Prevention of pre-eclampsia by low-molecular-weight heparin in addition to aspirin: a meta-analysis.

OBJECTIVE: To estimate the impact of adding low-molecular-weight heparin (LMWH) or unfractionated heparin to low-dose aspirin started ≤ 16 weeks' gestation on the prevalence of pre-eclampsia (PE) and the delivery of a small-for-gestational-age (SGA) neonate. METHODS: A systematic review and meta-analysis of randomized controlled trials (RCTs) was performed by searching the medical databases PubMed, EMBASE, Web of Science and Cochrane Central. Pregnant women randomized to receive LMWH or unfractionated heparin in addition to low-dose aspirin were compared with those who received low-dose aspirin alone. Outcome measures were PE, severe PE, early-onset PE and SGA. Pooled relative risks (RRs) with 95% CI were calculated using a random-effects model. RESULTS: Eight RCTs met the inclusion criteria; the indication for recruitment was previous recurrent miscarriage in five studies (three included women with thrombophilia) and a history of severe or early-onset PE in three studies (including women with thrombophilia in one). LMWH was administered in seven studies and unfractionated heparin in one. In women with a history of PE, treatment with LMWH and aspirin, compared with aspirin alone, was associated with a significant reduction in development of PE (three trials (n = 379); RR, 0.54 (95% CI, 0.31-0.92); P = 0.03) and in delivery of SGA neonates (two trials (n = 363); RR, 0.54 (95% CI, 0.32-0.91); P = 0.02). These outcomes were not significantly reduced in women with recurrent miscarriage who received LMWH and aspirin, compared with aspirin alone. The small number of studies precluded sensitivity analyses and the evaluation of publication biases. Blinding to the allocation treatment was absent in all RCTs. CONCLUSIONS: Based on limited evidence, the addition of LMWH to low-dose aspirin could reduce the prevalence of PE and SGA in women with a history of PE. This observation should be the basis of a well-conducted future trial rather than a recommendation for immediate clinical application. Copyright © 2015 ISUOG. Published by John Wiley & Sons Ltd.

Prevention of perinatal death and adverse perinatal outcome using low-dose aspirin: a meta-analysis.

OBJECTIVE: To compare early vs late administration of low-dose aspirin on the risk of perinatal death and adverse perinatal outcome. METHODS: Databases were searched for keywords related to aspirin and pregnancy. Only randomized controlled trials that evaluated the prophylactic use of low-dose aspirin (50-150 mg/day) during pregnancy were included. The primary outcome combined fetal and neonatal death. Pooled relative risks (RR) with their 95% CIs were compared according to gestational age at initiation of low-dose aspirin (≤ 16 vs > 16 weeks of gestation). RESULTS: Out of 8377 citations, 42 studies (27 222 women) were included. Inclusion criteria were risk factors for pre-eclampsia, including: nulliparity, multiple pregnancy, chronic hypertension, cardiovascular or endocrine disease, prior gestational hypertension or fetal growth restriction, and/or abnormal uterine artery Doppler. When compared with controls, low-dose aspirin started at ≤ 16 weeks' gestation compared with low-dose aspirin started at >16 weeks' gestation was associated with a greater reduction of perinatal death (RR = 0.41 (95% CI, 0.19-0.92) vs 0.93 (95% CI, 0.73-1.19), P = 0.02), pre-eclampsia (RR = 0.47 (95% CI, 0.36-0.62) vs 0.78 (95% CI, 0.61-0.99), P < 0.01), severe pre-eclampsia (RR = 0.18 (95% CI, 0.08-0.41) vs 0.65 (95% CI, 0.40-1.07), P < 0.01), fetal growth restriction (RR = 0.46 (95% CI, 0.33-0.64) vs 0.98 (95% CI, 0.88-1.08), P < 0.001) and preterm birth (RR = 0.35 (95% CI, 0.22-0.57) vs 0.90 (95% CI, 0.83-0.97), P < 0.001). CONCLUSION: Low-dose aspirin initiated at ≤ 16 weeks of gestation is associated with a greater reduction of perinatal death and other adverse perinatal outcomes than when initiated at >16 weeks.

Vascular morphogenesis and remodeling in a model of tissue repair: blood vessel formation and growth in the ovarian pedicle after ovariectomy.

