bluuebunny/crossref_metadata_2025 huggingface dataset:
Research with Children and Young People: The Issue of Parental (Proxy) Consent
Authors: Coyne Imelda
Published: February 2009

This study examines the potential problems with the parental consent requirement, substantiated with examples mainly from healthcare and social research studies. This will illustrate how the parental consent requirement, instead of promoting high ethical standards, may result in some instances of children’s rights and ethical considerations being ignored or receiving cursory attention. The ‘blanket’ requirement of parental consent for all research involving children under the age of 18 years needs to be challenged as it fails to recognise children’s capacities and accord children due respect as persons in their own right. Flexible ethical guidelines should be developed that take cognisance of children’s competence in contemporary society and at the same time protects children from inappropriate research and procedures.
A dynamic model of colonic concentrations of delayed‐release 5‐aminosalicylic acid (Asacol)
Authors: THORPE M. P. , EHRENPREIS E. D. , PUTT K. S. , HANNON B.
Published: February 2009

Backround 5‐ASA in a pH sensitive tablet (Asacol) is administered as three doses/day to treat ulcerative colitis. Once daily dosing may improve patient adherence. Simulation of colonic levels of 5‐ASA can be used to compare dosing regimens.

Aim To create a dynamic model of colonic concentrations of delayed‐release 5‐aminosalicylic acid (Asacol).

Methods Using published data, we created a computer model with STELLA software to simulate amounts of colonic 5‐ASA in the total colon, right, transverse, descending and sigmoid/rectum after daily and three time/day Asacol.

Results The model predicted similar total and regional amounts of 5‐ASA with both regimens. Distribution of 5‐ASA was 38% in the right colon, 33% in the transverse colon and 14% each in the descending and sigmoid/rectal colon. Simulated increases in colonic motility and defecation rate exaggerated this 5‐ASA distribution, resulting in negligible amounts of 5‐ASA in the sigmoid/rectal region.

Conclusions This computer model suggests that Asacol can be administered as a single daily dose. The model supports experimental and clinical observations that alternate dose or route of administration may be necessary to achieve adequate 5‐ASA amounts in the distal colon during acute exacerbations of ulcerative colitis. This simulation cannot account for all sources of variability in the clinical setting, but provides a rationale for further investigation.
The influence of symptom type and duration on the fate of the metaplastic columnar‐lined Barrett’s oesophagus
Authors: GATENBY P. A. C. , RAMUS J. R. , CAYGILL C. P. J. , FITZGERALD R. C. , CHARLETT A. , WINSLET M. C. , WATSON A.
Published: February 2009

Background  Prolonged gastro‐oesophageal reflux resulting in columnar metaplasia of the oesophagus is the main risk factor for oesophageal adenocarcinoma.

Aim  To examine the duration of symptoms and associations of different symptoms with the development of columnar‐lined oesophagus, dysplasia and adenocarcinoma.

Methods  UK multicentre cohort study of patients with columnar‐lined oesophagus whose date of symptom onset (1082 patients) and/or types of symptoms reported (1681 patients) were documented. Follow‐up was examined by analysis of histological reports from the registering centers.

Results  Symptoms of dysphagia/odynophagia and nausea/vomiting were associated with development of dysplasia. High‐grade dysplasia and adenocarcinoma were associated with dysphagia/odynophagia and weight loss. Median duration from symptom onset to detection of columnar‐lined oesophagus without intestinal metaplasia: 2.6 years, columnar‐lined oesophagus with intestinal metaplasia: 5.0 years, indefinite changes for dysplasia: 19.3 years and low‐grade dysplasia: 30.0 years. One tenth of patients had developed high‐grade dysplasia at 9.6 years and one tenth had developed adenocarcinoma at 13.8 years from symptom onset.

Conclusions  In patients with columnar‐lined oesophagus, symptoms of dysphagia/odynophagia and nausea/vomiting were associated with a higher risk of development of dysplasia and adenocarcinoma. There is a trend for longer duration of symptoms to the detection of dysplasia.
Treatment of ulcerative colitis from the patient’s perspective: a survey of preferences and satisfaction with therapy
Authors: GRAY J. R. , LEUNG E. , SCALES J.
Published: February 2009

Background  Data available regarding patient perspectives on ulcerative colitis (UC) and their preferences and satisfaction with therapy are limited.

Aims  To examine the preferences of UC patients to understand better what they look for in a therapy when managing their disease, as this may influence overall medication adherence.

Methods  The study surveyed 100 Canadian UC patients on topics including educational resources used to learn about the disease, medication attributes that are most valued and preferred by the patient and satisfaction with current therapy.

Results  Overall, efficacy‐ and safety‐related medication attributes were rated by patients to be more important than those related to dosing regimen (e.g. dosing frequency, number of pills), cost and formulary coverage. In pair‐wise comparisons of specific medication attributes, UC patients rated speed of symptom relief and few side effects as the most important factors when considering a UC medication (preferred on average 84% and 74% of the time respectively).

Conclusion  This study provides insight into UC patient preferences and satisfaction with therapy that may be important when counselling on treatment options, and generates relevant discussions on adherence. Larger studies may be warranted to examine further how these findings can be extrapolated to broader UC populations.
Natural course of treated and untreated chronic HCV infection: results of the nationwide Hepnet.Greece cohort study
Authors: MANESIS E. K. , PAPATHEODORIDIS G. V. , TOULOUMI G. , KARAFOULIDOU A. , KETIKOGLOU J. , KITIS G. E. , ANTONIOU A. , KANATAKIS S. , KOUTSOUNAS S. J. , VAFIADIS I. ,   for the HEPNET.Greece Cohort Study
Published: February 2009

Background  Interferon (IFN‐α)‐based regimens have been used with varying success in the treatment of chronic hepatitis C (CHC) for over two decades. The effect of such treatments on the natural course of CHC has been evaluated in small clinical trials with conflicting results.

