bluuebunny/crossref_metadata_2025 huggingface dataset:
Exploring the validity of estimating EQ-5D and SF-6D utility values from the health assessment questionnaire in patients with inflammatory arthritis
Authors: Harrison Mark J , Lunt Mark , Verstappen Suzanne MM , Watson Kath D , Bansback Nick J , Symmons Deborah PM
Published: February 2010

Background

Utility scores are used to estimate Quality Adjusted Life Years (QALYs), applied in determining the cost-effectiveness of health care interventions. In studies where no preference based measures are collected, indirect methods have been developed to estimate utilities from clinical instruments. The aim of this study was to evaluate a published method of estimating the EuroQol-5D (EQ-5D) and Short Form-6D (SF-6D) (preference based) utility scores from the Health Assessment Questionnaire (HAQ) in patients with inflammatory arthritis.

Methods

Data were used from 3 cohorts of patients with: early inflammatory arthritis (<10 weeks duration); established (>5 years duration) stable rheumatoid arthritis (RA); and RA being treated with anti-TNF therapy. Patients completed the EQ-5D, SF-6D and HAQ at baseline and a follow-up assessment. EQ-5D and SF-6D scores were predicted from the HAQ using a published method. Differences between predicted and observed EQ-5D and SF-6D scores were assessed using the paired t-test and linear regression.

Results

Predicted utility scores were generally higher than observed scores (range of differences: EQ-5D 0.01 - 0.06; SF-6D 0.05 - 0.10). Change between predicted values of the EQ-5D and SF-6D corresponded well with observed change in patients with established RA. Change in predicted SF-6D scores was, however, less than half of that in observed values (p < 0.001) in patients with more active disease. Predicted EQ-5D scores underestimated change in cohorts of patients with more active disease.

Conclusion

Predicted utility scores overestimated baseline values but underestimated change. Predicting utility values from the HAQ will therefore likely underestimate the QALYs of interventions, particularly for patients with active disease. We recommend the inclusion of at least one preference based measure in future clinical studies.
Short term hypothyroidism affects ovarian function in the cycling rat
Authors: Hapon María Belén , Gamarra-Luques Carlos , Jahn Graciela A
Published: February 2010

Background

Rats made hypothyroid with propilthyouracil start showing abnormal cycling on the second cycle after the start of the treatment, with a high proportion of spontaneous pseudopregnancies and reduced fertility.

Methods

To investigate some of the mechanisms involved in these reproductive abnormalities, hypothyroidism was induced in virgin rats by propilthyouracil (0.1 g/L in the drinking water) and we determined circulating hormones by radioimmunoassay and whole ovary expression of ovarian hormone receptors, growth factors and steroidogenic enzymes using semi-quantitative RT-PCR.

The study was performed on days 6 to 9 of treatment, corresponding to diestrus I (at 20.00-22.00 h), diestrus II (at 20.00-22.00 h), proestrus and estrus (both at 8.00-10.00 h and 20.00-22.00 h) of the second estrous cycle after beginning propilthyouracil treatment. Another group of rats was mated on day 8 and the treatment continued through the entire pregnancy to evaluate reproductive performance.

Results

Hypothyroidism increased circulating prolactin and estradiol on estrus 5 to 7-fold and 1.2 to 1.4-fold respectively. Growth hormone and insulin-like growth factor 1 diminished 60 and 20% respectively on proestrus morning. Hypothyroidism doubled the ovarian mRNA contents of estrogen receptor-beta on proestrus and estrus evenings, cyp19A1 aromatase mRNA on estrus evening and of growth hormone receptor on proestrus evening. Hypothyroidism did not influence ovulation rate or the number of corpora lutea at term, but a diminished number of implantation sites and pups per litter were observed (Hypothyroid: 11.7 +/- 0.8 vs. Control: 13.9 +/- 0.7).

Conclusions

Short term hypothyroidism alters normal hormone profile in the cycling rat increasing the expression of estrogen receptor-beta and cyp19A1 aromatase on estrus, which in turn may stimulate estradiol and prolactin secretion, favouring corpus luteum survival and the subsequent instauration of pseudopregnancy.
Effects of collagen membranes enriched with in vitro-differentiated N1E-115 cells on rat sciatic nerve regeneration after end-to-end repair
Authors: Amado Sandra , Rodrigues Jorge M , Luís Ana L , Armada-da-Silva Paulo AS , Vieira Márcia , Gartner Andrea , Simões Maria J , Veloso António P , Fornaro Michele , Raimondo Stefania , Varejão Artur SP , Geuna Stefano , Maurício Ana C
Published: February 2010

Peripheral nerves possess the capacity of self-regeneration after traumatic injury but the extent of regeneration is often poor and may benefit from exogenous factors that enhance growth. The use of cellular systems is a rational approach for delivering neurotrophic factors at the nerve lesion site, and in the present study we investigated the effects of enwrapping the site of end-to-end rat sciatic nerve repair with an equine type III collagen membrane enriched or not with N1E-115 pre-differentiated neural cells. After neurotmesis, the sciatic nerve was repaired by end-to-end suture (End-to-Endgroup), end-to-end suture enwrapped with an equine collagen type III membrane (End-to-EndMembgroup); and end-to-end suture enwrapped with an equine collagen type III membrane previously covered with neural cells pre-differentiatedin vitrofrom N1E-115 cells (End-to-EndMembCellgroup). Along the postoperative, motor and sensory functional recovery was evaluated using extensor postural thrust (EPT), withdrawal reflex latency (WRL) and ankle kinematics. After 20 weeks animals were sacrificed and the repaired sciatic nerves were processed for histological and stereological analysis. Results showed that enwrapment of the rapair site with a collagen membrane, with or without neural cell enrichment, did not lead to any significant improvement in most of functional and stereological predictors of nerve regeneration that we have assessed, with the exception of EPT which recovered significantly better after neural cell enriched membrane employment. It can thus be concluded that this particular type of nerve tissue engineering approach has very limited effects on nerve regeneration after sciatic end-to-end nerve reconstruction in the rat.
Dietary phytosterols and phytostanols decrease cholesterol levels but increase blood pressure in WKY inbred rats in the absence of salt-loading
Authors: Chen Qixuan , Gruber Heidi , Swist Eleonora , Coville Kara , Pakenham Catherine , Ratnayake Walisundera MN , Scoggan Kylie A
Published: February 2010

