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Evidence for pituitary regulation of placental lactogen in the rat.

The present studies examined whether the pituitary hormones, particularly prolactin, have any role in the regulation of rat placental lactogen (rPL) secretion. Total rPL was measured using a lymphoma cell bioassay. Hypophysectomy on day 13 of pregnancy increased total serum rPL levels above those of intact control rats and delayed for 3 days the decline in rPL usually seen by day 14. To test the effect of early hypophysectomy on rPL secretion, a delayed implantation model was used. Hypophysectomy was carried out on day 3, progesterone (2 mg) was given daily until day 8 and oestrone (1.0 micrograms) was given on day 8. This initiated implantation on day 9, which is 4 days later than normal. Rats hypophysectomized on day 3 had high serum levels of rPL (10-13 mg/l) 7 days after initiation of implantation compared with control values of 2-3 mg/l, and these levels remained raised for the duration of the experiment. Termination of maintenance injections of steroids did not affect the high levels of rPL for several days, even though there was fetal and placental resorption. When a pair of anterior pituitaries was transplanted under the kidney capsule on day 13 (day 9 of development) of rats hypophysectomized on day 3, serum rPL levels still increased for the next 3 days. However, unlike similarly treated rats not receiving transplants, rPL levels fell rapidly thereafter and were only 5% of those in control animals 1 week later.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Cytokine gene polymorphisms and susceptibility to recurrent pregnancy loss in Iranian women.

Recurrent pregnancy loss (RPL) is defined as three or more sequential abortions before the 20th week of gestation. There is increasing evidence to support an immunological mechanism for the occurrence of RPL. Defective production of T helper type 2 (Th2) and/or higher production of T helper type 1 (Th1) cytokines have been reported in RPL. As cytokine gene polymorphisms may be associated with different rates of cytokine production, the aim of the present study was to investigate the bi-allelic polymorphisms in TNF-alpha -308 G-->A, TNF-beta +252 G-->A, IFN-gamma +874 A-->T genes as Th1 or pro-inflammatory factors as well as IL-4 -590 C-->T, IL-10 -592 C-->A, -819 C-->T, -1082 A-->G genes as Th2 cytokines in women with RPL compared with healthy women. A total of 139 women with RPL and 143 control women with at least two successful pregnancies were included in the study. The allele-specific oligonucleotide polymerase chain reaction (ASO-PCR) or PCR-RFLP (restriction fragment length polymorphism) methods were used for genotyping. Results indicated a significant association between the presence of CC genotype of IL-10 -592 C-->A polymorphism and the occurrence of RPL in Iranian women (63% in women with RPL and 46% in controls; OR=0.51, 95% CI: 0.3-0.85; p<0.01). There was no significant association with other positions. It may be concluded that IL-10 polymorphism at position -592 could be a genetic factor for RPL.

Abortion, Habitual↗

Inverse relationship of prolactin and rat placental lactogen during pregnancy.

Serum prolactin, progesterone and rat placental lactogen (rPL) were measured in pregnant rats following removal of various numbers of conceptuses and their placentas on Day 8 of pregnancy. Blood samples were taken during the time of the expected nocturnal surge of prolactin on Days 8 and 9, and at the same time on Days 11 and 14 of pregnancy, when the surges are normally no longer present. As the number of conceptuses present decreased, the number of days the prolactin surge was present increased. Measurement of the early form of rPL (rPL-I) by lymphoma cell bioassay revealed a proportionate decrease in serum rPL-I levels on Days 9 and 11 as the number of conceptuses was decreased. This also was true for the late form of rPL (rPL-II) when measured by RIA on Day 14 of pregnancy. Thus, as rPL levels are reduced, prolactin surges remain present for increased numbers of days. This suggests that rPL may normally be an inhibitory factor to prolactin secretion during pregnancy, and is responsible for the termination of the prolactin surges at midpregnancy.

Animals↗

Long-term stability of clinical attachment following guided tissue regeneration and conventional therapy.

