bluuebunny/crossref_metadata_2025 huggingface dataset:
Genes involved in TGFβ1-driven epithelial-mesenchymal transition of renal epithelial cells are topologically related in the human interactome map
Authors: Campanaro Stefano , Picelli Simone , Torregrossa Rossella , Colluto Laura , Ceol Monica , Del Prete Dorella , D'Angelo Angela , Valle Giorgio , Anglani Franca
Published: October 2007

Background

Understanding how mesenchymal cells arise from epithelial cells could have a strong impact in unveiling mechanisms of epithelial cell plasticity underlying kidney regeneration and repair.

In primary human tubular epithelial cells (HUTEC) under different TGFβ1 concentrations we had observed epithelial-to-mesenchymal transition (EMT) but not epithelial-myofibroblast transdifferentiation. We hypothesized that the process triggered by TGFβ1 could be a dedifferentiation event. The purpose of this study is to comprehensively delineate genetic programs associated with TGFβ1-driven EMT in our in vitro model using gene expression profile on large-scale oligonucleotide microarrays.

Results

In HUTEC under TGFβ1 stimulus, 977 genes were found differentially expressed. Thirty genes were identified whose expression depended directly on TGFβ1 concentration. By mapping the differentially expressed genes in the Human Interactome Map using Cytoscape software, we identified a single scale-free network consisting of 2630 interacting proteins and containing 449 differentially expressed proteins. We identified 27 hub proteins in the interactome with more than 29 edges incident on them and encoded by differentially expressed genes. The Gene Ontology analysis showed an excess of up-regulated proteins involved in biological processes, such as "morphogenesis", "cell fate determination" and "regulation of development", and the most up-regulated genes belonged to these categories. In addition, 267 genes were mapped to the KEGG pathways and 14 pathways with more than nine differentially expressed genes were identified. In our model, Smad signaling was not the TGFβ1 action effector; instead, the engagement of RAS/MAPK signaling pathway seems mainly to regulate genes involved in the cell cycle and proliferation/apoptosis.

Conclusion

Our present findings support the hypothesis that context-dependent EMT generated in our model by TGFβ1 might be the outcome of a dedifferentiation. In fact: 1) the principal biological categories involved in the process concern morphogenesis and development; 2) the most up-regulated genes belong to these categories; and, finally, 3) some intracellular pathways are involved, whose engagement during kidney development and nephrogenesis is well known. These long-term effects of TGFβ1 in HUTEC involve genes that are highly interconnected, thereby generating a scale-free network that we named the "TGFβ1 interactome", whose hubs represent proteins that may have a crucial role for HUTEC in response to TGFβ1.
A systems-biology analysis of isogenic megakaryocytic and granulocytic cultures identifies new molecular components of megakaryocytic apoptosis
Authors: Chen Chi , Fuhrken Peter G , Huang Li Ting , Apostolidis Pani , Wang Min , Paredes Carlos J , Miller William M , Papoutsakis Eleftherios T
Published: October 2007

Background

The differentiation of hematopoietic stem cells into platelet-forming megakaryocytes is of fundamental importance to hemostasis. Constitutive apoptosis is an integral, yet poorly understood, facet of megakaryocytic (Mk) differentiation. Understanding Mk apoptosis could lead to advances in the treatment of Mk and platelet disorders.

Results

We used a Gene-ontology-driven microarray-based transcriptional analysis coupled with protein-level and activity assays to identify genes and pathways involved in Mk apoptosis. Peripheral blood CD34+ hematopoietic progenitor cells were induced to either Mk differentiation or, as a negative control without observable apoptosis, granulocytic differentiation. Temporal gene-expression data were analyzed by a combination of intra- and inter-culture comparisons in order to identify Mk-associated genes. This novel approach was first applied to a curated set of general Mk-related genes in order to assess their dynamic transcriptional regulation. When applied to all apoptosis associated genes, it revealed a decrease in NF-κB signaling, which was explored using phosphorylation assays for IκBα and p65 (RELA). Up-regulation was noted among several pro-apoptotic genes not previously associated with Mk apoptosis such as components of the p53 regulon and TNF signaling. Protein-level analyses probed the involvement of the p53-regulated GADD45A, and the apoptosis signal-regulating kinase 1 (ASK1). Down-regulation of anti-apoptotic genes, including several of the Bcl-2 family, was also detected.