Background

There are safety concerns regarding widespread consumption of phytosterol and phytostanol supplemented food products. The aim of this study was to determine, in the absence of excess dietary salt, the individual effects of excess accumulation of dietary phytosterols and phytostanols on blood pressure in Wistar Kyoto (WKY) inbred rats that have a mutation in the Abcg5 gene and thus over absorb phytosterols and phytostanols.

Methods

Thirty 35-day old male WKY inbred rats (10/group) were fed a control diet or a diet containing phytosterols or phytostanols (2.0 g/kg diet) for 5 weeks. The sterol composition of the diets, plasma and tissues were analysed by gas chromatography. Blood pressure was measured by the tail cuff method. mRNA levels of several renal blood pressure regulatory genes were measured by real-time quantitative PCR.

Results

Compared to the control diet, the phytosterol diet resulted in 3- to 4-fold increases in the levels of phytosterols in plasma, red blood cells, liver, aorta and kidney of WKY inbred rats (P < 0.05). The phytostanol diet dramatically increased (> 9-fold) the levels of phytostanols in plasma, red blood cells, liver, aorta and kidney of these rats (P < 0.05). The phytosterol diet decreased cholesterol levels by 40%, 31%, and 19% in liver, aorta and kidney, respectively (P < 0.05). The phytostanol diet decreased cholesterol levels by 15%, 16%, 20% and 14% in plasma, liver, aorta and kidney, respectively (P < 0.05). The phytostanol diet also decreased phytosterol levels by 29% to 54% in plasma and tissues (P < 0.05). Both the phytosterol and phytostanol diets produced significant decreases in the ratios of cholesterol to phytosterols and phytostanols in plasma, red blood cells, liver, aorta and kidney. Rats that consumed the phytosterol or phytostanol diets displayed significant increases in systolic and diastolic blood pressure compared to rats that consumed the control diet (P < 0.05). The phytosterol diet increased renal angiotensinogen mRNA levels of these rats.

Conclusion

These data suggest that excessive accumulation of dietary phytosterols and phytostanols in plasma and tissues may contribute to the increased blood pressure in WKY inbred rats in the absence of excess dietary salt. Therefore, even though phytosterols and phytostanols lower cholesterol levels, prospective clinical studies testing the net beneficial effects of dietary phytosterols and phytostanols on cardiovascular events for subgroups of individuals that have an increased incorporation of these substances are needed.
Maintenance of Sorafenib following combined therapy of three-dimensional conformal radiation therapy/intensity-modulated radiation therapy and transcatheter arterial chemoembolization in patients with locally advanced hepatocellular carcinoma: a phase I/II study
Authors: Zhao Jian-Dong , Liu Jin , Ren Zhi-Gang , Gu Ke , Zhou Zhen-Hua , Li Wen-Tao , Chen Zhen , Xu Zhi-Yong , Liu Lu-Ming , Jiang Guo-Liang
Published: February 2010

Background

Three-dimensional conformal radiation therapy (3DCRT)/intensity-modulated radiation therapy (IMRT) combined with or without transcatheter arterial chemoembolization (TACE) for locally advanced hepatocellular carcinoma (HCC) has shown favorable outcomes in local control and survival of locally advanced HCC. However, intra-hepatic spreading and metastasis are still the predominant treatment failure patterns. Sorafenib is a multikinase inhibitor with effects against tumor proliferation and angiogenesis. Maintenance Sorafenib would probably prevent or delay the intrahepatic and extrahepatic spread of HCC after radiotherapy, which provides the rationale for the combination of these treatment modalities.

Methods and design

Patients with solitary lesion (bigger than 5 cm in diameter) histologically or cytologically confirmed HCC receive TACE (1-3 cycles) plus 3DCRT/IMRT 4-6 weeks later. Maintenance Sorafenib will be administered only for the patients with non-progression disease 4 to 6 weeks after the completion of radiotherapy. The dose will be 400 mg, p.o., twice a day. Sorafenib will be continuously given for 12 months unless intolerable toxicities and/or tumor progression. If no more than 3 patients discontinue Sorafenib treatment who experience dose-limiting toxicity after necessary dose modification and delay and/or radiation-induced liver disease in the first 15 enrolled patients, the study will recruit second fifteen patients for further evaluating safety and efficacy of treatment. Hypothesis of the current study is that Sorafenib as a maintenance therapy after combined therapy of 3DCRT/IMRT and TACE is safe and superior to radiotherapy combined with TACE alone in terms of time to progression (TTP), progression-free survival (PFS) and overall survival (OS) in comparison to historical data.