Aim  To investigate the natural course of IFNα‐based ‐treated and untreated patients with CHC by analysing data from the HEPNET.GREECE study.

Methods  We retrospectively analysed 1738 patients from 25 Greek Centres (median age 40.1; males 57.6%; cirrhosis 9.2%), 734 untreated and 993 treated with IFNα‐based regimens [44.7% sustained viral response (SVR)], followed‐up for median 25.2 and 46.8 months, respectively.

Results During follow‐up, 48 patients developed liver decompensation and 24 HCC. Older age was significantly related to disease progression (HR = 2.6 per 10 years of increasing age). Stratified by baseline cirrhosis, Cox analysis showed that patients with SVR, but not without SVR, had significantly lower hazard for events compared with nontreated patients (HR = 0.16; P < 0.001), whereas the detrimental effect of older age remained highly significant. Separate group analysis demonstrated that in cirrhosis, the beneficial effect of treatment was evident even without SVR. Treatment effect interacted significantly with age, indicating that older patients, mainly noncirrhotic, gained the most benefit.

Conclusions IFNα‐based treatment does alter the natural course of CHC. A protective effect is mostly present in patients with SVR, but older patients, at higher risk of events, gain the greatest benefit. In established cirrhosis, treatment carries a protective effect even among those without SVR.
Phleum pratense alone is sufficient for allergen‐specific immunotherapy against allergy to Pooideae grass pollens
Authors: Hejl C. , Wurtzen P. A. , Kleine‐Tebbe J. , Johansen N. , Broge L. , Ipsen H.
Published: February 2009

Background
Specific immunotherapy is the only causal treatment of allergy available today. Traditionally, therapeutic products based on either a single grass species or a mix of such extracts are used for grass pollen immunotherapy. Investigations comparing the immunological response to these allergen preparations are needed to ensure optimal treatment. The objective of this study was to investigate patterns of T and B cell cross‐reactivity to Pooideae single‐species extracts and to extract mixes.

Methods
IgG4 induced by immunotherapy with Phleum pratense extract was investigated for cross‐reactivity using nine single‐species extracts and four mixes. For the mixes, studies of IgE cross‐reactivity were also performed. T cell cross‐reactivity was investigated in lines specific to nPhl p 1 or nPhl p 5 allergens, and the amounts of group 1 and 5 allergens in the extracts were quantified by a single radial immunodiffusion.

Results
The levels of treatment‐induced IgG4 detected by all the extracts displayed a clear correlation to that detected by the P. pratense pollen extract. The IgE studies confirmed the cross‐reactivity of P. pratense‐specific B cells towards the allergens contained in the mixes, and the T cell studies demonstrated cross‐reactivity towards group 1 and 5 major allergens in extracts of six temperate grass species.

Conclusion
Extensive T and B cell cross‐reactivity was observed towards the allergens of the Pooideae grasses, and the degree of B cell cross‐reactivity was independent of the number of species included in the extract mixes. This implies that treatment with pollen extract of just one Pooideae species will affect the allergic responses caused by any of the temperate grasses in this subfamily.
Taurine chloramine inhibits functional responses of human eosinophilsin vitro
Authors: Martinez‐Losa M. , Cortijo J. , Piqueras L. , Sanz M. J. , Morcillo E. J.
Published: February 2009

BackgroundEosinophils are prominent effectors of allergic inflammation. Taurine‐chloramine (TauCl), a derivative of the amino acid taurine, shows antioxidant properties in different cell systems but its effects on eosinophils have not been reported.

ObjectiveTo study the effects of TauCl and taurine on functional responses of isolated human eosinophils activated by different stimuli.

MethodsHuman eosinophils were purified from the blood of healthy donors by a magnetic bead separation system. The effects of TauCl and taurine (0.1–1 mm) were investigated on the generation of superoxide anion (ferricytochrome‐creduction microassay), calcium signal (fluorimetry), p47phox–p67phoxtranslocation (Western blot), leukotriene C4(LTC4) production (enzymeimmunoassay), eosinophil peroxidase (EPO) release (spectrophotometry), eosinophil cationic protein (ECP) release (radioimmunoassay), apoptosis (flow cytometry with annexin V‐propidium iodide), and nuclear factor‐κB (NF‐κB) activation (Western blot).

ResultsTauCl inhibited superoxide anion generation triggered byN‐formyl‐Met–Leu–Phe (fMLP; 30 nm), phorbol myristate acetate (1 nm) and serum opsonized zymosan (0.5 mg/mL) with similar potency (IC50∼200 μm) for the three stimuli, while taurine (0.1–1 mm) was scarcely effective. TauCl but not taurine inhibited p47phox–p67phoxtranslocation. TauCl (200 μm) and taurine (1 mm) did not modify the [Ca2+]iresponses to fMLP. TauCl inhibited the release of EPO (IC50∼200 μm) and reduced ECP and LTC4production from fMLP‐activated eosinophils while taurine was without significant effects. TauCl (1 mm) did not change constitutive apoptosis but significantly attenuated the ability of granulocyte‐monocyte colony‐stimulating factor (GM‐CSF) and IL‐5 to prevent apoptosis. The activation of eosinophil NF‐κB induced by GM‐CSF and IL‐5 was suppressed by TauCl.