To investigate mechanisms of vascular morphogenesis in tissue repair, we performed ovariectomy with resection of the corresponding branches of the ovarian vessels in nude mice. This induces a vascular network remodeling response in the healing ovarian pedicle. Reconstruction of 2000 histological serial sections demonstrated that a new vascular network composed of venous-venous loops forms in the wall of the dilated ovarian vein. Preexisting veins of all sizes, including a branch of the main artery, are subjected to segmentation. Loop formation and segmentation are based on intussusceptive microvascular growth. Loop formation is followed by elongation. Loop remodeling occurs also by intussusception and results in the formation of compound loop systems. All loop systems observed were completely patent. Blind-ending sprouts were extremely rare. Anastomoses between the preexisting vessels subjected to segmentation and the loop systems were established to include the newly formed vessels into the preexisting vascular network. The formation of an increasing number of patent loop systems likely decreases hypoxia and subsequently arrests angiogenesis with transformation of the granulation tissue into a scar. Loop formation also occurred inside a large thrombus that occluded a part of the lumen of the main vein.

Vascular morphogenesis and remodeling in a human tumor xenograft: blood vessel formation and growth after ovariectomy and tumor implantation.

To determine mechanisms of blood vessel formation and growth in solid tumors, we used a model in which LS174T human colon adenocarcinomas are grown in the isolated ovarian pedicle of nude mice. Reconstruction of 3500 histological serial sections demonstrated that a new vascular network composed of venous-venous loops of varying sizes grows inside the tumor from the wall of the adjacent main vein. Loops elongate and remodel to establish complex loop systems. The mechanisms of loop formation and remodeling correspond to intussusceptive microvascular growth (IMG). In the tissue surrounding the tumor segmentation, another mechanism of IMG is prevalent in venous vessels. Comparison to vascular morphogenesis in the ovariectomized pedicle not only confirms the existence of corresponding mechanisms in both systems, but also reveals numerous sprouts that are superimposed onto loop systems and pathological deviations of loop formation, remodeling, and segmentation in the tumor. These pathological mechanisms interfere with vessel patency that likely cause heterogenous perfusion and hypoxia thus perpetuating angiogenesis. Blood vessel formation based on IMG was also detected in a large thrombus that completely occluded a part of an ovarian artery branch.

Cellular membrane permeability of anthracyclines does not correlate with their delivery in a tissue-isolated tumor.

The clearance of anthracyclines from the vasculature was studied in perfused tissue-isolated tumors. Human tumor lines MCF-7, U87, and LS174T were implanted in the ovarian fat pad of immune-deficient mice and grown isolated from the surrounding tissue. The initial and continuous tissue uptakes of doxorubicin, daunorubicin, and idarubicin were measured. The clearance of these anthracyclines from the perfused vasculature of the tissue-isolated tumor was calculated using BSA as an intravascular marker. The measured clearances ranged from 50-200 microl/min/g tumor tissue, and the fractional clearances were between 0.30 and 0.70. On the basis of the in vitro cellular uptake rates of the anthracyclines, we expected a higher clearance of idarubicin than of doxorubicin. No significant differences were found among the clearances of the anthracyclines or among the tumor lines. The observed similarities in clearance of the anthracyclines was largely explained by differences in their protein binding and tissue diffusion gradients.

Effect of tumor necrosis factor alpha on vascular resistance, nitric oxide production, and glucose and oxygen consumption in perfused tissue-isolated human melanoma xenografts.

The effect of tumor necrosis factor alpha (TNF-alpha) on vascular resistance, nitric oxide production, and consumption of oxygen and glucose was examined in a perfused tissue-isolated tumor model in nude mice. One experimental group was perfused with heparinized Krebs-Henseleit buffer, a second one was perfused with TNF-alpha (500 microgram/kg) 5 h before perfusion. The vascular resistance increased significantly 5 h after TNF-alpha injection. The increase in vascular resistance did not seem to be mediated by a decrease in tumor nitric oxide production, as determined by perfusate nitrate/nitrite concentrations, but may be due to aggregation of leukocytes, platelets, and erythrocytes and/or endothelial consumption among the three experimental groups. The oxygen consumption was linearly dependent on the amount of available oxygen in the perfusate, whereas the glucose consumption was constant and independent of the glucose delivery rate. The present experiments provide new insights into physiological and metabolic mechanisms of action of TNF- alpha for optimization of future treatment schedules involving TNF-alpha.

Changes in vascularization of human breast cancer xenografts responding to antiestrogen therapy.

To elucidate the previously suggested vascular effect(s) of antiestrogen therapy, we studied the effect of estrogen withdrawal and tamoxifen on 1) vascular resistance, 2) glucose and oxygen consumption, and 3) vascular density in a perfused breast cancer line (ZR75-1). Furthermore, we examined ZR75-1 tumors by functional CT-scanning (fCT) to determine changes in parameters related to tumor capillary transfer constants and vascular volume fraction in response to antiestrogenic manipulations. The vascular resistance decreased significantly from 42.7 to 20.8 mmHg x min x g x ml(-1) (P< .03) on day 9 after estrogen withdrawal, but not after 9 days of tamoxifen treatment. The estrogen-depleted tumors were significantly smaller than controls on day 9. There was no difference in nutrient consumption or vascular density in any of the experimental groups compared to controls. fCT showed an increase (P < .03) in vascular volume fraction during tumor growth, and this parameter was significantly lower after estrogen withdrawal when compared to controls (P < .05). Vascular resistance correlated with tumor size (R = 0.7, P < .0001), indicating that vascular resistance increases during tumor growth. The changes in vascular parameters after estrogen withdrawal indicate a vascular remodeling effect. This inhibition of vascular development by hormone deprivation may have important implications for future planning of multimodal treatment regimens.

