bluuebunny/crossref_metadata_2025 huggingface dataset:
Redesigning a large-scale clinical trial in response to negative external trial results: the CAMUS study of Phytotherapy for Benign Prostatic Hyperplasia
Authors: Lee Jeannette , Andriole Gerald , Avins Andrew , Crawford E David , Foster Harris , Kaplan Steven , Kreder Karl , Kusek John , McCullough Andrew , McVary Kevin , Meleth Sreelatha , Naslund Michael , Nickel J Curtis , Nyberg Leroy , Roehrborn Claus , Dale Williams O. , Barry Michael
Published: December 2009

Background Benign prostatic hyperplasia (BPH), a common condition among older men, confers its morbidity through potentially bothersome lower urinary tract symptoms. Treatments for BPH include drugs such as alpha-adrenergic receptor blockers and 5-alpha reductase inhibitors, minimally invasive therapies that use heat to damage or destroy prostate tissue, and surgery including transurethral resection of the prostate. Complementary and alternative medicines are gaining popularity in the US. Two phytotherapies commonly used for BPH are extracts of the fruit of Serenoa repens, the Saw palmetto dwarf palm that grows in the Southeastern US, and extracts of the bark of Pygeum africanum, the African plum tree.

Purpose The objective of the Complementary and Alternative Medicines for Urological Symptoms (CAMUS) clinical trial is to determine if phytotherapy is superior to placebo in the treatment of BPH.

Methods CAMUS was originally designed as a 3300-participant, four-arm trial of S. repens, P. africanum, an alpha-adrenergic blocking drug, and placebo with time to clinical progression of BPH, a measure of long-term efficacy, as the primary endpoint. Before enrollment started, a randomized, double-blind, placebo-controlled, single institution clinical trial showed that S. repens at the usual dose did not demonstrate any benefit over placebo with respect to symptom relief at 1 year. Consequently, the focus of CAMUS shifted from evaluating long-term efficacy to determining if any short-term (6—18 months) symptom relief could be achieved with increasing doses of S. repens, the phytotherapy most commonly used in the US for BPH.

Results Results are anticipated in 2011.

Conclusions Trial design occurs in an environment of continually evolving information. In this case, emerging results from another trial suggested that a study of long-term efficacy was premature, and that an effective dose and preparation of S. repens had to be established before proceeding to a long-term clinical trial. Clinical Trials 2009; 6: 628—636. http://ctj.sagepub.com
Anomalous presentation of flexor carpi radialis brevis: a report of six cases
Authors: Mantovani G. , Lino W. , Fukushima W. Y. , Cho A. B. , Aita M. A.
Published: December 2009

An anomalous presentation of flexor carpi radialis brevis (FCRB) is reported in six patients. These findings occurred in a consecutive series of 172 distal radius fracture fixations using a volar approach between 2002 and 2007. This is the first report of this anomalous muscle from a clinical series.
Ulnar Variance And Scaphoid Fracture
Authors: Ramos-Escalona J. , García-Bordes L. , Martínez-Galarza P. , Yunta-Gallo A.
Published: December 2009

Between 1997 and 2006, radiographs of 66 scaphoid fractures were retrospectively reviewed to evaluate ulnar variance. Twenty-one (31.8%) patients had an ‘ulna neutral’ wrist, six (9.1%) had an ‘ulna plus’ and 39 (59.1%) had an ‘ulna minus’ wrist. The mean ulnar variance was −1.3 (SD 1.8) mm (range −5.5, 2.5). We observed a significant difference in the distribution of ulnar variance ( P < 0.00001) and in the proportion of cases with ulna minus (OR = 5.0; 95% CI: 2.7, 9.3) compared to previous publications.
A minimally invasive anterior approach to reduction and screw fixation of coronoid fractures
Authors: Kang L.-Q. , Ding Z.-Q. , Sha M. , Hong J.-Y. , Chen W.
Published: December 2009

We investigated a minimally invasive anterior approach to reduce and fix coronoid fractures with a screw in eight consecutive patients and evaluated seven patients at a minimum of 1 year (mean, 78 weeks; range, 61–89 weeks). It was only possible to repair seven of eight coronoid fractures. The average time to radiographic healing was 12 weeks and all healed without the screw loosening, migration or breakage. Endoscopy permitted an excellent view of coronoid fractures, allowing anatomic fixation, and as it avoided extensive soft tissue dissection, this method preserved soft tissue attachments of small coronoid fragments. This method was safe and minimally invasive.
The extensor tendon splitting approach to the proximal interphalangeal joint: do we need to reinsert the central slip?
Authors: Afifi A. M. , Richards A. , Medoro A. , Mercer D. , Moneim M.
Published: December 2009

Current approaches to the proximal interphalangeal (PIP) joint have potential complications and limitations. We present a dorsal approach that involves splitting the extensor tendon in the midline, detaching the insertion of the central slip and repairing the extensor tendon without reinserting the tendon into the base of the middle phalanx. A retrospective review of 16 digits that had the approach for a PIP joint arthroplasty with a mean follow up of 23 months found a postoperative PIP active ROM of 61° (range 25–90°). Fourteen digits had no extensor lag, while two digits had an extensor lag of 20° and 25°. This modified approach is fast and simple and does not cause an extensor lag.
The distribution of the types of thumb polydactyly in a middle eastern population: a study of 228 hands
Authors: Al-Qattan M. M.
Published: December 2009

