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1.
Mol Ecol Resour ; 15(2): 425-36, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25143045

ABSTRACT

This is the first de novo transcriptome and complete mitochondrial genome of an Antarctic sea urchin species sequenced to date. Sterechinus neumayeri is an Antarctic sea urchin and a model species for ecology, development, physiology and global change biology. To identify transcripts important to ocean acidification (OA) and thermal stress, this transcriptome was created pooling, and 13 larval samples representing developmental stages on day 11 (late gastrula), 19 (early pluteus) and 30 (mid pluteus) maintained at three CO2 levels (421, 652, and 1071 µatm) as well as four additional heat-shocked samples. The normalized cDNA pool was sequenced using emulsion PCR (pyrosequencing) resulting in 1.34M reads with an average read length of 492 base pairs. 40,994 isotigs were identified, averaging 1188 bp with a median coverage of 11×. Additional primer design and gap sequencing were required to complete the mitochondrial genome. The mitogenome of S. neumayeri is a circular DNA molecule with a length of 15 684 bp that contains all 37 genes normally found in metazoans. We detail the main features of the transcriptome and the mitogenome architecture and investigate the phylogenetic relationships of S. neumayeri within Echinoidea. In addition, we provide comparative analyses of S. neumayeri with its closest relative, Strongylocentrotus purpuratus, including a list of potential OA gene targets. The resources described here will support a variety of quantitative (genomic, proteomic, multistress and comparative) studies to interrogate physiological responses to OA and other stressors in this important Antarctic calcifier.


Subject(s)
DNA, Mitochondrial/genetics , Ecosystem , Genome , Sea Urchins/genetics , Transcriptome , Animals , Antarctic Regions , DNA, Complementary/chemistry , DNA, Complementary/genetics , DNA, Mitochondrial/chemistry , Molecular Sequence Data , Phylogeny , Sequence Analysis, DNA
2.
Biol Bull ; 201(3): 374-84, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11751249

ABSTRACT

Thermal stress has been considered to be among the most important determinants of organismal distribution in the rocky intertidal zone. Yet our understanding of how body temperatures experienced under field conditions vary in space and time, and of how these temperatures translate into physiological performance, is still rudimentary. We continuously monitored temperatures at a site in central California for a period of two years, using loggers designed to mimic the thermal characteristics of mussels, Mytilus californianus. Model mussel temperatures were recorded on both a horizontal and a vertical, north-facing microsite, and in an adjacent tidepool. We periodically measured levels of heat shock proteins (Hsp70), a measure of thermal stress, from mussels at each microsite. Mussel temperatures were consistently higher on the horizontal surface than on the vertical surface, and differences in body temperature between these sites were reflected in the amount of Hsp70. Seasonal peaks in extreme high temperatures ("acute" high temperatures) did not always coincide with peaks in average daily maxima ("chronic" high temperatures), suggesting that the time history of body temperature may be an important factor in determining levels of thermal stress. Temporal patterns in body temperature during low tide were decoupled from patterns in water temperature, suggesting that water temperature is an ineffective metric of thermal stress for intertidal organisms. This study demonstrates that spatial and temporal variability in thermal stress can be highly complex, and "snapshot" sampling of temperature and biochemical indices may not always be a reliable method for defining thermal stress at a site.


Subject(s)
Bivalvia/physiology , Ecology , Environment , HSP70 Heat-Shock Proteins/biosynthesis , Animals , Bivalvia/metabolism , Blotting, Western , Body Temperature , California , Climate , Protein Isoforms/biosynthesis , Seasons , Time Factors
3.
J Exp Biol ; 204(Pt 20): 3571-9, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11707506