Conclusion

Our comparative approach to analyzing dynamic large-scale transcriptional data, which was validated using a known set of Mk genes, robustly identified candidate Mk apoptosis genes. This led to novel insights into the molecular mechanisms regulating apoptosis in Mk cells.
Ecological study of Paracoccidioides brasiliensis in soil: growth ability, conidia production and molecular detection
Authors: Terçarioli Gisela Ramos , Bagagli Eduardo , Reis Gabriela Martins , Theodoro Raquel Cordeiro , Bosco Sandra De Moraes Gimenes , Macoris Severino Assis da Graça , Richini-Pereira Virgínia Bodelão
Published: October 2007

Background

Paracoccidioides brasiliensis ecology is not completely understood, although several pieces of evidence point to the soil as its most probable habitat. The present study aimed to investigate the fungal growth, conidia production and molecular pathogen detection in different soil conditions.

Methods

Soils samples of clayey, sandy and medium textures were collected from ground surface and the interior of armadillo burrows in a hyperendemic area of Paracoccidioidomycosis. P. brasiliensis was inoculated in soil with controlled humidity and in culture medium containing soil extracts. The molecular detection was carried out by Nested PCR, using panfungal and species specific primers from the ITS-5.8S rDNA region.

Results

The soil texture does not affect fungus development and the growth is more abundant on/in soil saturated with water. Some soil samples inhibited the development of P. brasiliensis, especially those that contain high values of Exchangeable Aluminum (H+Al) in their composition. Some isolates produced a large number of conidia, mainly in soil-extract agar medium. The molecular detection was positive only in samples collected from armadillo burrows, both in sandy and clayey soil.

Conclusion

P. brasiliensis may grow and produce the infectious conidia in sandy and clayey soil, containing high water content, mainly in wild animal burrows, but without high values of H+Al.
luxRI homologs are universally present in the genus Aeromonas
Authors: Jangid Kamlesh , Kong Richard , Patole Milind S , Shouche Yogesh S
Published: October 2007

Background

Aeromonas spp. have been regarded as "emerging pathogens". Aeromonads possess multifactorial virulence and the production of many of these virulence determinants is associated with high cell density, a phenomenon that might be regulated by quorum sensing. However, only two species of the genus are reported to possess the luxRI quorum sensing gene homologs. The purpose of this study was to investigate if the luxRI homologs are universally present in the Aeromonas strains collected from various culture collections, clinical laboratories and field studies.

Results

Of all the 73 Aeromonas strains used in the study, seventy-one strains elicited acyl-homoserine lactone-mediated response in multiple biosensor strains. However, dot blot hybridization revealed that the luxRI homologs are present in all the strains. PCR amplification and sequencing revealed that the luxRI homologs shared a very high percentage sequence similarity. No evidence for lateral gene transfer of the luxRI homologs between aeromonads and other genera was noted.

Conclusion

We propose that the luxRI quorum sensing gene homologs are universally present in the genus Aeromonas independently from their origin. This study is the first genus-wide report of the taxonomic distribution of the luxRI homologs.
Factors associated with good self-rated health of non-disabled elderly living alone in Japan: a cross-sectional study
Authors: Sun Wei , Watanabe Misuzu , Tanimoto Yoshimi , Shibutani Takahiro , Kono Rei , Saito Masahisa , Usuda Kan , Kono Koichi
Published: October 2007

Background

Self-rated health (SRH) is reported as a reliable predictor of disability and mortality in the aged population and has been studied worldwide to enhance the quality of life of the elderly. Nowadays, the elderly living alone, a particular population at great risk of suffering physical and mental health problems, is increasing rapidly in Japan and could potentially make up the majority of the aged population. However, few data are available pertaining to SRH of this population. Given the fact that sufficient healthcare is provided to the disabled elderly whereas there is little support for non-disabled elderly, we designed this population-based survey to investigate SRH of non-disabled elderly living alone and to identify the factors associated with good SRH with the purpose of aiding health promotion for the elderly.

