Adaptation of a trap door technique for the recovery of ovarian cortical biopsies from Cebus apella (capuchin monkey)

Zygote ◽  
2012 ◽  
Vol 21 (2) ◽  
pp. 158-161 ◽  
Author(s):  
Luana N. Santana ◽  
Adriel B. Brito ◽  
Danielle C. Brito ◽  
Julianne S. Lima ◽  
Sheyla F.S. Domingues ◽  
...  
Keyword(s):  
Ovarian Tissue ◽  
Capuchin Monkeys ◽  
Safe Procedure ◽  
New World Primates ◽  
Primordial Follicles ◽  
Trap Door ◽  
Ovarian Structure ◽  
Biopsy Method ◽  
Follicular Density

SummaryThere is a paucity of efficient cryopreservation protocols for primordial follicles enclosed in the ovarian tissue from non-human primates (NHP), in special New World primates. Our objective was to establish an optimal procedure for the recovery of ovarian biopsies from capuchin monkeys. To this end, we adapted a trap door biopsy method. Follicular density and quality of the biopsies were evaluated and ultrasound analysis was performed before and continuously after surgery to assess ovarian structure. Ovarian tissue biopsies recovered by the trap door technique allowed the successful harvesting of primordial follicles from capuchin monkeys, and no complication was recorded. The female cycle was not affected by surgery and no adherence was found thereafter. In conclusion, the adaptation of a trap door biopsy method is a safe procedure and allows recovery of healthy primordial follicles.

scholarly journals 203 ASSESSMENT OF OVARIAN RESERVE. IS THERE A ROLE FOR OVARIAN BIOPSY?

2005 ◽  
Vol 17 (2) ◽  
pp. 252
Author(s):  
R. De Roover ◽  
C. Hanzen
Keyword(s):  
Ovarian Reserve ◽  
Ovarian Tissue ◽  
Primordial Follicles ◽  
Ovarian Cortex ◽  
Human Fertility ◽  
Multiple Biopsies ◽  
Size And Morphology ◽  
Blind Sampling ◽  
Small Parts

The pool of primordial follicles in the ovary or ovarian reserve is a major factor in human fertility potential. In bovine medicine as well, this ovarian reserve has been linked to the results of superovulation procedures (Cushman et al. 1999 Biol. Reprod. 60, 349–354). These authors suggested a biopsy to assess the level of this reserve. Whether the biopsy(ies) is(are) a true reflection of the follicular distribution in the ovarian cortex, is (to the best of our knowledge) a factor never investigated until now in bovine medicine. In human medicine, this procedure has been critically examined for that particular use and found not to be suited (Lass et al. 2004 Hum. Reprod. 19, 467–469). Indeed, randomized or “blind” sampling of one biopsy is adequate only if follicles are evenly spread in the ovarian cortex; in any case they are not deeper than a few mm from the surface. Moreover, the quantitative counting of follicles does not provide any information about the quality of the oocytes embedded in them. Taking a biopsy of a bovine ovary in a minimally invasive way is technically feasible (Aerts 2004 Reprod. Fertil. Dev. 16, 229–230). Therefore, the aim of this study was to examine the natural distribution of primordial follicles in the ovarian cortex of bovine ovaries. Slaugtherhouse ovaries were collected at random. The volume (mL) was measured and the macroscopically visible follicles were counted. Then the ovaries were cut in slices of 5Âμm, and every 8th (8 × 5 = 40 μm interval) slice was subjected to fixation in formalin and hematoxylin-eosin staining. Before counting of the primordial follicles, the ovarian cortex was subdivided into 8 equal parts. These “parts” were supposed to mimick a (single) ovarian biopsy. The 8 parts of a slice represent here multiple biopsies. For each of these parts, the number of primordial follicles was counted; only follicles with a visible oocyte were included. The results of the parts containing the ligament of the ovary were excluded. Results are shown in Table 1. The results show that the distribution of primordial follicles between small parts of the bovine ovarian tissue was extremely uneven. A large variation was observed between samples obtained from the same ovary. Moreover, an extrapolation of follicle numbers found in biopsies to entire ovaries were hampered by the uneven size and morphology of these ovaries. Therefore, we conclude that the use of single biopsies of ovarian cortex for a quantitative evaluation of the ovarian reserve has limited value; an empty cortex or a cortex with very few follicles might be just incidental and meaningless. Even the use of multiple biopsies, although less variable, does not solve the problem of extrapolation of these data to entire ovaries. Table 1. Macroscopically visible follicles on 4 ovaries and primordial (“microscopical”) follicles on 4 slices of each of these ovaries

scholarly journals In-vitro fragmentation of ovarian tissue activates primordial follicles through the Hippo pathway