Discussion

A recent meta-analysis showed TACE in combination with radiotherapy, improved the survival and the tumor response of patients, and was thus more therapeutically beneficial. In this study, local therapy for HCC is the combination of TACE and radiotherapy. Radiation exposure as a kind of stress might induce the compensatory activations of multiple intracellular signaling pathway mediators, such as PI3K, MAPK, JNK and NF-kB. Vascular endothelial growth factor (VEGF) was identified as one factor that was increased in a time- and dose-dependent manner after sublethal irradiation of HCC cells in vitro, translating to enhanced intratumor angiogenesis in vivo. Therefore, Sorafenib-mediated blockade of the Raf/MAPK and VEGFR pathways might enhance the efficacy of radiation, when Sorafenib is followed sequentially as a maintenance modality. (ClinicalTrials.gov number, NCT00999843.)
Occult small bowel perforation in a patient with Ehlers Danlos syndrome: a case report and review of the literature
Authors: Leake Tessa Frances , Singhal Tarun , Chandra Aninda , Ashcroft Alexandra , Doddi Sudeendra , Hussain Abdulzahra , Smedley Frank
Published: February 2010

Patients who present with a co-existing connective tissue disorder add a degree of complexity to operative intervention. We present an unusual case of a 53-year-old Caucasian female patient with Ehlers Danlos syndrome who presented with an occult perforation of the distal ileum. The patient had known small bowel diverticulae yet the perforation occurred within the normal bowel wall. The pre-operative CT only showed malrotation of the large bowel and did not correlate with the intra-operative findings. Our case has highlighted that although small bowel perforation is a rare occurrence, it may be more common in Ehlers Danlos and may present with atypical features. Perforation may also occur alongside normal bowel as well as diverticulae within the bowel. Where diverticulae exists within a patient with Ehlers Danlos syndrome and there is some diagnostic uncertainty, there should be a lower threshold for operative intervention. We present in the discussion a number of salient features and learning points.
Differential expression level of cytokeratin 8 in cells of the bovine nucleus pulposus complicates the search for specific intervertebral disc cell markers
Authors: Gilson Audrey , Dreger Mathias , Urban Jill PG
Published: February 2010

Introduction

Development of cell therapies for repairing the intervertebral disc is limited by the lack of a source of healthy human disc cells. Stem cells, particularly mesenchymal stem cells, are seen as a potential source but differentiation strategies are limited by the lack of specific markers that can distinguish disc cells from articular chondrocytes.

Methods

We searched for markers using the differential in-gel electrophoresis proteomic technology to compare proteins of bovine nucleus pulposus cells, phenotypically similar to mature human nucleus cells, with those of bovine articular chondrocytes. In the cohort of the differentially expressed proteins identified by mass spectrometry, cytokeratin 8 (CK8) was further validated by immunostaining of freshly isolated cells and frozen tissue sections using monoclonal antibodies.

Results

We identified a set of 14 differentially expressed proteins. Immunohistochemistry showed that only a subset of cells (approximately 10%) was positive for one of these proteins, CK8, an intermediate filament protein present in epithelial but not mesenchymal cells. In tissue sections, CK8-positive cells were seen in all discs examined and appeared as small isolated clusters surrounded by gelatinous matrix. Notochordal nucleus pulposus cells from pig, phenotypically similar to human infant nucleus pulposus cells, were all CK8-positive. The mesenchymal intermediate filament protein vimentin was present in all bovine and porcine nucleus pulposus cells.

Conclusions

The notochordal cell population is reported to disappear from the nucleus pulposus of bovine discs before birth and from human discs in childhood. However our finding of the co-expression of vimentin and CK8 in small isolated clusters of the bovine nucleus pulposus cells indicates that a subpopulation of notochordal-like cells remains in the mature bovine disc. This finding agrees with reports in the literature on co-expression of cytokeratins and vimentin in adult human discs. As notochordal cells produce factors that promote matrix production, the CK8-positive subpopulation could have important implications for activity and survival of the nucleus pulposus, and should be considered in development of cell therapies for disc repair. In addition, the finding of differential expression of proteins in the cell population of nucleus pulposus has implications with regard to the search for specific markers.
Diffusion-weighted magnetic resonance imaging for predicting the clinical outcome of comatose survivors after cardiac arrest: a cohort study
Authors: Choi Seung Pill , Park Kyu Nam , Park Hae Kwan , Kim Jee Young , Youn Chun Song , Ahn Kook Jin , Yim Hyeon Woo
Published: February 2010

Introduction

The aim of this study was to examine whether the patterns of diffusion-weighted imaging (DWI) abnormalities and quantitative regional apparent diffusion coefficient (ADC) values can predict the clinical outcome of comatose patients following cardiac arrest.

Methods

Thirty-nine patients resuscitated from out-of-hospital cardiac arrest were prospectively investigated. Within five days of resuscitation, axial DWIs were obtained and ADC maps were generated using two 1.5-T magnetic resonance scanners. The neurological outcomes of the patients were assessed using the Glasgow Outcome Scale (GOS) score at three months after the cardiac arrest. The brain injuries were categorised into four patterns: normal, isolated cortical injury, isolated deep grey nuclei injury, and mixed injuries (cortex and deep grey nuclei). Twenty-three subjects with normal DWIs served as controls. The ADC and percent ADC values (the ADC percentage as compared to the control data from the corresponding region) were obtained in various regions of the brains. We analysed the differences between the favourable (GOS score 4 to 5) and unfavourable (GOS score 1 to 3) groups with regard to clinical data, the DWI abnormalities, and the ADC and percent ADC values.