Methods

A cross-sectional study was conducted in a metropolitan suburb in Japan. Questionnaires pertaining to SRH and physical conditions, lifestyle factors, psychological status, and social activities, were distributed in October 2005 to individuals aged ≥ 65 years and living alone. Response rate was 75.1%. Among these respondents, a total of 600 male and 2587 female respondents were identified as non-disabled elderly living alone and became our subjects. Multivariate logistic regression was used to identify the factors associated with good SRH and sex-specific effect was tested by stepwise logistic regression.

Results

Good SRH was reported by 69.8% of men and 73.8% of women. Multivariate logistic regression analysis showed that good SRH correlated with, in odds ratio sequence, "can go out alone to distant places", no depression, no weight loss, absence of self-rated chronic disease, good chewing ability, and good visual ability in men; whereas with "can go out alone to distant places", absence of self-rated chronic disease, no weight loss, no depression, no risk of falling, independent IADL, good chewing ability, good visual ability, and social integration (attend) in women.

Conclusion

For the non-disabled elderly living alone, sex-appropriate support should be considered by health promotion systems from the view point of SRH. Overall, the ability to go out alone to distant places is crucial to SRH of both men and women.
The structure of Mycobacteria 2C-methyl-D-erythritol-2,4-cyclodiphosphate synthase, an essential enzyme, provides a platform for drug discovery
Authors: Buetow Lori , Brown Amanda C , Parish Tanya , Hunter William N
Published: October 2007

Background

The prevalence of tuberculosis, the prolonged and expensive treatment that this disease requires and an increase in drug resistance indicate an urgent need for new treatments. The 1-deoxy-D-xylulose 5-phosphate pathway of isoprenoid precursor biosynthesis is an attractive chemotherapeutic target because it occurs in many pathogens, including Mycobacterium tuberculosis, and is absent from humans. To underpin future drug development it is important to assess which enzymes in this biosynthetic pathway are essential in the actual pathogens and to characterize them.

Results

The fifth enzyme of this pathway, encoded by ispF, is 2C-methyl-D-erythritol-2,4-cyclodiphosphate synthase (IspF). A two-step recombination strategy was used to construct ispF deletion mutants in M. tuberculosis but only wild-type double crossover strains were isolated. The chromosomal copy could be deleted when a second functional copy was provided on an integrating plasmid, demonstrating that ispF is an essential gene under the conditions tested thereby confirming its potential as a drug target. We attempted structure determination of the M. tuberculosis enzyme (Mt IspF), but failed to obtain crystals. We instead analyzed the orthologue M. smegmatis IspF (Ms IspF), sharing 73% amino acid sequence identity, at 2.2 Å resolution. The high level of sequence conservation is particularly pronounced in and around the active site. Ms IspF is a trimer with a hydrophobic cavity at its center that contains density consistent with diphosphate-containing isoprenoids. The active site, created by two subunits, comprises a rigid CDP-Zn2+ binding pocket with a flexible loop to position the 2C-methyl-D-erythritol moiety of substrate. Sequence-structure comparisons indicate that the active site and interactions with ligands are highly conserved.

Conclusion

Our study genetically validates Mt IspF as a therapeutic target and provides a model system for structure-based ligand design.
AhR transcriptional activity in serum of Inuits across Greenlandic districts
Authors: Long Manhai , Deutch Bente , Bonefeld-Jorgensen Eva C
Published: October 2007

Background

Human exposure to lipophilic persistent organic pollutants (POPs) including polychlorinated dibenzo-p-dioxins/furans (PCDDs/PCDFs), polychlorinated biphenyls (PCBs) and organochlorine pesticide is ubiquitous. The individual is exposed to a complex mixture of POPs being life-long beginning during critical developmental windows. Exposure to POPs elicits a number of species- and tissue-specific toxic responses, many of which involve the aryl hydrocarbon receptor (AhR). The aim of this study was to compare the actual level of integrated AhR transcriptional activity in the lipophilic serum fraction containing the actual POP mixture among Inuits from different districts in Greenland, and to evaluate whether the AhR transactivity is correlated to the bio-accumulated POPs and/or lifestyle factors.