ConclusionTaurine is without significantin vitroeffects on human eosinophil functions but its derivative TauCl inhibits oxidative burst and generation of inflammatory mediators, and reverses the survival effect produced by inflammatory cytokines. Therefore, endogenous TauCl may help to suppress excessive inflammatory response in eosinophils at inflammatory sites.
Glucocorticoid receptor gene variant is associated with increased body fatness in youngsters
Authors: Voorhoeve Paul G. , Van Den Akker Erica L. T. , Van Rossum Elisabeth F.C. , Koper Jan W. , Van Mechelen Willem , Lamberts Steven W. J. , Delemarre‐van de Waal Henriette A.
Published: February 2009

Objective  Sensitivity to glucocorticoids is known to be highly variable between individuals and is partly determined by polymorphisms in the glucocorticoid receptor (GR) gene. We investigated the relationship between four GR gene polymorphisms and body composition during puberty and at young adult age.

Design  An observational study with repeated measurements.

Patients  Two comparable young Dutch cohorts with a generational difference of about 20 years were investigated. The first cohort consisted of 284 subjects born between 1961 and 1965. Measurements were performed from 13 to 36 years of age. The second cohort consisted of 235 subjects born between 1981 and 1989. Measurements were performed from 8 to 14 years of age.

Measurements  Associations between height, weight, BMI, fat mass (FM) and fat‐free mass and four well‐known functional polymorphisms were investigated.

Results  In boys in the younger cohort, the G‐allele of the BclI polymorphism (haplotype 2) was associated with a higher body weight, weight‐SDS, BMI, BMI‐SDS and FM. These associations were not observed in the older cohort. Irrespective of genotype, the younger cohort showed a significantly higher total FM, body weight and BMI compared with the older cohort.

Conclusions  Because the associations between the G‐allele of the BclI polymorphism in the GR gene and body FM in boys were only found in a healthy young population, but not in a comparable, generally leaner cohort from an older generation, it is suggested that carriers of this polymorphism are likely to be more vulnerable to fat accumulation in today’s obesity promoting environment, than noncarriers.
How should a nonfunctioning pituitary macroadenoma be monitored after debulking surgery?
Authors: Greenman Yona , Stern Naftali
Published: February 2009

Transsphenoidal surgery is the treatment of choice for nonfunctioning pituitary macroadenomas but is seldom curative. Tumour progression rates are high in patients with postoperative remnants. Therefore, long‐term monitoring is necessary to detect tumour growth, which may be asymptomatic or manifest with visual field defects and/or pituitary dysfunction. In view of the generally slow‐growing nature of these tumours, yearly magnetic resonance imaging, neuro‐ophalmologic and pituitary function evaluation are appropriate during the first 3–5 years after surgery. If there is no evidence for tumour progression during this period, testing intervals may be extended thereafter.
Association between low bone mass and aortic valve sclerosis in Koreans
Authors: Choi Han S. , Rhee Yumie , Hur Nam W. , Chung Namsik , Lee Eun J. , Lim Sung‐Kil
Published: February 2009

Background  Osteoporosis or low bone mass has been associated with cardiovascular disease and calcification in several clinical studies. However, few studies have assessed the relationship between bone mass and valvular calcification. The aim of this study was to evaluate the relationship between low bone mass and aortic valve sclerosis in Korean men and women.

Methods  A total of 211 men and 117 women were included in this study. Each subject’s bone mineral density (BMD) was measured with dual‐energy X‐ray absorptiometry, and aortic valve sclerosis was assessed using transthoracic echocardiography. Association between low bone mass and aortic valve sclerosis was assessed with multivariate logistic regression analysis in this cross‐sectional study.

Results  Of 328 total subjects enrolled in this study, 50 men (23·7%) and 18 women (15·4%) were found to have aortic valve sclerosis. The mean (±SD) BMD T‐scores were −0·5 (±0·8) in men and −0·9 (±1·0) in women. After adjusting for covariates, only women with aortic valve sclerosis had significantly lower BMD T‐scores than those without it. Multivariate logistic regression analysis showed that age, smoking and hypertension were independently associated with increased risk of aortic valve sclerosis in men. In women, however, logistic regression analysis showed that BMD T‐score, as well as age, was an independent variable for aortic valve sclerosis. We also found that a T‐score of less than −1·5 was significantly associated with increased risk of aortic valve sclerosis in women compared to normal T‐scores.

Conclusion  Low bone mass might be independently associated with increased risk of aortic valve sclerosis in women, but not in men. Women with low bone mass should be further evaluated for the presence of aortic valve sclerosis and related cardiovascular diseases.
Regulatory T cells in Graves’ disease
Authors: Pan Deshun , Shin Young‐Ha , Gopalakrishnan Geetha , Hennessey James , De Groot Leslie J.
Published: February 2009

Context  Graves’ disease (GD) involves auto‐immunity against thyroid cell antigens, but the reasons for induction of auto‐immunity are uncertain. We wished to determine whether there was a deficiency of regulatory T cells in patients with active GD.

Design  Venous blood samples were obtained from patients with GD before and after treatment, and controls, and peripheral blood mononuclear cells were prepared.

Patients and measurements  Regulatory T cells were enumerated by Fluorescent Activated Cell sorting (FACS) in nineteen patients with untreated GD, 9 patients 6–8 weeks post RAI therapy, and 30 control subjects. Twenty‐one patients with active GD prior to control of hyperthyroidism, 23 euthyroid controls without known autoimmune thyroid disease, and 10 patients who were euthyroid 6–12 months after RAI treatment were studied for expression of genes found in regulatory T cells by real‐time Polymerase Chain reaction (PCR).