2020 ◽  
Vol 2020 (4) ◽  
Author(s):  
C De Roo ◽  
S Lierman ◽  
K Tilleman ◽  
P De Sutter
Keyword(s):  
Tissue Culture ◽  
Ovarian Tissue ◽  
Hippo Pathway ◽  
Primordial Follicle ◽  
Akt Pathway ◽  
Primordial Follicles ◽  
Primordial Follicle Activation ◽  
The Hippo Pathway ◽  
Follicle Activation

Abstract STUDY QUESTION What is the role of the Hippo and PI3K/Akt pathway in follicles during ovarian tissue culture in tissue derived from oncological patients and transgender men? SUMMARY ANSWER Results highlight a Hippo pathway driven primordial follicle activation in vitro, predominantly from Day 0 to Day 4. WHAT IS KNOWN ALREADY In-vitro ovarian tissue culture aims at activating and maturing primordial follicles for fertility restoration in patients with a threatened ovarian reserve. Not all patients are eligible for ovarian cortex transplantation and therefore several groups are attempting to culture ovarian tissue in-vitro. Cortex fragmentation disrupts the Hippo pathway, leading to increased expression of downstream growth factors and follicle growth. The PI3K/Akt pathway is considered the intracellular pathway to where different extracellular factors involved in primordial follicle activation in-vivo converge. In order to optimise current ovarian tissue culture models, information on progression of these pathways during tissue culture is mandatory. STUDY DESIGN, SIZE, DURATION The first step of a multistep cortex culture system was performed using 144 ovarian cortex pieces from a total of six patients. Per patient, 24 cortical strips were cultured for 6 days and six pieces per patient were collected for downstream analysis of follicle development and Hippo and PI3K/Akt pathway targets every second day. PARTICIPANTS/MATERIALS, SETTING, METHODS Ovarian tissue was obtained from oncological (N = 3; 28.67 ± 4.51 years) and transgender (N = 3; 23.33 ± 1.53 years) patients. Follicles were analysed using haematoxylin-eosin staining and pathways were studied using immunohistochemistry and precise follicle excision by laser capture micro-dissection for RT-qPCR analysis. MIQE guidelines for RT-qPCR were pursued. Reference gene selection (GAPDH, RPL3A, 18s rRNA) was performed using GeNorm Reference Gene Selection Kit. Statistical analysis was conducted with IBM SPSS Statistics 23 (Poisson regression, negative binomial regression, ANOVA and paired t-test). MAIN RESULTS AND THE ROLE OF CHANCE Immunohistochemical analysis confirmed a Hippo pathway driven primordial follicle activation due to mechanical manipulation of the cortical strips. Ovarian tissue preparation and culture induced the inhibitory phosphorylated Yes-associated protein (pYAP) to disappear in granulosa cells of primordial follicles on Day 2. The stimulatory YAP on the contrary appeared in primordial granulosa cells over increasing culture days. Looking at the YAP target connective tissue growth factor (CTGF), a significantly up-regulated CTGF was noted in primordial follicles when comparing Day 2 and Day 4 (ratio Day 2/4 = 0.082; P < 0.05), clearly showing an effect on the Hippo pathway in primordial follicles during tissue culture. Follicle classification showed a significant drop in estimated primordial follicle counts in the oncological cohort (−78%; P = 0.021) on Day 2 and in the transgender cohort on Day 4 (−634%; P = 0.008). Intermediate follicle counts showed a non-significant increasing trend to during culture and this follicle recruitment and growth resulted in a significant rise in estimated primary follicle counts on Day 6 in oncological patients (170%; P = 0.025) and, although limited in absolute numbers, a significant increase in secondary follicles on Day 4 (367%; P = 0.021) in the transgender cohort. Subsequent antral follicle development could not be observed. LIMITATIONS, REASONS FOR CAUTION A limitation is the small sample size, inherent to this study subject, especially as a large amount of tissue was needed per patient to reduce inter-patient variation in different downstream analysis techniques. A particular and specific weakness of this study is the inability to include an age-matched control group. WIDER IMPLICATIONS OF THE FINDINGS These findings support an adapted tissue preparation for Hippo pathway disruption and a shorter first phase of tissue culture. This work may also have implications for transplantation of cryopreserved tissue as larger strips (and thus slower burnout due to less Hippo pathway disruption) could be a benefit. STUDY FUNDING/COMPETING INTEREST(S) This research was financially supported by the Foundation Against Cancer (Stichting tegen Kanker, TBMT001816N), the Flemish Foundation of Scientific Research (FWO Vlaanderen, FWO G0.065.11N10) and the Gender Identity Research and Education Society (GIRES) foundation. The authors declare no competing interests. TRIAL REGISTRATION NUMBER N/A.

