bluuebunny/crossref_metadata_2025 huggingface dataset:
Measurement of Normal Pressures and Friction Forces Acting on Buried Pipes Subjected to Cyclic Axial Displacements in Laboratory Experiments
Authors: Weidlich I. , Achmus M.
Published: February 2008

The earth pressure acting on buried pipes is decisive for the pipe design. For pipes shifting in the ground, such as district heating pipes, which are subjected to cyclic temperature loading, the mobilizable friction force, which is also dependent on the earth pressure load, is a very important quantity. Friction force changes with cyclic axial displacement of a pipeline in operation. A special testing device was designed to investigate the friction forces and the normal pressures on a pipe buried in sand and how these change during cyclic axial displacement dependent on system and soil parameters. The experimental setup is presented, and in particular the application of the tactile pressure sensor, the special measuring technique used, is described. The dependence of the decrease in friction force on pipe diameter, soil overburden height, and relative density of the soil was investigated in the tests. The results are presented and the findings are discussed.
Long-term effects of Helicobacter pylori eradication on circulating ghrelin and leptin concentrations and body composition in prepubertal children
Authors: Pacifico Lucia , Anania Caterina , Osborn John F , Ferrara Eva , Schiavo Elisa , Bonamico Margherita , Chiesa Claudio
Published: February 2008

Background

Helicobacter pylori, and the chronic gastric inflammation that it causes, may compromise the function and survival of ghrelin-producing cells, resulting in a decrease of circulating ghrelin levels. This finding raises the possibility that the infection might affect growth in children by reducing the ghrelin production.

Aims

To determine baseline circulating levels of ghrelin and leptin in prepubertal children with and without H. pylori infection and to evaluate the long-term effects of H. pylori eradication on circulating levels of ghrelin and leptin as well as on body composition.

Patients

Thirty children with H. pylori-associated gastritis, 35 children with H. pylori-negative gastric mucosa, and 20 healthy controls were studied.

Results

At baseline, while leptin levels were significantly lower in H. pylori-positive patients, ghrelin concentrations did not differ among the three study groups. However, a significant inverse correlation between ghrelin concentrations and histological severity of gastritis was found. Eradication of the organism was associated with a progressive decrease in ghrelin concentrations over baseline at both 6- and 12-month follow-ups. SDS-body mass index (BMI), lean and fat mass, as well as leptin concentrations, significantly increased over baseline at both follow-ups.

Conclusions

In prepubertal children, serum ghrelin concentrations are inversely related to the severity of H. pylori-associated gastritis. In these youngsters, long-term eradication of H. pylori infection is associated with a significant increase in BMI, lean and fat mass along with a significant decrease in circulating ghrelin levels and an increase in leptin levels.
Combined immunostaining with galectin-3, fibronectin-1, CITED-1, Hector Battifora mesothelial-1, cytokeratin-19, peroxisome proliferator-activated receptor-γ, and sodium/iodide symporter antibodies for the differential diagnosis of non-medullary thyroid carcinoma
Authors: Liu Ying Y , Morreau Hans , Kievit Job , Romijn Johannes A , Carrasco Nancy , Smit Johannes W
Published: February 2008

Objectives

The microscopic distinction between benign and malignant thyroid lesions in clinical practice is still largely based on conventional histology. This study was performed to evaluate the diagnostic value of galectin-3 (Gal-3), Hector Battifora mesothelial-1 (HBME-1), cytokeratin (CK)-19, CBP P300-interacting transactivator with glutamic acid E- and aspartic acid D-rich C-terminal domain (CITED-1), fibronectin (FN)-1, peroxisome proliferator-activated receptor (PPAR)-γ, and intracellular sodium/iodide symporter (iNIS) immunostaining in a large panel of thyroid neoplasms. Our study differed from earlier ones with regard to the identification of optimal semiquantitative cut-off levels using receiver operator curve (ROC) analysis and hierarchical cluster analysis.

Methods

We used tissue arrays containing 177 thyroid tissues: 100 benign tissues (including normal thyroid, Graves disease, multinodular goiter, and follicular adenoma (FA)) and 77 thyroid carcinomas (including papillary thyroid carcinoma (PTC), follicular thyroid carcinoma, and follicular variant of PTC (FVPTC)). Antibody staining was scored semiquantitatively based on the ROC analyses and with hierarchical cluster analysis.

Results

In general, we found overexpression of FN-1, CITED-1, Gal-3, CK-19, HBME-1, and iNIS in malignant thyroid lesions. Gal-3, FN-1, and iNIS had the highest accuracy in the differential diagnosis of follicular lesions. A panel of Gal-3, FN-1, and iNIS, identified by hierarchical cluster analysis, had a 98% accuracy to differentiate between FA and malignant thyroid lesions. In addition, HBME-1 was found to be useful in the differentiation between FA and FVPTC (accuracy 88%).

Conclusion

We conclude that identifying optimal antibody panels with cluster analysis increases the diagnostic value in the differential diagnosis of thyroid neoplasms, the combination of FN-1, Gal-3, and iNIS having the best accuracy (98%).
Growth modelling of non-functioning pituitary adenomas in patients referred for surgery
Authors: Honegger Juergen , Zimmermann Sanna , Psaras Tsambika , Petrick Manfred , Mittelbronn Michel , Ernemann Ulrike , Reincke Martin , Dietz Klaus
Published: February 2008

Objective

Recent observational studies have established progression and recurrence rates of pituitary adenomas. However, it is still unknown how individual pituitary adenomas grow over years and whether growth kinetics follow a distinct growth model. The objective of this study was to define a growth model for non-functioning pituitary adenomas.