UNLABELLED: 44 patients (34% smokers) presenting with severe periodontitis were treated with full mouth root planing (RPL). In each patient, 1 intrabony defect was treated with guided tissue regeneration (GTR). After 1 year of monthly prophylaxis, full mouth plaque (FMPS) and bleeding (FMBS) scores were 8.3 +/- 4.1% and 5.6 +/- 3.8%. At 1 year, the GTR treated sites were matched, in each patient, with 1 RPL site, in terms of probing attachment level (PAL 6.8 +/- 2.4 mm GTR, and 6.5 +/- 2.3 mm RPL). At this point, 24 patients took part in a supportive periodontal care program. 20 patients did not participate, and received only sporadic care by general dentists. At 5 years, all patients were reexamined. FMPS was 10.5 +/- 6.8% and FMBS 7.7 +/- 6.4%. A significant PAL loss was observed in both sites (1.2 +/- 1.4 mm GTR, 1.3 +/- 1.3 mm RPL, p < 0.0001) between 1 and 5 years. Differences in PAL loss between GTR and RPL sites were not statistically significant. Only a minority of sites (34%), however, lost PAL, while 66% remained stable. 75% of the matched sites (GTR and RPL) within the same patients were concordant in terms of PAL stability. The 23 patients in which both sites remained stable, had good oral hygiene, complied with the recall system, and did not smoke. The 10 patients in which both sites lost PAL showed deteriorating oral hygiene, did not comply with the recall system, and smoked. PAL loss in the GTR and/or RPL sites was consistently observed in patients (losers) showing PAL loss in other teeth. Losers had, in general, negative subjects characteristics, and showed a higher prevalence of tooth loss. IN CONCLUSION: (i) GTR and RPL sites showed comparable susceptibility to periodontal breakdown; (ii) stability of outcomes was consistently associated with good oral hygiene, compliance with a supportive periodontal care program, and no cigarette smoking.

Adult↗

Characterization of a recombinant pneumolysin and its use as a protein carrier for pneumococcal type 18C conjugate vaccines.

Pneumolysin from Streptococcus pneumoniae was expressed in Escherichia coli as a glutathione S-transferase fusion protein and purified by affinity and hydroxylapatite chromatography. The purified recombinant pneumolysin (rPL), with a molecular mass of 53 kDa, had a specific activity of 3 x 10(5) hemolytic units per mg of protein on rabbit erythrocytes and reacted identically in immunodiffusion with the antisera against native pneumolysin. The rPL was used as a protein carrier to prepare conjugate vaccine with pneumococcal type 18C polysaccharide (PS18C). The PS18C was directly coupled to rPL by reductive animation or was indirectly coupled to rPL via a spacer molecule, adipic acid dihydrazide. The conjugates were nontoxic for mice and guinea pigs at 100 micrograms per dose. The immunogenicity and protective efficacy of both conjugates were tested in mice. A single dose of either of the vaccines elicited a rise in immunoglobulin G antibody production; after two booster injections of the vaccines, statistically significant booster responses (P < 0.001) to both rPL and PS18C were produced. The sera containing the antibodies to rPL were capable of neutralizing the hemolytic activity of rPL to rabbit erythrocytes and the cytotoxicity of rPL to bovine pulmonary endothelial cells. Immunization with the conjugate vaccines conferred statistically significant protection in mice against lethal challenge with type 18C pneumococci.

Animals↗

Rat placental lactogen-I binds to the choroid plexus and hypothalamus of the pregnant rat.

Recent findings suggest that placental lactogen has a role in the regulation of hypothalamic function during pregnancy. To explore the mechanisms by which placental hormones may exert effects in the maternal central nervous system, we have examined the binding of rat placental lactogen-I (rPL-I) to brain slices from pregnant rats at mid- and late gestation. The binding of rPL-I to maternal rat brain was compared with that of human GH (hGH). Radiolabelled rPL-I bound specifically to ependymal cells of the choroid plexus in the lateral ventricles and in the roof of the third ventricle. The binding of 125I-labelled rPL-I was inhibited by unlabelled rPL-I, hGH or rat prolactin but not by rat GH, indicating that rPL-I and rat prolactin interact with a common binding site in maternal rat brain. Radiolabelled hGH bound to the choroid plexus and to ependymal cells lining the third ventricle in the region of the arcuate nucleus. In addition, hGH bound specifically to the ventromedial nuclei and to the medial preoptic area of the hypothalamus. The binding of radiolabelled hGH to all brain regions was inhibited by unlabelled rPL-I as well as hGH, indicating that rPL-I competes for lactogenic binding sites in the hypothalamus as well as the choroid plexus of the pregnant rat. These findings suggest potential mechanisms by which placental hormones may exert direct effects on the maternal central nervous system during pregnancy. The precise functions and roles of the PL-I binding sites in maternal choroid plexus and hypothalamus remain to be explored.