scholarly journals Subcutaneous Quadrantectomy Is A Safe Procedure in Management of Early Stage Breast Cancer.

2021 ◽  
Author(s):  
Eva Lieto ◽  
Annamaria Auricchio ◽  
Silvia Erario ◽  
Giovanni Del Sorbo ◽  
Francesca Cardella
Keyword(s):  
Breast Cancer ◽  
Quality Of Life ◽  
Axillary Dissection ◽  
Early Stage ◽  
Length Of Hospital Stay ◽  
Oncologic Outcome ◽  
Conservative Surgery ◽  
Early Stage Breast Cancer ◽  
Safe Procedure

Abstract Background: Surgical treatment of breast cancer, the most frequent cancer in women, is deeply changed in the last years turning towards a progressive minimally invasion, both in extend of demolition and in axillary dissection completeness. This is due to the flexibility of the concept of radicality that today is taylored on the patient, rather than on the disease; If oncologic radicality is preserved, a less invasive operation on the breast is mandatory. In the era of mini-invasive surgery, a patient may ask for an aesthetic care without any additional health risk.Methods: In this article, we compared two groups of 75 consecutive patients undergoing conservative surgery for early stage breast cancer; the two groups were randomized for standard quadrantectomy and totally subcutaneous quadrantectomy. Statistical analysis was carried out for comparing data.Results: No difference in oncologic outcome was found with the different surgical procedures; the length of hospital stay and the incidence of late breast deformities were significantly less in subcutaneous quadrantectomy group. Conclusions: We conclude that, in early breast cancer, a totally subcutaneous surgical procedure of resection is feasible and safe and ensures an absolutely better aesthetical result, that involves patient’s quality of life.

scholarly journals POSSIBILITIES OF ULTRASONIC MONITORING OF ENDOMETRIUM AND OVARIES IN PATIENTS AFTER CANCELLATION OF LONG-TERM RECEPTION OF COMBINED ORAL CONTRACEPTIVES WHEN PLANNING PREGNANCY

2019 ◽  
Vol 6 (4) ◽  
pp. 215-220
Author(s):  
Sh. N Bortsvadze ◽  
Evgeniya A. Svidinskaya ◽  
T. A Dzhibladze ◽  
I. D Khokhlova ◽  
Yan Van
Keyword(s):  
Menstrual Cycle ◽  
Oral Contraceptives ◽  
Ovarian Tissue ◽  
Hormonal Contraception ◽  
Reproductive Function ◽  
Good Health ◽  
Automatic Assessment ◽  
Normal Limits ◽  
Ultrasound Diagnostics