Methods

Fifteen patients who had five or more serial high-quality examinations with magnetic resonance images or computerized tomography scans were identified among 216 patients with non-functioning pituitary adenomas. Tumour volumes were assessed using a stereological method based on the Cavalieri principle. Tumour growth during the observation period was analysed and different growth models were fitted to the data.

Results

Fifteen pituitary adenomas (12 recurrent tumours and 3 newly diagnosed tumours) were longitudinally observed during a median observation period of 7.4 years (range: 2.3–11.9 years). Growth kinetics could be described either by an exponential growth model (nine patients) or by a logistic model (five patients) with initial exponential growth followed by deceleration of growth. One tumour remained unchanged in size during the observation period. None of the adenomas showed accelerated growth during the observation period. Overall, the linear growth model was not suitable to describe the growth kinetics of non-functioning pituitary adenomas.

Conclusions

Our study shows that growth of pituitary adenomas can be described by distinct growth models. Knowledge of growth dynamics has implications for clinical practice and helps to adjust scanning protocols for follow-up investigations.
Hypoactive sexual desire in transsexual women: prevalence and association with testosterone levels
Authors: Elaut Els , De Cuypere Griet , De Sutter Petra , Gijs Luk , Van Trotsenburg Michael , Heylens Gunter , Kaufman Jean-Marc , Rubens Robert , T'Sjoen Guy
Published: February 2008

Objective

An unknown proportion of transsexual women (defined as post-operative male-to-female transsexuals on oestrogen replacement) experience hypoactive sexual desire disorder (HSDD). It has been suggested that the absence of ovarian androgen production together with oestrogen treatment-related increase in sex hormone-binding globulin (SHBG) levels could be leading to HSDD, due to low levels of biologically available testosterone. This study wishes to document the HSDD prevalence among transsexual women and the possible association to androgen levels.

Design

Cross-sectional study.

Methods

Transsexual women (n=62) and a control group of ovulating women (n=30) participated in this study. Questionnaires measuring sexual desire (sexual desire inventory) and relationship and sexual satisfaction (Maudsley Marital Questionnaire) were completed. Serum levels of total testosterone, LH and SHBG were measured in blood samples obtained at random in transsexual women and in the early follicular phase in ovulating women.

Results

The transsexual group had lower levels of total and calculated free testosterone (both P<0.001) than the ovulating women. HSDD was reported in 34% of the transsexual and 23% of the ovulating women (P=0.30). Both groups reported similar levels of sexual desire (P=0.97). For transsexual women, no significant correlation was found between sexual desire and total (P=0.64) or free testosterone (P=0.82). In ovulating women, these correlations were significant (P=0.006, resp. P=0.003).

Conclusions

HSDD is reported in one-third of transsexual women. This prevalence is not substantially different from controls, despite markedly lower (free) testosterone levels, which argues against a major role of testosterone in this specific group.
Acute changes in serum osteoprotegerin and receptor activator for nuclear factor-κB ligand levels in women with established osteoporosis treated with teriparatide
Authors: Anastasilakis Athanasios D , Goulis Dimirtios G , Polyzos Stergios A , Gerou Spiridon , Pavlidou Vasiliki , Koukoulis George , Avramidis Avraam
Published: February 2008

Objective

The mechanisms regulating the anabolic response of the skeleton to intermittent exogenous parathyroid hormone (PTH) administration are not fully elucidated. The aim of this prospective study was to evaluate the acute effect (up to 1 month) of teriparatide (TPTD; human recombinant PTH 1–34) on serum levels of osteoprotegerin (OPG) and receptor activator for nuclear factor-κB ligand (RANKL) in women with established osteoporosis.

Design

Twenty-three postmenopausal Caucasian women with established osteoporosis (mean age 66.7±1.6 years) received daily injections of 20 μg TPTD for 12 months.

Methods

Serum samples for total calcium (Ca), phosphate, alkaline phosphatase, N-terminal propeptide of type I collagen, intact PTH (iPTH), OPG, and RANKL were obtained at baseline, 1 h, 1 day, and 1 month after initiation of therapy. Lumbar spine bone mineral density (BMD) was measured before and 12 months after TPTD treatment.

Results

Serum total Ca increased and iPTH gradually decreased with TPTD treatment. Serum OPG levels remained unchanged, while RANKL increased gradually during the study (P<0.001). There was no correlation between OPG or RANKL and BMD changes or iPTH levels.

Conclusions

TPTD therapy in women with postmenopausal osteoporosis results in acute increase in serum RANKL levels but does not affect serum OPG. These changes may reflect an increase in the number of active osteoblasts with therapy and might be responsible for the acceleration of bone turnover rate that characterizes TPTD.
Quantitative analysis of somatostatin receptor subtypes (1–5) gene expression levels in somatotropinomas and correlation to in vivo hormonal and tumor volume responses to treatment with octreotide LAR
Authors: Taboada Giselle F , Luque Raul M , Neto Leonardo Vieira , Machado Evelyn de O , Sbaffi Bruna C , Domingues Romeu C , Marcondes Jorge B , Chimelli Leila M C , Fontes Rosita , Niemeyer Paulo , de Carvalho Denise P , Kineman Rhonda D , Gadelha Mônica R
Published: February 2008

Objective

To determine whether the somatostatin receptor subtype (SSTR) expression profile correlates with hormonal and tumor volume responses to postsurgical octreotide long acting repeatable (OCT LAR) treatment.

Design and methods

Quantitative real-time RT-PCR was used to evaluate the absolute mRNA copy numbers for all five SSTR subtypes in 22 somatotropinomas. Response to OCT LAR was studied by hormone levels (GH and IGF-I) and tumor volume (sella turcica magnetic resonance imaging).