Results

The restricted diffusion abnormalities in the cerebral cortex, caudate nucleus, putamen and thalamus were significantly different between the favourable (n = 13) and unfavourable (n = 26) outcome groups. The cortical pattern of injury was seen in one patient (3%), the deep grey nuclei pattern in three patients (8%), the cortex and deep grey nuclei pattern in 21 patients (54%), and normal DWI findings in 14 patients (36%). The cortex and deep grey nuclei pattern was significantly associated with the unfavourable outcome (20 patients with unfavourable vs. 1 patient with favourable outcomes, P < 0.001). In the 22 patients with quantitative ADC analyses, severely reduced ADCs were noted in the unfavourable outcome group. The optimal cutoffs for the mean ADC and the percent ADC values determined by receiver operating characteristic (ROC) curve analysis in the cortex, caudate nucleus, putamen, and thalamus predicted the unfavourable outcome with sensitivities of 67 to 93% and a specificity of 100%.

Conclusions

The patterns of brain injury in early diffusion-weighted imaging (DWI) (less than or equal to five days after resuscitation) and the quantitative measurement of regional ADC may be useful for predicting the clinical outcome of comatose patients after cardiac arrest.
The Incidence of Ischemic Heart Disease and Mortality in People with Subclinical Hypothyroidism: Reanalysis of the Whickham Survey Cohort
Authors: Razvi Salman , Weaver Jola U. , Vanderpump Mark P. , Pearce Simon H. S.
Published: February 2010

Context: The Whickham Survey evaluated vascular events over 20 yr in community-dwelling subjects stratified by thyroid function and thyroid autoantibody status. No association between ischemic heart disease (IHD) and a composite autoimmune thyroid disease group, comprising individuals with subclinical hypothyroidism (SCH), with positive thyroid antibodies or those using levothyroxine, was found. This result appears to be at odds with the findings of other cohort studies.

Objective: The objective of the study was to evaluate incident IHD and mortality in participants in relation to their thyroid status.

Outcomes, Design, and Participants: Data were reanalyzed assessing incident IHD events and mortality during 20 yr of follow-up in individuals with endogenous SCH (n = 97; TSH 6.0–15 mIU/liter) vs. the euthyroid group (n = 2279), who were IHD free at baseline.

Results: Incident IHD was significantly higher in the SCH group [adjusted hazard ratio 1.76 (95% confidence interval 1.15–2.71); P = 0.01]. IHD mortality was also increased in the SCH group [hazard ratio of 1.79 (1.02–3.56); P = 0.05]. These findings lost their significance when subsequent treatment with levothyroxine was excluded from the regression model. There was no difference in all-cause mortality between the groups.

Conclusion: In the Whickham Survey, there is an association between incident IHD events and IHD-related mortality with SCH over the 20 yr of follow-up. Furthermore, subsequent treatment of SCH with levothyroxine appears to attenuate IHD-related morbidity and mortality, and this may explain why some other longitudinal studies of SCH have not shown such an association; properly designed controlled trials of treatment of SCH are required to answer this question definitively.
The Association of Mitochondrial Content with Prevalent and Incident Type 2 Diabetes
Authors: Reiling Erwin , Ling Charlotte , Uitterlinden André G. , van't Riet Esther , Welschen Laura M. C. , Ladenvall Claes , Almgren Peter , Lyssenko Valeriya , Nijpels Giel , van Hove Els C. , Maassen Johannes A. , de Geus Eco J. C. , Boomsma Dorret I. , Dekker Jacqueline M. , Groop Leif , Willemsen Gonneke , 't Hart Leen M.
Published: February 2010

Context: It has been shown that mitochondrial DNA (mtDNA) content is associated with type 2 diabetes (T2D) and related traits. However, empirical data, often based on small samples, did not confirm this observation in all studies. Therefore, the role of mtDNA content in T2D remains elusive.

Objective: In this study, we assessed the heritability of mtDNA content in buccal cells and analyzed the association of mtDNA content in blood with prevalent and incident T2D.

Design and Setting: mtDNA content from cells from buccal and blood samples was assessed using a real-time PCR-based assay. Heritability of mtDNA content was estimated in 391 twins from the Netherlands Twin Register. The association with prevalent T2D was tested in a case control study from The Netherlands (n = 329). Incident T2D was analyzed using prospective samples from Finland (n = 444) and The Netherlands (n = 238).

Main Outcome Measures: We measured the heritability of mtDNA content and the association of mtDNA content in blood with prevalent and incident T2D.

Results: A heritability of mtDNA content of 35% (19–48%) was estimated in the twin families. We did not observe evidence of an association between mtDNA content and prevalent or incident T2D and related traits. Furthermore, we observed a decline in mtDNA content with increasing age that was male specific (P = 0.001).