The article aims to assess the possibility of ultrasound scanning and dopplerstudy in the assessment of the condition of the ovaries and endometrial in patients after long intake of COC, predicting the restoration of reproductive function after the abolition of hormonal contraception. Material and methods. In 2018-2019, 37 women were examined after the abolition of oral contraceptives, which they took for a long period of time. At the time of the examination, all the patients stopped taking COC at least 2 months ago due to the fact that they were planning a pregnancy. UW the study was conducted on the apparat Voluson E8 Expert (General Electric), improving the quality of diagnostics used Automatic assessment of follicle condition based on ultrasonic echography (Sono AVCTM follicle) and tomographic ultrasound (TUI). Results. According to the study significant changes recorded 17 patients whose menstrual cycle did not recover within the first 6 months after the cancellation of COC. In a group of 20 patients with a regular menstrual cycle, the results of the study were within normal limits, pregnancy within 6 months occurred on their own in 8 of them. Conclusion. The possibilities of 3 dimensional reconstruction and programs for evaluating the follicular apparatus significantly improve the quality of ultrasound diagnostics, give more information about the presence of small follicles, blood flow in the ovarian tissue, the structure of the cortical apparatus substances and stroma, whichis the ability of a clinician to obtain a complete understanding of the condition of the ovaries and endometrial and predicting a good health of patients. The article may be of interest to obstetricians-gynecologists, ultrasound specialists, endocrinologists, reproductive specialists.

scholarly journals Breast-conserving treatment in oncoplastic times: indications, cosmesis, and quality of life

Mastology ◽  
2021 ◽  
Vol 31 ◽  
Author(s):  
Idam de Oliveira-Junior ◽  
Raphael Luiz Haikel ◽  
René Aloísio da Costa Vieira
Keyword(s):  
Breast Cancer ◽  
Quality Of Life ◽  
Breast Conserving Surgery ◽  
Safe Procedure ◽  
Breast Conserving Treatment ◽  
Oncological Treatment ◽  
Life Data ◽  
Generic Term ◽  
Cosmetic Outcomes

Breast-conserving treatment was established as an oncologically safe procedure for breast cancer. However, the cosmetic outcomes of breast-conserving treatments are often unsatisfactory. In this scenario, oncoplastic breast-conserving surgery incorporated plastic surgery concepts and techniques into the oncological treatment in order to ensure better cosmesis, thus increasing the indications for breast-conserving treatment. At the same time, oncoplastic breast-conserving surgery is usually presented as a generic term, which should be evaluated taking many aspects into account: indication, patient selection, the surgery itself, cosmetic quality, and quality of life — data that are still scarce in the literature.

scholarly journals In vitro follicle culture in the context of IVF

Reproduction ◽  
2018 ◽  
Vol 156 (1) ◽  
pp. F59-F73 ◽  
Author(s):  
Anamaria C Herta ◽  
Francesca Lolicato ◽  
Johan E J Smitz
Keyword(s):  
Fertility Preservation ◽  
Polycystic Ovary ◽  
Ovarian Tissue ◽  
Primordial Follicle ◽  
Assisted Reproduction Techniques ◽  
Realistic Potential ◽  
Primordial Follicles ◽  
Primordial Follicle Activation ◽  
Extracellular Matrix Components

The currently available assisted reproduction techniques for fertility preservation (i.e.in vitromaturation (IVM) andin vitrofertilization) are insufficient as stand-alone procedures as only few reproductive cells can be conserved with these techniques. Oocytes in primordial follicles are well suited to survive the cryopreservation procedure and of use as valuable starting material for fertilization, on the condition that these could be grown up to fully matured oocytes. Our understanding of the biological mechanisms directing primordial follicle activation has increased over the last years and this knowledge has paved the way toward clinical applications. New multistepin vitrosystems are making use of purified precursor cells and extracellular matrix components and by applying bio-printing technologies, an adequate follicular niche can be built. IVM of human oocytes is clinically applied in patients with polycystic ovary/polycystic ovary syndrome; related knowhow could become useful for fertility preservation and for patients with maturation failure and follicle-stimulating hormone resistance. The expectations from the research on human ovarian tissue and immature oocytes cultures, in combination with the improved vitrification methods, are high as these technologies can offer realistic potential for fertility preservation.

P–449 Evaluation of ovarian reserve in female children and adolescents with non-iatrogenic primary ovarian insufficiency to establish criteria for ovarian tissue cryopreservation

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
A Volodarsky-Perel ◽  
M Zajicek ◽  
D Shai ◽  
H Raanani ◽  
N Gruber ◽  
...  
Keyword(s):  
Fertility Preservation ◽  
Ovarian Reserve ◽  
Predictive Value ◽  
Ovarian Function ◽  
Ovarian Tissue ◽  
Antral Follicle ◽  
Ovarian Tissue Cryopreservation ◽  
Positive Parameter ◽  
Primordial Follicles