Animals↗

Moderate food restriction reduces serum IGF-I and alters circulating IGF-binding protein profiles in lactating rats.

The role of somatogenic and lactogenic hormones in the adaptative mechanisms which occur in response to nutrient restriction during lactation is unknown. To characterize the effect of food restriction during lactation on serum IGF-I, GH and prolactin concentrations and serum IGF-binding protein (IGFBP) profiles, lactating dams had free access to food (control) or were restricted to 60% of control intake during pregnancy and lactation (RPL) or only during lactation (RL). Serum, milk and mammary gland samples were collected throughout lactation. RL dams lost body weight, control dams gained weight, while RPL dams maintained body weight during lactation. By day 20, body and mammary gland weights of RL and RPL dams did not differ and were lower than control (P < 0.05). Serum IGF-I concentrations in restricted groups were lower than control (P < 0.05), however, hepatic expression of IGF-I mRNA did not differ between groups in early (day 1) or mid-lactation (day 8) and was increased on day 20 in RL dams compared with RPL or control. These data suggest that serum IGF-I and hepatic IGF-I mRNA expression are not co-ordinately regulated in the food-restricted lactating rat. In early lactation, serum IGFBP-3 was lower in RPL dams than control (P < 0.05), whereas IGFBP-1 and -2 were increased in RL and RPL dams in late lactation compared with control. The decrease in IGFBP-3 and increase in lower molecular weight IGFBP may have contributed to the reduction in serum IGF-I by increasing IGF-I clearance from the circulation. Serum GH and prolactin were measured in samples obtained between 0900 and 1200 h. Serum GH did not differ with the exception of an increase on day 1 in control relative to RPL dams and on day 20 in RL dams relative to RPL and control. Serum prolactin was higher in the RL dams than controls on day 4. In summary, food restriction during pregnancy and lactation or solely during lactation results in similar reductions in serum IGF-I and alterations in serum IGFBP despite differences in body weight responses to food restriction during lactation.

Animals↗

Failure of injection of rat placental lactogen to inhibit prolactin in vivo.

In an attempt to demonstrate a negative feedback of rat placental lactogen (rPL) on prolactin secretion, pregnant rats were hysterectomized and injected intraperitoneally with placental extracts. Hysterectomy alone prolonged the incidence of nocturnal prolactin surges and injection of placental extracts did not alter this response. However, the absence of rPL in the serum following the injections indicated a primary reason why no inhibition was seen. Only when rPL was given intravenously was there detectable amounts found in the blood. The slow disappearance of rPL from the circulation following hysterectomy in Day 11 pregnant rats suggests that the lack of rPL in the blood following ip injection of placental extracts is not due to rapid clearance of rPL from blood. The failure to show a negative feedback of rPL on prolactin in vivo may be due primarily to the lack of appearance of rPL in the circulation following an ip injection of placental extracts.

Animals↗

Evaluation of the association between hereditary thrombophilias and recurrent pregnancy loss: a meta-analysis.

BACKGROUND: Recurrent pregnancy loss (RPL) is a significant clinical problem. Recently, thrombophilias have been implicated as a possible cause. Factor V Leiden (FVL) and prothrombin gene (G20210A) mutations are the most common types of hereditary thrombophilias, but are usually undiagnosed because most carriers are asymptomatic. The relationship between FVL, G20210A, and RPL has been investigated with conflicting results. This study analyzed existing data to determine whether an association exists. METHODS: A systematic review of the literature was performed. Only case-control studies that defined RPL as 2 or more pregnancy losses in the first or second trimester and that confirmed mutations by DNA analysis were included. Sixteen studies were selected for the FVL meta-analysis and 7 for the G20210A analysis. Stratified and multivariate logistic regression analyses were performed with the use of aggregate data. Results were confirmed by means of fixed- and random-effects meta-analyses models. RESULTS: The combined odds ratios for the association between RPL and FVL and between RPL and G20210A were 2.0 (95% confidence interval, 1.5-2.7; P<.001) and 2.0 (95% confidence interval, 1.0-4.0; P =.03), respectively. Similar results were produced by the logistic regression and both fixed- and random-effects meta-analysis models. CONCLUSIONS: Carriers of FVL or prothrombin gene mutations have double the risk of experiencing 2 or more miscarriages compared with women without thrombophilias. Hereditary thrombophilias may be an unrecognized cause of RPL. We recommend testing for these mutations in women with RPL.