Several series have reported the distribution of the types of thumb polydactyly in the Caucasian and Far Eastern populations. No data are available for the Middle East. A total of 196 Saudi patients (228 hands) with thumb polydactyly were reviewed. The most common type was Wassel type IV (33.8%) and the least common was Wassel type I (0.4%). A total of 26 hands (11.4%) did not fit into the classic Wassel types including 18 cases of rudimentary duplications, two cases of thumb triplication, and five cases of thumb duplication with symphalangism. Concurrent ipsilateral little finger triplication (one case) and contralateral thumb aplasia/hypoplasia (two cases) were seen and the developmental biology of these cases are discussed. After reviewing our results and other series in the literature, we concluded that Wassel types IV and I remain to be the most and least common types respectively in almost all races. However, the genetic pools of various races greatly affect the distribution of other non-classic duplications (unclassifiable by Wassel classification) such as rudimentary duplication in the Far East, triplication in the Southwestern region of the Netherlands, and symphalangism in Asia.
The association between intraoperative correction of Dupuytren’s disease and residual postoperative contracture
Authors: Donaldson O. W. , Pearson D. , Reynolds R. , Bhatia R. K.
Published: December 2009

The purpose of this study was to determine whether preoperative contracture and the amount of intraoperative correction can be used to predict the postoperative outcome of fasciectomy for Dupuytren’s disease. A prospective study of 52 patients undergoing primary fasciectomy during an 18 month period was undertaken. The contracture of each joint was measured preoperatively, after fasciectomy during the operation and 6 months after surgery. Forty-two metacarpophalangeal (MCPJ) and 58 proximal interphalangeal (PIPJ) joints were treated surgically. Full intraoperative correction was achieved in 41 MCPJs. Thirty-seven had full correction at follow-up. Full intraoperative correction was obtained in 35 PIPJs and 13 had complete correction at follow-up. The extent of the preoperative deformity was a significant predictor of complete intraoperative correction. The extent of both preoperative deformity and intraoperative correction were significant predictors of loss of surgical correction after operation.
Warm or refrigerated local anaesthetic for open carpal tunnel release: a single blind randomized controlled study
Authors: Tomlinson P. J. , Field J.
Published: December 2009

The aim of this randomized controlled study was to determine whether administration of lignocaine with adrenaline is less painful when injected at room temperature compared to refrigerated temperature. A cohort of 50 patients undergoing unilateral carpal tunnel decompression was randomized to room temperature or refrigerated local anaesthetic. Pain scores were assessed using a 10 mm visual analogue scale. Mean pain scores were 4.0 (SD ± 1.5) for room temperature and 6.5 (SD ± 1.7) for refrigerated local anaesthetic ( P < 0.001). This study demonstrates that patients experience greater pain levels with administration of local anaesthetic at refrigerated temperatures prior to open carpal tunnel release.
Mechanical properties and microstructures of magnesium alloy AZ31B joint fabricated by resistance spot welding with cover plates
Authors: Qiu R. F. , Satonaka S. , Iwamoto C.
Published: December 2009

The authors welded magnesium alloy AZ31B sheets using the technique of resistance spot welding with cover plates, and investigated the effects of welding parameters on the tensile shear strength of joints and shape characteristic of nugget. The joints with high tensile shear strength were obtained under relatively low welding current. The equiaxed grains with the many intragranularly precipitated particles Mg17Al12 in the nugget were observed.
New technique to control welding buckling distortion and residual stress with non-contact electromagnetic impact
Authors: Xu D. , Liu X. S. , Wang P. , Yang J. G. , Fang H. Y.
Published: December 2009

A new technique using non-contact electromagnetic forces has been proposed for controlling welding buckling distortion and residual stresses in welded thin plates. The experimental results show that the method can successfully eliminate the buckling distortion and reduce the residual stresses. Three-dimensional finite element modelling has been developed to study the evolution of the stress and strain throughout the welding and electromagnetic impacts. The predicted welding distortion and residual stresses are in good agreement with the experimental results. The numerical analyses show that the reduction in distortion and stress is a result of the change of the plastic strain field in the weld region: electromagnetic impacts reduce longitudinal compressive plastic strain in the local region near the weld, and even produce the tensile plastic strain. Moreover, it is found that the residual stress can promote the changes of the longitudinal plastic strain state under electromagnetic impact.
Ferrite to austenite reverse transformation process in B containing 9%Cr heat resistant steel HAZ
Authors: Shirane T. , Tsukamoto S. , Tsuzaki K. , Adachi Y. , Hanamura T. , Shimizu M. , Abe F.
Published: December 2009

Creep properties of the high Cr heat resistant steel welded joint can be improved by adding B due to prevention of the grain refinement in heat affected zone (HAZ). In the present study, phase transformation behaviour of the B steel HAZ has been investigated to understand suppression mechanism of the grain refinement. During reverse transformation, fine austenite was formed through diffusional transformation at the prior austenite grain boundary in the first stage, and then coarse austenite was formed at the same location of the original austenite. The volume fraction of the fine austenite increased with increasing perk temperature of the weld thermal cycle. This phenomenon can be explained if the coarse austenite contains high density of dislocations. Clear surface relief was observed during the reverse transformation by a confocal laser microscope. These results indicate that martensitic or displacive reverse transformation takes place during welding and it prevents the grain refinement in HAZ.
Weld shape variation and electrode protection under Ar–(Ar–O2) double shielded GTA welding
Authors: Lu S. P. , Fujii H. , Nogi K.
Published: December 2009

