In situ identification of follicles in ovarian cortex as a tool for quantifying follicle density, viability and developmental potential in strategies to preserve female fertility

Posted on by

BACKGROUND

Ovarian tissue cryopreservation, in combination with autotransplantation or long-term culture, has been proposed as a means of fertility preservation. However follicle density within ovarian cortex has a profound impact on the success of in vivo and in vitro systems designed to support follicle growth and restore fertility. The objective of this study was to investigate the dye neutral red (NR) as a tool to quantify follicle density in situ, without compromising follicle viability and developmental potential.

METHODS

In the first experimental series thin slices of cryopreserved and fresh ovine cortical tissue were incubated in 50 µg/ml NR and assessed for the presence of red colouration. Slices were then used for follicular structure isolation and viability evaluation using 5-(and 6)-carboxyfluoresceindiacetate succinimidylester (CFDA-SE), or prepared histologically for follicle counting or evaluation of apoptosis via terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labelling (TUNEL). An additional subset of slices were cultured for 8 days, followed by re-evaluation of follicle viability. NR staining was further assessed in a pilot study using thin slices of cryopreserved human ovarian tissue donated by 17 patients undergoing laparoscopic sterilisation or elective Caesarean section.

RESULTS

In both ovine and human ovarian cortex NR concentrated in follicular structures within weakly stained stroma. NR colouration was observed in 41.7 ± 4.6% of cryopreserved and 49.3 ± 6.5% of the fresh ovine tissue slices, and NR staining was consistently predictive of the presence of follicles. Non-stained ovine slices contained highly apoptotic follicles, while lower levels of apoptosis were observed in NR positive slices, indicating preferential detection of viable follicles by NR. Following culture the majority of ovine slices re-stained with NR, no significant increases in the levels of apoptosis were observed and 94.6 ± 3.1% of follicles were viable by CFDA-SE. In the human study, NR identified follicles in 19.3 ± 3.7% of tissue slices, and follicle density tended to decrease with advancing patient age.

CONCLUSIONS

NR predicts viable follicle density in situ in slices of ovine and human ovarian cortex. Furthermore incubation of tissue in NR prior to culture does not compromise subsequent follicle survival in vitro, indicating the potential suitability of this approach in fertility preservation regimes.

Size and spatial orientation of uterine tissue transplants on the peritoneum crucially determine the growth and cyst formation of endometriosis-like lesions in mice

Posted on by

BACKGROUND

In many studies in rodents, intraperitoneal endometriosis-like lesions are surgically induced by syngeneic or autologous transplantation of uterine tissue samples, which are sutured to the abdominal wall. However, until now the surgical techniques have not been standardized, and we address this issue here.

METHODS

Uterine tissue samples were transplanted to the peritoneum of C57BL/6 mice (four study groups, n = 7 each). Using non-invasive high-resolution ultrasound imaging over a period of 4 weeks, we analyzed growth characteristics and cyst formation of the endometriosis-like lesions which developed, in relation to mode of transplantation (syngeneic versus autologous), type of tissue fixed adjacent to the peritoneum (endometrium versus perimetrium), and size of tissue transplanted (2 versus 3 mm). Immunohistochemical analysis was also performed.

RESULTS

When the perimetrium, with underlying myometrium, was sutured next to the host peritoneum the endometriosis-like lesions which developed exhibited a higher growth rate (P< 0.05 versus endometrium), and contained more proliferating cell nuclear antigen (PCNA)-positive cells and an increased microvessel density (both P< 0.05 versus endometrium). In the group with 3 mm uterine tissue grafts, lesion growth was significantly decreased when compared with 2 mm samples (P< 0.05). However, the larger grafts developed more cysts throughout the observation period than the smaller ones. There was no difference between syngeneic and autologous endometriosis-like lesions.

CONCLUSIONS

Our study demonstrates that size and spatial orientation of peritoneally fixed uterine tissue samples crucially determine growth and cyst formation of endometriotic lesions in mice. These findings should improve the standardization and reliability of future studies, performed in the frequently used mouse model of surgically induced endometriosis.

High risk men’s perceptions of pre-implantation genetic diagnosis for hereditary breast and ovarian cancer

Posted on by

BACKGROUND

Pre-implantation genetic diagnosis (PGD) is an assisted reproductive technology procedure which provides parents with the option of conducting genetic analyses to determine if a mutation is present in an embryo. Though studies have discussed perceptions of PGD from a general population, couples or high-risk women, no studies to date have specifically examined PGD usage among men. This study sought to explore perceptions and attitudes towards PGD among males who either carry a BRCA mutation or have a partner or first degree relative with a BRCA mutation.

METHODS

A cross-sectional survey was conducted among 228 men visiting the Facing Our Risk of Cancer Empowered or Craigslist website. Eligibility criteria included men who self-reported they had been tested for a BRCA mutation or had a partner or first degree relative tested for a BRCA mutation. A 41-item survey assessed socio-demographic, clinical characteristics, PGD knowledge and attitudinal factors and consideration of the use of PGD. Differences in proportions of subgroups were tested using the Monte Carlo exact test for categorical data. A multiple logistic regression model was then built through a backward elimination procedure.

RESULTS

Although 80% of men reported being previously unfamiliar with PGD, after learning the definition of PGD, 34% of the 228 respondents then said they would ‘ever consider the use of PGD’. Respondents who thought of PGD only in terms of ‘health and safety’ were almost three times more likely (OR = 2.82; 95% 1.19–6.71) to ‘ever consider the use of PGD’ compared with respondents who thought of PGD in terms of both ‘health and safety’, and ‘religion and morality’.

CONCLUSIONS

As with other anonymous web-based surveys, we cannot verify clinical characteristics that may impact consideration of PGD use. Our findings indicate high-risk men need more information about PGD and may benefit from educational materials to assist them in reproductive decision-making.

Glial cell line-derived neurotrophic factor is constitutively produced by human testicular peritubular cells and may contribute to the spermatogonial stem cell niche in man

Posted on by

BACKGROUND

Testicular peritubular cells form an ill-characterized cellular compartment of the human testis, which forms a border with Sertoli cells and spermatogonial stem cells (SSCs). A recently developed culture method has identified parts of the secretory repertoire of human testicular peritubular cells (HTPCs), which includes nerve growth factor. Whether peritubular cells produce glial cell line-derived neurotrophic factor (GDNF) and may thus contribute to the stem cell niche is not known.