Abstract Study question What is the predictive value of ovarian reserve evaluation in patients with non-iatrogenic primary ovarian insufficiency (NIPOI) for follicle detection in ovarian tissue harvested for cryopreservation? Summary answer Ovarian tissue cryopreservation (OTCP) should be considered if patients present at least one of the following parameters: detectable AMH, FSH≤20mIU/ml, detection of ≥ 1 antral follicle. What is known already In pre-pubertal girls suffering from NIPOI, which majorly has a genetic etiology, fertility preservation using OTCP is commonly practiced. When OTCP was performed in an unselected group of children and adolescents with NIPOI, only 26% of them had follicles in ovarian tissue while 74% did not benefit from the surgery. The role of preoperative evaluation of anti-müllerian hormone (AMH) serum level, follicular stimulating hormone (FSH) serum level, and trans-abdominal ultrasound for the antral follicle count to predict the detection of primordial follicles in the harvested ovarian tissue is unclear. Study design, size, duration We conducted a retrospective analysis of all patients ≤ 18 years old who were referred for fertility preservation counseling due to NIPOI at a single tertiary hospital between 2010 and 2020. If initial evaluation suggested a diminished ovarian reserve and at least one positive parameter indicating a follicular activity (AMH > 0.16ng/ml, FSH ≤ 20mIU/ml, detection of ≥ 1 antral follicle by transabdominal sonography), OTCP was offered. Patients with 46XY gonadal dysgenesis were excluded. Participants/materials, setting, methods OTCP was performed laparoscopically in all cases. A fresh sample of cortical tissue was fixed in buffered formaldehyde for histological analysis. The rest of the ovarian tissue was cut into small cuboidal slices 1–2 mm in thickness and cryopreserved. After the serial sections, the histological slides were evaluated for the presence of follicles by a certified pathologist. Follicles were counted and categorized as primordial, primary, and secondary. Main results and the role of chance During the study period, 39 patients with suspected NIPOI were referred to the fertility preservation center. Thirty-seven patients included in the study were diagnosed with Turner’s syndrome (n = 28), Galactosemia (n = 3), Blepharophimosis-Ptosis-Epicanthus Inversus syndrome (n = 1), and idiopathic NIPOI (n = 6). Of 28 patients with Turner’s syndrome, 6 had 45X monosomy, 15 had mosaicism and 7 had structural anomalies in X-chromosome. One patient with gonadal dysgenesis and one with the presence of Y-chromosome in 20% of somatic cells were excluded from the study. OTCP was conducted in 14 patients with at least one positive parameter suggesting ovarian function. No complications of the surgical procedure or the anesthesia were observed. Primordial follicles were found in all patients with two or three positive parameters (100%) and in three of six cases with one positive parameter (50%). In total, of the 14 patients who underwent OTCP with at least one positive parameter, 11 (79%) had primordial follicles at biopsy (mean 23.9, range 2–47). This study demonstrates a positive predictive value of 79% for the detection of primordial follicles in patients who had at least one positive parameter of ovarian reserve evaluation. If two or three parameters were positive, the positive predictive value increased to 100%. Limitations, reasons for caution This study did not examine the negative predictive value of our protocol as OTCP was not recommended in the absence of positive parameters. The future fertility potential of cryopreserved tissue in the population with NIPOI is unclear and should be discovered in further studies. Wider implications of the findings: We suggest the evaluation of ovarian reserve by antral follicles count, AMH, and FSH serum levels prior to OTCP in patients with NIPOI. By recommendation of OTCP only if ≥ 1 parameter suggesting the ovarian function is positive, unnecessary procedures can be avoided. Trial registration number Not applicable

scholarly journals Intranasal Ectopic Tooth in Adult and Pediatric Patients: A Report of Two Cases

2019 ◽  
Vol 2019 ◽  
pp. 1-5
Author(s):  
Isabela Polesi Bergamaschi ◽  
Bernardo Olsson ◽  
Aline Monise Sebastiani ◽  
Guilherme dos Santos Trento ◽  
Nelson Luis Barbosa Rebellato ◽  
...  
Keyword(s):  
Cleft Lip ◽  
Cleft Lip And Palate ◽  
Case Reports ◽  
Facial Trauma ◽  
Surgical Removal ◽  
Safe Procedure ◽  
Main Complaint ◽  
First Case ◽  
Ectopic Tooth