Double shielded gas tungsten arc welding (GTA welding or TIG welding) of an SUS304 stainless steel with pure inert argon as the inner layer shielding and the Ar–O2 active gas as the outer layer shielding is proposed in this study in order to investigate its effect on the tungsten electrode protection and the weld shape variation. The experimental results show that the inner inert argon gas can successfully prevent the outer layer active gas from contacting and oxidising the tungsten electrode during the welding process. The active gas, oxygen, in the outer layer shielding is decomposed in the arc and dissolves in the liquid pool, which effectively adjusts the active element, oxygen, content in the weld metal. When the weld metal oxygen content is over 70 ppm, the surface tension induced Marangoni convection changes from outward into inward, and the weld shape varies from a wide shallow one to a narrow deep one. The effect of the inner layer gas flowrate on the weld bead morphology and the weld shape is investigated systematically. The results showed that when the flowrate of the inner argon shielding gas is too low, the weld bead is easily oxidised and the weld shape is wide and shallow. A heavy continuous oxide layer on the liquid pool is a barrier to the liquid pool movement.
Transient Epstein-Barr virus reactivation in CD3 monoclonal antibody-treated patients
Authors: Keymeulen Bart , Candon Sophie , Fafi-Kremer Samira , Ziegler Anette , Leruez-Ville Marianne , Mathieu Chantal , Vandemeulebroucke Evy , Walter Markus , Crenier Laurent , Thervet Eric , Legendre Christophe , Pierard Denis , Hale Geoff , Waldmann Herman , Bach Jean-François , Seigneurin Jean Marie , Pipeleers Daniel , Chatenoud Lucienne
Published: December 2009

Here we report a unique situation in which an early and synchronized Epstein-Barr virus (EBV) reactivation was induced by a 6-day course of treatment with a humanized CD3-specific monoclonal antibody in patients with recent onset of type 1 diabetes. The virologic and immunologic analysis demonstrated that this reactivation was transient, self-limited, and isolated, associated with the rapid advent of an EBV-specific T-cell response. The anti-CD3 antibody administration induced short-lasting immunosuppression and minor yet clear-cut signs of T-cell activation that preceded viral reactivation. Early posttransplant monitoring of renal and islet allograft recipients showed that no comparable phenomenon was observed after the administration of full-dose immunosuppressive therapy. This EBV reactivation remains of no apparent clinical concern over the long term and should not preclude further development of therapeutic anti-CD3 antibodies. This phenomenon may also direct new research avenues to understand the still ill-defined nature of stimuli triggering EBV reactivation in vivo.
EF-hand domains of MCFD2 mediate interactions with both LMAN1 and coagulation factor V or VIII
Authors: Zheng Chunlei , Liu Hui-hui , Zhou Jiahai , Zhang Bin
Published: December 2009

Combined deficiency of factor V and factor VIII (F5F8D) is a bleeding disorder caused by mutations in either LMAN1 or MCFD2. LMAN1 (ERGIC-53) and MCFD2 form a Ca2+-dependent cargo receptor that cycles between the endoplasmic reticulum (ER) and the ER-Golgi intermediate compartment for efficient transport of FV/FVIII from the ER to the Golgi. Here we show that the C-terminal EF-hand domains are both necessary and sufficient for MCFD2 to interact with LMAN1. MCFD2 with a deletion of the entire N-terminal non-EF hand region still retains the LMAN1-binding function. Deletions that disrupt core structure of the EF-hand domains abolish LMAN1 binding. Circular dichroism spectroscopy studies on missense mutations localized to different structural elements of the EF-hand domains suggest that Ca2+-induced folding is important for LMAN1 interaction. The EF-hand domains also mediate the interaction with FV and FVIII. However, mutations in MCFD2 that disrupt the tertiary structure and abolish LMAN1 binding still retain the FV/FVIII binding activities, suggesting that this interaction is independent of Ca2+-induced folding of the protein. Our results suggest that the EF-hand domains of MCFD2 contain separate binding sites for LMAN1 and FV/FVIII that are essential for cargo receptor formation and cargo loading in the ER.
The P2X7-nonmuscle myosin membrane complex regulates phagocytosis of nonopsonized particles and bacteria by a pathway attenuated by extracellular ATP
Authors: Gu Ben J. , Saunders Bernadette M. , Jursik Claudia , Wiley James S.
Published: December 2009