METHODS

We studied GDNF production in isolated peritubular cells from men with normal spermatogenesis (HTPCs) and impaired spermatogenesis and testicular fibrosis (HTPC-Fs). Human testicular biopsies and peritubular cells in culture were evaluated using immunohistochemistry, laser microdissection (LMD), RT–PCR and measurement of GDNF and cAMP by enzyme-linked immunosorbent assay. We also tested whether GDNF production is regulated by tumour necrosis factor- (TNF-) or tryptase, the products of mast cells or macrophages.

RESULTS

Peritubular wall cells are in close proximity to cells expressing the GDNF family co-receptor-1. GDNF mRNA was detected in LMD samples of the peritubular and tubular but not interstitial compartments. HTPCs and HTPC-Fs lack FSH- and LH-receptors but express receptors for TNF- and tryptase. Importantly, peritubular cells express GDNF and constitutively released GDNF into the medium in comparably high amounts. TNF- and tryptase had no effect on the secretion of GDNF by HTPCs or HTPC-Fs.

CONCLUSIONS

Peritubular cells in testes of normal and sub-/infertile men produce GDNF and are likely constitutive contributors of the SSC niche in the human testis.

Why the Medical Research Council refused Robert Edwards and Patrick Steptoe support for research on human conception in 1971

Posted on by

BACKGROUND

In 1971, Cambridge physiologist Robert Edwards and Oldham gynaecologist Patrick Steptoe applied to the UK Medical Research Council (MRC) for long-term support for a programme of scientific and clinical ‘Studies on Human Reproduction’. The MRC, then the major British funder of medical research, declined support on ethical grounds and maintained this policy throughout the 1970s. The work continued with private money, leading to the birth of Louise Brown in 1978 and transforming research in obstetrics, gynaecology and human embryology.

METHODS

The MRC decision has been criticized, but the processes by which it was reached have yet to be explored. Here, we present an archive-based analysis of the MRC decision.

RESULTS

We find evidence of initial support for Edwards and Steptoe, including from within the MRC, which invited the applicants to join its new directly funded Clinical Research Centre at Northwick Park Hospital. They declined the offer, preferring long-term grant support at the University of Cambridge, and so exposed the project to competitive funding mode. Referees and the Clinical Research Board saw the institutional set-up in Cambridge as problematic with respect to clinical facilities and patient management; gave infertility a low priority compared with population control; assessed interventions as purely experimental rather than potential treatments, and so set the bar for safety high; feared fatal abnormalities and so wanted primate experiments first; and were antagonized by the applicants’ high media profile. The rejection set MRC policy on IVF for 8 years, until, after the birth of just two healthy babies, the Council rapidly converted to enthusiastic support.

CONCLUSIONS

This analysis enriches our view of a crucial decision, highlights institutional opportunities and constraints and provides insight into the then dominant attitudes of reproductive scientists and clinicians towards human conception research.

Human oocyte maturation is dependent on LH-stimulated accumulation of the epidermal growth factor-like growth factor, amphiregulin

Posted on by

BACKGROUND

The LH surge promotes ovulation via activation of multiple signaling networks in the ovarian follicle. Studies in animal models have shown the importance of LH-induced activation of the epidermal growth factor (EGF)signaling network in critical peri-ovulatory events. We investigated the biological significance of regulatory mechanisms mediated by EGF-like growth factors during LH stimulation in humans.

METHODS

We characterized the EGF signaling network in mature human ovarian follicles using in vivo and in vitro approaches. Amphiregulin (AREG) levels were measured in 119 follicular fluid (FF) samples from IVF/ICSI patients. Biological activity of human FF was assessed using in vitro oocyte maturation, cumulus expansion and cell mitogenic assays.

RESULTS

AREG is the most abundant EGF-like growth factor accumulating in the FF of mature follicles of hCG-stimulated patients. No AREG was detected before the LH surge or before hCG stimulation of granulosa cells in vitro, demonstrating that the accumulation of AREG requires gonadotrophin stimulation. Epiregulin and betacellulin mRNA were detected in both human mural and cumulus granulosa cells, although at significantly lower levels than AREG. FF from stimulated follicles causes cumulus expansion and oocyte maturation in a reconstitution assay. Immunodepletion of AREG abolishes the ability of FF to stimulate expansion (P < 0.0001) and oocyte maturation (P < 0.05), confirming the biological activity of AREG. Conversely, mitogenic activity of FF remained after depletion of AREG, indicating that other mitogens accumulate in FF. FF from follicles yielding an immature germinal vesicle oocyte or from an oocyte that develops into an aberrant embryo contains lower AREG levels than that from follicles yielding a healthy oocyte (P = 0.008).

CONCLUSIONS

EGF-like growth factors play a role in critical peri-ovulatory events in humans, and AREG accumulation is a useful marker of gonadotrophin stimulation and oocyte competence.

An innovative virtual reality technique for automated human embryonic volume measurements

Posted on by

BACKGROUND

The recent introduction of virtual reality (VR) enables us to use all three dimensions in a three-dimensional (3D) image. The aim of this prospective study was to evaluate an innovative VR technique for automated 3D volume measurements of the human embryo and yolk sac in first trimester pregnancies.

METHODS

We analysed 180 3D first trimester ultrasound scans of 42 pregnancies. Scans were transferred to an I-Space VR system and visualized as 3D ‘holograms’ with the V-Scope volume-rendering software. A semi-automatic segmentation algorithm was used to calculate the volumes. The logarithmically transformed outcomes were analysed using repeated measurements ANOVA. Interobserver and intraobserver agreement was established by calculating intraclass correlation coefficients (ICCs).

RESULTS

Eighty-eight embryonic volumes (EVs) and 118 yolk sac volumes (YSVs) were selected and measured between 5+5 and 12+6 weeks of gestational age (GA). EV ranged from 14 to 29 877 mm3 and YSV ranged from 33 to 424 mm3. ANOVA calculations showed that when the crown-rump length (CRL) doubles, the mean EV increases 6.5-fold and when the GA doubles, the mean EV increases 500-fold (P < 0.001). Furthermore, it was found that a doubling in GA results in a 3.8-fold increase of the YSV and when the CRL doubles, the YSV increases 1.5-fold (P < 0.001). Interobserver and intraobserver agreement were both excellent with ICCs of 0.99.