Results  Percent distribution of CD4+, CD4+CD25+ and CD4+ CD25+int‐hi CD127+lo regulatory T cells was similar in active GD patients and control subjects. The number of CD25+ and CD4+ CD25+int‐hi CD127+lo cells was similar in GD patients and control subjects, but was lower in recently treated patients. Messenger RNA was prepared from PBMC, and reverse transcribed. Copy DNA abundance was evaluated by Real Time PCR using appropriate primers, for GAPDH (glyceraldehyde phosphate dehydrogenase) as a control housekeeping gene, and 5 genes related to function of regulatory T cells. Message RNA for Gadd45 alpha, Gadd45beta (growth arrest and damage inducible proteins), GITR (glucocorticoid inducible TNF receptor) and CD25 (IL‐2R subunit) was more abundant in patients with active GD than in normal controls, and FoxP3 mRNA level was equal to that in controls. Message RNA levels in patients treated and euthyroid for 6 months were also greater than or equal to values in controls.

Conclusion  This study provides evidence that there is no deficit in T regulatory cells during active GD, or during the months post therapy.
Testicular dysgenesis syndrome: foetal origin of adult reproductive problems
Authors: Wohlfahrt‐Veje Christine , Main Katharina M. , Skakkebæk Niels Erik
Published: February 2009

The evidence for the existence of testicular dysgenesis syndrome (TDS) is presented in this review. Several epidemiological studies have shown that conditions like cryptorchidism, impaired spermatogenesis, hypospadias and testicular cancer can be associated as risk factors for each other. Thus, the risk of testis cancer is significantly increased in men with cryptorchidism and/or infertility. Several recent studies point towards early dysgenesis of the foetal testis as the biological link between these disorders. Dysgenesis has been demonstrated in biopsies of the contralateral testis of men with testis cancer and in infertile men. The histological evidence includes immature seminiferous tubules with undifferentiated Sertoli cells, microliths and Sertoli‐cell only tubules. Dysgenetic testes often have an irregular ultrasound pattern, where microliths may also be visible. Our current hypothesis is that maternal exposure to endocrine disrupting chemicals may contribute to the pathogenesis of TDS. Animal experiments have shown that all TDS symptoms, except testicular cancer, can be induced by foetal exposure to anti‐androgenic chemicals. However, the cause of TDS in humans remains to be determined.
Plasma visfatin concentrations increase in both hyper and hypothyroid subjects after normalization of thyroid function and are not related to insulin resistance, anthropometric or inflammatory parameters
Authors: Caixàs A. , Tirado R. , Vendrell J. , Gallart L. , Megía A. , Simón I. , Llauradó G. , González‐Clemente J.M. , Giménez‐Palop O.
Published: February 2009

Objective  The objective of this study was to evaluate plasma visfatin levels in thyroid dysfunction and its relationship with inflammatory, anthropometric and insulin resistance parameters.

Design and patients  Twenty‐four hyperthyroid and 27 hypothyroid patients were studied before and after treatment. Forty‐five euthyroid subjects were used as control group.

Measurements  Fasting plasma visfatin, IL‐6, C reactive protein, adiponectin, thyroid hormones, waist‐to‐hip ratio, BMI, percentage of body fat and homeostasis model insulin resistance index (HOMA‐IR) were measured.

Results  Hyperthyroid patients showed increased insulin resistance, IL‐6 and visfatin levels compared with controls (3·21 ± 3·0 vs. 1·67 ± 0·75, P = 0·022; 3·35 ± 0·41 vs. 2·10 ± 0·25 pg/ml, P = 0·016; and 37·4 ± 5·81 vs. 23·79 ± 4·2 ng/ml, P = 0·061 respectively). After normalization of thyroid function, IL‐6 levels and HOMA‐IR decreased (2·35 ± 0·37 vs. 2·10 ± 0·25 pg/ml, P = 0·045 and 3·21 ± 0·60 vs. 2·28 ± 0·38, P = 0·032 respectively), while body weight, adiposity and visfatin levels increased (26·1 ± 1·2 vs. 26·7 ± 1·2 kg/m2, P = 0·049; 30·9 ± 1·6 vs. 32·2 ± 1·6%, P = 0·007; and 37·4 ± 5·81 vs. 63·13 ± 8·72 ng/ml, P = 0·047 respectively). C reactive protein and adiponectin levels were similar to those of the control group. Hypothyroid patients showed high visfatin levels (40·59 ± 3·07 vs. 29·34 ± 4·9 ng/ml, P = 0·049) that increased after treatment (81·4 ± 9·2 ng/ml, P = 0·001) without changes in anthropometric or insulin resistance parameters. C reactive protein, IL‐6 and adiponectin levels were similar to those of the control group. No correlations between visfatin and any analysed parameter were found in either hyper‐ or hypothyroidism.