Results

SSTR5 was present at the highest level followed by SSTR2, SSTR3, SSTR1, and SSTR4 (2327 (1046–5555), 2098 (194–23 954), 97 (0–460), 14 (0–29 480), and 0 (0–652) copies respectively). Positive correlations were found between SSTR2 levels and the percentage decrease of GH and IGF-I after 3 (r=0.49, P<0.027 and r=0.49, P<0.029 respectively) and 6 (r=0.59, P<0.006 and r=0.58, P<0.008 respectively) months of OCT LAR. A negative correlation was found between SSTR5 mRNA levels and the percentage decrease of GH after 3 months of OCT LAR (r=−0.52, P=0.016, n=21). A higher SSTR2/SSTR5 ratio was observed among patients who obtained hormonal control with OCT LAR, when compared with those uncontrolled (2.4 (0.7–10) vs 0.3 (0.1–7.7), P=0.001). A ROC curve analysis showed a SSTR2/SSTR5 ratio of 1.3 as the best predictor of disease control, with a sensitivity of 88% and a specificity of 92% – area under curve, 0.9. A positive correlation was also found between SSTR2 mRNA levels and the percentage decrease in tumor volume after 6 months of OCT LAR (r=0.79, P=0.002, n=12).

Conclusions

Somatostatin receptor subtype 2 mRNA expression levels in somatotropinomas correlate positively with in vivo hormonal and tumor volume responses to OCT LAR.
Smoking is negatively associated with the presence of thyroglobulin autoantibody and to a lesser degree with thyroid peroxidase autoantibody in serum: a population study
Authors: Pedersen Inge Bülow , Laurberg Peter , Knudsen Nils , Jørgensen Torben , Perrild Hans , Ovesen Lars , Rasmussen Lone Banke
Published: February 2008

Background

Autoimmune thyroid diseases are common and the prevalence of circulating thyroid antibodies (thyroid peroxidase antibody, TPO-Ab and thyroglobulin antibody, Tg-Ab) is high in the population. The knowledge of a possible association between lifestyle factors and circulating thyroid antibodies is limited.

Aim

To evaluate the correlation between smoking habits and the presence of circulating TPO-Ab and Tg-Ab.

Material and methods

In a cross-sectional comparative population study performed in two areas of Denmark with moderate and mild iodine deficiency, 4649 randomly selected subjects from the population in some predefined age groups between 18 and 65 years were examined. Blood tests were analysed for TPO-Ab and Tg-Ab using assays based on the RIA technique. The participants answered questionnaires, were clinically examined and blood and urine samples collected.

Results

Data were analysed in multivariate logistic regression models. There was a negative association between smoking and the presence of thyroid autoantibodies in serum. This association was observed for the presence of TPO-Ab and/or Tg-Ab, TPO-Ab (without respect to Tg-Ab status), Tg-Ab (without respect to TPO-Ab status) and both antibodies together. The association between smoking and thyroid autoantibodies was stronger for Tg-Ab than for TPO-Ab. There was no association between smoking and TPO-Ab measured alone or between smoking and TPO-Ab when Tg-Ab was included in the model as an explanatory variable.

Conclusion

Smoking was negatively associated with the presence of thyroid autoantibodies with the strongest association between smoking and Tg-Ab. The study design precludes any conclusions as to the cause of the negative association between smoking thyroid autoantibodies.
Uncommon association of germline mutations of RET proto-oncogene and CDKN2A gene
Authors: Foppiani L , Forzano F , Ceccherini I , Bruno W , Ghiorzo P , Caroli F , Quilici P , Bandelloni R , Arlandini A , Sartini G , Cabria M , Del Monte P
Published: February 2008

Introduction

Calcitonin measurement is advised in the diagnosis of thyroid nodules, as it is an accurate marker of medullary thyroid carcinoma (MTC). C-cell hyperplasia (CCH)-induced hypercalcitoninemia cannot be distinguished from that induced by MTC, unless surgery is performed.

Case

We report the clinical and biological features of a patient with a family history of cancer, including melanoma and pancreatic cancer, who had previously undergone surgery for melanoma. He presented the unusual association of papillary thyroid carcinoma (PTC), normocalcemic hyperparathyroidism, and hypercalcitoninemia with a pathological response to pentagastrin, which was histologically deemed secondary to CCH. Multiple endocrine neoplasia (MEN) 2A was diagnosed. RET gene analysis showed a p.V804M missense mutation in exon 14, a low- but variably penetrant defect found in both sporadic and MEN2A-associated MTC/CCH, and a p.G691S polymorphism in exon 11. Furthermore, the germline P48T mutation was found in the CDKN2A gene exon 1, which is known to be associated with melanoma and pancreatic cancer. The patient showed the uncommon coexistence of a germline mutation in two suppressor genes, RET and CDKN2A; this finding, deemed to be a mere coincidence, did not modify the phenotype expected by each single mutation. CCH associated with V804M RET mutation is a precancerous condition and surgery is recommended. In order to exclude MTC, surgery is advised in patients with a pathological calcitonin response to pentagastrin, in the absence of thyroid autoimmunity. CCH-induced hypercalcitoninemia can be associated with thyroid cancers other than MTC (e.g., PTC). Family history is important in scheduling specific genetic screening in high-risk patients and their relatives.
The association between serum parathyroid hormone and bone mineral density, and the impact of smoking: the Tromsø Study
Authors: Sneve Monica , Emaus Nina , Joakimsen Ragnar Martin , Jorde Rolf
Published: February 2008

Objective

To explore the relation between serum parathyroid hormone (PTH) and bone mineral density (BMD), adjusted for lifestyle factors including smoking.

Design

Cross-sectional study.