ABSTRACT

Spatio-temporal variation in heat-shock gene expression gives organisms the ability to respond to changing thermal environments. The temperature at which heat-shock genes are induced, the threshold induction temperature, varies as a function of the recent thermal history of an organism. To elucidate the mechanism by which this plasticity in gene expression is achieved, we determined heat-shock protein (Hsp) induction threshold temperatures in the intertidal mussel Mytilus trossulus collected from the field in February and again in August. In a separate experiment, threshold induction temperatures, endogenous levels of both the constitutive and inducible isoforms of Hsps from the 70 kDa family and the quantity of ubiquitinated proteins (a measure of cellular protein denaturation) were measured in M. trossulus after either 6 weeks of cold acclimation in the laboratory or acclimatization to warm, summer temperatures in the field over the same period. In addition, we quantified levels of activated heat-shock transcription factor 1 (HSF1) in both groups of mussels (HSF1 inducibly transactivates all classes of Hsp genes). Lastly, we compared the temperature of HSF1 activation with the induction threshold temperature in the congeneric M. californianus. It was found that the threshold induction temperature in M. trossulus was 23 degrees C in February and 28 degrees C in August. This agreed with the acclimation/acclimatization experiment, in which mussels acclimated in seawater tables to a constant temperature of 10-11 degrees C for 6 weeks displayed a threshold induction temperature of 20-23 degrees C compared with 26-29 degrees C for individuals that were experiencing considerably warmer body temperatures in the intertidal zone over the same period. This coincided with a significant increase in the inducible isoform of Hsp70 in warm-acclimatized individuals but no increase in the constitutive isoform or in HSF1. Levels of ubiquitin-conjugated protein were significantly higher in the field mussels than in the laboratory-acclimated individuals. Finally, the temperature of HSF1 activation in M. californianus was found to be approximately 9 degrees C lower than the induction threshold for this species.


Subject(s)
Bivalvia/physiology , Heat-Shock Proteins/biosynthesis , Hot Temperature , Acclimatization , Animals , Cold Temperature , DNA/metabolism , DNA-Binding Proteins/analysis , DNA-Binding Proteins/metabolism , Gills/metabolism , HSP70 Heat-Shock Proteins/biosynthesis , Heat Shock Transcription Factors , Temperature , Transcription Factors , Ubiquitin/metabolism
4.
J Exp Biol ; 204(Pt 15): 2675-82, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11533117

ABSTRACT

Molecular chaperones participate in many aspects of protein biogenesis. Mechanistically, they recognize and bind to non-native proteins, prevent the aggregation of unfolded proteins and also, in some cases, facilitate refolding. Although a great deal is known about the cellular function of molecular chaperones in general, very little is known about the effect of temperature on molecular chaperones in non-model organisms, particularly in ectotherms that fold proteins under variable-temperature conditions in nature. To address this issue, we studied the temperature interactions of a major cytosolic molecular chaperone, Hsc70, from the eurythermal marine goby Gillichthys mirabilis. Using in vitro assays, we measured the intrinsic activity, unfolded-protein-stimulated activity, temperature sensitivity and heat stability of the ATPase activity of native Hsc70 purified from G. mirabilis white muscle. Similar to other chaperones in the 70kDa heat-shock protein family, G. mirabilis Hsc70 exhibited a low intrinsic ATPase activity that was stimulated in vitro by the addition of unfolded protein. Across the environmentally relevant temperature range (10-35 degrees C), the ATPase activity of G. mirabilis Hsc70 displayed differential thermal sensitivity, with the greatest sensitivity occurring between 10 and 15 degrees C and the least sensitivity between 15 and 25 degrees C. In addition, the activity of Hsc70 was not significantly different between the unstimulated and unfolded-protein-stimulated treatments, suggesting that the ATPase activity and the peptide-binding domain of Hsc70 have similar thermal sensitivities in vitro. Finally, the thermal stability of Hsc70 ATPase activity greatly exceeded environmental temperatures for G. mirabilis, with activity up to 62.5 degrees C. Overall, the biochemical characterization of the ATPase activity suggests that, although Hsc70 is not an extraordinarily thermally stable protein, it is capable of protein chaperoning cycles even at the extremes of environmental temperatures encountered by G. mirabilis in nature.


Subject(s)
Fishes/metabolism , HSP70 Heat-Shock Proteins/metabolism , Temperature , Adenosine Triphosphatases/metabolism , Animals , Drug Stability , HSC70 Heat-Shock Proteins , HSP70 Heat-Shock Proteins/chemistry , HSP70 Heat-Shock Proteins/isolation & purification , Hot Temperature , Luciferases/metabolism
5.
Fertil Steril ; 74(6): 1192-5, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11119749