Conclusion: In this study, we show that mtDNA content has a heritability of 35% in Dutch twins. There is no association between mtDNA content in blood and prevalent or incident T2D and related traits in our study samples.
Mixed Lineage Kinase-3/JNK1 Axis Promotes Migration of Human Gastric Cancer Cells following Gastrin Stimulation
Authors: Mishra Prajna , Senthivinayagam Subramanian , Rangasamy Velusamy , Sondarva Gautam , Rana Basabi
Published: February 2010

Gastrin is a gastrointestinal peptide hormone, secreted by the gastric G cells and can exist as a fully processed amidated form (G17) or as unprocessed forms. All forms of gastrin possess trophic properties towards the gastrointestinal mucosa. An understanding of the signaling pathways involved is important to design therapeutic approaches to target gastrin-mediated cellular events. The studies described here were designed to identify the signaling pathways by which amidated gastrin (G17) mediates cancer cell migration. These studies indicated a time- and dose-dependent increase in gastric cancer cell migration after G17 stimulation, involving cholecystokinin 2 receptor. G17-induced migration was preceded by activation of MAPK pathways and was antagonized after pretreatment with SP600125, a pharmacological inhibitor of c-Jun-NH2-terminal kinase (JNK) pathway. Knockdown of endogenous JNK1 expression via small interference RNA (JNK1-siRNA) inhibited G17-induced phosphorylation of c-Jun and migration, and overexpression of wild-type JNK1 or constitutive active JNK1 promoted G17-induced migration. Studies designed to identify the MAPK kinase kinase member mediating JNK activation indicated the involvement of mixed lineage kinase-3 (MLK3), which was transiently activated upon G17 treatment. Inhibition of MLK3 pathway via a pan-MLK inhibitor or knockdown of MLK3 expression by MLK3-siRNA antagonized G17-induced migration. Incubation with G17 also resulted in an induction of matrix metalloproteinase 7 promoter activity, which is known to mediate migration and invasion pathways in cancer cells. Modulation of MLK3, JNK1, and c-Jun pathways modulated G17-induced matrix metalloproteinase 7 promoter activation. These studies indicate that the MLK3/JNK1 axis mediates G17-induced gastric cancer cell migration, which can be targeted for designing novel therapeutic strategies for treating gastric malignancies.
Ligand-Mediated Activation of an Engineered Gs G Protein-Coupled Receptor in Osteoblasts Increases Trabecular Bone Formation
Authors: Hsiao Edward C. , Millard Susan M. , Louie Alyssa , Huang Yong , Conklin Bruce R. , Nissenson Robert A.
Published: February 2010

Age-dependent changes in skeletal growth play important roles in regulating skeletal expansion and in the course of many diseases affecting bone. How G protein-coupled receptor (GPCR) signaling affects these changes is poorly understood. Previously, we described a mouse model expressing Rs1, an engineered receptor with constitutive Gs activity. Rs1 expression in osteoblasts from gestation induced a dramatic age-dependent increase in trabecular bone with features resembling fibrous dysplasia; however, these changes were greatly minimized if Rs1 expression was delayed until after puberty. To further investigate whether ligand-induced activation of the Gs-GPCR pathway affects bone formation in adult mice, we activated Rs1 in adult mice with the synthetic ligand RS67333 delivered continuously via an osmotic pump or intermittently by daily injections. We found that osteoblasts from adult animals can be stimulated to form large amounts of bone, indicating that adult mice are sensitive to the dramatic bone- forming actions of Gs signaling in osteoblasts. In addition, our results show that intermittent and continuous activation of Rs1 led to structurally similar but quantitatively different degrees of trabecular bone formation. These results indicate that activation of a Gs-coupled receptor in osteoblasts of adult animals by either intermittent or continuous ligand administration can increase trabecular bone formation. In addition, osteoblasts located at the bone epiphyses may be more responsive to Gs signaling than osteoblasts at the bone diaphysis. This model provides a powerful tool for investigating the effects of ligand-activated Gs-GPCR signaling on dynamic bone growth and remodeling.
The cytolinker Pigs is a direct target and a negative regulator of Notch signalling
Authors: Pines Mary K. , Housden Ben E. , Bernard Fred , Bray Sarah J. , Röper Katja
Published: February 2010

Gas2-like proteins harbour putative binding sites for both the actin and the microtubule cytoskeleton and could thus mediate crosstalk between these cytoskeletal systems. Family members are highly conserved in all metazoans but their in vivo role is not clear. The sole Drosophila Gas2-like gene, CG3973 (pigs), was recently identified as a transcriptional target of Notch signalling and might therefore link cell fate decisions through Notch activation directly to morphogenetic changes. We have generated a null mutant in CG3973 (pigs): pigs1 mutants are semi-viable but adult flies are flightless, showing indirect flight muscle degeneration, and females are sterile, showing disrupted oogenesis and severe defects in follicle cell differentiation, similar to phenotypes seen when levels of Notch/Delta signalling are perturbed in these tissues. Loss of Pigs leads to an increase in Notch signalling activity in several tissues. These results indicate that Gas2-like proteins are essential for development and suggest that Pigs acts downstream of Notch as a morphogenetic read-out, and also as part of a regulatory feedback loop to relay back information about the morphogenetic state of cells to restrict Notch activation to appropriate levels in certain target tissues.
In situcardiac function in Atlantic cod (Gadus morhua): effects of acute and chronic hypoxia
Authors: Petersen L. H. , Gamperl A. K.
Published: February 2010