Granulosa cells of the cumulus oophorus are different from mural granulosa cells in their response to gonadotrophins and IGF-I.

There are two types of granulosa cells: those which surround the oocyte are cumulus cells (CC) and those which surround the antrum are mural granulosa cells (MGC). These cells are under the influence of several hormones and growth factors, the most important of which are gonadotrophins and IGF-I. In this article, we report novel observations on the differences between these two types of granulosa cells and their interaction with gonadotrophins and IGF-I. We were able to conduct physiological studies on the role of IGF-I by using an analogue of IGF-I which does not bind to IGF-I-binding proteins (LR3-IGF-I). Immature rats received saline, equine chorionic gonadotrophin (eCG), LR3-IGF-I or eCG plus LR3-IGF-I by infusion using a pump from 24-29 days of age. The rats were killed and the ovaries removed. Surface follicles were punctured and MGC and oocyte cumulus complexes were removed. These were cultured in saline (control) and in three different doses of FSH. Cell replication was assessed by 3H-thymidine incorporation and differentiation was evaluated by the measurement of progesterone secretion. It was noted that CC replicated ten times more than MGC. Similarly, progesterone secretion by CC was six times more than by MGC. In vivo exposure to gonadotrophins (eCG) positively influenced in vitro treatment with FSH in both cell types. This phenomenon was observed in both cell replication and progesterone secretion. The IGF-I analogue had a positive effect on cell replication of MGC but a negative effect on the cell replication of CC. With respect to progesterone secretion, the IGF-I analogue had a negative effect on CC but a positive effect on MGC. In conclusion, CC behaved differently from MGC in response to gonadotrophins and the IGF-I analogue. IGF-I and FSH acted additively, synergistically or antagonistically in different circumstances.

Production of anti-idiotypic antibodies resembling bovine somatotropin by active immunization of lactating cows.

Anti-idiotypic antibodies produced in lactating cows by immunization with rabbit antibodies raised against bovine somatotropin (bST) were evaluated for bST-like immunoactive and biological activities. Twenty New Zealand White rabbits were immunized against bST. Serum was purified using protein A-Sepharose CL-4B and bST-Sepharose 4B affinity chromatography to yield specific anti-bST immunoglobulin G (IgG). Twenty-four lactating cows were allocated to a randomized complete block design and were immunized with either: (1) rabbit anti-bST (n = 12) or (2) non-specific rabbit IgG (n = 12, control). Cows were immunized starting at day 68 of lactation with 800 micrograms antigen emulsified with Freund's complete adjuvant and 1 mg Quil A administered by i.m. injections. Repeated immunizations of 800 micrograms antigen in Freund's incomplete adjuvant and 1 mg Quil A followed after 3 weeks and then every 5 weeks throughout lactation. Serum samples were collected twice weekly and were analysed for anti-rabbit IgG titre and immunoactive bST concentration by radioimmunoassay. All cows developed antibodies against rabbit IgG by the second and third immunizations. With rising titres, immunoactive bST concentrations increased in cows treated with anti-bST following each of the six repeated immunizations to 4.7 +/- 3.3, 10.0 +/- 4.2, 12.0 +/- 4.8, 17.3 +/- 6.7, 24.0 +/- 10.0 and 36.3 +/- 20.4 ng/ml (mean +/- S.E.M.) compared with controls (0.81 +/- 0.09 ng/ml, overall mean +/- S.E.M.). To assess the biological activity of these bST anti-idiotypic antibodies, milk yield and serum insulin-like growth factor-I (IGF-I) concentrations were analysed in all cows and a subset of six cows per treatment respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Regulation of testicular differentiation and testosterone production in the fetal mouse gonad in vitro.

The objective of the present study was to develop a chemically-defined medium in which early stages of testicular differentiation can be investigated in an organ culture system. Mouse gonadal primordia were explanted before and after initiation of morphological sex differentiation, i.e. 11 and 12 day of gestation (d.g.), respectively. We found that a combination of human albumin fraction, insulin (or IGF-I), and sodium pyruvate promoted testicular organization of gonadal explants of 11 d.g., but not those of 12 d.g. Insulin also increased the production of testosterone from testicular explants of 11 d.g., but not those of 12 d.g. For the younger explants, progesterone was more efficient than pregnenolone as a steroid precursor during the first day of culture, but the maximum effect of pregnenolone was much higher than that of progesterone in later stages. The responsiveness to human chorionic gonadotropin increased gradually along with testicular organization. The addition of either serum or pregnenolone prominently increased the activity of delta 5-3 beta hydroxysteroid dehydrogenase in testicular explants of 11 d.g., but not the number of positive cells as demonstrated by histochemical staining. These results suggest that insulin (or IGF-I) is required during the initial phase of testicular organization, which is reflected by an increase in testosterone production and sensitivity to gonadotropins.