CONCLUSION

We measured the human EV and YSV in early pregnancy using a VR system. This innovative technique allows us to obtain unique information about the size of the embryo using all dimensions, which may be used to differentiate between normal and abnormal human development.

Intravenous immunoglobulin and idiopathic secondary recurrent miscarriage: a multicentered randomized placebo-controlled trial

Posted on by

BACKGROUND

Idiopathic secondary recurrent miscarriage may be associated with an abnormal maternal immune response to subsequent pregnancies. Intravenous immunoglobulin (IVIG) has been studied in randomized controlled trials (RCTs) with conflicting results. Therefore, a definitive trial was proposed.

METHODS

We conducted an investigator-initiated, multicentered, randomized, double-blinded, placebo-controlled trial comparing IVIG with saline in women with idiopathic secondary recurrent miscarriage, defined as a history of at least one prior ongoing pregnancy followed by three or more consecutive unexplained miscarriages. Subjects received either IVIG 500 mg/kg or the equivalent volume of normal saline. Preconception infusions were administered 14–21 days from the projected next menstrual period. With documentation of pregnancy, the subject received the same infusion every 4 weeks until 18–20 weeks of gestation. The primary outcome was an ongoing pregnancy of at least 20 weeks of gestation.

RESULTS

A total of 82 patients enrolled, of whom 47 had an index pregnancy. All ongoing pregnancies resulted in live births. Therefore, the live birth rates were 70% (16/23) in the IVIG group and 63% (15/24) in the control group (P = 0.760); odds ratio (OR) 1.37 [95% confidence interval (CI) 0.41–4.61]. Including only clinical pregnancies (embryo with cardiac activity at 6 weeks of gestation), the live birth rates were equivalent, 94% (16/17) and (15/16), respectively (P > 0.999); OR 1.07 (95% CI 0.06–18.62). Meta-analysis of randomized controlled trials (RCTs) evaluating IVIG for idiopathic secondary recurrent miscarriage revealed live birth rates of 70% (31/44) in the IVIG group and 62% (28/45) in the control group (P = 0.503); common OR 1.44 (95% CI 0.59–3.48).

CONCLUSIONS

This is the largest RCT to date in which IVIG was evaluated in women with idiopathic secondary recurrent miscarriage; no treatment benefit was found. The meta-analysis, which combined our study results with two prior RCTs, also showed no significant effect of treatment with IVIG.

ClinicalTrials.gov NCT00606905.

Expression of a truncated form of KIT tyrosine kinase in human spermatozoa correlates with sperm DNA integrity

Posted on by

BACKGROUND

TR-KIT, a truncated form of KIT (the KITL receptor), corresponding to the c-terminal half of the intracellular split tyrosine kinase domain, is expressed during the haploid stages of mouse spermatogenesis, and is one of the candidate sperm factors possibly involved in egg activation at fertilization.

METHODS

Immunocytochemistry of adult human testis, and studies of human semen samples from volunteer donors through immunofluorescence, confocal microscopy, flow cytometry, western blot and RT–PCR analyses were performed.

RESULTS

We show that the TR-KIT is expressed during spermiogenesis in the human testis, and that it is maintained in human ejaculated spermatozoa. TR-KIT is localized both in the equatorial segment and in the sub-acrosomal region of the human sperm head. The equatorial localization of the TR-KIT persists after the spontaneous acrosome reaction. Cytometric analysis of several sperm samples from volunteer donors, showed variable degrees of the TR-KIT-specific immunolabeling, and a significant inverse correlation (Pearson's coefficient, r = –0.76, P < 0.0001, n = 23) of the TR-KIT positivity with markers of sperm damage, i.e. DNA fragmentation, as revealed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-nick end labeling (TUNEL) analysis and the intense clusterin positivity. We also found less significant inverse correlation with altered head morphology (r = –0.47, P < 0.05, n = 23) and direct correlation with sperm forward motility parameters (r = 0.59, P < 0.01, n = 23).

CONCLUSIONS

The TR-KIT is present in the equatorial region of human spermatozoa, which is the first sperm component entering into the oocyte cytoplasm after fusion with the egg. This localization is consistent with the function previously proposed for this protein in mice. In addition, the TR-KIT represents a potential predictive parameter of human sperm quality.

Use of cryo-banked oocytes in an ovum donation programme: a prospective, randomized, controlled, clinical trial

Posted on by

BACKGROUND

An efficient oocyte cryopreservation method is mandatory to establish a successful egg-banking programme. Although there are increasing reports showing good clinical outcomes after oocyte cryopreservation, there is still a lack of large controlled studies evaluating the effectiveness of oocyte cryo-banking. In this study, we aimed to compare the outcome of vitrified-banked oocytes with the gold standard procedure of employing fresh oocytes.

METHODS

A randomized, prospective, triple-blind, single-centre, parallel-group controlled-clinical trial (NCT00785993), including 600 recipients ( = 0.05 and power of 80% for sample-size calculation) selected among 1032 eligible patients from November 2008 to September 2009, was designed to compare the outcome of vitrified-banked oocytes with the gold standard procedure of employing fresh oocytes. The study was designed to establish the superiority of the ongoing pregnancy rate (OPR) of fresh oocytes over that of vitrified oocytes, by performing a likelihood ratio test in a logistic regression analysis expressed as odds ratio (OR) with 95% confidence interval (CI). A limit of 0.66 for OR of vitrified versus fresh groups was defined to set up a possible conversion from superiority to non-inferiority. Randomization was performed 1:1 based on a computer randomization list in vitrification (n = 300) or fresh groups (n = 300). The primary end-point was the OPR per randomized patient i.e. intention-to-treat population (ITT). Secondary end-points were clinical pregnancy (CPR), implantation (IR) and fertilization rates, respectively. Additionally, embryo developmental characteristics were recorded.