Conclusion  Visfatin exhibits a marked increase after normalization of thyroid function in both hyper and hypothyroid patients. We suggest that visfatin may play a role in the hormone stabilization process independent of anthropometric, inflammatory or insulin resistance variables.
Organic carbon additions: effects on soil bio‐physical and physico‐chemical properties
Authors: Bhogal A. , Nicholson F. A. , Chambers B. J.
Published: February 2009

The effects of organic carbon (OC) additions from farm manures and crop residues on selected soil bio‐physical and physico‐chemical properties were measured at seven experimental sites, on contrasting soil types, with a history of repeated applications of farm manure or differential rates of inorganic fertilizer nitrogen (N). Repeated (> 7 years annual additions) and relatively large OC inputs (up to 65 t OC ha−1) were needed to produce measurable changes in soil properties, particularly physical properties. However, over all the study sites, there was a positive relationship between OC inputs and changes in total soil OC and ‘light’ fraction OC (LFOC), with LFOC providing a more sensitive indicator of changes in soil organic matter status. Total soil OC increased by an average of 3% for every 10 t ha−1manure OC applied, whereas LFOC increased byc. 14%. The measured soil OC increases were equivalent toc. 23% of the manure OC applied (up to 65 t OC ha−1applied over 9 years) andc. 22% of the crop residue OC applied (up to 32 t OC ha−1over 23 years). The manure OC inputs (but not crop residue OC inputs) increased topsoil porosity and plant available water capacity, and decreased bulk density by 0.6%, 2.5% and 0.5% with every 10 t ha−1manure OC applied, respectively. Both OC sources increased the size of the microbial biomass (11% increase in biomass C with 10 t OC ha−1input), but only manure OC increased its activity (16% increase in the soil respiration rate with 10 t OC ha−1input). Likewise, the potentially mineralizable N pool only increased with manure N inputs (14% increase with 1 t manure total N ha−1). However, these soil quality benefits need to be balanced with any potential environmental impacts, such as excessive nutrient accumulation, increased nitrate leaching and phosphorus losses and gaseous emissions to the atmosphere.
Molecular changes in organic matter of a compost‐amended soil
Authors: Spaccini R. , Sannino D. , Piccolo A. , Fagnano M.
Published: February 2009

The molecular changes of organic matter in a cultivated soil after compost amendments was followed by off‐line‐pyrolysis‐TMAH GC‐MS. Thermochemolysis of soil and compost provided a detailed molecular characterization of soil organic matter (SOM) by releasing a large amount of different molecules mainly derived from plant biopolymers such as lignin, waxes and bio‐polyesters. No significant differences were found before and after 1 year of cultivation in the pyrolytic products released by control soil, which were mainly fatty acids, oxidized forms of lignins, and minor amounts of microbial bio‐products and biopolyesters derivatives. Conversely, significant qualitative and quantitative variations were found in the molecular characteristics of SOM between control and compost‐amended soils after 1 year of cultivation. Increasing amounts and diversified components of fatty acids, n‐alkanes and various biopolyesters derivatives such as hydroxy‐alkanoic and alkandioic acids were found in the compost‐amended soil. These results indicate that a significant amount of exogenous compost‐derived organic molecules were incorporated into SOM after 1 year of cultivation. The organic structural indexes derived from these results indicated direct inputs of undecomposed lignin residues and hydrocarbon waxes from compost material. When compared with the control soil, small but significant amounts of plant biomarkers, such as cyclic di‐ and triterpenes derivatives, were found only in the compost‐amended soil. These findings suggest that the molecular changes of SOM brought about by amendment with biomass residues can be followed by using thermochemolysis of bulk soil samples.
Expression of embryonic stem cell markers in cultured JKT‐1, a cell line derived from a human seminoma
Authors: Bouskine Adil , Vega Aurélie , Nebout Marielle , Benahmed Mohamed , Fénichel Patrick
Published: February 2009

Testicular germ cell tumours (TGCTs), the most frequent solid tumour of the young men, originate from the primitive germ cells. They share some pluripotency stem‐cell markers which may help to distinguish between seminoma, the most frequent TGCTs and non‐seminoma tumours, such as embryonal carcinoma, teratocarcinoma or choriocarcinoma. Due probably to the propensity of seminoma to apoptosis, only two cell lines originated from pure testicular seminoma, TCam‐2 and JKT‐1 have been up to now, established, maintained and proposed as representative models of human testicular seminoma. However, both seem, following recent reports, to be able to drift. Thus, the molecular signature of embryonic stem‐cell markers of the JKT‐1 cells cultured in our laboratory, were studied by RT‐PCR, Western blot and immunofluorescence (IF). JKT‐1 cells analysed after 30 passages, expressed placenta alkaline phosphatase but not alphafoetoprotein (αFP) nor beta‐human chorionic gonadotropin. JKT‐1 cells also expressed markers of pluripotency such as NANOG and OCT3/4 and more specific seminoma markers, such as AP2γ and HIWI. However, protein expression of OCT3/4 and AP2y was weak and these JKT‐1 cells expressed SOX2, a marker of embryonal carcinoma and did not express c‐KIT usually expressed in most seminoma. Possible derivation through in vitro culture conditions was supported by looking at later passages (61) which showed a decrease of NANOG and HIWI protein expression. JKT‐1 cells express a signature of markers which is still near from the one express by seminoma cells, allowing carcinogenetic studies. However, because of their great ability to drift as shown for TCam‐2, it is recommended to verify and to precise this molecular signature before reporting functional results.
The age‐related decline of testosterone is associated with different specific symptoms and signs in patients with sexual dysfunction
Authors: Corona G. , Mannucci E. , Ricca V. , Lotti F. , Boddi V. , Bandini E. , Balercia G. , Forti G. , Maggi M.
Published: February 2009