Abortion, Habitual↗

Carbohydrate-binding site of a novel mannose-specific lectin from fugu (Takifugu rubripes) skin mucus.

Pufflectin-s, identified in the skin mucus of the fugu Takifugu rubripes, is a novel mannose-specific lectin with similar structure to monocotyledonous plant lectins. In the present study, mutational analysis was used to reveal the mannose-binding sites of pufflectin-s. Putative binding sites were mutated as follows: binding site 1; rPL-D32E (Asp32-->Glu32), rPL-N34S (Asn34-->Ser34) and rPL-V36A (Val36-->Ala36) whereas binding site 2; rPL-D61E (Asp61-->Glu61), rPL-N63S (Asn63-->Ser63) and rPL-V65A (Val65-->Ala65). All recombinant proteins were expressed in Escherichia coli, purified with two chromatographic steps, and then subjected to mannose-binding assay by affinity chromatography. Recombinant wild-type pufflectin-s (rPL-wt) as well as three mutants with changes in binding site 2 could bind to mannose, in contrast to the three mutants with changes in binding site 1 in which mannose-binding activity was completely lost. These results clearly demonstrate that, at the least, binding site 1 is critical to mannose-binding activity in pufflectin-s.

Amino Acid Sequence↗

Predictive value of the presence of an embryonic heartbeat for live birth: comparison of women with and without recurrent pregnancy loss.

OBJECTIVE: To determine the predictive value of an embryonic heart rate (EHR) for a live birth in women with and without a history of recurrent pregnancy loss (RPL). DESIGN: Prospective cohort study with concurrent controls. SETTING: A subspecialty clinic for couples with RPL. PATIENT(S): Three hundred pregnant women who previously had been diagnosed with RPL, followed prospectively compared with 300 age-, race-, and gestational age-matched pregnant control women. INTERVENTION(S): Transvaginal sonography between 6 to 8 weeks of gestation. MAIN OUTCOME MEASURE(S): The EHR was determined between 6 and 8 weeks of gestation by transvaginal sonography. Obstetrical history and current pregnancy outcome were evaluated. RESULT(S): Data were analyzed by using the two-tailed t test and Fisher's exact test. In women with RPL, an EHR predicted a successful live birth in 246 (82%) of 300, compared with 294 (98%) of 300 in control women. The mean (+/- SD) EHR from successful pregnancies in the control group (143.2 +/- 20.8 beats per minute) was significantly higher than the mean in women with a history of RPL (131.4 +/- 22.9 beats per minute). CONCLUSION(S): An EHR in women with RPL is associated with a live birth rate of 82% and is significantly lower than EHR in controls. Clinicians should use this information to counsel patients with RPL.

Abortion, Habitual↗

Evidence-based investigations and treatments of recurrent pregnancy loss.

OBJECTIVE: To give an overview of currently used investigations and treatments offered to women with recurrent pregnancy loss (RPL) and, from an evidence-based point of view, to evaluate the usefulness of these interventions. DESIGN: Ten experts on epidemiologic, genetic, anatomic, endocrinologic, thrombophilic, immunologic, and immunogenetic aspects of RPL discussed methodologic problems threatening the validity of research in RPL during and after an international workshop on the evidence-based management of RPL. CONCLUSION(S): Most RPL patients have several risk factors for miscarriage, and an extensive investigation for all major factors should always be undertaken. There is an urgent need for agreement concerning the thresholds for detecting what is normal and abnormal, irrespective of whether laboratory tests or uterine abnormalities are concerned. A series of lifestyle factors should be reported in future studies of RPL because they might modify the effect of laboratory or anatomic risk factors. More and larger randomized controlled trials, including trials of surgical procedures, are urgently needed, and to achieve this objective multiple centers have to collaborate. Current meta-analyses evaluating the efficacy of treatments of RPL are generally pooling very heterogeneous patient populations and treatments. It is recommended that future meta-analyses look at subsets of patients and treatment protocols that are more combinable.