Methods

The Tromsø Study is a population-based study performed for the fifth time in 2001. Serum PTH was measured and the subjects filled in a questionnaire covering lifestyle factors. BMD at the hip, distal and ultradistal forearm was measured.

Results

Complete datasets were available in 1442 men and 1368 women. Age, body mass index and serum PTH were strong predictors of BMD level at the hip in both genders. No significant relation was seen between serum PTH and BMD at the distal or ultradistal forearm. When smokers and non-smokers were analysed separately, the relation between PTH and BMD at the hip was significant in current non-smokers only. In males, current non-smokers had significantly higher BMD at all three measurement sites compared with current smokers. Male former smokers had values in between current and never smokers. There was a significant and negative relation between number of years smoked and BMD at the hip. In male former smokers, there was an increase in BMD with increasing years since smoking cessation.

Conclusion

Serum PTH is negatively associated with BMD at the hip, and the relation seems to be masked, or diminished, by smoking. Smoking reduces BMD at the hip, distal and ultradistal forearm in males, and the effect appears to be mainly time and not dose dependent.
Impaired quality of life of patients with acromegaly: control of GH/IGF-I excess improves psychological subscale appearance
Authors: Matta Maria P , Couture Elisabeth , Cazals Laurent , Vezzosi Delphine , Bennet Antoine , Caron Philippe
Published: February 2008

Introduction

Acromegaly, a chronic disease caused by GH/IGF-I excess, has a major impact on quality of life (QoL).

Objective

To evaluate QoL of acromegalic patients in relation to control status of the disease.

Design and methods

Single center observational study including 93 patients with acromegaly recruited to complete QoL questionnaire (AcroQol). QoL was evaluated at least 3 months after surgery and/or medical treatment. Patients were divided into two groups: controlled (I) and uncontrolled (II) according to the latest consensus acromegaly ‘control’ criteria and further subdivided into four subgroups according to the previous pituitary adenoma surgery (Ib and IIb) or without surgery (Ia and IIa).

Results

Mean GH (0.81±0.47 ng/ml) and IGF-I (195±71 ng/ml) values in group I were significantly lower than in group II (GH, 7.01±12.05 ng/ml and IGF-I, 513±316 ng/ml; P<0.001). There was no difference in total AcroQol score, physical, or psychological scales between groups I and II. However, when adjusted to age and disease duration since diagnosis, patients of group I (63±20%) showed an improved psychological subscale appearance than those of group II (58±17%; P=0.035). In group II, IGF-I level was lower after surgery (IIa=588±353, IIb=410±225 ng/ml; P<0.038), and psychological subscale appearance was significantly better in subgroup IIb (64.9±18.1%) than in subgroup IIa who had medical treatment (53.9±14.3%; P=0.009).

Conclusion

QoL is severely impaired in acromegalic patients. Control of GH/IGF-I excess by surgery or medical treatment seems to have a positive impact on psychological subscale appearance.
Urinary messenger RNA expression of podocyte-associated molecules in patients with diabetic nephropathy treated by angiotensin-converting enzyme inhibitor and angiotensin receptor blocker
Authors: Wang Gang , Lai Fernand Mac-Moune , Lai Ka-Bik , Chow Kai-Ming , Kwan Bonnie Ching-Ha , Li Philip Kam-Tao , Szeto Cheuk-Chun
Published: February 2008

Background

Podocyte injury and its subsequent loss in urine play an important role in the pathogenesis of diabetic nephropathy; blockade of the renin–angiotensin system may ameliorate the damage.

Methods

In a non-randomized setting, we studied 71 patients with diabetic nephropathy on a stable dose of angiotensin-converting enzyme inhibitor (ACEI). In 37 patients, angiotensin receptor blocker (ARB) was added (the combination group); ACEI alone was continued in the other 34 (the control group). The mRNA expressions of nephrin, podocin, and synaptopodin in urinary sediment were measured at 0 and 12 weeks.

Results

Baseline glomerular filtration rate (GFR) correlated with the urinary expression of nephrin (r=0.320, P=0.007), podocin (r=0.336, P=0.004), and synaptopodin (r=0.350, P=0.003). After adjusting for the baseline expression, the combination group had a significantly lower urinary synaptopodin expression (7.49 (95% confidence interval CI, 0.62–115.29) vs 14.83 (95% CI, 1.03–241.43), P=0.026) than the control group after 12 weeks of treatment. The percentage change in urinary podocin expression over 12 weeks of treatment had a modest correlation with the rate of GFR decline in 1 year (r=−0.243, P=0.041).

Conclusion

In patients with diabetic nephropathy, urinary mRNA expression of podocyte markers correlated with baseline renal function. Urinary expression of synaptopodin was lower after 12 weeks of ACEI and ARB combination therapy. Our result suggests that serial measurement of urinary podocyte markers may have a value for the monitoring of therapeutic response.
Metabolic evidence for impaired 17α-hydroxylase activity in a kindred bearing the E305G mutation for isolate 17,20-lyase activity
Authors: Tiosano Dov , Knopf Carlos , Koren Ilana , Levanon Nurit , Hartmann Michaela F , Hochberg Ze'ev , Wudy Stefan A
Published: February 2008

Context

The CYP17A1 gene encodes many enzymatic reactions including 17α-hydroxylase and 17,20-lyase activities. Mutations that selectively ablate the 17,20-lyase activity, causing isolated 17,20-lyase deficiency, are exceedingly rare and may belong to the rarest of all disorders of steroidogenesis. We have previously reported an E305G mutation in the active site of CYP17A1 that apparently causes isolated 17,20-lyase deficiency. Expression studies suggested intact 17α-hydroxylase activity which was at odds with subnormal tetracosactrin stimulated cortisol in the patients.