ABSTRACT

OBJECTIVE: To determine the incidence of diminished ovarian reserve (OR) in patients with recurrent pregnancy loss (RPL). DESIGN: Retrospective chart review. SETTING: Tertiary fertility center. PATIENT(S): Six hundred ninety-two women undergoing a fertility evaluation. INTERVENTION(S): Clomiphene citrate challenge test (CCCT). MAIN OUTCOME MEASURE(S): FSH concentrations measured on menstrual days 3 and 10. RESULT(S): Forty-four women were diagnosed with RPL (+RPL), and 648 women had non-RPL diagnoses (-RPL). Compared with -RPL women, women with +RPL were younger (following statistics are listed as +RPL vs. -RPL, respectively; 34 +/- 5 vs. 35 +/- 4 y) but had similar menstrual cycle length (29 +/- 4 vs. 28 +/- 4 d), and lower day 3 FSH levels (8.9 + 7 vs. 11 +/- 9 mIU/mL) and similar day 10 FSH levels (11 +/- 8 vs. 12 +/- 11 mIU/mL). Eight of 44 women with +RPL (18%) had an abnormal CCCT, compared with 117/648 (18%) of women in the -RPL group. For women with normal OR, delivery rates were similar for -RPL and +RPL patients. For women with an abnormal CCCT, delivery rates were < 5%. CONCLUSION(S): Women with RPL have a similar incidence of diminished OR as the general infertile population. Reproductive outcome for patients with an abnormal CCCT is equally poor for both groups. Ovarian reserve screening should be considered in the work-up of RPL before initiation of anticoagulant or immunotherapy.


Subject(s)
Abortion, Habitual/physiopathology , Ovary/physiopathology , Abortion, Spontaneous/complications , Adult , Birth Rate , Clomiphene , Female , Humans , Infertility, Female/complications , Infertility, Female/physiopathology , Pregnancy , Retrospective Studies
7.
J Exp Biol ; 203(Pt 15): 2331-9, 2000 Aug.
Article in English | MEDLINE | ID: mdl-10887071

ABSTRACT

The heat-shock response, the enhanced expression of one or more classes of molecular chaperones termed heat-shock proteins (hsps) in response to stress induced by high temperatures, is commonly viewed as a 'universal' characteristic of organisms. We examined the occurrence of the heat-shock response in a highly cold-adapted, stenothermal Antarctic teleost fish, Trematomus bernacchii, to determine whether this response has persisted in a lineage that has encountered very low and stable temperatures for at least the past 14-25 million years. The patterns of protein synthesis observed in in vivo metabolic labelling experiments that involved injection of (35)S-labelled methionine and cysteine into whole fish previously subjected to a heat stress of 10 degrees C yielded no evidence for synthesis of any size class of heat-shock protein. Parallel in vivo labelling experiments with isolated hepatocytes similarly showed significant amounts of protein synthesis, but no indication of enhanced expression of any class of hsp. The heavy metal cadmium, which is known to induce synthesis of hsps, also failed to alter the pattern of proteins synthesized in hepatocytes. Although stress-induced chaperones could not be detected under any of the experimental condition used, solid-phase antibody (western) analysis revealed that a constitutively expressed 70 kDa chaperone was present in this species, as predicted on the basis of requirements for chaperoning during protein synthesis. Amounts of the constitutively expressed 70 kDa chaperone increased in brain, but not in gill, during 22 days of acclimation to 5 degrees C. The apparent absence of a heat-shock response in this highly stenothermal species is interpreted as an indication that a physiological capacity observed in almost all other organisms has been lost as a result of the absence of positive selection during evolution at stable sub-zero temperatures. Whether the loss of the heat-shock response is due to dysfunctional genes for inducible hsps (loss of open reading frames or functional regulatory regions), unstable messenger RNAs, the absence of a functional heat-shock factor or some other lesion remains to be determined.


Subject(s)
Heat-Shock Proteins/biosynthesis , Perciformes/metabolism , Acclimatization , Animals , Antarctic Regions , Cold Climate , Female , Gills/metabolism , HSP70 Heat-Shock Proteins/biosynthesis , In Vitro Techniques , Liver/metabolism , Male
8.
Article in English | MEDLINE | ID: mdl-10825695

ABSTRACT

The cold and constant water temperature of the Southern Ocean surrounding Antarctica provides a natural laboratory to address questions of temperature adaptation in marine organisms. In this study, endogenous levels and the number of isoforms of the 70 kDa heat shock protein multigene family (hsp70) of Antarctic and cold temperate notothenioid fishes were determined by SDS-polyacrylamide gel electrophoresis and Western blotting. Tissues from three Antarctic Trematomus congeners had significantly lower levels of 70 kDa Hsp isoforms than their temperate confamilial from New Zealand waters. However, these two thermally disparate sets of fish did not differ in number or pattern of 70 kDa Hsp isoforms expressed under normal physiological conditions. Additionally, levels of 70 kDa Hsp isoforms in specimens of one Antarctic species, Trematomus bernacchii, acclimated to 4 degrees C were significantly higher than non-acclimated conspecifics, indicating a direct effect of temperature on Hsp expression in this species. This study shows that constitutive expression of some members of the 70 kDa Hsp multigene family have been maintained, despite the absence of environmental heat stress for at least 2.5 million years.