In males, testosterone (T) levels decline with ageing. Several symptoms characteristic of the ageing process are similar to those related to hypogonadism. The aim of the present study was to evaluate the specific association among hypogonadism‐related symptoms and signs and the ageing process. A consecutive series of 1647 (mean age 52.4 ± 13.1 years) male patients with sexual dysfunction were investigated. Several hormonal and biochemical, instrumental and psychological parameters were studied. The parameters significantly associated with total levels in the entire cohort, after adjustment for confounders, were studied as a function of age and T quartiles. In all age quartiles, low T was associated with higher waist circumference and triglyceride levels and with an increased prevalence of metabolic syndrome. The prevalence of hypoactive sexual desire decreased as a function of T only in the youngest (17‐ to 42‐year old) age quartile as well as the reported reduction in nocturnal erections. In the oldest age quartile, we found an inverse relationship between T levels and the prevalence of severe erectile dysfunction and a positive relationship with intercourse frequency. Accordingly, in the oldest age quartile, subjects with higher T levels showed better penile flow at penile colour doppler ultrasound as well as a better lipid profile. Finally, an inverse association between somatized anxiety and T levels was observed only in the oldest age quartile. In conclusion, our study shows for the first time that in subjects with sexual dysfunction, some hypogonadism‐related symptoms can be age‐specific. In particular, low T is associated with sexual dysfunction more often in the oldest subjects.
Hepatic steatosis is associated with low serum testosterone and high serum DHEAS levels in men
Authors: Völzke Henry , Aumann Nicole , Krebs Alexander , Nauck Matthias , Steveling Antje , Lerch Markus M. , Rosskopf Dieter , Wallaschofski Henri
Published: February 2009

Obesity and metabolic syndrome are associated with low serum testosterone levels. Hepatic steatosis contributes to the metabolic syndrome and might be regarded as its hepatic manifestation. In this study, we sought to investigate the relationship between hepatic steatosis, serum testosterone and dehydroepiandrosterone sulphate (DHEAS) levels in men. This is a cross‐sectional population‐based study. We used data of 1912 men recruited for the population‐based Study of Health in Pomerania, which was conducted in a region with high prevalence of metabolic syndrome and related diseases. Hepatic steatosis was defined according to sonographic criteria. The relationship of hepatic steatosis with serum testosterone and DHEAS levels was analysed by multivariable logistic regression. Men with low serum testosterone levels had a higher risk of hepatic steatosis than men with high serum testosterone levels. Adjustment for age and further confounders attenuated this association, but did not affect statistical significance (odds ratio 2.36; 95% confidence interval 1.66–3.37; p < 0.05). In the full model, the highest risk of hepatic steatosis was found in subjects with the highest serum DHEAS levels (odds ratio 1.59; 95% confidence interval 1.04–2.43; p < 0.05). Exclusion of men with high alcohol consumption did not affect these results substantially. Hepatic steatosis is associated with low serum testosterone and high serum DHEAS levels. These associations are independent of alcohol consumption.
Gonadal Steroid Action and Brain Sex Differentiation in the Rat
Authors: Sakuma Y.
Published: February 2009

Gonadal steroids that establish sexually dimorphic characteristics of brain morphology and physiology act at a particular stage of ontogeny. Testosterone secreted by the testes during late gestational and neonatal periods causes significant brain sexual dimorphism in the rat. This results in both sex‐specific behaviour and endocrinology in adults. Sexual differentiation may be due to neurogenesis, migration or survival. Each mechanism appears to be uniquely regulated in a site‐specific manner. Thus, the volume of an aggregate of neurones in the rat medial preoptic area (POA), termed the sexually dimorphic nucleus of the POA (SDN‐POA), is larger in males than in females. The anteroventral periventricular nucleus (AVPV) is packed with neurones containing oestrogen receptor (ER)β in female rats but, in males, ERβ‐positive neurones scatter into the more lateral portion of the POA. POA neurones are born up to embryonic days 16–17 and not after parturition. Therefore, neurogenesis is unlikely to contribute to the larger SDN‐POA in males. DNA microarray analysis for oestrogen‐responsive genes and western blotting demonstrated site‐specific regulation of apoptosis‐ and migration‐related genes in the SDN‐POA and AVPV.
Sagittalis Nucleus: A Novel Hypothalamic Nucleus
Authors: Mori H. , Matsuda K.‐I. , Pfaff D. W. , Kawata M.
Published: February 2009

The sagittalis nucleus (SGN) of the hypothalamus is a newly‐identified nucleus that is located in the interstitial area between the arcuate and ventromedial nuclei of the rat hypothalamus and for which the long axis of the nucleus is oriented sagittally. Interestingly, the SGN exhibits structural and physiological sex differences, as defined by Nissl staining and oestrogen receptor (ER)α immunoreactivity (‐ir), being larger in males than females. The structural sex difference is established by sex steroid action in neonates because the treatment of female pups with testosterone propionate masculinised the SGN. The phenotypical sex difference in ERα‐ir is mediated hormonally in adulthood. Ovariectomy of female rats caused a significant increase in ERα‐ir in the SGN, and eliminated the physiological sex difference, but with recovery to the level of gonad‐intact females when given oestradiol replacement. Adult females have oestrous cycle‐related variations in ERα‐ir in the SGN, with levels at a nadir during the evening of pro‐oestrous. The discovery of the SGN, a target of sex steroid action, provides a new opportunity for explaining hormonal regulation of sexually‐differentiated behavioural and endocrine functions.
Occult hepatitis B virus infection increases hepatocellular carcinogenesis by eight times in patients with non‐B, non‐C liver cirrhosis: a cohort study
Authors: Ikeda K. , Kobayashi M. , Someya T. , Saitoh S. , Hosaka T. , Akuta N. , Suzuki F. , Suzuki Y. , Arase Y. , Kumada H.
Published: February 2009

