Patient demographics RNA Extraction. A standard extraction protocol included with the RNAzol kit was used. The sample tubes 0. Control RNA [ bp chloramphenicol acetyltransferase derived from previous experiments in the lab] was included to verify the PCR reaction. Primers The sequence for the forward and reverse PCR primers were as follows: The specific activity obtained for the probes was , to , cpm per 10 pmol. After electrophoresis, the gels were denatured and blotted to Zeta probe Nylon membrane Bio-Rad , transfer was done by alkaline transfer method using 0. A final wash was done with buffer II at room temp for 15 min. The relative abundance of hybridization was measured using ImageQuant software Amersham Biosciences under uniform parameters of equal area, pixels, color intensity values etc.
Dating the Endometrial Biopsy
A pelvic or gynaecologic ultrasound is an ultrasound of the female pelvis. Most pelvic ultrasounds are performed using both the transabdominal and transvaginal approaches. Transabdominal ultrasound involves scanning through your lower abdomen.
It is asserted that examination of the endometrium during the secretory phase yields more information about the time of ovulation, degree of progestational change, and normality of the endometrium than any other test used in sterility studies.
In this context, it was felt to be of interest whether or not the hysteroscopic appearance of the endometrium is associated with early pregnancy loss as well as with implantation failure of fertilized ovum. The maturation process of the endometrium is mediated by ovarian sex steroid hormones. In clinical practice, morphological information on endometrial maturation is available from histological evaluation and ultrasound assessment.
Patients with chemical or subclinical abortion, i. The endometrium of the secretory phase is morphologically and functionally well-prepared for implantation and the early development of the fertilized ovum. In the mid-secretory phase days post-ovulation or days of the cycle , the glands remain dilated, but become more irregular with a papillary or saw-tooth appearence.
First Trimester / Dating Ultrasound
Myomectomy can be conducted in various ways: The gross appearance of these specimens is particularly important with regard to the question of possible leiomyosarcoma. Conversely, however, most grossly peculiar myomectomy specimens ultimately prove to be benign when their histologic appearances have been evaluated. The preoperative administration of gonadotropin-releasing hormone has been shown to reduce the size of myomas, and these agents may modify the appearance of benign smooth muscle tumors 6 — 9 ; however, the gross and histologic correlates of this clinical shrinkage have been difficult to establish with certainty
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Can the TruTest capture enough tissue if the patient is bleeding heavily? Bleeding is an indication for endometrial biopsy so this is a common occurrence. We have seen many examples of adequate tissue and cytology during menstrual phase, even with menorrhagia. Also, remember that Gynecor’s unique fixative allows us to deliver a result on cytology from every specimen returned to us, regardless of what the tissue yields.
While there are no reports of complications, theoretically the brush could entangle in the IUD, and it would be prudent to inform the patient of this occurrence. It is recommended that an endometrial biopsy be performed prior to the insertion of an IUD. Can the TruTest be used for endometrial dating for infertility testing? Secretory endometrium can be dated from the cell block but not the cytology. The client would also have to specifically request dating on the Gynecor requisition.
Why is a specimen obtained with the Tao Brush taken at the fundus and not in the lower section of the uterus. Would a tumor in the lower section of the uterus be missed with this method? Placing the bead of the brush of the fundus is the best method for collecting samples. Endocervical contamination is a problem and adherence to proper technique is needed to keep it at a minimum.
Inserting the brush all the way to the fundus before retracting the sheath is part of that technique.
To reassess endometrial morphological criteria of normality identifying the best To date endometrium , should see surface endometrium , but date based on most advanced area; Must biopsy uterine corpus above the level of the isthmus; must. Endometrial maturation, important in the diagnosis of infertile couples , has been evaluated since using the Noyes criteria.
secretory phase- Progesterone modulated- date specific features. Features for Endometrial Dating.
Uterus Uterus The uterus is made up of an external layer of smooth muscle called the myometrium, and an internal layer called the endometrium. The endometrium has three layers: The Stratum compactum and stratum spongialis develop into the stratum functionalis during the first half of the menstrual cycle proliferative phase The wall of the uterus changes during the menstrual cycle, as shown diagramatically here.
Proliferative Phase In the proliferative phase, facilitated by FSH, the endometrium thickens, connective tissue is renewed, along with glandular structures and ehlicrine arteries. Oestrogen causes the endometrial stroma to become deep and richly vascularised. Simple tubular glands in the stratum functionalis open out onto the surface, and the endometrium thickens. Can you recognise the lumen, stratum compactum, stratum spongiosum stratum basalis and myometrium in this photograph?