Subject(s)
Adaptation, Physiological , Cold Temperature , Fishes/physiology , HSP70 Heat-Shock Proteins/metabolism , Animals , Antarctic Regions , HSP70 Heat-Shock Proteins/genetics , New Zealand , Protein Isoforms/genetics , Protein Isoforms/metabolism
9.
Annu Rev Physiol ; 61: 243-82, 1999.
Article in English | MEDLINE | ID: mdl-10099689

ABSTRACT

Molecular chaperones, including the heat-shock proteins (Hsps), are a ubiquitous feature of cells in which these proteins cope with stress-induced denaturation of other proteins. Hsps have received the most attention in model organisms undergoing experimental stress in the laboratory, and the function of Hsps at the molecular and cellular level is becoming well understood in this context. A complementary focus is now emerging on the Hsps of both model and nonmodel organisms undergoing stress in nature, on the roles of Hsps in the stress physiology of whole multicellular eukaryotes and the tissues and organs they comprise, and on the ecological and evolutionary correlates of variation in Hsps and the genes that encode them. This focus discloses that (a) expression of Hsps can occur in nature, (b) all species have hsp genes but they vary in the patterns of their expression, (c) Hsp expression can be correlated with resistance to stress, and (d) species' thresholds for Hsp expression are correlated with levels of stress that they naturally undergo. These conclusions are now well established and may require little additional confirmation; many significant questions remain unanswered concerning both the mechanisms of Hsp-mediated stress tolerance at the organismal level and the evolutionary mechanisms that have diversified the hsp genes.


Subject(s)
Heat-Shock Proteins/physiology , Molecular Chaperones/physiology , Stress, Physiological/physiopathology , Animals , Biological Evolution , Ecology , Heat-Shock Proteins/genetics , Heat-Shock Response/physiology , Humans
10.
Fertil Steril ; 69(3): 474-7, 1998 Mar.
Article in English | MEDLINE | ID: mdl-9531880

ABSTRACT

OBJECTIVE: To determine inhibin-B concentrations during ovarian reserve screening in women with normal and diminished ovarian reserve as determined by the clomiphene citrate challenge test. DESIGN: Retrospective. SETTING: Tertiary fertility center. PATIENT(S): Women undergoing ovarian reserve screening for a routine fertility evaluation. INTERVENTION(S): Clomiphene citrate challenge test. MAIN OUTCOME MEASURE(S): Inhibin-B concentrations on menstrual days 3 and 10. RESULT(S): Nineteen patients with normal ovarian reserve and 15 with diminished ovarian reserve had serum inhibin-B concentrations determined during ovarian reserve screening. For all patients, day 10 inhibin-B concentrations were higher than day 3. Women with normal ovarian reserve had higher inhibin-B concentrations on both days 3 and 10 than women with diminished ovarian reserve. Inhibin-B concentrations demonstrated a negative correlation with FSH levels on both cycle days 3 and 10 and a positive correlation with E2 on cycle day 10. CONCLUSION(S): Women with diminished ovarian reserve during ovarian reserve screening had reduced granulosa cell inhibin-B production compared with women with normal ovarian reserve. The lower inhibin-B concentrations may be responsible for the elevated FSH concentrations and may be indicative of the aging follicular apparatus.


Subject(s)
Clomiphene , Infertility, Female/diagnosis , Inhibins/blood , Ovary/physiopathology , Adult , Female , Follicle Stimulating Hormone/blood , Granulosa Cells/metabolism , Humans , Infertility, Female/physiopathology , Inhibins/biosynthesis , Kinetics , Retrospective Studies
11.
Am J Obstet Gynecol ; 177(3): 568-72, 1997 Sep.
Article in English | MEDLINE | ID: mdl-9322625