Objectives

To investigate the in vivo activity of the adrenal enzymes, we used the metabolomics approach with urinary steroid profiling by gas chromatography–mass spectrometry.

Patients

Of the 11 subjects investigated, 6 patients in the kindred were found to be homozygous, 4 members were asymptomatic heterozygous, and 1 was homozygous for the wild-type allele.

Results

In the homozygous patients for E305G, both serum and urinary steroids showed a severe lack of androgens (C19-steroids) pointing to the absence of 17,20-lyase activities. Furthermore, precursor/product ratios of urinary steroid metabolites characterizing 17α-hydroxylase activity showed variable decreases in 17α-hydroxylase activities.

Conclusions

The results confirm the complete absence of 17,20-lyase activity in vivo, as in the in vitro expression studies. On the other hand, in vivo 17α-hydroxylase activity was partially impaired. Thus, the in vivo metabolic data seem to be more sensitive than the expression study and suggests that this mutation also impairs 17α-hydroxylase activity.
GH responses to two consecutive bouts of neuromuscular electrical stimulation in healthy adults
Authors: Sartorio A , Jubeau M , Agosti F , De Col A , Marazzi N , Lafortuna C L , Maffiuletti N A
Published: February 2008

Background

It is well established that repeated GHRH administration or repeated voluntary exercise bouts are associated with a complete blunting of GH responsiveness when the administration of the second stimulus follows the first one after a 2-h interval.

Aim

To evaluate GH responses to neuromuscular electrical stimulation (NMES) in healthy adults.

Methods

Six volunteers (mean age±s.d. 31.7±5.5 years) were studied before and after two consecutive bouts of NMES exercise (a series of 20 contractions at the maximum of individual tolerance, frequency: 75 Hz, pulse duration: 400 μs, on–off ratio: 6.25–20 s) administered at a 2-h interval.

Results

Baseline GH levels (mean: 0.3±0.2 ng/ml) significantly increased after the first NMES (peak: 4.2±3.7 ng/ml), with a complete normalization after 120 min (0.3±0.3 ng/ml). The administration of the second bout of NMES of comparable characteristics also resulted in a significant GH increase (peak: 5.2±3.2 ng/ml), which was comparable with that observed after the previous one. GH net incremental area under the curve after the first and second bouts of NMES were not significantly different (155.1±148.5 and 176.9±123.3 ng/ml per h, P=0.785).

Conclusions

Unlike repeated pharmacological stimuli and voluntary exercise bouts, subsequent sessions of NMES administered at a 2-h interval appear to circumvent feedback mechanisms and to re-induce the GH responses, thus indicating a possible different underlying mechanism elicited by different GH-releasing stimuli.
Cyclosporin A improves murine pregnancy outcome in abortion-prone matings: involvement of CD80/86 and CD28/CTLA-4
Authors: Zhou Wen-Hui , Dong Lin , Du Mei-Rong , Zhu Xiao-Yong , Li Da-Jin
Published: February 2008

Immune regulation during pregnancy is complex, and thus an optimal therapy for pregnancy complications is always a big challenge to reproductive medicine. Cyclosporin A (CsA), a potent immunosuppressant, prevents rejection of allografts by hosts, but little is known about the modulating effect of CsA on the materno-fetal relationship. Here, pregnant CBA/J females mated with DBA/2 males as an abortion-prone model were administered with CsA on day 4.5 of gestation, and the pregnant CBA/J females mated with BALB/c males were established as successful pregnancy control. It was demonstrated that administration of CsA at the window of implantation significantly up-regulated the expression of CTLA-4, while down-regulating the levels of CD80, CD86, and CD28 at the materno-fetal interface in the CBA/J×DBA/2 abortion-prone matings, and the embryo resorption rate of the abortion-prone matings reduced significantly after CsA treatment, implying that modulation of costimulatory molecule expression by CsA might contribute to preventing the fetus from maternal immune attack. In addition, treatment with CsA induced enhanced growth and reduced cell apoptosis of the murine trophoblast cells. Together, these findings indicate that CsA has a beneficial effect on the materno-fetal interface in abortion-prone matings, leading to a pregnancy outcome improvement, which might provide new therapeutics for spontaneous pregnancy wastage.
The effect of glycosaminoglycans on rat gametes in vitro and the associated signal pathway
Authors: Borg Neil , Holland Michael
Published: February 2008

The effects of adding the extracellular glycosaminoglycans (GAGs), hyaluronic acid (HA) and chondroitin sulphate (CS) to ratin vitrofertilisation (IVF) media were assessed. Metaphase II (MII) oocytes were also incubated in GAG-supplemented modified rat 1-cell embryo culture medium (mR1ECM+BSA) for 3 days. Cytoplasmic fragmentation was significantly reduced in mR1ECM+BSA with HA (39.0–48.0%) compared with the control (82.0%). In IVF experiments, neither HA (8.0–30.8%) nor CS (9.7–42.5%) improved fertilisation rates compared with controls fertilised in M16 (47.2%) or enriched Krebs–Ringer bicarbonate solution (61.5%). RT-PCR and Western blot were used to probe for CD44 mRNA and protein in Sprague–Dawley gametes and cumulus cells. CD44 was identified in cumulus cells, suggesting a role for oocyte maturation and cumulus expansion. The CD44 protein was also present on caudal epididymal spermatozoa that were highly stimulated by CSin vitroimplicating a role in fertilisation for CS and CD44.
Associations between tumor necrosis factor-α and lymphotoxin-α polymorphisms and idiopathic recurrent miscarriage
Authors: Zammiti W , Mtiraoui N , Khairi H , Gris J-C , Almawi W Y , Mahjoub T
Published: February 2008