Abortion, Habitual↗

Do women with unexplained recurrent pregnancy loss have higher day 3 serum FSH and estradiol values?

OBJECTIVE: To determine the potential role of diminished ovarian reserve in unexplained habitual abortion. DESIGN: Retrospective comparative analysis. SETTING: University-based practice. PATIENT(S): Fifty-seven women who presented for evaluation of recurrent pregnancy loss (RPL). INTERVENTION(S): The test group (n = 36) comprised women with unexplained RPL. The control group (n = 21) comprised women with a known cause of RPL. Mean age, parity, day 3 serum FSH and E(2) levels, and presence or absence of a history of infertility were compared between groups. MAIN OUTCOME MEASURE(S): Day 3 serum FSH and E(2) levels. RESULT(S): Both day 3 FSH and E(2) levels were elevated in the unexplained group compared with the control group. When combined, FSH or E(2) levels, or both, were elevated in 58% of the unexplained RPL group and 19% of the control group (odds ratio, 5.95 [95% CI, 1.7-21.3]; P<.004). Age, parity, and presence of infertility did not differ between groups. CONCLUSION(S): Women with unexplained RPL have a greater incidence of elevated day 3 serum FSH and E(2) levels than do women with a known cause of RPL. Therefore, diminished ovarian reserve may contribute to recurrent pregnancy loss and should be considered part of the work-up for RPL.

Abortion, Habitual↗

Increased prevalence of insulin resistance in women with a history of recurrent pregnancy loss.

OBJECTIVE: To determine whether insulin resistance is associated with recurrent pregnancy loss (RPL). DESIGN: Single center, case-controlled, prospective study. SETTING: University-associated reproductive endocrinology clinical practice. PATIENT(S): Seventy-four nonpregnant, nondiabetic women with RPL. Controls were 74 fertile, nonpregnant, nondiabetic women without RPL who had at least one live infant, and were matched by age, race, and body mass index (BMI). INTERVENTION(S): Both groups consented to obtaining fasting insulin and glucose levels. MAIN OUTCOME MEASURE(S): Insulin resistance was defined as a fasting insulin level >20 microU/mL or a fasting glucose to insulin ratio of <4.5. RESULT(S): Among the 74 women with RPL, 20 (27.0%) demonstrated insulin resistance, whereas only 7 of 74 (9.5%) of the matched controls were insulin resistant (odds ratio 3.55; 95% confidence interval 1.40-9.01). The RPL and control groups were similar with respect to age, ethnicity, and BMI. The RPL and control groups had similar fasting glucose levels and glucose-to-insulin ratios. However, fasting insulin levels > or =20 microU/mL were statistically different between the two groups (odds ratio 3.92). CONCLUSION(S): Women with RPL have a significantly increased prevalence of insulin resistance when compared with matched fertile controls.

Abortion, Habitual↗

beta2-Glycoprotein 1 as a marker of antiphospholipid syndrome in women with recurrent pregnancy loss.

OBJECTIVE: To determine if beta2-glycoprotein 1 (beta2-GP1) antibodies are a better marker of the antiphospholipid antibody syndrome (APS) in women with recurrent pregnancy loss (RPL). DESIGN: Evaluation and testing of sera from women with RPL. SETTING: A university-affiliated reproductive endocrinology practice. PATIENT(S): 90 women with RPL; 45 women met criteria for APS and 45 women met criteria for RPL without antiphospholipid antibodies (APA). Both groups were of similar age and had a similar history of RPL. INTERVENTION(S): Patient sera were obtained from women with RPL and were tested for APA and beta2-GP1. MAIN OUTCOME MEASURE(S): A standard antiphospholipid antibody assay was employed to detect the presence of immunoglobulin (Ig)G, IgM, and IgA antibodies in serum against cardiolipin, phosphatidyl inositol, phosphatidyl glycerol, phosphatidyl serine, and phosphatidyl ethanolamine. Samples were also assayed with a commercial beta2-GP1 assay for IgG antibodies. RESULT(S): Among the 45 women with APS, 10 (22.2%) had positive IgG antibodies for beta2-GP1. Only 1 woman (2.2%) of 45 was positive for beta2-GP1 among the control group of women with RPL but negative APA. There was no correlation noted among the beta2-GP1-positive patients for a specific phospholipid antibody or isotype. CONCLUSION(S): These data suggest that IgG beta2-GP1 antibodies are less sensitive than antiphospholipid antibodies for the diagnosis of APS.