Phagocytosis of nonopsonized bacteria is central to innate immunity, but its regulation is less defined. We show that overexpression of the P2X7 receptor greatly augments the phagocytosis of nonopsonized beads and heat-killed bacteria by transfected HEK-293 cells, whereas blocking P2X7 expression by siRNA significantly reduces the phagocytic ability of human monocytic cells. An intact P2X7-nonmuscle myosin complex is required for phagocytosis of nonopsonized beads because activation of P2X7 receptors by adenosine triphosphate (ATP), which dissociates myosin IIA from the P2X7 complex, inhibits this phagocytic pathway. Fresh human monocytes rapidly phagocytosed live and heat-killed Staphylococcus aureus and Escherichia coli in the absence of serum, but the uptake was reduced by prior incubation with ATP, or P2X7 monoclonal antibody, or recombinant P2X7 extracellular domain. Injection of beads or bacteria into the peritoneal cavity of mice resulted in their brisk phagocytosis by macrophages, but injection of ATP before particles markedly decreased this uptake. These data demonstrate a novel pathway of phagocytosis of nonopsonized particles and bacteria, which operate in vivo and require an intact P2X7-nonmuscle myosin IIA membrane complex. The inhibitory effect of ATP on particle uptake by the macrophage is regulated by the P2X7 receptor and defines this phagocytic pathway.
The emerging role of cardiovascular risk factor-induced mitochondrial dysfunction in atherogenesis
Authors: Puddu Paolo , Puddu Giovanni M , Cravero Eleonora , De Pascalis Susanna , Muscari Antonio
Published: December 2009

An important role in atherogenesis is played by oxidative stress, which may be induced by common risk factors. Mitochondria are both sources and targets of reactive oxygen species, and there is growing evidence that mitochondrial dysfunction may be a relevant intermediate mechanism by which cardiovascular risk factors lead to the formation of vascular lesions. Mitochondrial DNA is probably the most sensitive cellular target of reactive oxygen species. Damage to mitochondrial DNA correlates with the extent of atherosclerosis. Several cardiovascular risk factors are demonstrated causes of mitochondrial damage. Oxidized low density lipoprotein and hyperglycemia may induce the production of reactive oxygen species in mitochondria of macrophages and endothelial cells. Conversely, reactive oxygen species may favor the development of type 2 diabetes mellitus, mainly through the induction of insulin resistance. Similarly - in addition to being a cause of endothelial dysfunction, reactive oxygen species and subsequent mitochondrial dysfunction - hypertension may develop in the presence of mitochondrial DNA mutations. Finally, other risk factors, such as aging, hyperhomocysteinemia and cigarette smoking, are also associated with mitochondrial damage and an increased production of free radicals. So far clinical studies have been unable to demonstrate that antioxidants have any effect on human atherogenesis. Mitochondrial targeted antioxidants might provide more significant results.
An improved empirical bayes approach to estimating differential gene expression in microarray time-course data: BETR (Bayesian Estimation of Temporal Regulation)
Authors: Aryee Martin J , Gutiérrez-Pabello José A , Kramnik Igor , Maiti Tapabrata , Quackenbush John
Published: December 2009

Background

Microarray gene expression time-course experiments provide the opportunity to observe the evolution of transcriptional programs that cells use to respond to internal and external stimuli. Most commonly used methods for identifying differentially expressed genes treat each time point as independent and ignore important correlations, including those within samples and between sampling times. Therefore they do not make full use of the information intrinsic to the data, leading to a loss of power.

Results

We present a flexible random-effects model that takes such correlations into account, improving our ability to detect genes that have sustained differential expression over more than one time point. By modeling the joint distribution of the samples that have been profiled across all time points, we gain sensitivity compared to a marginal analysis that examines each time point in isolation. We assign each gene a probability of differential expression using an empirical Bayes approach that reduces the effective number of parameters to be estimated.

Conclusions

Based on results from theory, simulated data, and application to the genomic data presented here, we show that BETR has increased power to detect subtle differential expression in time-series data. The open-source R packagebetris available through Bioconductor. BETR has also been incorporated in the freely-available, open-source MeV software tool available fromhttp://www.tm4.org/mev.html.
Disruption of occludin function in polarized epithelial cells activates the extrinsic pathway of apoptosis leading to cell extrusion without loss of transepithelial resistance
Authors: Beeman Neal E , Baumgartner Heidi K , Webb Patricia G , Schaack Jerome B , Neville Margaret C
Published: December 2009

Background

Occludin is a tetraspanin protein normally localized to tight junctions. The protein interacts with a variety of pathogens including viruses and bacteria, an interaction that sometimes leads to its extrajunctional localization.

Results

Here we report that treatment of mammary epithelial monolayers with a circularized peptide containing a four amino acid sequence found in the second extracellular loop of occludin, LHYH, leads to the appearance of extrajunctional occludin and activation of the extrinsic apoptotic pathway. At early times after peptide treatment endogenous occludin and the LYHY peptide were co-localized in extrajunctional patches, which were also shown to contain components of the death inducing signaling complex (DISC), caspases 8 and 3, the death receptor FAS and the adaptor molecule FADD. After this treatment occludin could be immunoprecipitated with FADD, confirming its interaction with the DISC. Extrusion after LYHY treatment was accomplished with no loss of epithelial resistance.

Conclusion

These observations provide strong evidence that, following disruption, occludin forms a complex with the extrinsic death receptor leading to extrusion of apoptotic cells from the epithelial monolayer. They suggest that occludin has a protective as well as a barrier forming role in epithelia; pathogenic agents which utilize this protein as an entry point into the cell might set off an apoptotic reaction allowing extrusion of the infected cell before the pathogen can gain entry to the interstitial space.
An experimental assessment of in silico haplotype association mapping in laboratory mice
Authors: Burgess-Herbert Sarah L , Tsaih Shirng-Wern , Stylianou Ioannis M , Walsh Kenneth , Cox Allison J , Paigen Beverly
Published: December 2009

Background

To assess the utility of haplotype association mapping (HAM) as a quantitative trait locus (QTL) discovery tool, we conducted HAM analyses for red blood cell count (RBC) and high density lipoprotein cholesterol (HDL) in mice. We then experimentally tested each HAM QTL using published crosses or new F2 intercrosses guided by the haplotype at the HAM peaks.