Heightened expression of tumor necrosis factor (TNF)-α and lymphotoxin-α (LT-α) was associated with pregnancy complications, including idiopathic recurrent miscarriage (RM). Whereas TNF-α and LT-α gene polymorphisms affect serum cytokine concentrations, their contribution to RM is controversial. The single nucleotide polymorphisms (SNPs) TNF-α (−238G/A, −308G/A) and LT-α (+252A/G) were investigated in 350 RM women and 200 control women. Higher frequency of the TNF-α −238A, but not the TNF-α −308A or the LT-α+252G, allele was seen in patients, with comparable frequencies of TNF-α −238G/A, TNF-α −308G/A, and LT-α+252A/G genotypes seen between both groups, except for TNF-α −238G/G, which was lower in patients. Regression analysis confirmed the association of the TNF-α −238G/A SNP with idiopathic RM, and both TNF-α −308A/TNF −238G/LT-α+252Gand TNF-α −308G/TNF-α −238A/LT-α+252Ghaplotypes played a susceptible role in idiopathic RM. TNF-α −238G/A and −238A/A, and LT-α+252G/G genotypes were positively associated only with exclusively early RM. This supports the concept of the association of TNF-α (−238G/A) and LT-α (+252A/G) polymorphic variants in idiopathic RM.
Postnatal uterine development in Inverdale ewe lambs
Authors: Hayashi Kanako , O'Connell Anne R , Juengel Jennifer L , McNatty Ken P , Davis George H , Bazer Fuller W , Spencer Thomas E
Published: February 2008

Postnatal development of the uterus involves, particularly, development of uterine glands. Studies with ovariectomized ewe lambs demonstrated a role for ovaries in uterine growth and endometrial gland development between postnatal days (PNDs) 14 and 56. The uterotrophic ovarian factor(s) is presumably derived from the large numbers of growing follicles in the neonatal ovary present after PND 14. The Inverdale gene mutation (FecXI) results in an increased ovulation rate in heterozygous ewes; however, homozygous ewes (II) are infertile and have ‘streak’ ovaries that lack normal developing of preantral and antral follicles. Uteri were obtained on PND 56 to determine whether postnatal uterine development differs between wild-type (++) and II Inverdale ewes. When compared with wild-type ewes, uterine weight of II ewes was 52% lower, and uterine horn length tended to be shorter, resulting in a 68% reduction in uterine weight:length ratio in II ewes. Histomorphometrical analyses determined that endometria and myometria of II ewes were thinner and intercaruncular endometrium contained 38% fewer endometrial glands. Concentrations of estradiol in the neonatal ewes were low and not different between ++ and II ewes, but II ewes had lower concentrations of testosterone and inhibin-α between PNDs 14 and 56. Receptors for androgen and activin were detected in the neonatal uteri of both ++ and II ewes. These results support the concept that developing preantral and/or antral follicles of the ovary secrete uterotrophic factors, perhaps testosterone or inhibin-α, that acts in an endocrine manner to stimulate uterine growth and endometrial gland development in the neonatal ewes.
Cryptorchidism induced in normal rats by the relaxin-like factor inhibitor
Authors: Büllesbach Erika E , Boockfor Fredric R , Fullbright George , Schwabe Christian
Published: February 2008

Cryptorchidism is a serious problem, which affects 2–5% of the male population. Failure of the testes to descend into the scrotal region impairs germ cell development and is associated with a greater incidence of testicular cancer. The relaxin-like factor (RLF or insulin-like-3) has been shown to be critically important for the timely descent of the testicles in mice. We have discovered that the signal initiation site of the RLF can be eliminated without measurable effects on hormone binding to its receptor and that the resulting RLF derivative is a competitive inhibitor of RLF called RLFi. RLFi administered to pregnant rats causes dose-dependent gonadal retention in the offspring. The ability to control the severity of the syndrome by altering the concentration of RLFi and the timing of administration enables us to study in detail the structural changes that are associated with the action of RLF during critical stages of development. Targeted inhibition of the physiological migration pattern of testicles by RLFi lets one dissect the physiological process such as to find a window for clinical application of RLF and to search for ancillary factors that might play a role during normal development.
C-kit receptor in the human vas deferens: distinction of mast cells, interstitial cells and interepithelial cells
Authors: Metzger Roman , Rolle Udo , Fiegel Henning C , Franke Folker E , Muenstedt Karsten , Till Holger
Published: February 2008

The molecular mechanisms underlying the regulation of vas deferens (VD) motility and semen emission are still poorly understood. Interstitial cells of Cajal (ICC), which harbour the c-kit receptor (CD117), provide the basis of coordinated gut motility. We investigated whether c-kit receptor-positive cells also exist in the normal human VD. Enzyme and fluorescence immunohistochemical techniques were applied on serial sections of human proximal, middle, and distal VD segments (n=49) employing 13 different monoclonal and polyclonal antibodies recognizing the c-kit receptor. The c-kit receptor was detected in either round- or spindle-shaped cells. On account of their antigenic profile, the round- and oval-shaped c-kit receptor-positive cells were identified as mast cells (MC) occurring in all layers of the VD except the epithelium. In contrast, two distinct populations of exclusively c-kit receptor-positive spindle-shaped cells were found within the lamina propria and, rarely, in the inner and outer smooth muscle layers, as well as within the epithelium. Different shaped c-kit receptor-positive MC and IC were present in all layers of the human VD. Our findings demonstrate the presence of different c-kit receptor-positive cells also in the human VD. Their rather ubiquitous distribution within the lamina propria and muscle layers suggests that IC and MC may modulate the neuromuscular transmission and the propagation of electrical signals in multiple systems involved in the draining of fluids. The importance of the c-kit receptor-positive interepithelial cells remains unclear.
Semen from scrapie-infected rams does not transmit prion infection to transgenic mice
Authors: Sarradin Pierre , Melo Sandrine , Barc Céline , Lecomte Céline , Andréoletti Olivier , Lantier Frédéric , Dacheux Jean-Louis , Gatti Jean-Luc
Published: February 2008