The aim of this study was to report two rare cases of ectopic tooth into the nasal cavity. The first case reports a 32-year-old female patient with the main complaint of having a tooth inside her nose. According to her, this condition causes pain and sporadic nosebleed. The patient had a facial trauma when she was 6 years old. The second case refers to an 8-year-old girl with left incomplete unilateral cleft lip and palate. The main complaint was left-sided nasal obstruction by a white hard mass. The treatment for both cases was surgical removal of the ectopic erupted tooth under general anesthesia. In conclusion, we can state that the surgical removal of intranasal tooth is a safe procedure and improves patient’s quality of life.

scholarly journals Effect of Previous Chemotherapy on the Quality of Cryopreserved Human Ovarian Tissue In Vitro

PLoS ONE ◽  
2015 ◽  
Vol 10 (7) ◽  
pp. e0133985 ◽  
Author(s):  
Babak Asadi Azarbaijani ◽  
Mona Sheikhi ◽  
Irma C. Oskam ◽  
Mirja Nurmio ◽  
Tiina Laine ◽  
...  
Keyword(s):  
Ovarian Tissue ◽  
Human Ovarian Tissue

202 EFFECTS OF ASCORBIC ACID ON IN VITRO CULTURE OF CATTLE PREANTRAL FOLLICLES

2010 ◽  
Vol 22 (1) ◽  
pp. 259
Author(s):  
E. R. Andrade ◽  
R. van den Hurk ◽  
L. A. Lisboa ◽  
M. F. Hertel ◽  
F. A. Melo-Sterza ◽  
...  
Keyword(s):  
Ascorbic Acid ◽  
In Vitro Culture ◽  
Ovarian Tissue ◽  
Immunohistochemical Analysis ◽  
Development Stage ◽  
Nuclear Antigen ◽  
Primordial Follicles ◽  
Preantral Follicles ◽  
Ovarian Cortex

The mechanisms that regulate the gradual exit of ovarian follicles from the nongrowing, primordial pool are poorly understood. The objective of this study was to evaluate the effects of adding ascorbic acid to the media for in vitro culture of cattle ovarian fragments and to determine the effects of this addition on the growth activation and viability of preantral follicles. The ovarian cortex was divided into small fragments; 1 fragment was immediately fixed in Bouin’s solution (control). The other fragments were cultured for 2, 4, 6, or 8 days on culture plates in minimum essential medium (MEM) supplemented with insulin-transferrin-selenium (ITS), pyruvate, glutamine, hypoxantine, BSA, and antibiotics (MEM+) or in MEM+ plus ascorbic acid (5, 25, 50, 100, or 200 μg mL-1). Ovarian tissue was processed for classical histology, TEM, and immunohistochemical demonstration of proliferating cell nuclear antigen (PCNA). Preantral follicles were classified according to their development stage (primordial, intermediate, primary, and secondary) and on the basis of morphological features (normal or degenerated). Pair-wise comparisons were done using Tukey’s procedure. Chi-square test was used to compare percentages of follicles with PCNA-positive granulosa cells. All analyses were done with Statistical Analysis System (SAS Institute, Cary, NC, USA); P ≤ 0.05 was considered significant. Compared with control fragments, the percentage of primordial follicles was reduced (P ≤ 0.05) and the percentage of growing follicles was increased (P ≤ 0.05) in cultured cortical fragments, independent of the tested medium or incubation time. Furthermore, compared with control tissue, culture of ovarian cortex for 8 days reduced the percentages of healthy, viable follicles (P ≤0.05), but not when cultures were supplemented with 25, 50, and 100 μg mL-1 of ascorbic acid. Ultrastructural and immunohistochemical analysis of ovarian cortical fragments cultured for 8 days, however, showed the integrity and viability of follicles only when fragments were cultured in the presence of 50 μg mL-1 of ascorbic acid. In conclusion, this study demonstrated that addition of ascorbic acid to MEM at a concentration of 50 μg mL-1 not only stimulates the activation and subsequent growth of cattle primordial follicles that are cultured in vitro for 8 days but also safeguards the viability of these preantral follicles. E. R. Andrade and A. A. Alfieri are recipients of the PRODOC/CAPES fellowship.

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