Results

The HAM for RBC, using 33 classic inbred lines, revealed 8 QTLs; 2 of these were true positives as shown by published crosses. A HAM-guided (C57BL/6J × CBA/J)F2 intercross we carried out verified 2 more as true positives and 4 as false positives. The HAM for HDL, using 81 strains including recombinant inbred lines and chromosome substitution strains, detected 46 QTLs. Of these, 36 were true positives as shown by published crosses. A HAM-guided (C57BL/6J × A/J)F2 intercross that we carried out verified 2 more as true positives and 8 as false positives. By testing each HAM QTL for RBC and HDL, we demonstrated that 78% of the 54 HAM peaks were true positives and 22% were false positives. Interestingly, all false positives were in significant allelic association with one or more real QTL.

Conclusion

Because type I errors (false positives) can be detected experimentally, we conclude that HAM is useful for QTL detection and narrowing. We advocate the powerful and economical combined approach demonstrated here: the use of HAM for QTL discovery, followed by mitigation of the false positive problem by testing the HAM-predicted QTLs with small HAM-guided experimental crosses.
Identifying sexual differentiation genes that affect Drosophila life span
Authors: Shen Jie , Ford Daniel , Landis Gary N , Tower John
Published: December 2009

Background

Sexual differentiation often has significant effects on life span and aging phenotypes. For example, males and females of several species have different life spans, and genetic and environmental manipulations that affect life span often have different magnitude of effect in males versus females. Moreover, the presence of a differentiated germ-line has been shown to affect life span in several species, including Drosophila and C. elegans.

Methods

Experiments were conducted to determine how alterations in sexual differentiation gene activity might affect the life span of Drosophila melanogaster. Drosophila females heterozygous for the tudor[1] mutation produce normal offspring, while their homozygous sisters produce offspring that lack a germ line. To identify additional sexual differentiation genes that might affect life span, the conditional transgenic system Geneswitch was employed, whereby feeding adult flies or developing larvae the drug RU486 causes the over-expression of selected UAS-transgenes.

Results

In this study germ-line ablation caused by the maternal tudor[1] mutation was examined in a long-lived genetic background, and was found to increase life span in males but not in females, consistent with previous reports. Fitting the data to a Gompertz-Makeham model indicated that the maternal tudor[1] mutation increases the life span of male progeny by decreasing age-independent mortality. The Geneswitch system was used to screen through several UAS-type and EP-type P element mutations in genes that regulate sexual differentiation, to determine if additional sex-specific effects on life span would be obtained. Conditional over-expression of transformer female isoform (traF) during development produced male adults with inhibited sexual differentiation, however this caused no significant change in life span. Over-expression of doublesex female isoform (dsxF) during development was lethal to males, and produced a limited number of female escapers, whereas over-expression of dsxF specifically in adults greatly reduced both male and female life span. Similarly, over-expression of fruitless male isoform A (fru-MA) during development was lethal to both males and females, whereas over-expression of fru-MA in adults greatly reduced both male and female life span.

Conclusion

Manipulation of sexual differentiation gene expression specifically in the adult, after morphological sexual differentiation is complete, was still able to affect life span. In addition, by manipulating gene expression during development, it was possible to significantly alter morphological sexual differentiation without a significant effect on adult life span. The data demonstrate that manipulation of sexual differentiation pathway genes either during development or in adults can affect adult life span.
Expression of HIF-1alpha and VEGF in colorectal cancer: association with clinical outcomes and prognostic implications
Authors: Cao Dan , Hou Mei , Guan Yong-song , Jiang Ming , Yang Yu , Gou Hong-feng
Published: December 2009

Background

Hypoxia-inducible factor 1 alpha (HIF-1α) and vascular endothelial growth factor (VEGF) are frequently overexpressed in numerous types of cancers and are known to be important regulators of angiogenesis. Until now, few studies have been carried out to investigate the prognostic role of these factors in solid tumors, especially in colorectal cancer (CRC). The purpose of this study was to evaluate the expression of HIF-1α and VEGF in CRC tissues, and to analyze the association of these two factors with several clinical and pathological characteristics, and patients' survival.

Methods

Paraffin-embedded tissue samples were retrospectively collected from 71 CRC patients, who received surgical resection between 2001 and 2002, with a median follow-up of 5 years. We examined the patterns of expression of HIF-1α and VEGF by immunohistochemistry method. Statistical analysis was performed with univariate tests and multivariate Cox proportional hazards model to evaluate the differences.

Results

Expression of HIF-1α and VEGF was positively observed in 54.93% and 56.34% among the patients, respectively. HIF-1α and VEGF status were significantly associated with tumor stage, lymph nodes and liver metastases (P < 0.05). Expression of both HIF-1α and VEGF remained significantly associated with overall survival (OS) (P < 0.01), and HIF-1α was positively correlative to VEGF in CRC (r = 0.72, P < 0.001).