Recent in vivo experiments on Atlantic cod (Gadus morhua) acclimated to chronic hypoxia (6-12 weeks at 10°C; PwO2 ~8-9 kPa) revealed a considerable decrease in the pumping capacity of the heart. To examine whether this diminished cardiac performance was due to the direct effects of chronic moderate hypoxia on the myocardium (as opposed to alterations in neural and/or hormonal control), we measured the resting and maximum in situ function of hearts from normoxia- and hypoxia-acclimated cod: (1) when initially perfused with oxygenated saline; (2) at the end of a 15 min exposure to severe hypoxia (PO2 ~0.6 kPa); and (3) 30 min after the hearts had been reperfused with oxygenated saline. Acclimation to hypoxia did not influence resting (basal) in situ cardiac performance during oxygenated or hypoxic conditions. However, it caused a decrease in maximum cardiac output () under oxygenated conditions (from 49.5 to 40.3 ml min−1 kg−1; by 19%), that was due to diminished values for maximum stroke volume (VS) and scope for VS. Severe hypoxia reduced in both groups to ~20 ml min−1 kg−1, yet, the hearts of hypoxia-acclimated fish were better able to sustain this level of under hypoxia, and the recovery of (as compared with initial values under oxygenated conditions) was significantly improved (94% vs 83%). These data show that acclimation to hypoxia has a direct effect on cod myocardial function and/or physiology, and suggest that the cod heart shows some adaptations to prolonged hypoxia.
Changes in rabbit jaw-muscle activity parameters in response to reduced masticatory load
Authors: Grünheid T. , Brugman P. , Zentner A. , Langenbach G. E. J.
Published: February 2010

Mechanical food properties influence the neuromuscular activity of jaw-closing muscles during mastication. It is, however, unknown how the activity profiles of the jaw muscles are influenced by long-term alterations in masticatory load. In order to elucidate the effect of reduced masticatory load on the daily habitual activity profiles of three functionally different jaw muscles, the electromyograms of the masseter, temporalis and digastric muscles were recorded telemetrically in 16 male rabbits between seven and 20 weeks of age. Starting at eight weeks of age the experimental animals were fed significantly softer pellets than the control animals. Daily muscle activity was quantified by the relative duration of muscle use (duty time), burst number and burst length in relation to multiple activity levels. The daily duty time and burst number of the masseter muscle were significantly lower in the experimental group than in the control group at 5% and 10% of the maximum activity during the two weeks following the change in food hardness. By contrast, altered food hardness did not significantly influence the activity characteristics of the temporalis and digastric muscles. The findings suggest that a reduction in masticatory load decreases the neuromuscular activity of the jaw-closing muscles that are primarily responsible for force generation during mastication. This decrease is most pronounced in the weeks immediately following the change in food hardness and is limited to the activity levels that reflect muscle contractions during chewing. These findings support the conclusion that the masticatory system manifests few diet-specific long-term changes in the activity profiles of jaw muscles.
Understanding the vertebrate immune system: insights from the reptilian perspective
Authors: Zimmerman L. M. , Vogel L. A. , Bowden R. M.
Published: February 2010

Reptiles are ectothermic amniotes, providing the key link between ectothermic anamniotic fishes and amphibians, and endothermic amniotic birds and mammals. A greater understanding of reptilian immunity will provide important insights into the evolutionary history of vertebrate immunity as well as the growing field of eco-immunology. Like mammals, reptile immunity is complex and involves innate, cell-mediated and humoral compartments but, overall, there is considerably less known about immune function in reptiles. We review the current literature on each branch of the reptilian immune system, placing this information in context to other vertebrates. Further, we identify key areas that are prime for research as well as areas that are lagging because of lack of reagents in non-model systems.
Fitness costs may explain the post-colonisation erosion of phenotypic plasticity
Authors: Aubret F. , Shine R.
Published: February 2010

Many organisms can adjust their phenotypes to match local environmental conditions via shifts in developmental trajectories, rather than relying on changes in gene frequencies wrought by natural selection. Adaptive developmental plasticity confers obvious benefits in terms of rapid response and higher mean fitness, so why is it not more common? Plausibly, adaptive plasticity also confers a cost; reshaping the phenotype takes time and energy, so that canalised control of trait values enhances fitness if the optimal phenotype remains the same from one generation to the next. Although this idea is central to interpreting the fitness consequences of adaptive plasticity, empirical data on costs of plasticity are scarce. In Australian tiger snakes, larger relative head size enhances maximal ingestible prey size on islands containing large prey. The trait arises via adaptive plasticity in snake populations on newly colonised islands but becomes genetically canalised on islands where snakes have been present for much longer periods. We experimentally manipulated relative head size in captive neonatal snakes to quantify the costs of adaptive plasticity. Although small-headed snakes were able to increase their head sizes when offered large prey, the delay in doing so, and their inability to consume large prey at the outset, significantly reduced their growth rates relative to conspecifics with larger heads at the beginning of the experiment. This study describes a proximate cause to the post-colonisation erosion of developmental plasticity recorded in tiger snake populations.
Urinary proteases degrade albumin: implications for measurement of albuminuria in stored samples
Authors: Kania Kasia , Byrnes Elizabeth A , Beilby John P , Webb Steve A R , Strong Kimberley J
Published: February 2010

Background

Previous studies have shown that albumin in stored urine samples degrades over time, and that albumin losses are greatest in samples with low pH conditions (pH < 5). Furthermore, the high-performance liquid chromatography (HPLC) assay for urinary albumin has been shown to be particularly susceptible to the effects of prolonged storage.

Methods

Frozen urine samples, stored for 12 months at −70 and −20°C, were analysed for albumin fragmentation. Urinary protease activity was investigated in vitro in urine adjusted to pH 2.3–2.5. Albumin was measured by nephelometry, HPLC and sodium dodecyl sulphate-polyacrylamide gel electrophoresis.

Results

In the unadjusted samples, albumin was degraded in 11 out of 40 samples stored at −20°C. In the in vitro experiments, both endogenous albumin and exogenous albumin added to urine were rapidly degraded into large fragments within minutes after adjustment to low pH. The fragments produced were consistent with those produced during digestion with pepsin and urinary degradation was completely inhibited by pepstatin. Albumin concentration measured by HPLC was most dramatically affected, with near-complete loss of albumin-sized material within one hour of incubation at pH 2.3–2.5. Sample reactivity with antiserum in a nephelometry assay initially declined then increased, possibly due to exposure of internal epitopes during albumin digestion.