RESULTS

There were no differences in donor ovarian stimulation parameters, demographic baseline characteristics for donors and recipients, ovum donation indications or male factor distribution between groups (NS). The OPR per ITT was 43.7 and 41.7% in the vitrification and fresh groups, respectively. The OR of OPR was 0.921 in favour of the vitrification group. Nevertheless, the 95% CI was 0.667–1.274, thus the superiority of fresh group with respect to OPR was not proven (P = 0.744). Non-inferiority of the vitrified group compared with the fresh group was shown with a margin of 0.667, which was above the pre-established non-inferiority limit of 0.66. CPR per cycle (50.2 versus 49.8%; P = 0.933) or per embryo-transfer (55.4 versus 55.6% ; P = 0.974), and IR (39.9 versus 40.9%; P = 0.745) were similar for patients receiving either vitrified or fresh oocytes. The proportion of top-quality embryos obtained either by inseminated oocyte (30.8 versus 30.8% for Day-2; and 36.1 versus 37.7% for Day-3, respectively) or by cleaved embryos (43.6 versus 43.8% for Day-2 and 58.4 versus 60.7% for Day-3, respectively) was similar between groups (NS).

CONCLUSIONS

This controlled-randomized, clinical trial confirmed the effectiveness of oocyte cryo-storage in an ovum donation programme, failing to demonstrate the superiority of using fresh oocytes with respect to the use of vitrified egg-banked ones in terms of OPR. Instead, the non-inferiority of vitrified oocytes was confirmed. These findings involve highly relevant issues that may open a new range of possibilities in ART.

Clinical Trials identifier: http://www.clinicaltrials.gov: NCT 00785993.

Possible role of the exchange protein directly activated by cyclic AMP (Epac) in the cyclic AMP-dependent functional differentiation and syncytialization of human placental BeWo cells

Posted on by

BACKGROUND

The mononuclear villous cytotrophoblast (CTB) differentiates and fuses to the multinucleated syncytiotrophoblast (STB), which produces hCG and progesterone. cAMP-mediated intracellular pathways are involved in the process of endocrine differentiation and fusion (syncytialization). The exchange protein directly activated by cAMP (Epac) is a mediator of cAMP signaling. We examined the differential roles of Epac and protein kinase A (PKA) signaling in the cell fusion and differentiation of trophoblast-derived BeWo cells.

METHODS

Epac1 and Epac2 were localized in human placental tissue (n = 9) by immunohistochemistry. The PKA-selective cAMP analog (N6-phenyl-cAMP, Phe) or Epac-selective cAMP analog (CPT) was tested for effects on hCG and progesterone production, and syncytialization in BeWo cells. The effect of knockdown of Epac or its downstream target molecule (Rap1) on syncytialization was evaluated.

RESULTS

Epac1 and Epac2 proteins were expressed in villous CTB, STB, stroma, blood vessels and extravillous CTB of the placenta. Phe increased the expression of hCG/β mRNA and secretion of hCG protein in BeWo cells (P < 0.01 versus control). CPT-stimulated production of hCG (P < 0.05), albeit to a lesser extent than Phe. Progesterone production was also enhanced by Phe or CPT (P < 0.01 and P < 0.05, respectively). CPT or a stable cAMP analog (dibutyryl-cAMP: Db) increased the number of syncytialized BeWo cells (P < 0.01), whereas Phe did not stimulate fusion. CPT- or Db-induced syncytialization was observed, even in the presence of a PKA inhibitor. Knockdown of Epac1 or Rap1 repressed the Db-, CPT- or forskolin-induced cell fusion.

CONCLUSIONS

The Epac signaling pathway may be associated with the cAMP-mediated functional differentiation and syncytialization of human trophoblasts.

Activation of Toll-like receptor 5 decreases the attachment of human trophoblast cells to endometrial cells in vitro

Posted on by

BACKGROUND

Embryo implantation in the uterus involves the trophoblast cells apposing and adhering to, then invading across the epithelium lining of the endometrium. However, ethical concerns regarding experimentation with primary human tissue during this period of life necessitates creation of in vitro models for understanding the basic mechanisms involved. Toll-like receptors (TLRs) play a crucial role in defence against pathogens invading the female reproductive tract. The objective of this study is to establish and optimize an in vitro model for studying human endometrial embryonic interactions and to understand the effect of TLR5 stimulation on the attachment of trophoblast cells to endometrial cells.

METHODS

By using a human telomerase immortalized endometrial epithelial cell line (hTERT-EECs) and choriocarcinoma human trophoblast cells (JAr cells), an in vitro assay of human implantation was established. In order to investigate the impact of TLR5 stimulation on attachment in this assay, bacterial flagellin was applied to the endometrial and trophoblast cells. In order to block TLR5 in the endometrial and trophoblast cells, TLR5 function-blocking antibody was applied to the cells prior to flagellin treatment.

RESULTS

The results demonstrated that JAr spheroids attached to hTERT-EECs in a time and concentration-dependent manner. Our results also demonstrated that treatment of endometrial cells with flagellin, suppressed the attachment of JAr spheres to the endometrial cells. Application of TLR5 function-blocking antibody significantly restored the attachment of JAr spheres to the endometrium.

CONCLUSIONS

These data suggest a novel mechanism by which the presence of intrauterine infection through TLR5 activation may result in implantation failure. These data may provide a new opportunity in the management of infertility cases.

The value of pre-operative treatment with GnRH analogues in women with submucous fibroids: a double-blind, placebo-controlled randomized trial

Posted on by

BACKGROUND

Submucous fibroids are common benign tumours responsible for menorrhagia, subfertility and miscarriage. They can be readily removed by hysteroscopic transcervical resection of myoma (TCRM). To facilitate resection, pre-operative GnRH analogues have been suggested, but the value of this treatment is uncertain. Our aim was to assess the value of pre-operative GnRH analogues for the resection of submucous fibroids.

METHODS

This was a prospective, double-blind, placebo-controlled, randomized trial. Women found to have submucous fibroids on three-dimensional saline infusion sonohysterography (3D SIS) were randomized to receive GnRH or placebo. Following treatment patients underwent TCRM by a single operator blinded to the group allocation. Women were followed up 6 weeks after their operation to ascertain resolution of symptoms. The primary outcome measure of the study was completeness of fibroid resection. Secondary outcome measures included the duration of the TCRM, the fluid deficit recorded at TCRM, the resolution of symptoms post-operatively and the number of subsequent fibroid related operations.

RESULTS

Forty-seven women were randomized to GnRH or placebo. On the basis of intention-to-treat analysis, there was no significant difference in the number of complete fibroid resections between women who received GnRH analogues [14/24, 58.3% (95% CI 38.6–78.1)] and those who received placebo [16/23, 69.6% (50.8–88.4)] (RR 0.84, 95% CI 0.54–1.29; P = 0.43). Similarly there was no significant difference between the groups in any of the secondary outcome measures.