Summary.  An impact of serum hepatitis B virus (HBV) DNA on hepatocarcinogenesis has not been investigated in a cohort of patients with non‐B, non‐C cirrhosis. Eighty‐two consecutive Japanese patients with cirrhosis, who showed negative hepatitis B surface antigen and negative anti‐hepatitis C virus, were observed for a median of 5.8 years. Hepatitis B virus core (HBc) region and HBx region were assayed with nested polymerase chain reaction. Both of HBc and HBx DNA were positive in 9 patients (11.0%) and both were negative in 73. Carcinogenesis rates in the whole patients were 13.5% at the end of the 5th year and 24.6% at the 10th year. The carcinogenesis rates in the patients with positive DNA group and negative DNA group were 27.0% and 11.8% at the end of the 5th year, and 100% and 17.6% at the 10th year, respectively (P = 0.0078). Multivariate analysis showed that men (P = 0.04), presence of HBc and HBx DNA (hazard ratio: 8.25, P = 0.003), less total alcohol intake (P = 0.010), older age (P = 0.010), and association of diabetes (P = 0.005) were independently associated with hepatocellular carcinogenesis. Existence of serum HBV DNA predicted a high hepatocellular carcinogenesis rate in a cohort of patients with non‐B, non‐C cirrhosis.
Microsatellite variability reveals the necessity for genetic input from wild giant pandas (Ailuropoda melanoleuca) into the captive population
Authors: SHEN FUJUN , ZHANG ZHIHE , HE WEI. , YUE BISONG , ZHANG ANJU. , ZHANG LIANG , HOU RONG. , WANG CHENGDONG , WATANABE TOSHI
Published: February 2009

Recent success in breeding giant pandas in captivity has encouraged panda conservationists to believe that the ex situ population is ready to serve as a source for supporting the wild population. In this study, we used 11 microsatellite DNA markers to assess the amount and distribution of genetic variability present in the two largest captive populations (Chengdu Research Base of Giant Panda Breeding, Sichuan Province and the China Research and Conservation Center for the Giant Panda at Wolong, Sichuan Province). The data were compared with those samples from wild pandas living in two key giant panda nature reserves (Baoxing Nature Reserve and Wanglang Nature Reserve). The results show that the captive populations have retained lower levels of allelic diversity and heterozygosity compared to isolated wild populations. However, low inbreeding coefficients indicate that captive populations are under careful genetic management. Excessive heterozygosity suggests that the two captive populations have experienced a genetic bottleneck, presumably caused by founder effects. Moreover, evidence of increased genetic divergence demonstrates restricted breeding options within facilities. Based on these results, we conclude that the genetic diversity in the captive populations is not optimal. Introduction of genetic materials from wild pandas and improved exchange of genetic materials among institutions will be necessary for the captive pandas to be representative of the wild populations.
Conservation of the introgressed European water frog complex using molecular tools
Authors: HOLSBEEK G. , MAES G. E. , DE MEESTER L. , VOLCKAERT F. A. M.
Published: February 2009

In Belgium, the Pelophylax esculentus complex has recently been subjected to multiple introductions of non‐native water frogs, increasing the occurrence of hybridisation events. In the present study, we tested the reliability of morphometric and recently developed microsatellite tools to identify introgression and to determine the origin of exotic Belgian water frogs. By analysing 150 individuals of each taxon of the P. esculentus complex and an additional 60 specimens of the introduced P. cf. bedriagae, we show that neither of the currently available tools appears to have sufficient power to reliably distinguish all Belgian water frog species. We therefore aimed at increasing the discriminatory power of a microsatellite identification tool by developing a new marker panel with additional microsatellite loci. By adding only two new microsatellite loci (RlCA5 and RlCA1b20), all taxa of the P. esculentus complex could be distinguished from each other with high confidence. Three more loci (Res3, Res5 and Res17) provided a powerful discrimination of the exotic species.
Ancient isolation and independent evolution of the three clonal lineages of the exotic sudden oak death pathogen Phytophthora ramorum
Authors: GOSS E. M. , CARBONE I. , GRÜNWALD N. J.
Published: February 2009

The genus Phytophthora includes some of the most destructive plant pathogens affecting agricultural and native ecosystems and is responsible for a number of recent emerging and re‐emerging infectious diseases of plants. Sudden oak death, caused by the exotic pathogen P. ramorum, has caused extensive mortality of oaks and tanoaks in Northern California, and has brought economic losses to US and European nurseries as well due to its infection of common ornamental plants. In its known range, P. ramorum occurs as three distinct clonal lineages. We inferred the evolutionary history of P. ramorum from nuclear sequence data using coalescent‐based approaches. We found that the three lineages have been diverging for at least 11% of their history, an evolutionarily significant amount of time estimated to be on the order of 165 000 to 500 000 years. There was also strong evidence for historical recombination between the lineages, indicating that the ancestors of the P. ramorum lineages were members of a sexually reproducing population. Due to this recombination, the ages of the lineages varied within and between loci, but coalescent analyses suggested that the European lineage may be older than the North American lineages. The divergence of the three clonal lineages of P. ramorum supports a scenario in which the three lineages originated from different geographic locations that were sufficiently isolated from each other to allow independent evolution prior to introduction to North America and Europe. It is thus probable that the emergence of P. ramorum in North America and Europe was the result of three independent migration events.
Multiple gene genealogies reveal asymmetrical hybridization and introgression among strongylocentrotid sea urchins
Authors: ADDISON JASON A. , POGSON GRANT H.
Published: February 2009