Conclusions

HIF-1α and VEGF could be used as biomarkers indicating tumors in advanced stage and independently implied poor prognosis in patients with CRC. Treatment that inhibits HIF-1α might be a promising targeted approach in CRC to exhibit its potential to improve outcomes in future perspective, just as VEGF targeting has proved to be.
Transcriptional profile of isoproterenol-induced cardiomyopathy and comparison to exercise-induced cardiac hypertrophy and human cardiac failure
Authors: Galindo Cristi L , Skinner Michael A , Errami Mounir , Olson L Danielle , Watson David A , Li Jing , McCormick John F , McIver Lauren J , Kumar Neil M , Pham Thinh Q , Garner Harold R
Published: December 2009

Background

Isoproterenol-induced cardiac hypertrophy in mice has been used in a number of studies to model human cardiac disease. In this study, we compared the transcriptional response of the heart in this model to other animal models of heart failure, as well as to the transcriptional response of human hearts suffering heart failure.

Results

We performed microarray analyses on RNA from mice with isoproterenol-induced cardiac hypertrophy and mice with exercise-induced physiological hypertrophy and identified 865 and 2,534 genes that were significantly altered in pathological and physiological cardiac hypertrophy models, respectively. We compared our results to 18 different microarray data sets (318 individual arrays) representing various other animal models and four human cardiac diseases and identified a canonical set of 64 genes that are generally altered in failing hearts. We also produced a pairwise similarity matrix to illustrate relatedness of animal models with human heart disease and identified ischemia as the human condition that most resembles isoproterenol treatment.

Conclusion

The overall patterns of gene expression are consistent with observed structural and molecular differences between normal and maladaptive cardiac hypertrophy and support a role for the immune system (or immune cell infiltration) in the pathology of stress-induced hypertrophy. Cross-study comparisons such as the results presented here provide targets for further research of cardiac disease that might generally apply to maladaptive cardiac stresses and are also a means of identifying which animal models best recapitulate human disease at the transcriptional level.
Trends in healthcare utilization among older Americans with colorectal cancer: A retrospective database analysis
Authors: Lang Kathleen , Lines Lisa M , Lee David W , Korn Jonathan R , Earle Craig C , Menzin Joseph
Published: December 2009

Background

Analyses of utilization trends (cost drivers) allow us to understand changes in colorectal cancer (CRC) costs over time, better predict future costs, identify changes in the use of specific types of care (eg, hospice), and provide inputs for cost-effectiveness models. This retrospective cohort study evaluated healthcare resource use among US Medicare beneficiaries diagnosed with CRC between 1992 and 2002.

Methods

Cohorts included patients aged 66+ newly diagnosed with adenocarcinoma of the colon (n = 52,371) or rectum (n = 18,619) between 1992 and 2002 and matched patients from the general Medicare population, followed until death or December 31, 2005. Demographic and clinical characteristics were evaluated by cancer subsite. Resource use, including the percentage that used each type of resource, number of hospitalizations, and number of hospital and skilled nursing facility days, was evaluated by stage and subsite. The number of office, outpatient, and inpatient visits per person-year was calculated for each cohort, and was described by year of service, subsite, and treatment phase. Hospice use rates in the last year of life were calculated by year of service, stage, and subsite for CRC patients who died of CRC.

Results

CRC patients (mean age: 77.3 years; 44.9% male) used more resources than controls in every category (P < .001), with the largest differences seen in hospital days and home health use. Most resource use (except hospice) remained relatively steady over time. The initial phase was the most resource intense in terms of office and outpatient visits. Hospice use among patients who died of CRC increased from 20.0% in 1992 to 70.5% in 2004, and age-related differences appear to have evened out in later years.

Conclusion

Use of hospice care among CRC decedents increased substantially over the study period, while other resource use remained generally steady. Our findings may be useful for understanding CRC cost drivers, tracking trends, and forecasting resource needs for CRC patients in the future.
Development of multiplex real-time PCR assays for identification of members of the Anopheles funestus species group
Authors: Vezenegho Samuel B , Bass Chris , Puinean Mirel , Williamson Martin S , Field Linda M , Coetzee Maureen , Koekemoer Lizette L
Published: December 2009

Background

The malaria vector and non-vector species of the Anopheles funestus group are morphologically very similar and accurate identification is required as part of effective control strategies. In the past, this has relied on morphological and cytogenetic methods but these have been largely superseded by a robust allele-specific PCR (AS-PCR). One disadvantage of AS-PCR is the requirement for post-PCR processing by gel electrophoresis of PCR products. In this study, three new high-throughput 'closed-tube' assays were developed and compared with the previously described AS-PCR technique.

Methods

Protocols for three fluorescence-based assays based on Melt Curve Analysis (MCA), High Resolution Melt (HRM) and TaqMan SNP genotyping were developed to detect and discriminate Anopheles parensis, Anopheles leesoni, Anopheles vaneedeni, Anopheles rivulorum and An. funestus s.s. The sensitivity and specificity of these assays were compared with the widely used AS-PCR in a blind trial using DNA extracted from wild-caught mosquitoes.

Results

The TaqMan assay proved to be the most sensitive and specific of the three new assays. The MCA and HRM assays initially gave promising results, but were more sensitive to both DNA quality and quantity and consequently showed a higher rate of incorrect identifications.