CONCLUSIONS

Our study does not support routine administration of GnRH analogues before transcervical resection of fibroid as we did not identify any benefit in such treatment.

Controlled-trials.com: ISRCTN06560767.

Immediate pre-ovulatory administration of 30 mg ulipristal acetate significantly delays follicular rupture

Posted on by

BACKGROUND

Current methods of hormonal emergency contraception (EC) are ineffective in preventing follicular rupture when administered in the advanced pre-ovulatory phase. This study was designed to determine the capacity of ulipristal acetate (UPA), a selective progesterone receptor modulator developed for EC, to block follicular rupture when administered with a follicle of ≥18 mm.

METHODS

This was a double-blind, crossover, randomized, placebo-controlled study. Thirty-five women contributed with UPA (30 mg. oral) and a placebo cycle. Serial blood sampling for luteinizing hormone (LH), estradiol and progesterone measurements and follicular monitoring by ultrasound were performed before and for 5 days following treatment. Follicular rupture inhibition was assessed in the overall study population and in subgroups of women stratified by when treatment was administered in relation to LH levels (before the onset of the LH surge, after the onset of the surge but before the LH peak or after the LH peak).

RESULTS

Follicular rupture failed to occur for at least 5 days following UPA administration in 20/34 cycles [59%; 95% confidence interval (CI) (40.7–75.4%)], whereas rupture took place in all cycles within 5 days of placebo intake. When UPA was administered before the onset of the LH surge, or after the onset but before the LH peak, follicle rupture had not occurred within 5 days in 8/8 (100%) and 11/14 [78.6%; 95% CI (49.2–95.3)] cycles, respectively. In contrast, when UPA was given after the LH peak, follicle rupture inhibition was only observed in 1/12 [8.3%; 95% CI (0.2–38.5)] cycles.

CONCLUSIONS

This study demonstrates that UPA can significantly delay follicular rupture when given immediately before ovulation. This new generation EC compound could possibly prevent pregnancy when administered in the advanced follicular phase, even if LH levels have already begun to rise, a time when levonorgestrel EC is no longer effective in inhibiting ovulation.

NCT01107093: Comparison of CDB-2914 versus placebo in the prevention of follicular rupture post-LH surge.

Expression of adiponectin receptors and effects of adiponectin isoforms in mouse preimplantation embryos

Posted on by

BACKGROUND

Adiponectin, a pleiotropic hormone secreted from adipose tissue, can mediate some negative effects of obesity on female health, and can participate in the impaired reproductive performance of obese women. Using a mouse model, we investigated expression of adiponectin receptors in ovulated oocytes and in vivo derived preimplantation embryos, and tested effects of different adiponectin isoforms on development of preimplantation embryos in vitro.

METHODS AND RESULTS

Using RT–PCR and immunohistochemistry, we found expression of adiponectin receptors AdipoR1 and AdipoR2, at the mRNA and protein level, in mouse ovulated oocytes and preimplantation embryos. Quantitative real-time RT–PCR analysis showed a decrease in the amount of AdipoR1 and AdipoR2 mRNA after fertilization, which was followed by an increase in mRNA at the morula and blastocyst stage; mRNA for adiponectin was detected only at the blastocyst stage. Administration of full-length adiponectin significantly changed the distribution in numbers of cells of cultured preimplantation embryos, increasing the proportion of embryos with high cell numbers (>128 cells) and decreasing the proportion of embryos with lower cell numbers (<65 cells). Blastocysts possessed significantly higher cell numbers after full-length adiponectin treatment. Mutated trimeric adiponectin had the opposite effect, a significant decrease in the proportion of embryos with higher cell numbers (>96 cells) and increase in the proportion of embryos with lower cell numbers (<65 cells). Trimeric adiponectin also significantly decreased the cell number and increased cell death in blastocysts. Truncated globular adiponectin had no significant effect on development of mouse preimplantation embryos.

CONCLUSIONS

Our results indicate that adiponectin can directly influence the development of the preimplantation embryo, and the effects are isoform dependent.

Adnexal torsion: a predictive score for pre-operative diagnosis

Posted on by

BACKGROUND

Adnexal torsion (AT) is difficult to diagnose and requires immediate surgery. The aim of this study was to develop a simple score for assisting in the pre-operative diagnosis of AT in women with acute pelvic pain.

METHODS

Using data from a retrospective cohort of 142 patients with acute pelvic pain, we developed a score based on multiple logistic regression after a jackknife procedure. We validated the score in a prospective cohort of 35 women with acute pelvic pain.

RESULTS

Five criteria were independently associated with AT confirmed by surgery: unilateral lumbar or abdominal pain [adjusted odds ratio (aOR), 4.1; 95% confidence interval (95% CI), 1.2–14.0]; pain duration <8 h at first presentation (aOR, 8.0; 95% CI, 1.7–37.5), vomiting (aOR, 7.9; 95% CI, 2.3–27.0), absence of leucorrhoea and metrorrhagia (aOR, 12.6; 95% CI, 2.3–67.6) and ovarian cyst larger than 5 cm by ultrasonography (aOR, 10.6; 95% CI, 2.9–38.8). The torsion score was based on these five criteria. Low-risk and high-risk groups were derived from values of the score [probability of AT, 3.7% (95% CI, 0–7.8) and 69% (95% CI, 53–84), respectively]. Application of these criteria to the prospective cohort confirmed the diagnostic accuracy of the score [probability of AT, 0% (95% CI, 0–16) and 75% (95% CI, 26–100) in the low-risk and high-risk groups, respectively].

CONCLUSIONS

This easy-to-calculate score may prove useful for diagnosing AT in patients with acute pelvic pain seen at general or gynaecology emergency departments.

Up-regulation of apoptosis by gonadotrophin-releasing hormone agonist in cultures of endometrial cells from women with symptomatic myomas

Posted on by

BACKGROUND

The aim of the present study was to evaluate the effect of gonadotrophin-releasing hormone agonist (GnRH-a), which is widely used in the medical treatment of symptomatic myomas, on the rate of endometrial cell apoptosis in cultures from women with symptomatic myomas.