ABSTRACT

OBJECTIVE: Our goal was to evaluate the effect of intrauterine diethylstilbestrol exposure on ovarian reserve. STUDY DESIGN: Four hundred two women underwent ovarian reserve screening. Twenty had intrauterine diethylstilbestrol exposure, and 382 women did not have exposure. The groups were compared regarding age, the incidence of diminished ovarian reserve, and cycle length. Nine women with and 204 women without diethylstilbestrol exposure underwent comparison of the total human menopausal gonadotropin dose, the day of human chorionic gonadotropin administration, the peak estradiol level, and the number of mature follicles. RESULTS: The diethylstilbestrol-exposed women were similar in age (37 +/- 3.4 years) to the non-diethylstilbestrol-exposed women (35 +/- 4.4 years, p > 0.05). Three of 20 exposed women (15.8%) and 57 of 382 nonexposed women (15.3%) had diminished ovarian reserve (p = 0.41). When the exposed women were compared with the nonexposed subjects, the amounts of human menopausal gonadotropin (30 +/- 10 vs 33.7 +/- 10.6 ampules) required to achieve peak estradiol levels (633 +/- 323 vs 817 +/- 518 pg/ ml) with comparable numbers of follicles (5.7 +/- 2.7 vs 5.4 +/- 2.8) on the day of human chorionic gonadotropin administration were similar (p > 0.05). CONCLUSIONS: The diethylstilbestrol-exposed and nonexposed women had a similar incidence of diminished ovarian reserve and a similar follicular response to gonadotropins.


Subject(s)
Diethylstilbestrol/pharmacology , Estrogens, Non-Steroidal/pharmacology , Infertility, Female/diagnosis , Ovarian Follicle/drug effects , Ovary/drug effects , Adult , Chorionic Gonadotropin/pharmacology , Clomiphene/pharmacology , Diethylstilbestrol/adverse effects , Estradiol/blood , Estrogens, Non-Steroidal/adverse effects , Female , Fertility Agents, Female/pharmacology , Follicle Stimulating Hormone/blood , Humans , Incidence , Infertility, Female/chemically induced , Infertility, Female/physiopathology , Mass Screening , Menotropins/pharmacology , Middle Aged , Oocytes/cytology , Oocytes/drug effects , Oocytes/physiology , Ovarian Follicle/physiology , Ovary/physiology , Prognosis
12.
Z Gastroenterol ; 35(4): 247-53, 1997 Apr.
Article in German | MEDLINE | ID: mdl-9221610

ABSTRACT

INTRODUCTION: The exact source of phospholipase A2 is unknown. In this context the liver is discussed because the secretion of phospholipase A2 could be initiated during an acute phase response. PATIENTS AND METHODS: Three groups of patients were prospectively established: A liver resection (n = 12); B control (n = 22; oesophageal resection, gastrectomy, rectum resection); C sepsis (n = 5). Blood was collected preoperatively, each day postoperatively for seven days and at the day of discharge. Biochemical procedures: Phospholipase A2, PMN-elastase, C-reactive protein, GPT, GOT, GLDH, cholinesterase. RESULTS: Postoperatively phospholipase A2 is unchanged in the liver resection and control group, whereas it increases significantly in septic patients. Only following liver resection GPT, GOT and GLDH increase. C-reactive protein is increased in all groups. DISCUSSION: Unchanged phospholipase A2 after liver surgery despite an acute phase response indicated by C-reactive protein does not support the view of phospholipase A2 being of hepatic origin.


Subject(s)
Acute-Phase Proteins/metabolism , Acute-Phase Reaction/diagnosis , Hepatectomy , Liver Neoplasms/secondary , Phospholipases A/blood , Postoperative Complications/diagnosis , Rectal Neoplasms/surgery , Acute-Phase Reaction/enzymology , Adult , Aged , C-Reactive Protein/metabolism , Female , Humans , Leukocyte Elastase/blood , Liver Function Tests , Liver Neoplasms/surgery , Male , Middle Aged , Phospholipases A2 , Postoperative Complications/enzymology , Reference Values
13.
Biol Bull ; 192(2): 309-320, 1997 Apr.
Article in English | MEDLINE | ID: mdl-28581871

ABSTRACT

Heat-shock protein (hsp) expression was examined in gill of field-acclimatized and laboratory-acclimated mussels (Mytilus californianus) from the Oregon coast. Endogenous levels of heat-shock proteins in the 70-kDa class (hsp70 isoforms) and profiles of induction temperature for newly synthesized hsp 70 were measured in freshly field-collected specimens as functions of location height in the intertidal and season, and in mussels after 7 weeks of laboratory thermal acclimation. There were significant differences in endogenous levels of hsp70 as functions of season and collection height. Strong induction of new hsp70 synthesis occurred at body temperatures within the range measured in field specimens. Profiles of hsp70 thermal induction varied significantly with season, but not with height of collection. In contrast to the large differences in hsp70 expression between winter- and summer-acclimatized mussels, no differences related to temperature occurred in the differently acclimated mussels. The differences found between the effects of field acclimatization and laboratory thermal acclimation suggest that the stress response is modulated by environmental factors in addition to body temperature. Thus, caution is required in extrapolating from laboratory acclimation studies to acclimatization effects in field populations. The seasonal and tidal-height variations in the heat-shock response are discussed in the context of energy costs of protein turnover.