The evolution of incompatibilities between eggs and sperm is thought to play important roles in establishing and maintaining reproductive isolation among species of broadcast‐spawning marine invertebrates. However, the effectiveness of gametic isolation in initiating the speciation process and/or in limiting the introgression of genes among species at later stages of divergence remains largely unknown. In the present study, we collected DNA sequence data from five loci in four species ofStrongylocentrotussea urchins (S. droebachiensis,S. pallidus,S. purpuratus, andS. franciscanus) to test whether the susceptibility ofS. droebachiensiseggs to fertilization by heterospecific sperm results in gene flow between species. Despite the potential for introgression, a small but statistically significant signal of introgression was observed only between the youngest pair of sister taxa (S. pallidusandS. droebachiensis) that was strongly asymmetrical (from the former into the latter). No significant gene flow was observed for eitherS. purpuratusorS. franciscanusdespite the ability of their sperm to readily fertilize the eggs ofS. droebachiensis. Our results demonstrate that asymmetrical gamete compatibilities in strongylocentrotids can give rise to asymmetrical patterns of introgression but suggest that gamete traits alone cannot be responsible for maintaining species integrities. The genetic boundaries between strongylocentrotid urchin species in the northeast Pacific appear to be related to postzygotic isolating mechanisms that scale with divergence times and not intrinsic gametic incompatibilitiesper se.
Evidence of parasite‐mediated disruptive selection on genetic diversity in a wild fish population
Authors: BLANCHET SIMON , REY OLIVIER , BERTHIER PAULINE , LEK SOVAN , LOOT GERALDINE
Published: February 2009

Identifying the processes maintaining genetic variability in wild populations is a major concern in conservation and evolutionary biology. Parasite‐mediated selection may strongly affect genetic variability in wild populations. The inbreeding depression theory predicts that directional selection imposed by parasites should act against the most inbred hosts, thus favouring genetic diversity in wild populations. We have tested this prediction by evaluating the strength and shape of the relationship between the load of a harmful fin‐feeder ectoparasite (Tracheliastes polycolpus) and the genome‐wide genetic diversity (i.e. heterozygosity measured at a set of 15 microsatellites) of its fish host, the rostrum dace (Leuciscus leuciscus). Contrary to expectation, we found a nonlinear relationship between host genetic diversity and ectoparasite load, with hosts that were either homozygous or heterozygous harbouring significantly fewer parasites than hosts with an intermediate level of heterozygosity. This relationship suggests that parasites could increase the variance of global heterozygosity in this host population through disruptive selection on genetic diversity. Moreover, when genetic diversity was measured at each locus separately, we found two very strong positive associations between host genetic diversity and the ectoparasite load. This latter result has three main implications: (i) genome‐wide effect cannot alone explain the nonlinear relationship between global heterozygosity and ectoparasite load, (ii) negative non‐additive allelic interactions (i.e. underdominance) may be a mechanism for resisting ectoparasite infection, and (iii) ectoparasites may favour homozygosity at some loci in this host population.
Two small c‐type cytochromes affect virulence gene expression in Bacillus anthracis
Authors: Wilson Adam C. , Hoch James A. , Perego Marta
Published: February 2009

Regulated expression of the genes for anthrax toxin proteins is essential for the virulence of the pathogenic bacterium Bacillus anthracis. Induction of toxin gene expression depends on several factors, including temperature, bicarbonate levels, and metabolic state of the cell. To identify factors that regulate toxin expression, transposon mutagenesis was performed under non‐inducing conditions and mutants were isolated that untimely expressed high levels of toxin. A number of these mutations clustered in the haem biosynthetic and cytochrome c maturation pathways. Genetic analysis revealed that two haem‐dependent, small c‐type cytochromes, CccA and CccB, located on the extracellular surface of the cytoplasmic membrane, regulate toxin gene expression by affecting the expression of the master virulence regulator AtxA. Deregulated AtxA expression in early exponential phase resulted in increased expression of toxin genes in response to loss of the CccA‐CccB signalling pathway. This is the first function identified for these two small c‐type cytochromes of Bacillus species. Extension of the transposon screen identified a previously uncharacterized protein, BAS3568, highly conserved across many bacterial and archeal species, as involved in cytochrome c activity and virulence regulation. These findings are significant not only to virulence regulation in B. anthracis, but also to analysis of virulence regulation in many pathogenic bacteria and to the study of cytochrome c activity in Gram‐positive bacteria.
Novel promoter and alternate transcription start site of the human serotonin reuptake transporter in intestinal mucosa
Authors: Linden D. R. , White S. L. , Brooks E. M. , Mawe G. M.
Published: February 2009

Abstract  Selective serotonin‐reuptake inhibitors are therapies for psychological and bowel disorders, but produce adverse effects in the non‐targeted system. To determine whether human serotonin‐selective reuptake transporter (SERT) transcripts in the intestine are different from the brain, rapid amplification of cDNA ends, primer extension and RT‐PCR assays were used to evaluate SERT transcripts from each region. Potential SLC6A4 gene promoter constructs were evaluated with a secreted alkaline phosphatase reporter assay. A novel transcript of the human SLC6A4 gene was discovered that predominates in the intestine, and differs from previous transcripts in the 5′‐untranslated region. The distinct transcriptional start site and alternate promoter suggest that gastrointestinal SERT can be differentially regulated from brain SERT, may explain why the polymorphism in the previously identified promoter is associated with affective disorders, but not associated with gastrointestinal dysfunction, and suggest the intriguing possibility of the development of site‐specific therapeutics for SERT regulation in the treatment of multiple disorders.