METHODS

The study included 36 women with symptomatic myomas without endometrial hyperplasia or endometrial carcinoma, and 22 controls. Endometrial biopsy specimens were obtained from all subjects. Levels of apoptosis were examined in epithelial endometrial cell cultures before and after incubation with GnRH-a (triptorelin). The percentage of apoptotic cells was evaluated using the terminal deoxynucleotidyl transferase-mediated d-UTP nick end labeling assay and flow cytometry was used to evaluate Annexin V levels.

RESULTS

Levels of spontaneous apoptosis were significantly lower in endometrial cultures from patients with symptomatic myomas than in those from control subjects (P < 0.01). Concentrations as low as 10–7 M GnRH-a enhanced apoptosis in endometrial cultures from patients with symptomatic myomas (3.48% ± 0.27% apoptotic cells in untreated samples and 25.45 ± 0.95% in cells treated with 10–7 M GnRH-a; P <0.01). The percentage of apoptotic cells also increased when cultures from control women were treated with GnRH-a (8.10 ± 0.18% in untreated samples and 15.29 ± 2.30% in treated samples; P <0.01). Levels of apoptosis were dependent on both dose of GnRH-a and time of treatment.

CONCLUSIONS

GnRH-a stimulates apoptosis in endometrial cells from patients with symptomatic myomas and this could, at least in part, account for the therapeutic action of GnRH-a.

How to Accurately Measure Blood Pressure, According to Dr. Peter … – Men’s Health

Posted on by

Peter Attia, M.D., is a longevity expert and creator of

BLOOD-PRESSURE MANAGEMENT isnt a sexy topic like anti-aging drugs or DNA-based diets, but it has enormous implications for health and life span. And because lifestyle factors have such a profound impact, everyone has the power to take steps toward controlling and improving their BP. Its bread-and-butter primary-care medicinebut a critical part of the longevity playbook.

High blood pressure is a silent killer. There are no symptoms, yet over time the effects of high blood pressure can compound, elevating risk for heart attacks and other serious health concerns, including stroke and dementia. For this reason, managing blood pressure relatively early in life is critical, but many people arent even aware of their BP status. Though nearly half of all adults in the United States have hypertensiondefined as a blood pressure with a systolic reading at or above 130 mm Hg and a diastolic reading at or above 80 mm Hg (see below)only about one in four among this group have their condition under control.

While standard medical wisdom once advised treating to a target of <140/<90, the landmark SPRINT study found that bringing BP down to <120/<80 resulted in huge reductions in the risk of heart attack, stroke, and death from all causes. So when it comes to keeping BP in check, an aggressive approach provides the most benefitsbut where do we begin?

Blood Pressure Categories

NORMAL: < 120 mm Hg (Systolic); < 80 mm Hg (Diastolic)

ELEVATED: 120129 mm Hg (Systolic); < 80 mm Hg (Diastolic)

STAGE 1 HYPERTENSION: 130139 mm Hg (Systolic); 140 mm Hg (Diastolic)

STAGE 2 HYPERTENSION: 8089 mm Hg (Systolic); 90 mm Hg

To manage your BP, you first need to know where youre starting. While a one-time measurement might provide a rough estimate, blood pressure is subject to acute fluctuations based on hydration, stress level, posture, caffeine intake, sleep deprivation, and other variables, so a more accurate picture requires multiple measurements over a span of days. I advise patients to use a home device to check BP twice a day, at the same time each morning and evening, for at least two weeks. The average of those readings will provide a much more accurate reference value than any one-time check. From there, you can monitor changes with weekly or even monthly checks, each time taking one measurement in the morning and one in the evening. The goal in checking frequently is to recognize increases as quickly as possible to facilitate faster intervention.

The gold standard for measuring BP is with a cuff and stethoscope, but this manual method can be intimidating to learn. In my experience, automated cuffs can overestimate BP because they work on an algorithm that estimates systolic and diastolic blood pressure from a measured mean arterial pressure (average BP, essentially), but they are generally good alternatives. Ive used both the Withings (withings.com; BPM Connect, $130) and Omron (amazon.com; M3 model, $80) brand cuffs.

Because so many variables affect BP, its important to follow the same protocol for every measurement:

Sit with your back supported for five minutes before measuring. (This can feel like a long time, since you should not be looking at your phone or speaking.)

Do not cross your legs.

Check that the cuff is against your skin and fits well.

Avoid talking during the check.

Note: Having a full bladder can bump up your BP by 10 to 15 mm Hg.

This story appears in the November 2023 issue of Men's Health.

Author

Peter Attia, MD, is the author of the New York Times Bestseller, Outlive: The Science and Art of Longevity and is host of the popular health and medicine podcast, The Drive. He received his medical degree from the Stanford University School of Medicine and trained at the Johns Hopkins Hospital in general surgery. Hes the founder of Early Medical, a medical practice that helps patients lengthen their lifespan and simultaneously improve their healthspan.

See original here:
How to Accurately Measure Blood Pressure, According to Dr. Peter ... - Men's Health

Cheap longevity drug? Researchers aim to test if metformin can slow down aging : Shots – Health News – NPR

Posted on by

Venca-Stastny/Getty Images

Venca-Stastny/Getty Images

A drug taken by millions of people to control diabetes may do more than lower blood sugar.

Research suggests metformin has anti-inflammatory effects that could help protect against common age-related diseases including heart disease, cancer, and cognitive decline.

Scientists who study the biology of aging have designed a clinical study, known as The TAME Trial, to test whether metformin can help prevent these diseases and promote a longer healthspan in healthy, older adults.

Michael Cantor, an attorney, and his wife Shari Cantor, the mayor of West Hartford, Connecticut both take metformin. "I tell all my friends about it," Michael Cantor says. "We all want to live a little longer, high-quality life if we can," he says.

Michael Cantor started on metformin about a decade ago when his weight and blood sugar were creeping up. Shari Cantor began taking metformin during the pandemic after she read that it may help protect against serious infections.

Shari and Michael Cantor both take metformin. They are both in their mid-60s and say they feel healthy and full of energy. Theresa Oberst/Michael Cantor hide caption

Shari and Michael Cantor both take metformin. They are both in their mid-60s and say they feel healthy and full of energy.

The Cantors are in their mid-60s and both say they feel healthy and have lots of energy. Both noticed improvements in their digestive systems feeling more "regular" after they started on the drug,

Metformin costs less than a dollar a day, and depending on insurance, many people pay no out-of-pocket costs for the drug.