Abortion, Habitual↗

Chronic hypoxia attenuates ischemia-reperfusion-induced increase in pulmonary vascular resistance.

This study explored the effect of chronic hypoxia on the elevation of pulmonary vascular resistance caused by ischemia-reperfusion (IR) in anesthetized rats. Experiments were separated into five parts. In Part 1, we examined the increase in left pulmonary vascular resistance (Rpl) after ischemia of the left lung and localized the major site for the increased resistance of the left pulmonary vasculature in both the normoxic and chronic hypoxia groups. Here, IR induced a significant increase in Rpl in the normoxic but not the chronic hypoxia group. This increased Rpl in the normoxic group was attributed to contraction of pulmonary arterial segments. Part 2 and Part 3 were focused on the changes in plasma nitrate/nitrite (NOx) and thromboxane B(2) (TxB(2)) levels. TxB(2) increased significantly in the normoxic group, whereas NOx increased significantly in the chronic hypoxia group, following ischemia. Indomethacin (Part 4) prevented IR-induced increase in Rpl in the normoxic group, whereas the IR-induced increase in Rpl appeared in the chronic hypoxia group after N(G)-nitro-L-arginine methyl ester treatment (Part 5). We conclude that IR elicited increases in the cyclooxygenase products such as TxB(2), which in turn caused an increase in Rpl. However, this increased Rpl was attenuated by elevated NOx in the chronic hypoxia group.

Animals↗

Research methodology and epidemiology of relevance in recurrent pregnancy loss.

With respect to recurrent pregnancy loss (RPL), unfortunately there is very little consensus about which investigations are useful for identifying causes and evaluating the prognosis, and also about which treatments are effective. In this review, arguments are given for the claim that this lack of consensus may mainly be because studies in the field of RPL have yielded very heterogeneous results. This heterogeneity, in the authors' belief, is caused by the scientists' lack of appreciation, when they do research, of important epidemiological knowledge about RPL (e.g., about the multifactorial background for most of the RPL cases and the importance of matching/adjusting for a series of prognostic variables when groups are mutually compared). Furthermore, many studies in RPL contain methodological flaws that are sometimes severe. A series of important epidemiological features of RPL is highlighted in the review and the most important methodological pitfalls, many of them specific for RPL research, are discussed. Advice is given about to how to avoid the pitfalls in order that the validity of the studies can improve for the benefit of the patients.

Abortion, Habitual↗

Amount and type of dietary fat regulate pancreatic lipase gene expression in rats.

Both amount and type of dietary triglycerides regulate pancreatic lipase, but the mechanism is not fully understood. This study investigated the effects of type (safflower oil and lard) and amount [low (50 g/kg diet) or moderate (174 g/kg diet)] of fat on rat pancreatic lipase (rPL) activity and mRNA levels. Polyunsaturated safflower oil resulted in 80% greater lipase activity compared with the saturated lard at moderate levels, whereas safflower oil resulted in 50% lower lipase activity compared with lard at low levels. The rPL-3 mRNA levels were greater in rats fed the moderate safflower oil diet (163%) or the moderate lard diet (212%) than in those fed the respective low fat diets and were 45% greater in those fed safflower oil than in those fed lard. The rPL-1 mRNA levels were greater in rats fed moderate safflower oil (50%) or lard (135%) than in those fed the respective low fat diet, but these levels were not affected by type of fat, in contrast to rPL-3 mRNA levels. The amount of fat independent of its type regulates pancreatic lipase pre-translationally, because increasing dietary saturated or polyunsaturated fat resulted in parallel changes in rPL-3 and rPL-1 mRNA levels. However, type of fat may regulate pancreatic lipase at other levels such as translational or post-translational, because the 212% increase in rPL-3 mRNA in rats fed the moderate lard diet compared with the low lard diet did not result in greater pancreatic lipase activity.

Animals↗