Conclusions

This study demonstrated that proteases are present and active in stored human urine samples. Urinary albumin digestion occurred in a manner consistent with activity of endogenous urinary proteases. Adjustment to neutral pH or addition of protease inhibitors may be useful techniques for sample preservation.
Molecular Diversity of Hemoglobin H Disease in India
Authors: Nadkarni Anita H. , Nair Sona B. , Italia Khushnooma Y. , Warang Prashant , Dalvi Madhura , Ghosh Kanjaksha , Colah Roshan B.
Published: February 2010

This study was undertaken to evaluate the variable clinical expression of hemoglobin (Hb) H disease in India. For the study, α genotyping was done in 8 patients with Hb H disease using multiplex polymerase chain reaction and DNA sequencing.

The study revealed that 4 genotypes (– –SEA/ –α3.7, – –SA/–α3.7, – –SEA/–α3.7 Sallanches, – –α3.7/–α3.7 Sallanches) were responsible for Hb H disease, the α+ thalassemia mutation (–α3.7 deletion) being the most common defect. The nondeletional mutation Hb Sallanches (α 2 codon 104 G → A) was seen in 3 cases. Two unique and novel genotypes leading to Hb H disease were characterized (– –SEA/–α3.7 Sallanches and –α3.7/–α3.7 Sallanches).

Because a majority of patients with Hb H disease do not have severe manifestations, prenatal diagnosis is usually unwarranted in India.
A Rapid and Simple High-Performance Liquid Chromatography Assay for the Leflunomide Metabolite, Teriflunomide (A77 1726), in Renal Transplant Recipients
Authors: Sobhani Kimia , Garrett Danette A. , Liu Dong-Pei , Rainey Petrie M.
Published: February 2010

Leflunomide (Arava), a drug with immunosuppressive and antiviral effects, is being used in renal transplant recipients, primarily for its action against BK polyomavirus (BKV), which affects 1% to 10% of renal transplant recipients and often causes failure of grafted kidneys. Leflunomide effects are solely due to an active metabolite, teriflunomide (formerly A77 1726). Trough blood concentrations of teriflunomide exceeding 40 μg/mL (148 μmol/L) are associated with progressive clearance of BKV. Toxic effects become increasingly apparent at higher concentrations.

We have developed a rapid, simple, and robust high-performance liquid chromatography (HPLC) method for therapeutic monitoring of teriflunomide in renal transplant recipients. Sample preparation is rapid, and each HPLC separation takes about 7 minutes. Intraday and interday coefficients of variation were 1.5% or less and 5.6% or less, respectively. The method was linear to 200 μg/mL (740 μmol/L), which is well above teriflunomide concentrations that are likely to be observed.
Recurrent Capillary Leak Syndrome Following Bortezomib Therapy in a Patient with Relapsed Myeloma
Authors: Hsiao Shu-Chen , Wang Ming-Chung , Chang Hung , Pei Sung-Nan
Published: February 2010

Objective:

To describe the first case of bortezomib-induced capillary leak syndrome (CLS), a rare but potentially life-threatening condition characterized by the shift of intravascular fluid and protein to the interstitial space.

Case Summary:

A 65-year-old female with relapsed multiple myeloma developed fluid retention, ascites, and general anasarca following bortezomib administration (1.3 mg/m2 on days 1, 4, 8, and 11). Aggressive albumin infusion and loop diuretics did not lead to improvement and the patient received 2 sessions of hemodialysis for pulmonary edema. Although the bortezomib dose was reduced (0.7 mg/m2 on days 1, 4, and 11) for the second cycle, CLS recurred after treatment. Dexamethasone was given along with bortezomib in the third cycle and subsequent CLS was prevented. The patient's multiple myeloma responded partially to the treatment, but the patient later died from cardiac amyloidosis.

Discussion:

Bortezomib is associated with several well-known adverse effects, such as peripheral neuropathy, thrombocytopenia, and gastrointestinal complications. CLS has not previously been reported to be associated with bortezomib. In this case, CLS developed twice after the patient received bortezomib treatment. The severity of CLS was dose-dependent and this adverse effect was preventable by concomitant use of steroids; this clearly demonstrated the close relationship between CLS and bortezomib in this patient. Using the Naranjo probability scale, the occurrence of CLS related to bortezomib treatment was probable.

Conclusions:

Our report demonstrates CLS as an unusual adverse effect of bortezomib. As bortezomib use may become more common, clinicians should be aware of this novel but potentially life-threatening adverse effect. Based on our experience, timely management with steroids is important in dealing with this complication.
Effects of elevated CO2 and phosphorus supply on growth, photosynthesis and nutrient uptake in the marine macroalga Gracilaria lemaneiformis (Rhodophyta)
Authors: Xu Zhiguang , Zou Dinghui , Gao Kunshan
Published: February 2010