Secretory Phase In the secretory phase, facilitated by LH, the endometrial glands become cork-screw shaped, and filled with glycogen. They secrete a glycogen rich secretion during the secretory phase after ovulation. You should be able to recognise the glands, and glycogen secretions in this high magnification photo of a secretory phase uterus. Menses Decreased levels of LH and progesterone result in the menstrual phase, or menses. During menses shedding of the uterine lining, which occurs if the egg is not fertilised the spiral arterioles in the stratum functionalis layer contract, resulting in ischaemia, and degeneration of the functionalis layer.
The arteries rupture, and the rapid blood flow dislodges the necrotic functional layer, which is lost. The basal layer is unaffected, because it is supplied by straight arteries.
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References Progesterone Support in Pregnancy [Figure , p. Furthermore, most of the assessment of progesterone in pregnancy as it relates to various complications of pregnancy was accomplished from the early s through the early s. In spite of improvements in the accuracy and precision of progesterone assays since that time and a better ability to date pregnancy and establish more accurate gestational ages, very little subsequent work has been accomplished in this area. However, data on the level of progesterone in normal pregnancy, and as it relates to a variety of pregnancy-related complications and features of previous reproductive history has been generated in a study which was conducted from the years through at the Pope Paul VI Institute.
Modern means of progesterone assessment with improved accuracy and precision were used along with more precise means of dating the pregnancies.
Finally, it should be stressed that the emphasis of these articles should be on the “histological dating” and not on the “endometrial biopsy.” These studies and the one by Murray et al. (6) put to rest the use of histological dating of secretory endometrium in the routine evaluation of the infertile couple.
E2 is the most potent, whereas E1 is less potent in binding to estrogen receptors. The regulation of the enzymatic activity to oxidize E2 to E1 in the endometrium was studied extensively in the s by Tseng et al. The activity was approximately 10 times greater in the secretory endometrium than in the proliferative endometrium, and elevation of the enzyme activity was induced by progesterone that was secreted from the corpus luteum during the secretory phase via the progesterone receptor-mediated action.
Similar findings have been reported by several other investigators 11 , 12 , Although histologically the eutopic endometria of patients with benign uterine diseases resemble the endometria of disease-free uterus, the estrogen metabolism may be remarkably different 18 , Materials and Methods Section:
Dating the endometrial biopsy.
The discipline has benefited greatly from substantial recent advances in reproductive biology and allied fields and technologic improvements in computers, ultrasonography, and surgical instrumentation. All of these developments have been applied to clinical practice at an unprecedented rate. The first full-term pregnancies were achieved with oocytes from unstimulated cycles 2.
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Rock Abstract It is asserted that examination of the endometrium during the secretory phase yields more information about the time of ovulation degree of progestational change and normality of the endometrium than any other test used in sterility studies. Attention to qualitative changes in 8 morphological factors is most useful in dating the endometrial biopsy. During the 1st week of luteal activity attention should be focused on changes occurring in gland epithelium: During the 2nd week stromal changes including edema predecidual reaction stromal mitosis and leukocytic infiltration are the key criteria.
Design. Duplicate endometrial biopsies from 25 women were dated by five evaluators on two separate occasions to evaluate the overall precision of the measure.
The methods that have been used in the past were indirect, assumptive and not reproducible. Researchers in Spain have created a new tool which has been shown to be promising for identifying molecular markers for uterine receptivity. Remarkably, as the blastocyst floats within the uterine cavity looking for a place to land, a dialog takes place between the blastocyst and the endometrium.
The hormonal preparation of the uterus plays a critical role each month in creating this environment in which the blastocyst can adhere to the endometrium in the hope that implantation will take place. The uterine lining undergoes changes during the two phases of the menstrual cycle that prepare it for blastocyst implantation. During the proliferative phase, it grows due to the increasing production of estrogen by the ovaries.
The second phase is called the secretory phase where the production of progesterone, produced by the corpus luteum, converts the endometrial lining to a secretory one, changing the cells to prepare for implantation a process called decidualization. Should implantation not take place, the hormone levels will fall, resulting in a shedding of the lining, which results in menses.
Studying the mid-secretory phase is of great importance since the window of implantation WOI takes place then. The sweet spot of WOI is approximately a 2-day period when the uterus is prepared to accept the implantation of a blastocyst. Conventionally, it was assumed that every woman had the same WIO, approximately days after ovulation so embryo transfers would be scheduled to take place during this time.
This theory has recently been challenged, with researchers proposing that the WOI can vary among women. In the past, this was done using histological criteria, i.