Methods

The study included 357 serum samples from the Greenlandic districts: Nuuk and Sisimiut (South West Coast), Qaanaaq (North Coast) and Tasiilaq (East Coast). The bio-accumulated serum POPs were extracted by ethanol: hexane and clean-up on Florisil columns. Effects of the serum extract on the AhR transactivity was determined using the Hepa 1.12cR mouse hepatoma cell line carrying an AhR-luciferase reporter gene, and the data was evaluated for possible association to the serum levels of 14 PCB congeners, 10 organochlorine pesticide residues and/or lifestyle factors.

Results

In total 85% of the Inuit samples elicited agonistic AhR transactivity in a district dependent pattern. The median level of the AhR-TCDD equivalent (AhR-TEQ) of the separate genders was similar in the different districts. For the combined data the order of the median AhR-TEQ was Tasiilaq > Nuuk ≥ Sisimiut > Qaanaaq possibly being related to the different composition of POPs. In overall, the AhR transactivity was inversely correlated to the levels of sum POPs, age and/or intake of marine food.

Conclusion

i) We observed that the proportion of dioxin like (DL) compounds in the POP mixture was the dominating factor affecting the level of serum AhR transcriptional activity even at very high level of non DL-PCBs; ii) The inverse association between the integrated serum AhR transactivity and sum of POPs might be explained by the higher level of compounds antagonizing the AhR function probably due to selective POP bioaccumulation in the food chain.
Warning: Anti-tobacco activism may be hazardous to epidemiologic science
Authors: Phillips Carl V
Published: October 2007

This commentary accompanies two articles submitted to Epidemiologic Perspectives & Innovations in response to a call for papers about threats to epidemiology or epidemiologists from organized political interests. Contrary to our expectations, we received no submissions that described threats from industry or government; all were about threats from anti-tobacco activists. The two we published, by James E. Enstrom and Michael Siegel, both deal with the issue of environmental tobacco smoke. This commentary adds a third story of attacks on legitimate science by anti-tobacco activists, the author's own experience. These stories suggest a willingness of influential anti-tobacco activists, including academics, to hurt legitimate scientists and turn epidemiology into junk science in order to further their agendas. The willingness of epidemiologists to embrace such anti-scientific influences bodes ill for the field's reputation as a legitimate science.
Radical stereotactic radiosurgery with real-time tumor motion tracking in the treatment of small peripheral lung tumors
Authors: Collins Brian T , Erickson Kelly , Reichner Cristina A , Collins Sean P , Gagnon Gregory J , Dieterich Sonja , McRae Don A , Zhang Ying , Yousefi Shadi , Levy Elliot , Chang Thomas , Jamis-Dow Carlos , Banovac Filip , Anderson Eric D
Published: October 2007

Background

Recent developments in radiotherapeutic technology have resulted in a new approach to treating patients with localized lung cancer. We report preliminary clinical outcomes using stereotactic radiosurgery with real-time tumor motion tracking to treat small peripheral lung tumors.

Methods

Eligible patients were treated over a 24-month period and followed for a minimum of 6 months. Fiducials (3–5) were placed in or near tumors under CT-guidance. Non-isocentric treatment plans with 5-mm margins were generated. Patients received 45–60 Gy in 3 equal fractions delivered in less than 2 weeks. CT imaging and routine pulmonary function tests were completed at 3, 6, 12, 18, 24 and 30 months.

Results

Twenty-four consecutive patients were treated, 15 with stage I lung cancer and 9 with single lung metastases. Pneumothorax was a complication of fiducial placement in 7 patients, requiring tube thoracostomy in 4. All patients completed radiation treatment with minimal discomfort, few acute side effects and no procedure-related mortalities. Following treatment transient chest wall discomfort, typically lasting several weeks, developed in 7 of 11 patients with lesions within 5 mm of the pleura. Grade III pneumonitis was seen in 2 patients, one with prior conventional thoracic irradiation and the other treated with concurrent Gefitinib. A small statistically significant decline in the mean % predicted DLCO was observed at 6 and 12 months. All tumors responded to treatment at 3 months and local failure was seen in only 2 single metastases. There have been no regional lymph node recurrences. At a median follow-up of 12 months, the crude survival rate is 83%, with 3 deaths due to co-morbidities and 1 secondary to metastatic disease.