Conclusion

The TaqMan assay proved to be the most robust of the three protocols tested in this study. This assay very effectively identified all five members of the An. funestus group using fluorescently-labeled probes with distinct emission and excitation spectra allowing their independent detection in a single reaction. This method is at least as sensitive and specific as the gold standard AS-PCR approach and because it has no requirement for post-PCR processing is simpler and more rapid to run. The one disadvantage of the TaqMan assay is the cost of this assay, both in terms of initial capital outlay and running cost per sample, which is higher than AS-PCR. However, the cost of both the real-time PCR machine and fluorescently labelled probes required is falling and in the future the cost of this assay is likely to become closer to that of standard PCR.
Multiplex real-time quantitative PCR, microscopy and rapid diagnostic immuno-chromatographic tests for the detection of Plasmodium spp: performance, limit of detection analysis and quality assurance
Authors: Khairnar Krishna , Martin Donald , Lau Rachel , Ralevski Filip , Pillai Dylan R
Published: December 2009

Background

Accurate laboratory diagnosis of malaria species in returning travelers is paramount in the treatment of this potentially fatal infectious disease.

Materials and methods

A total of 466 blood specimens from returning travelers to Africa, Asia, and South/Central America with suspected malaria infection were collected between 2007 and 2009 at the reference public health laboratory. These specimens were assessed by reference microscopy, multipex real-time quantitative polymerase chain reaction (QPCR), and two rapid diagnostic immuno-chromatographic tests (ICT) in a blinded manner. Key clinical laboratory parameters such as limit of detection (LOD) analysis on clinical specimens by parasite stage, inter-reader variability of ICTs, staffing implications, quality assurance and cost analysis were evaluated.

Results

QPCR is the most analytically sensitive method (sensitivity 99.41%), followed by CARESTART (sensitivity 88.24%), and BINAXNOW (sensitivity 86.47%) for the diagnosis of malaria in returning travelers when compared to reference microscopy. However, microscopy was unable to specifically identifyPlasmodia spp. in 18 out of 170 positive samples by QPCR. Moreover, the 17 samples that were negative by microscopy and positive by QPCR were also positive by ICTs. Quality assurance was achieved for QPCR by exchanging a blinded proficiency panel with another reference laboratory. The Kappa value of inter-reader variability among three readers for BINAXNOW and CARESTART was calculated to be 0.872 and 0.898 respectively. Serial dilution studies demonstrated that the QPCR cycle threshold correlates linearly with parasitemia (R2= 0.9746) in a clinically relevant dynamic range and retains a LOD of 11 rDNA copies/μl forP. falciparum, which was several log lower than reference microscopy and ICTs. LOD for QPCR is affected not only by parasitemia but the parasite stage distribution of each clinical specimen. QPCR was approximately 6-fold more costly than reference microscopy.

Discussion

These data suggest that multiplex QPCR although more costly confers a significant diagnostic advantage in terms of LOD compared to reference microscopy and ICTs for all four species. Quality assurance of QPCR is essential to the maintenance of proficiency in the clinical laboratory. ICTs showed good concordance between readers however lacked sensitivity for non-falciparumspecies due to antigenic differences and low parasitemia.

Conclusion

Multiplex QPCR but not ICTs is an essential adjunct to microscopy in the reference laboratory detection of malaria species specifically due to the superior LOD. ICTs are better suited to the non-reference laboratory where lower specimen volumes challenge microscopy proficiency in the non-endemic setting.
A microarray-based system for the simultaneous analysis of single nucleotide polymorphisms in human genes involved in the metabolism of anti-malarial drugs
Authors: Hodel Eva Maria , Ley Serej D , Qi Weihong , Ariey Frédéric , Genton Blaise , Beck Hans-Peter
Published: December 2009

Background

In order to provide a cost-effective tool to analyse pharmacogenetic markers in malaria treatment, DNA microarray technology was compared with sequencing of polymerase chain reaction (PCR) fragments to detect single nucleotide polymorphisms (SNPs) in a larger number of samples.

Methods

The microarray was developed to affordably generate SNP data of genes encoding the human cytochrome P450 enzyme family (CYP) andN-acetyltransferase-2 (NAT2) involved in anti-malarial drug metabolisms and with known polymorphisms, i.e.CYP2A6, CYP2B6,CYP2C8,CYP2C9,CYP2C19,CYP2D6,CYP3A4,CYP3A5, andNAT2.

Results

For some SNPs, i.e.CYP2A6*2,CYP2B6*5,CYP2C8*3,CYP2C9*3/*5,CYP2C19*3,CYP2D6*4andNAT2*6/*7/*14, agreement between both techniques ranged from substantial to almost perfect (kappa index between 0.61 and 1.00), whilst for other SNPs a large variability from slight to substantial agreement (kappa index between 0.39 and 1.00) was found, e.g.CYP2D6*17(2850C>T),CYP3A4*1Band CYP3A5*3.