14.
Fertil Steril ; 66(6): 980-6, 1996 Dec.
Article in English | MEDLINE | ID: mdl-8941065

ABSTRACT

OBJECTIVE: To determine if elevated serum P concentration on the day of hCG administration or P area under the curve (AUC) impacts on pregnancy rates (PRs) in IVF-ET cycles. DESIGN: Retrospective study. PATIENT(S): One hundred thirty-three couples underwent IVF-ET using luteal GnRH-agonist suppression followed by hyperstimulation with gonadotropins. Patient cycles were controlled for female age, ovarian reserve, male factor with intracytoplasmic sperm injection (ICSI), endometrial thickness and pattern, number of embryos, and number of high-quality embryos transferred, and cell stage at transfer. Pregnancy was defined as an ongoing pregnancy > 20 weeks. RESULT(S): Patients and cycle parameters did not differ significantly based on the control parameters. Similar numbers of severe male factors treated by ICSI were in each group with similar PRs. Pregnancy rates and implantation rates did not differ based on P levels < or = 0.9, < or = 1.1, and < or = 1.4 ng/mL (conversion factor to SI unit, 3.18). Serum P concentrations per total eggs retrieved or per mature egg retrieved did not influence pregnancy outcome. Efficiency curves for serum P levels and P AUC did not demonstrate any differences in PR. At very high serum P (P > 2.0 ng/mL) samples size limits the ability of these data to rule out a negative effect. CONCLUSION(S): Serum P levels (and P AUC) commonly encountered during cycles of IVF-ET have no impact on pregnancy outcome.


Subject(s)
Chorionic Gonadotropin/pharmacology , Corpus Luteum/physiology , Embryo Transfer , Fertilization in Vitro , Ovary/physiopathology , Pregnancy Rate , Adult , Area Under Curve , Female , Humans , Male , Ovary/drug effects , Pregnancy , Progesterone/blood , Retrospective Studies
15.
Fertil Steril ; 66(3): 380-3, 1996 Sep.
Article in English | MEDLINE | ID: mdl-8751733

ABSTRACT

OBJECTIVE: To determine if ultrasonographic endometrial pattern or thickness is predictive of histologic endometrial maturation in women undergoing hormone replacement for ovum donation. DESIGN: Ultrasonographic endometrial thickness and pattern were determined and compared with histologic assessment of endometrial maturation. PATIENTS: Forty-six women underwent 52 preparatory cycles for ovum donation. Transvaginal ultrasound (US) was performed after 14 days of E2 replacement and, after 12 days of P, an endometrial biopsy was performed. In 12 cycles, a continuous dose of 2 mg/d E2 was administered. In cycles with out-of-phase biopsies (dated earlier than day 24) and in the last 34 cycles, all women received an escalating dose of E2 before initiation of P. Additionally, the 46 women underwent 55 ETs with USs performed on cycle day 15. RESULTS: Six women had abnormal biopsies in their first preparatory cycle on the continuous E2 protocol, which normalized with the escalating protocol. All other women had normal biopsies. Women with abnormal biopsies had significantly thinner endometrium (< or = 6 mm) but similar endometrial patterns compared with women with normal biopsies. In women having US in preparatory and transfer cycles, there were no differences in endometrial thickness or pattern between examinations. CONCLUSIONS: Endometrial thickness > or = 7 mm in hormone replacement cycles predicts in phase endometrial histology and can replace the endometrial biopsy.


Subject(s)
Endometrium/cytology , Estrogen Replacement Therapy , Oocyte Donation/methods , Biopsy , Endometrium/diagnostic imaging , Endometrium/pathology , Estrogens/pharmacology , Female , Humans , Ovarian Follicle/cytology , Ovarian Follicle/diagnostic imaging , Predictive Value of Tests , Progesterone/pharmacology , Retrospective Studies , Ultrasonography
16.
Fertil Steril ; 66(1): 49-53, 1996 Jul.
Article in English | MEDLINE | ID: mdl-8752610