Scrapie is the most common transmissible spongiform encephalopathy (TSE) in livestock. Natural contamination in sheep flocks is presumed to occur by maternal transmission to offspring. However, horizontal prion transmission from animal to animal exists and may be significant in sustaining and spreading contagion in the field. Artificial insemination is widely used in modern farming, and as large amounts of prion protein have been found in sheep sperm membrane, epididymal fluid and seminal plasma, horizontal transmission by this route was hypothesized since no clear information has been obtained on possible sexual transmission of TSE. We therefore tested the contamination levels of semen from scrapie-infected rams at different stages of incubation, including the clinical phase of the disease. We report here that under our experimental conditions ram semen did not transmit infectivity to scrapie-susceptible transgenic mice overexpressing the V136R154Q171 allele of the sheep prion (PRNP) gene. These results suggest that artificial insemination and natural mating have a very low or negligible potential for the transmission of scrapie in sheep flocks.
Development of a reliable in vitro maturation system for zebrafish oocytes
Authors: Seki Shinsuke , Kouya Toshimitsu , Tsuchiya Ryoma , Valdez Delgado M , Jin Bo , Hara Takao , Saida Naoya , Kasai Magosaburo , Edashige Keisuke
Published: February 2008

In zebrafish oocytes, it has been reported that a 60 or 75% Leibovitz L-15 medium or simple balanced saline solution containing 17α, 20β-dihydroxy-4-pregnen-3-one (DHP) is effective for nuclear maturation. However, most of the oocytes that matured under these conditions were not fertilized and did not hatch. Thus, thesein vitromaturation methods could not support the cytoplasmic maturation of zebrafish oocytes. Therefore, we tried to develop a reliablein vitromaturation method for zebrafish oocytes, which supports their ability to be fertilized and to develop till hatching. When zebrafish oocytes at stage III were cultured in 50–100% Leibovitz L-15 medium supplemented with DHP, the highest rates of cleavage (24%) and hatching (12%) were obtained from oocytes matured in 90% Leibovitz L-15 medium. When we examined the suitable pH (7.5–9.5) of the 90% medium, higher rates of cleavage (45%) and hatching (33%) were obtained in oocytes matured at pH 9.0 than at pH 7.5, 8.5, or 9.5 (cleavage rate, 16–29%; hatching rate, 8–21%). In oocytes matured in 90% Leibovitz L-15 medium at pH 9.0, high rates of cleavage (70%) and hatching (63%) were obtained when oocytes were cultured for 270 min with 0.5 mg/ml BSA. Thus, 90% Leibovitz L-15 medium at pH 9.0 containing 0.5 mg/ml BSA was effective for normal maturation of zebrafish oocytes. This method will become a powerful tool for understanding the mechanism ofin vitromaturation in zebrafish oocytes and for the practical use of immature oocytes.
Sperm competition and sperm cooperation: the potential role of diploid and haploid expression
Authors: Immler Simone
Published: February 2008

Sperm competition is a powerful selective force driving the evolution of sperm shape and function. Recent findings suggest that sperm cooperation is a potential evolutionary response to sperm competition. Sperm cooperation may enhance the performance of the ejaculate increasing a male's chance to outcompete rival males in competition for fertilisation. Whether and how sperm cooperation may evolve is the focal point of this review. The relative importance of haploid and diploid gene expression for the evolution of sperm cooperation and the potential conflict of interest between (i) haploid sperm and diploid male and (ii) among sibling sperm, since sibling sperm only share an average of 50% of their genes in a diploid organism, are discussed. Furthermore, sperm cooperation is defined and the literature for empirical evidence of sperm cooperation is reviewed in light of the author's definitions.
Pretreatment with Albuterol versus Montelukast for Exercise‐Induced Bronchospasm in Children
Authors: Raissy Hengameh H. , Harkins Michelle , Kelly Franceska , Kelly H. William
Published: February 2008

Study Objectives. To compare pretreatment with albuterol versus montelukast added to the current asthma regimen for protection against exercise‐induced bronchospasm in children with mild‐to‐moderate asthma, and to determine whether cysteinyl leukotriene (Cys‐LT) concentrations measured in the exhaled breath condensate correlated with response to montelukast.

Design. Prospective, randomized, double‐blind, double‐dummy, crossover study

Setting. Asthma clinic at a university‐affiliated medical center.

Patients. Eleven children aged 7–17 years with physician‐diagnosed mild‐to‐moderate asthma for at least 6 months and with self‐reported exercise‐induced bronchospasm (defined as ≤ 15% decrease in forced expiratory volume in 1 sec [FEV1] at screening and baseline visit).

Intervention. Patients were randomly assigned to receive 3–7 days of oral montelukast 5–10 mg/day or 2 puffs of an albuterol metered‐dose inhaler just before an exercise challenge and then were crossed over to the alternate therapy for the last visit.