Conclusion

The major limit of the microarray technology for this purpose was lack of robustness and with a large number of missing data or with incorrect specificity.
A malaria outbreak in Naxalbari, Darjeeling district, West Bengal, India, 2005: weaknesses in disease control, important risk factors
Authors: Sharma Puran K , Ramanchandran Ramakrishnan , Hutin Yvan J , Sharma Raju , Gupte Mohan D
Published: December 2009

An outbreak of malaria in Naxalbari, West Bengal, India, in 2005 was investigated to understand determinants and propose control measures. Malaria cases were slide-confirmed. Methods included calculation of annual blood examination rates (ABER, number of slides examined/population), collection of water specimens from potential vector-breeding sites, sorting of villages in categories depending on the number of abandoned wells within two kilometers radius and review of the DDT spray coverage. Cases were compared with matched neighbourhood controls in terms of personal protection using matched odds ratios (MOR). 7,303 cases and 17 deaths were reported between April 2005 and March 2006 with a peak during October rains (Attack rate: 50 per 1,000, case fatality: 0.2%). The attack rate increased according to the number of abandoned wells within 2 kilometres radius (P < 0.0001, Chi-square for trend). Abandoned wells were Anopheles breeding sites. Compared with controls, cases were more likely to sleep outdoors (MOR: 3.8) and less likely to use of mosquito nets and repellents (MOR: 0.3 and 0.1, respectively). DDT spray coverage and ABER were 39% and 3.5%, below the recommended 85% and 10%, respectively. Overall, this outbreak resulted from weaknesses in malaria control measures and a combination of factors, including vector breeding, low implementation of personal protection and weak case detection.
CENPA overexpression promotes genome instability in pRb-depleted human cells
Authors: Amato Angela , Schillaci Tiziana , Lentini Laura , Di Leonardo Aldo
Published: December 2009

Background

Aneuploidy is a hallmark of most human cancers that arises as a consequence of chromosomal instability and it is frequently associated with centrosome amplification. Functional inactivation of the Retinoblastoma protein (pRb) has been indicated as a cause promoting chromosomal instability as well centrosome amplification. However, the underlying molecular mechanism still remains to be clarified.

Results

Here we show that pRb depletion both in wild type and p53 knockout HCT116 cells was associated with the presence of multipolar spindles, anaphase bridges, lagging chromosomes and micronuclei harbouring whole chromosomes. In addition aneuploidy caused by pRb acute loss was not affected by p53 loss.

Quantitative real-time RT-PCR showed that pRB depletion altered expression of genes involved in centrosome duplication, kinetochore assembly and in the Spindle Assembly Checkpoint (SAC). However, despite MAD2 up-regulation pRb-depleted cells seemed to have a functional SAC since they arrested in mitosis after treatments with mitotic poisons. Moreover pRb-depleted HCT116 cells showed BRCA1 overexpression that seemed responsible for MAD2 up-regulation.

Post-transcriptional silencing of CENPA by RNA interference, resulting in CENP-A protein levels similar to those present in control cells greatly reduced aneuploid cell numbers in pRb-depleted cells.

Conclusion

Altogether our findings indicate a novel aspect of pRb acute loss that promotes aneuploidy mainly by inducing CENPA overexpression that in turn might induce micronuclei by affecting the correct attachment of spindle microtubules to kinetochores.
Direct rosiglitazone action on steroidogenesis and proinflammatory factor production in human granulosa-lutein cells
Authors: Chen Qiuju , Sun Xiaoxi , Chen Junling , Cheng Linan , Wang Jian , Wang Yongwei , Sun Zhaogui
Published: December 2009

Background

Ovarian granulosa cells are the predominant source of estradiol and progesterone biosynthesis in vivo. Rosiglitazone, a synthetic agonist of the peroxisome proliferator-activated receptor gamma (PPAR gamma), is applied as the treatment of insulin resistance including women with PCOS. The aim of the study was to investigate the direct effects of rosiglitazone on steroidogenesis and proinflammatory factor production in human granulosa-lutein cells (GLCs).

Methods

Primary human GLCs were separated during in vitro fertilization and cultured in the presence of rosiglitazone, GW9662 (an antagonist of PPAR gamma) and hCG. The mRNA expression of key steroidogenic factors including 3beta- hydroxysteriod dehydrogenase (3beta-HSD), cytochrome P-450 scc (CYP11A1), cytochrome P-450 aromatase (CYP19A1), and steroidogenic acute regulatory protein (StAR) were detected by quantitative real-time PCR. Estradiol and progesterone levels in GLCs cultures were measured by chemiluminescence immunoassay, and the proinflammtory factors (TNFalpha and IL-6) in conditioned culture media were measured by ELISA.

Results

PPAR gamma mRNA levels increased up to 3.24 fold by rosiglitazone at the concentration of 30 microM compared to control (P < 0.05). hCG alone or hCG with rosiglitazone had no significant effects on PPAR gamma mRNA levels. The CYP19A1 mRNA level at exposure to rosiglitazone alone showed a drop, but was not significantly reduced comparing to control. The expression levels of enzymes 3beta-HSD and CYP11A1 in all treatments did not alter significantly. The StAR mRNA expression at exposure to rosiglitazone was significantly increased comparing to control (P < 0.05). The media concentrations of E2 and progesterone by rosiglitazone treatment showed a declining trend comparing to control or cotreatment with hCG, which did not reach significance. Most importantly, treatment with rosiglitazone decreased TNFalpha secretion in a statistically significant manner compared with control (P < 0.05). The concentration of IL-6 following rosiglitazone exposure did not significantly decrease comparing to control.

Conclusion

In cultured GLCs, rosiglitazone stimulated StAR expression, but did not significantly affect steroidogenic enzymes, as well as E2 and progesterone production. Moreover, rosiglitazone significantly decreased the production of TNFalpha in human GLCs, suggesting that PPAR gamma may play a role in the regulation of GLCs functions through inhibiting proinflammatory factors.