ABSTRACT

OBJECTIVE: To examine selection criteria for ovarian reserve screening. DESIGN: Retrospective study. PATIENTS: Two hundred nineteen women underwent testing for ovarian reserve for woman's age > 35 years, any age with unexplained infertility, one ovary, or a poor response to hMG. INTERVENTIONS: Clomiphene citrate challenge test. MAIN OUTCOME MEASURES: Frequency of abnormal ovarian reserve screening, menstrual cycle parameters, response to hMG, and pregnancy outcome by screening criteria. RESULTS: One hundred eighty-four (84.0%) women had a normal ovarian reserve screening test; 35 (16.0%) had an abnormal ovarian reserve screening test. Twenty-six had abnormal ovarian reserve screening when screened by age, 14 for unexplained infertility, 5 for poor response to hMG, and 6 for one ovary. Fifteen women with abnormal ovarian reserve screening had more than one indication for screening. For women attempting pregnancy (n = 182), 49 of 148 (33.1%) with normal ovarian reserve screening became pregnant compared with 2 of 34 (5.9%) with abnormal ovarian reserve screening. Within each screening category, women with abnormal ovarian reserve had menstrual cycle parameters associated with a short follicular phase, required more hMG, and responded poorly to hMG. CONCLUSIONS: One of six women undergoing ovarian reserve screening had an abnormal test, which was associated a poor reproductive outcome. Age was the most important single criteria. Selected ovarian reserve screening is simple and inexpensive and should be offered to all fertility patients meeting the specific screening criteria listed above.


Subject(s)
Ambulatory Care Facilities , Clomiphene , Fertility , Infertility, Female/prevention & control , Mass Screening , Ovary/physiology , Adult , Age Factors , Female , Follicular Phase , Hormones/blood , Humans , Infertility, Female/drug therapy , Menotropins/therapeutic use , Menstrual Cycle/blood , Patient Selection , Pregnancy , Pregnancy Rate , Retrospective Studies
17.
Fertil Steril ; 65(5): 1021-5, 1996 May.
Article in English | MEDLINE | ID: mdl-8612828

ABSTRACT

OBJECTIVE: To examine intraobservational and interobservational variation in sperm critical morphology analysis between trained andrologists and a computer over the range of 0% to 15% normal forms. DESIGN: Retrospective. SETTING: Fertility center. PATIENTS: Twenty-four semen specimen slides were read five times in a randomized, blinded fashion by two andrologists and a computerized semen analyzer. Twenty-five samples were analyzed using a different stain and a centrifugation step. MAIN OUTCOME MEASURE: Sperm strict morphology. RESULTS: Mean average intraobserver deviation of normal forms was small (1.3% to 2.7%). Intraobserver deviation did not differ between andrologists, but both differed from the computer. Interobserver deviation for percent normal forms was similar for andrologists and computer. Mean average deviations were small over the variation was independent of the initial percent normal forms. The computer frequently evaluated significantly fewer cells than the andrologists. Updated software increased the number of cells read by the computer requiring more time than the andrologist. CONCLUSIONS: Trained andrologists and a computer determine sperm critical morphology similarly over the range of 0% to 15% normal forms. A single evaluation is highly predictive of multiple evaluations. Improvements in the speed of computer assessment are still necessary.


Subject(s)
Spermatozoa/abnormalities , Diagnosis, Computer-Assisted , Humans , Male , Observer Variation , Retrospective Studies , Software
18.
Biol Reprod ; 53(4): 899-904, 1995 Oct.
Article in English | MEDLINE | ID: mdl-8547486

ABSTRACT

Human cyto- and synchtiotrophoblasts have recently been shown to contain functional hCG/LH receptors. The present study investigated whether human intermediate trophoblasts also contain these receptors. Six implantation sites from 4-8 wk of pregnancy were processed for in situ hybridization for hCG/LH receptors coupled with immunocytochemistry for human placental lactogen (hPL), an endocrine marker for intermediate trophoblasts. In addition, the sections were also double-immunostained for hCG/LH receptor and hPL proteins. The results showed that some but not all the intermediate trophoblasts contained hCG/LH receptors. The intermediate trophoblasts in endovascular locations contained more receptors than those in perivascular locations. Villus syncytiotrophoblasts contained the highest level of receptors, followed by intermediate trophoblasts in endovascular and perivascular locations and then villus cytotrophoblasts. These data, demonstrating the presence of hCG/LH receptors in intermediate trophoblasts, suggest a possible role for hCG in trophoblast invasion and maternal recognition of pregnancy.