Conclusion

Radical stereotactic radiosurgery with real-time tumor motion tracking is a promising well-tolerated treatment option for small peripheral lung tumors.
Traumatic pulmonary pseuodocysts: two case reports
Authors: Kocer Bulent , Gulbahar Gultekin , Gunal Nesimi , Dural Koray , Sakinci Unal
Published: October 2007

Traumatic pulmonary pseudocyst (TPP) is a rare complication, sometimes encountered after blunt thoracic trauma and even more rarely following penetrating injuries. It is more common among pediatric and young adult patients. Although TPP is usually benign in nature, complications associated with hemoptysis and secondary infection may develop. The treatment is conservative. In this report, we present two rare cases of TPP occuring after a high-speed accident and a stab wound injury, where conservative treatment provided good outcomes.
Steering control of six-wheeled vehicles using linear quadratic regulator techniques
Authors: Chen B-C , Yu C-C , Hsu W-F
Published: October 2007

The middle- and rear-wheel steering angles of a six-wheeled vehicle need to be coordinated with the front-wheel steering angle to obtain the maximum manoeuvrability. A steering control strategy using the linear quadratic regulator technique with integral control is proposed in this paper such that both zero side-slip angle and target yaw rate following can be achieved simultaneously. An estimator to be used with the control law is also designed to provide the estimate of side-slip angle. AutoSim is used to establish a complex vehicle model with tyre dynamics in MATLAB/Simulink. Both open-loop and closed-loop manoeuvres are performed to evaluate the control performance of the proposed strategy.
Pseudo-transient computational fluid dynamics analysis of an underbonnet compartment during thermal soak
Authors: Franchetta M , Suen K O , Bancroft T G
Published: October 2007

Underbonnet simulations are proving to be crucially important within a vehicle development programme, reducing test work and time-to-market. While computational fluid dynamics (CFD) simulations of steady forced flows have been demonstrated to be reliable, studies of transient convective flows in engine compartments are not yet carried out owing to high computing demands and lack of validated work. The present work assesses the practical feasibility of applying the CFD tool at the initial stage of a vehicle development programme for investigating the thermally driven flow in an engine bay under thermal soak.

A computation procedure that enables pseudo time-marching CFD simulations to be performed with significantly reduced central processing unit (CPU) time usage is proposed. The methodology was initially tested on simple geometries and then implemented for investigating a simplified half-scale underbonnet compartment. The numerical results are compared with experimental data taken with thermocouples and with particle image velocimetry (PIV).

The novel computation methodology is successful in efficiently providing detailed and time-accurate time-dependent thermal and flow predictions. Its application will extend the use of the CFD tool for transient investigations, enabling improvements to the component packaging of engine bays and the refinement of thermal management strategies with reduced need for in-territory testing.
Bilateral hip reconstruction in severe whole-body cerebral palsy
Authors: Barakat M. J. , While T. , Pyman J. , Gargan M. , Monsell F.
Published: October 2007

The results of a functional, clinical and radiological study of 30 children (60 hips) with whole-body cerebral palsy were reviewed at a mean follow-up of 10.2 years (9.5 to 11). Correction of windsweep deformity of the hips was performed by bilateral simultaneous combined soft-tissue and bony surgery at a mean age of 7.7 years (3.1 to 12.2).

We were able to recall 22 patients; five had died of unrelated causes and three were lost to follow-up. Evaluation involved interviews with patients/carers and clinical and radiological examination.

The gross motor functional classification system was used to assess overall motor function and showed improvement in seven patients. Of the 12 patients thought to have pain pre-operatively, only one had pain post-operatively. Improved handling was reported in 18 of 22 patients (82%). Those with handling problems were attributed by the carers to growth of the patients. All patients/carers considered the procedure worthwhile. The range of hip movements improved, and the mean windsweep index improved from 50 pre-operatively to 36 at follow-up.

The migration percentage and centre-edge angle were assessed on plain radiographs. Radiological containment improved, the mean migration percentage improved from 50 pre-operatively to 20 at follow-up and the mean centre-edge angle improved from −5° to 29°.