"I don't know if metformin increases lifespan in people, but the evidence that exists suggests that it very well might," says Steven Austad, a senior scientific advisor at the American Federation for Aging Research who studies the biology of aging.

Metformin was first used to treat diabetes in the 1950s in France. The drug is a derivative of guanidine, a compound found in Goat's Rue, an herbal medicine long used in Europe.

The FDA approved metformin for the treatment of type 2 diabetes in the U.S. in the 1990s. Since then, researchers have documented several surprises, including a reduced risk of cancer. "That was a bit of a shock," Austad says. A meta-analysis that included data from dozens of studies, found people who took metformin had a lower risk of several types of cancers, including gastrointestinal, urologic and blood cancers.

Austad also points to a British study that found a lower risk of dementia and mild cognitive decline among people with type 2 diabetes taking metformin. In addition, there's research pointing to improved cardiovascular outcomes in people who take metformin including a reduced risk of cardiovascular death.

As promising as this sounds, Austad says most of the evidence is observational, pointing only to an association between metformin and the reduced risk. The evidence stops short of proving cause and effect. Also, it's unknown if the benefits documented in people with diabetes will also reduce the risk of age-related diseases in healthy, older adults.

"That's what we need to figure out," says Steve Kritchevsky, a professor of gerontology at Wake Forest School of Medicine, who is a lead investigator for the Tame Trial.

The goal is to better understand the mechanisms and pathways by which metformin works in the body. For instance, researchers are looking at how the drug may help improve energy in the cells by stimulating autophagy, which is the process of clearing out or recycling damaged bits inside cells.

Researchers also want to know more about how metformin can help reduce inflammation and oxidative stress, which may slow biological aging.

"When there's an excess of oxidative stress, it will damage the cell. And that accumulation of damage is essentially what aging is," Kritchevsky explains.

When the forces that are damaging cells are running faster than the forces that are repairing or replacing cells, that's aging, Kritchevsky says. And it's possible that drugs like metformin could slow this process down.

By targeting the biology of aging, the hope is to prevent or delay multiple diseases, says Dr. Nir Barzilai of Albert Einstein College of Medicine, who leads the effort to get the trial started.

Back in 2015, Austad and a bunch of aging researchers began pushing for a clinical trial.

"A bunch of us went to the FDA to ask them to approve a trial for metformin,' Austad recalls, and the agency was receptive. "If you could help prevent multiple problems at the same time, like we think metformin may do, then that's almost the ultimate in preventative medicine," Austad says.

The aim is to enroll 3,000 people between the ages of 65 and 79 for a six-year trial. But Dr. Barzilai says it's been slow going to get it funded. "The main obstacle with funding this study is that metformin is a generic drug, so no pharmaceutical company is standing to make money," he says.

Barzilai has turned to philanthropists and foundations, and has some pledges. The National Institute on Aging, part of the National Institutes of Health, set aside about $5 million for the research, but that's not enough to pay for the study which is estimated to cost between $45 and $70 million.

The frustration over the lack of funding is that if the trial points to protective effects, millions of people could benefit. "It's something that everybody will be able to afford," Barzilai says.

Currently the FDA doesn't recognize aging as a disease to treat, but the researchers hope this would usher in a paradigm shift from treating each age-related medical condition separately, to treating these conditions together, by targeting aging itself.

For now, metformin is only approved to treat type 2 diabetes in the U.S., but doctors can prescribe it off-label for conditions other than its approved use.

Michael and Shari Cantor's doctors were comfortable prescribing it to them, given the drug's long history of safety and the possible benefits in delaying age-related disease.

"I walk a lot, I hike, and at 65 I have a lot of energy," Michael Cantor says. I feel like the metformin helps," he says. He and Shari say they have not experienced any negative side effects.

Research shows a small percentage of people who take metformin experience GI distress that makes the drug intolerable. And, some people develop a b12 vitamin deficiency. One study found people over the age of 65 who take metformin may have a harder time building new muscle.

"There's some evidence that people who exercise who are on metformin have less gain in muscle mass, says Dr. Eric Verdin, President of the Buck Institute for Research on Aging. That could be a concern for people who are under-muscled.

But Verdin says it may be possible to repurpose metformin in other ways "There are a number of companies that are exploring metformin in combination with other drugs," he says. He points to research underway to combine metformin with a drug called galantamine for the treatment of sarcopenia, which is the medical term for age-related muscle loss. Sarcopenia affects millions of older people, especially women.

The science of testing drugs to target aging is rapidly advancing, and metformin isn't the only medicine that may treat the underlying biology.

"Nobody thinks this is the be all and end all of drugs that target aging," Austad says. He says data from the clinical trial could stimulate investment by the big pharmaceutical companies in this area. "They may come up with much better drugs," he says.

Michael Cantor knows there's no guarantee with metformin. "Maybe it doesn't do what we think it does in terms of longevity, but it's certainly not going to do me any harm," he says.

Cantor's father had his first heart attack at 51. He says he wants to do all he can to prevent disease and live a healthy life, and he thinks Metformin is one tool that may help.

For now, Dr. Barzilai says the metformin clinical trial can get underway when the money comes in.

This story was edited by Jane Greenhalgh

See original here:
Cheap longevity drug? Researchers aim to test if metformin can slow down aging : Shots - Health News - NPR

Medical Aesthetics Market Size Worth USD 43.2 Billion in 2032 … – GlobeNewswire

Posted on by

Vancouver, Nov. 22, 2023 (GLOBE NEWSWIRE) -- The global medical aesthetics market size was USD 15.3 Billion in 2022 and is expected to register rapid revenue CAGR of 11.0% during the forecast period. Key factors propelling this growth include the increasing awareness of aesthetic procedures, a surge in the adoption of minimally invasive devices, a rising demand for home-based aesthetic procedures, and continuous technological advancements in aesthetic devices.

Minimally invasive and non-surgical aesthetic treatments are gaining popularity, offering practical and risk-free alternatives to traditional surgery. Procedures such as Botox, dermal fillers, laser therapy, chemical peels, and microdermabrasion are on the rise due to their minimal downtime, making them optimal for quick recovery and the resumption of routine activities. The total number of non-surgical cosmetic operations increased by 44% in 2021 compared to surgical cosmetic procedures, with a substantial growth of 63% in the body procedures category. This trend is driven by accessibility, adaptability, and reduced scarring concerns associated with non-surgical treatments.