The red alga Gracilaria lemaneiformis was cultured under different CO2 and phosphorus conditions for 16 days, and its growth, photosynthesis and uptake of nitrate and phosphate were examined in order to establish the longer-term impacts of elevated CO2 and phosphorus supplies on this economically important seaweed. Enrichment with either CO2 or phosphorus in culture markedly increased the growth of G. lemaneiformis compared to the control. Light-saturated photosynthetic rate was enhanced significantly by phosphorus enrichment, but hardly affected by the elevation of CO2 when G. lemaneiformis was grown under low phosphorus conditions. High phosphorus stimulated photosynthetic inorganic carbon utilization and nitrogen uptake. Under low phosphorus conditions, the thalli grown at the high level of CO2 had a lower carbon utilization capacity and a higher nitrogen uptake rate compared to those grown under ambient CO2. Reversed results were found when the algae were grown under high phosphorus conditions. Hence, available phosphorus may regulate inorganic carbon utilization of G. lemaneiformis grown at different CO2 levels, and growth reflected a balance between carbon and nutrient metabolism.
Procalcitonin measurement in routine emergency medicine practice: comparison between two immunoassays
Authors: Hausfater Pierre , Brochet Christine , Freund Yonathan , Charles Vanessa , Bernard Maguy
Published: February 2010

Background: Accurate identification of bacterial infections in patients presenting at the emergency department is crucial for early and rational antibiotic treatment. In this situation, using a cut-off of 0.25 μg/L for procalcitonin allows for carefully monitoring of febrile patients. Most previous studies have been performed with the reference B·R·A·H·M·S PCT KRYPTOR® assay. The goal of this study was to compare this test with the VIDAS B·R·A·H·M·S PCT® assay and to validate clinically relevant cut-off thresholds.

Methods: This prospective study was conducted in adults presenting to the emergency departments of a tertiary hospital. We included 305 consecutive patients that had procalcitonin requested. Procalcitonin was measured first with the KRYPTOR, then with the VIDAS systems. Statistical analysis consisted in Passing and Bablok and Bland-Altman plots.

Results: In the overall cohort, 176 patients had procalcitonin concentrations measured using both methods and were well correlated. The Bland-Altman plot exhibited a bias of 0.108 [95% confidence interval: –0.044 to 0.260]. The concordance at different procalcitonin cut-off thresholds, respectively of 0.1, 0.25, 0.5 and 2 μg/L, indicated that above 0.25 μg/L, the κ coefficient was >0.80.

Conclusions: A highly significant correlation was observed between the two automated assays. Procalcitonin concentrations obtained from both methods led to the same clinical interpretation.

Clin Chem Lab Med 2010;48:501–4.
Beware of carryover in modern chemistry analyzers
Authors: Kyle Patrick B.
Published: February 2010

Background: Random-access analyzers that employ dedicated probes may be affected by carryover. Sample carryover, involving analyte from one sample that is measured in a subsequent sample, is most often detected in measurement procedures that have wide reportable ranges. However, reagent carryover can be more difficult to detect as it may involve only one pair of measurement procedures.

Methods: Our laboratory noted several patient samples with total cholesterol <2.58 mmol/L (100 mg/dL) during the initial months after a new chemistry analyzer was installed. The problem seemed to occur intermittently, approximately once per week.

Results: Immediate reanalysis of affected samples resulted in measured values that were 1.03–2.58 mmol/L (40–100 mg/dL) higher. Evaluation of reagent carryover revealed a significant decrease in total cholesterol after analysis of creatine kinase (EC 2.7.3.2). Carryover disappeared when an additional reagent probe wash was applied, whereas the root of the problem was eliminated with replacement of the reagent probes. The insidious nature of reagent carryover made it difficult to initially detect the problem, which affected two of the three instruments in our laboratory.

Conclusions: Laboratorians should be aware of the potential for carryover in random access analyzers, and should be mindful of appropriate troubleshooting techniques.

Clin Chem Lab Med 2010;48:519–21.
Comparison of different depletion strategies for improving resolution of the human urine proteome
Authors: Magagnotti Cinzia , Fermo Isabella , Carletti Rose Mary , Ferrari Maurizio , Bachi Angela
Published: February 2010

Background: Urine, being an ultrafiltrate of plasma, is a rich source for biomarker discovery. Since potential new disease markers are often present in low concentrations, a prefractionation/enrichment step could be useful in the discovery process. To enhance the detection of low-abundance proteins, three immuno-affinity depletion approaches were evaluated.

Methods: To remove the most abundant proteins from a human urine sample, GenWay™ Spin IgY-12 kit, HPLC Agilent Hu-PL7 and a home-made column vs. human serum albumin [immuno-affinity column (IAC)] were compared. Quantification of total proteins, 2-D gel electrophoresis (2-DE), Progenesis gel images analysis and mass spectrometric proteins identification were applied to evaluate these strategies.

Results: Reproducibility of depletion columns, by estimating protein content of unbound fractions, were: 343±20.0 μg, 5.8%; 186.3±13.3 μg, 7.2%; 292±20.6 μg, 8.8% [mean± standard deviation (SD), CV%], for GenWay™, Agilent and IAC methods, respectively. To isolate urinary protein after depletion, ethanol precipitation provided the highest recovery (80%). Applying 2-DE and Progenesis analysis, the number of spots visualized on the gels was 468±21, 331±7, 368±22 and 304±7 (mean±SD) for GenWay™, Agilent, IAC, and the undepleted urine pool sample, respectively, with a significant difference p<0.001 compared to the GenWay procedure.

Conclusions: The sequential procedure of urine samples using multi-protein immuno-affinity depletion represents a valid tool for simplifying 2-DE analysis of the urine proteome. Particularly, the GenWay™ kit followed by ethanol precipitation was found to be the most efficient method for exploring the urine proteome.

Clin Chem Lab Med 2010;48:531–5.