Subject(s)
Gene Expression , Receptors, LH/genetics , Trophoblasts/metabolism , Embryo Implantation , Female , Humans , Immunohistochemistry , In Situ Hybridization , Placental Lactogen/analysis , Pregnancy , RNA, Messenger/analysis
19.
Fertil Steril ; 63(6): 1267-71, 1995 Jun.
Article in English | MEDLINE | ID: mdl-7750599

ABSTRACT

OBJECTIVE: To determine if premature luteinization (serum P levels > 1.1 ng/mL on or before the day of hCG administration) during controlled ovarian hyperstimulation (COH) is associated with elevated levels of serum hCG. SETTING: Tertiary fertility center. DESIGN: Retrospective evaluation of ovum donors undergoing COH. PATIENTS: Forty-four women underwent COH. Comparisons of serum hCG levels and hormonal and cycle characteristics were made between cycles with premature luteinization (group I) and without premature luteinization (group II). RESULTS: Group I (16 women) were similar to women in group II in age, amount of hMG, and the ratio of FSH:hMG received. Both groups received hCG on similar days, but women in group I had higher peak E2 levels. Serum hCG levels increased and correlated with serum P levels in group I only and were higher on the day of hCG administration (group I 1.8 +/- 0.9 mIU/mL versus group II 1.2 +/- 0.45 mIU/mL; conversion factor to SI unit, 1.00). Peak E2 and LH levels, ampules of hMG and the FSH:LH ratio, and day of hCG administration did not correlate with hCG levels. Human chorionic gonadotropin exposure, as measured by area under the curve, was significantly higher in group I compared with group II. CONCLUSION: Higher serum levels of hCG and integrated hCG exposure are found in COH cycles with premature luteinization compared with cycles without premature luteinization. Higher hCG levels may be due to decreased clearance of hCG from the circulation and/or the hCG content of hMG.


Subject(s)
Chorionic Gonadotropin/blood , Corpus Luteum/physiology , Oocyte Donation , Ovulation Induction , Adult , Chorionic Gonadotropin/administration & dosage , Estradiol/blood , Female , Follicle Stimulating Hormone/administration & dosage , Follicle Stimulating Hormone/therapeutic use , Humans , Luteinizing Hormone/blood , Menotropins/administration & dosage , Menotropins/therapeutic use , Progesterone/blood , Retrospective Studies
20.
Fertil Steril ; 63(5): 979-83, 1995 May.
Article in English | MEDLINE | ID: mdl-7720942

ABSTRACT

OBJECTIVES: To evaluate the relationship of elevated day 10 P levels (> or = 1.1 ng/mL, conversion factor to SI unit, 3.18) during ovarian reserve screening and reproductive performance. DESIGN: Prospective screening with longitudinal follow-up. INTERVENTIONS: One hundred seven women underwent ovarian reserve screening with a clomiphene citrate challenge test. MAIN OUTCOME MEASURES: Serum FSH, LH, and E2 levels were determined on cycle day 3 and FSH, LH, and P levels were determined on day 10. A fertility evaluation was completed and a treatment plan was instituted. RESULTS: Twenty-two of 107 (20.6%) women had day 10 P levels > or = 1.1 ng/mL. Women with elevated day 10 P levels were similar in age to women with normal day 10 P levels (< or = 0.9 ng/mL) but had significantly shorter menstrual cycles, higher day 3 and day 10 FSH levels, higher day 3 E2 levels, and higher day 10 LH levels than women with normal day 10 P levels. During ovarian hyperstimulation, women with elevated day 10 P levels required more ampules of hMG, had lower peak E2 levels, and had fewer mature follicles than women with a normal day 10 P level. Sixteen of 85 (18.8%) women with normal day 10 P levels became pregnant, but none of the 22 women with elevated day 10 P levels became pregnant. The incidence of diminished ovarian reserve was higher in women with elevated day 10 P levels (13/22; 59%) when compared with women with a normal day 10 P levels (9/85; 10.6%). CONCLUSIONS: Elevated day 10 P levels (> or = 1.1 ng/mL) during ovarian reserve screening is associated with diminished ovarian reserve and correlates with menstrual cycle parameters associated with a short follicular phase and poor reproductive performance.


Subject(s)
Ovary/physiology , Progesterone/blood , Reproduction , Adult , Clomiphene , Estradiol/blood , Female , Follicle Stimulating Hormone/blood , Follicular Phase , Humans , Luteinizing Hormone/blood , Menotropins/administration & dosage , Menstrual Cycle , Pregnancy , Prospective Studies , Time Factors
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