The rising demand for home-based cosmetic procedures is a significant driver, fueled by the increasing prevalence of dermatological diseases like acne, psoriasis, and skin cancer. However, poor reimbursement conditions pose a restraint to market growth, as cosmetic procedures, equipment, and devices are often not covered by insurance providers. Recent trends include the growth of medical aesthetics tourism, the destigmatization of aesthetic procedures, and the use of the internet to find the best providers.

Request Free Sample Copy (To Understand the Complete Structure of this Report [Summary + TOC]) @ https://www.emergenresearch.com/request-sample/2500

The facial aesthetic product segment led the market in 2022, driven by the increasing number of individuals seeking a youthful appearance and disposable financial resources. Notably, Allergan Aesthetics introduced the SkinMedica Even & Correct Collection, focusing on lightening treatments and dark-spot reduction creams. The cosmetic implant segment is expected to experience moderate growth due to clear regulatory requirements, the acceptance of bio-implants, and an increase in target diseases. Noteworthy approvals include Allergan's NATRELLE INSPIRA Soft Touch breast implants and Sientra Inc.'s gel breast implants.

The non-surgical procedures segment is anticipated to hold a significant revenue share, driven by the increasing popularity of lipolysis. Technological advancements, product introductions like Lumenis Ltd.'s NuEra Tight, and growing awareness among the elderly contribute to this segment's growth. Hospitals and clinics are expected to register fast revenue growth, driven by automated systems, early disease diagnosis, and the digitization of patient workflows. Beauty centers and medical spas, offering non-invasive skin rejuvenation procedures, are projected to account for the largest revenue share.

North America dominated the market in 2022, driven by experienced cosmetic surgeons, increased adoption of cosmetic operations, and advanced healthcare infrastructure. The Cleveland Clinic Organization estimates over 15 million cosmetic surgery treatments annually in the U.S.

The Asia Pacific region is poised for the fastest revenue growth, attributed to rising awareness of anti-aging surgeries, medical tourism, improved healthcare infrastructure, and affordable treatment costs. Government policies promoting cosmetic medicine standardization contribute to market growth.

Europe is expected to account for a substantial revenue share, driven by the demand for high-quality aesthetic procedures, established healthcare infrastructure, and an effective reimbursement system. The rising number of aesthetic operations is also a contributing factor. The global medical aesthetics market is experiencing significant growth driven by a shift towards minimally invasive treatments, increasing demand for home-based procedures, and ongoing technological advancements. Market players are responding with innovative products, contributing to the expansion of the industry.

Emergen Research is Offering Limited Time Discount (Grab a Copy at Discounted Price Now) @ https://www.emergenresearch.com/request-discount/2500

Medical Aesthetics Report Summary

Medical Aesthetics Top Companies and Competitive Landscape

The global medical aesthetic market is fairly fragmented with many large and medium-sized players accounting for majority of market revenue. Major players are deploying various strategies, entering into mergers & acquisitions, strategic agreements & contracts, developing, testing, and introducing more effective medical aesthetic solutions. Some major players included in the global medical aesthetic market report are:

Medical Aesthetics Latest Industry News

Direct Order Can Be Placed Through This Link [Exclusive Copy] @ https://www.emergenresearch.com/select-license/2500

Medical Aesthetics Market Segment Analysis

For the purpose of this report, Emergen Research has segmented global medical aesthetic market on the basis of product, procedure, end-use, and region:

Browse Full Report Description + Research Methodology + Table of Content + Infographics@ https://www.emergenresearch.com/industry-report/medical-aesthetics-market

Curated Reports You Shouldn't Miss: Dive In Now!

Patient Registry Software Market By Delivery, By Database, By Registry Type, By Function, By Software Type (Integrated, Standalone), By End-use (Government & Third-Party Administrators, Pharmaceutical Companies, Hospitals, Research Centers, Others), and By Region, Forecast to 2027

Operating Room Management Solutions Market, By Component (Software and Services), By Solution, By Delivery Mode (On-Premises and Cloud-based), By End-Use, and By Region Forecast to 2030

Medical Smart Textiles Market By Technology (Textile Sensors, Wearable Technology), By Application (Surgery, Bio-Monitoring, Therapy, and Wellness), By End-use (Hospitals and Clinics, Medical Academic and Research Center), and By Region Forecast to 2027

Topical Drug Delivery Market By Product Form (Semi-Solid, Solid Formulations, Transdermal Products, and Liquid Formulations), By Route (Dermal, Ophthalmic, Nasal, Others), By End-Use (Hospitals, Home Healthcare, Clinics, Diagnostic Centers, Burn Center), and By Region Forecast to 2027

Advanced Prosthetics and Exoskeletons Market By Product Type (Lower Limb, Upper Limb, Full Body), By Application (Hospitals, Clinics, Orthotic and Prosthetic Centers, Others), By Mobility (Stationary, Mobile), and By Region, Forecast to 2028

Robotics Surgical Simulation Systems Market By Product & Service Type (Conventional Surgery Simulation, Minimally Invasive Surgery Simulation, Others), By Type of Surgery (General Surgery, Endoscopic Surgery, Neurological Surgery, Gynecological Surgery, Cardiac Surgery), By End-use (Hospitals, Surgical Centers, Ambulatory Surgical Centers, Others), and By Region, Forecast to 2028

About Emergen Research

Emergen Research is a market research and consulting company that provides syndicated research reports, customized research reports, and consulting services. Our solutions purely focus on your purpose to locate, target, and analyze consumer behavior shifts across demographics, across industries, and help clients make smarter business decisions. We offer market intelligence studies ensuring relevant and fact-based research across multiple industries, including Healthcare, Touch Points, Chemicals, Types, and Energy.

Contact Us:

Eric Lee

Corporate Sales Specialist

Emergen Research | Web: https://www.emergenresearch.com/

Direct Line: +1 (604) 757-9756

E-mail: sales@emergenresearch.com

Explore Our Blogs and Insights Section: https://www.emergenresearch.com/insights

See the rest here:
Medical Aesthetics Market Size Worth USD 43.2 Billion in 2032 ... - GlobeNewswire