Measurements and Main Results. Serial spirometry was performed before and at 0, 5, 10, 15, 30, 45, and 60 minutes after the exercise challenge at each visit. Measurement of exhaled breath condensate was performed at the screening visit and study visits 1 and 2. The primary outcome was the maximum change in FEV1 after exercise. Secondary outcomes were the area under the curve for FEV1 (expressed as percentage decrease from baseline) during the first 60 minutes (AUC0–60) after exercise and the proportion of patients in whom exercise‐induced bronchospasm was prevented (defined as < 15% decrease in FEV1 after exercise challenge). The mean ± SD maximum decrease in FEV1 was 27.5 ± 7.9% at baseline. Patients receiving montelukast had an 18.3 ± 13.7% decrease in FEV1 compared with 0.7 ± 1.6% in patients receiving albuterol (p=0.002, paired t test). Exercise‐induced bronchospasm was prevented in 100% of the patients receiving albuterol compared with 55% receiving montelukast (p<0.05, McNemar's test). The AUC0–60 was significantly smaller with albuterol compared with montelukast (p<0.001, Wilcoxon signed rank test). No correlations were found between Cys‐LT concentration and the severity of exercise‐induced bronchospasm or the response to montelukast.

Conclusion. Pretreatment with albuterol is more effective than montelukast for prevention of exercise‐induced bronchospasm in children with asthma.
In Vitro Activity of Cefepime, Imipenem, Tigecycline, and Gentamicin, Alone and in Combination, Against Extended‐Spectrum β‐Lactamase‐Producing Klebsiella pneumoniae and Escherichia coli
Authors: Cha Raymond
Published: February 2008

Study Objective. To evaluate the activity of cefepime, imipenem, tigecycline, and gentamicin, alone and in combination, against extended‐spectrum β‐lactamase (ESBL)‐producing Klebsiella pneumoniae and Escherichia coli.

Design. In vitro susceptibility and time‐kill analysis.

Setting. University‐affiliated research laboratory.

Isolates. Ten K. pneumoniae and 10 E. coli clinical strains known to produce ESBL.

Measurements and Main Results. Minimum inhibitory concentration (MIC) testing was performed at 5.5 and 7.0 log10 colony‐forming units (cfu)/ml. Time‐kill studies were performed over 24 hours with a high inoculum of 7.0 log10 cfu/ml for cefepime, imipenem, tigecycline, and gentamicin at 1 times MIC. Combination studies were tested for cefepime plus tigecycline, cefepime plus imipenem, imipenem plus tigecycline, and for adjunctive gentamicin with cefepime, imipenem, or tigecycline. At the high inoculum, the MIC ranges for cefepime, imipenem, tigecycline, and gentamicin were 0.25–256 (MIC for 90% of tested strains [MIC90] = 32), 0.125‐2 (MIC90 = 1), 0.25‐16 (MIC90 = 4), and 0.25‐4 mg/L (MIC90 =1), respectively, for all isolates. At the higher inoculum, MICs shifted for cefepime, imipenem, and tigecycline. Change in log10 cfu/ml from baseline to 24 hours for cefepime, imipenem, and tigecycline alone ranged from 4.85‐0.71, 5‐4.22, and 3.5‐1.01, respectively, for all isolates. Bactericidal activity was observed for cefepime alone (10 [50%] of 20 isolates), imipenem alone (20 [100%] of 20), and tigecycline alone (3 [15%] of 20). Combination studies with cefepime plus tigecycline resulted in synergy against 4 (20%) of 20 isolates. Combination studies with gentamicin resulted in synergy against 6 isolates (30%) with cefepime and 4 isolates (20%) with tigecycline. No synergy was observed with imipenem combinations. No antagonism was observed. With the exception of cefepime, correlations were observed between MIC and terminal densities for studied agents.

Conclusions. Cefepime exhibited bactericidal activity but was unrelated to susceptibility. Tigecycline exhibited predictable bacteriostatic activity and synergy in combination against a subset of study isolates. Imipenem exhibited predictable bactericidal activity against all isolates. The utility of combination regimens with novel agents requires further exploration.
Greater Immediate Gastric Acid Suppression with Lansoprazole 30 mg Administered as a 2‐Minute Intravenous Bolus Injection versus a 30‐Minute Infusion
Authors: Metz David C. , Devlin John W. , Vakily Majid , Atkinson Stuart , Lloyd Eric
Published: February 2008

Study Objective. To compare the pharmacokinetics, pharmacodynamics, and safety of lansoprazole administered as a 2‐minute intravenous bolus injection versus a 30‐minute continuous infusion.

Design. Phase I, open‐label, randomized, crossover, single‐center trial.

Setting. Clinical research facility.

Subjects. Thirty‐eight healthy volunteers aged 18–55 years.

Intervention. Each subject received one of three regimens. Each regimen contained the same three treatments but in a different sequence: an intravenous injection of lansoprazole 30 mg/10 ml of normal saline over 2 minutes, an infusion of lansoprazole 30 mg/60 ml of normal saline over 30 minutes, and an intravenous injection of 10 ml of normal saline (placebo) over 2 minutes. Each treatment was administered once/day for 7 days, with a washout period of at least 5 days separating each one.

Measurements and Main Results. Blood samples were assayed for lansoprazole concentrations by using liquid chromatography with tandem mass spectrometry. After the 2‐minute administration, mean peak lansoprazole concentrations were about 2‐fold higher than those after the 30‐minute administration on days 1 and 7. Lansoprazole area under the concentration versus time curve met criteria for bioequivalence on both days. Two‐minute administration resulted in a greater percentage of time that the 24‐hour pH was above 4 compared with the 30‐minute administration (53% vs 47%, p=0.045), with comparable 24‐hour integrated gastric acidity (114.4 vs 91.6 mmol • hr/L for 2‐min vs 30‐min, p=0.335). Significantly greater acid suppression occurred during the first hour after the 2‐minute administration compared with that after the 30‐minute administration (p≥0.001). Safety profiles were similar among the regimens.

Conclusion. Greater immediate gastric acid suppression occurred after administration of lansoprazole 30 mg over 2 minutes than over 30 minutes, with other pharmacokinetic, pharmacodynamic, and safety profiles being similar.