Differential effects of insulin-like growth factor-I and gonadotropins on the proliferative activity of two subgroups of granulosa cells: cumulus oophorus and mural granulosa cells.

OBJECTIVE: To evaluate the physiological role of insulin-like growth factor-I (IGFI) and its interaction with gonadotropins in cell replication of two types of granulosa cells (cumulus oophorus, CC) and mural granulosa cells (MGC). DESIGN: Controlled randomized study of the action and interaction of gonadotropins and IGFI on granulosa cell replication in the rat. SETTING: A university reproductive biology laboratory. INTERVENTION(S): The study examined the in vivo treatment with gonadotropins or an analogue of IGFI, long Arg3-insulin-like growth factor (LR3-IGFI), which does not bind to IGFI-binding proteins. MAIN OUTCOME MEASURE(S): Granulosa cell replication was evaluated by the use of 3H-thymidine incorporation. RESULT(S): It was noted that the CC replicate much faster than the MGC. These two types of granulosa cells have very different dose response curves to IGFI. Differential responses were seen in animal cells exposed to long Arg3-insulin-like growth factor in vivo and then exposed to FSH and LH in vitro. CONCLUSION(S): Although murine granulosa cells show proliferative activity when they are exposed to IGFI, the two types, CC and MGC, respond differently. IGFI is not the sole mediator of the action of FSH, and these two chemicals may act independently or in concert.

Intracytoplasmic sperm injection increased fertilization and good-quality embryo formation in patients with non-male factor indications for in vitro fertilization: a prospective randomized study.

OBJECTIVE: To compare the fertilization rate and formation of good-quality embryos with conventional IVF and ICSI in patients with non-male factor infertility. DESIGN: Prospective controlled study. SETTING: Infertility clinic. PATIENT(S): Thirty-five patients with non-male factor infertility. INTERVENTION(S): Retrieved sibling oocytes were randomly assigned to conventional IVF or ICSI. Of sibling oocytes assigned to ICSI, only metaphase II oocytes were injected with sperm. MAIN OUTCOME MEASURE(S): Fertilization rate and formation of good-quality embryos per retrieved oocyte. RESULT(S): Per retrieved oocyte, ICSI resulted in better fertilization rate compared with conventional IVF (71.3% [134 of 188] vs. 57.2% [107 of 187]). Per retrieved oocyte, ICSI also resulted in better formation of good-quality embryos at 48 hours after retrieval compared with conventional IVF (64.4% [121 of 188] vs. 47.1% [88 of 187]). CONCLUSION(S): In IVF patients with non-male factor infertility, subjecting some sibling oocytes to ICSI increased the fertilization rate and formation of good-quality embryos per retrieved oocyte. It also avoided the problem of total fertilization failure in almost all cases.

Pulmonary barotrauma in congenital diaphragmatic hernia: experimental study in lambs.

A left diaphragmatic hernia was created surgically in 20 fetal lambs between 93 and 110 days of gestation. Ten animals were alive with defects at cesarean section near term (135 to 140 days). These animals and two controls were submitted to various transpulmonary pressure gradients (inspiratory pressure minus pleural pressure). Hemodynamic and ventilatory studies were performed after the correction of the hernia. Morphometric analysis of the lung was carried out in all cases. The results showed a highly significant linear correlation between the transpulmonary pressure gradient employed and the pulmonary interstitial emphysema found at morphometry. Our data suggest that using low ventilatory pressures and not draining the pleural cavity results in less trauma to both lungs and may prevent one of the components of the pulmonary hypertension so often seen in newborns with congenital diaphragmatic hernia.

Superovulatory and endocrine responses in holstein heifers treated with either prostaglandin F(2alpha), cloprostenol or fenprostalene.

Fifty Holstein heifers were each superovulated three times with FSH-P. At 60 h after the first injection of FSH-P, the animals received either prostaglandin F(2alpha), cloprostenol or fenprostalene in random order. A significant decrease in serum progesterone and a significant increase in serum estradiol-17beta were observed within 24 h of prostaglandin injection, but there were no significant differences among the three treatments. Neither were there any significant differences among the treatments with respect to the frequency of nonresponse to FSH-P treatment, nor the total number of ova/embryos collected between Days 6 and 8 of gestation.