition of fat and changes in metabolism, um, growth of the sexual organs.
These all happen during puberty.
Menopause
What happens at the other end is also something we need to very briefly touch on.
So menopause happens usually between the ages of 40 and 60, um,
where menstrual cycle becomes a lot less regular so if you don’t have a period for six months, then you’re considered to be in menopause.
Okay. And this is where the oestrogen and progesterone levels will drop really really low.
and there’s lots and lots of research going on that says as mind fog and all sorts of things like
that that are really affected by oestrogen because it’s not just something that acts locally,
okay? It acts in all sorts of places.
And we’ll see next week that it also has effects on the skeleton and things like that as well.
Okay. So. The primordial follicles are remaining um and producing oestrogen at this particular time.
They aren’t producing very much, and it decreases to a very low level.
Um, and there’s not enough concentration of oestrogens to inhibit LH and FSH.
So of course this means that you get hot flushes, irritability, fatigue, um, also decreased strength is also observed.
Hormonal regulation of the Ovarian cycle
So this slide just and the next one just sort of summarise everything in timing order.
It’s got lots of text on there to help you know what’s happening at each step.
Okay. So the gonadotropin releasing hormone.
It’s up here in the hypothalamus is released.
And that causes the release of FSH and LH.
These will support follicle growth and luteinizing hormone peaks to cause ovulation.
This is also happening at the same time as we’ve got an increase in the proliferation in the endometrium
And as ovulation occurs in response to that high luteinizing hormone, there’s an increase in the secretion of progesterone and oestrogen,
which increase the blood supply to those growing cells of the endometrium
There’s also stimulation after this event of luteinizing hormone and of the corpus luteum by luteinizing hormone.
And that means that we continue to generate, uh, progesterone.
There is an increase in the size that continues until there is no, production of a fertilised ovum,
in which case the corpus luteum, disappears, it becomes the corpus albicans, and you start to induce menstruation.
Remember that we have got negative feedback that happens through inhibin and oestrogen and progesterone.
These are highlighted by the red lines.
We also need to bear in mind that we do have a feed back to the anterior pituitary.
That happens through inhibin. And the low levels of progesterone and oestrogen can actually
stimulate the release of gonadotropin releasing hormone from the hypothalamus.
*GnRH stimulates production of Luteinising Hormone (LH) and Follicle Stimulating Hormone (FSH) by gonadotrophcells in anterior pituitary.
*LH & FSH stimulate production of Oestrogen by cells inside the follicle (follicle maturation).
*Low levels of progesterone and oestrogen stimulate GnRH release and LH and FSH production in the anterior pituitary.
*Cells around the follicle secrete androgens in response to LH, driving cells inside follicle to multiply.
*Increase in cell number within the follicle causes an increase in overall oestrogen levels.
*Inhibin is produced by secondary follicles to prevent release of LH & FSH from anterior pituitary →build up of packaged hormones.
Hormonal regulation of the Ovarian cycle
So this slide just and the next one just sort of summarise everything in timing order.
It’s got lots of text on there to help you know what’s happening at each step.
Okay. So the gonadotropin releasing hormone.
It’s up here in the hypothalamus is released.
And that causes the release of FSH and LH.
These will support follicle growth and luteinizing hormone peaks to cause ovulation.
This is also happening at the same time as we’ve got an increase in the proliferation in the endometrium
And as ovulation occurs in response to that high luteinizing hormone, there’s an increase in the secretion of progesterone and oestrogen,
which increase the blood supply to those growing cells of the endometrium
There’s also stimulation after this event of luteinizing hormone and of the corpus luteum by luteinizing hormone.
And that means that we continue to generate, uh, progesterone.
There is an increase in the size that continues until there is no, production of a fertilised ovum,
in which case the corpus luteum, disappears, it becomes the corpus albicans, and you start to induce menstruation.
Remember that we have got negative feedback that happens through inhibin and oestrogen and progesterone.
These are highlighted by the red lines.
We also need to bear in mind that we do have a feed back to the anterior pituitary.
That happens through inhibin. And the low levels of progesterone and oestrogen can actually
stimulate the release of gonadotropin releasing hormone from the hypothalamus.
amounts of hormones
So I’ve given you. These.
And I’m going to say it very loud. Do not remember then, but come back and look at them if needed.
Okay, I’m going to set one more time. Do not remember them, but come back here and look at them if it’s needed.
Okay, if we’re asking you in some data question to look for these, this is where to come for them.