No statistical difference was noted between the three-year and ten-year follow-up results, indicating that the improvements in clinical and radiological outcome had been maintained.
Ten-year life expectancy after primary total hip replacement
Authors: Ramiah R. D. , Ashmore A. M. , Whitley E. , Bannister G. C.
Published: October 2007

We determined the ten-year life expectancy of 5831 patients who had undergone 6653 elective primary total hip replacements at a regional orthopaedic centre between April 1993 and October 2004. Using hospital, general practitioner and the local health authority records, we recorded the dates of death for those who died following surgery.

The mean age at operation was 67 years (13 to 96) with a male:female ratio of 2:3. Of 1154 patients with a ten-year follow-up 340 (29.5%) had died a mean of 5.6 years (0 to 10) after surgery. Using Kaplan-Meier curves, the ten-year survival was 89% in patients under 65 years at surgery, 75% in patients aged between 65 and 74 years, and 51% in patients over 75.

The standardised mortality rates were considerably higher for patients under 45 years, 20% higher for those between 45 and 64 years, and steadily reduced in patients aged 65 and over.

The survival of cemented hip replacement derived from the Swedish Hip Arthroplasty Register Annual Report 2004 exceeds the life expectancy of patients over the age of 60 in our area, suggesting that cemented hip replacement is the procedure of choice in this population.
Viability and apoptosis of human chondrocytes in osteochondral fragments following joint trauma
Authors: Hembree W. C. , Ward B. D. , Furman B. D. , Zura R. D. , Nichols L. A. , Guilak F. , Olson S. A.
Published: October 2007

Post-traumatic arthritis is a frequent consequence of articular fracture. The mechanisms leading to its development after such injuries have not been clearly delineated. A potential contributing factor is decreased viability of the articular chondrocytes. The object of this study was to characterise the regional variation in the viability of chondrocytes following joint trauma. A total of 29 osteochondral fragments from traumatic injuries to joints that could not be used in articular reconstruction were analysed for cell viability using the fluorescence live/dead assay and for apoptosis employing the TUNEL assay, and compared with cadaver control fragments.

Chondrocyte death and apoptosis were significantly greater along the edge of the fracture and in the superficial zone of the osteochondral fragments. The middle and deep zones demonstrated significantly higher viability of the chondrocytes. These findings indicate the presence of both necrotic and apoptotic chondrocytes after joint injury and may provide further insight into the role of chondrocyte death in post-traumatic arthritis.
Bone and joint tuberculosis
Authors: Sandher D. S. , Al-Jibury M. , Paton R. W. , Ormerod L. P.
Published: October 2007

We report 79 cases of bone and joint tuberculosis between 1988 and 2005, eight of which were in the Caucasian population and 71 in the non-white population.

The diagnosis was made in the majority (73.4%) by positive bacteriology and/or histology. The mean age at the time of diagnosis was higher in the Caucasian group at 51.5 years (28 to 66) than in the South Asian group at 36.85 years (12 to 93). Only one patient had previous BCG immunisation.

The spine was the site most commonly affected (44.3%). Surgical stabilisation and/or decompression was performed in 23% of these cases because of cord compression on imaging or the presence of neurological signs.

A six-month course of chemotherapy comprising of an initial two months of rifampicin, isoniazide, pyrazinamide and sometimes ethambutol followed by four months treatment with rifampicin and isoniazide, was successful in all cases without proven drug resistance.
The anterolateral approach leads to less disruption of the femoral head-neck blood supply than the posterior approach during hip resurfacing
Authors: Steffen R. , O’Rourke K. , Gill H. S. , Murray D. W.
Published: October 2007

In 12 patients, we measured the oxygen concentration in the femoral head-neck junction during hip resurfacing through the anterolateral approach. This was compared with previous measurements made for the posterior approach. For the anterolateral approach, the oxygen concentration was found to be highly dependent upon the position of the leg, which was adjusted during surgery to provide exposure to the acetabulum and femoral head. Gross external rotation of the hip gave a significant decrease in oxygenation of the femoral head. Straightening the limb led to recovery in oxygen concentration, indicating that the blood supply was maintained. The oxygen concentration at the end of the procedure was not significantly different from that at the start.

The anterolateral approach appears to produce less disruption to the blood flow in the femoral head-neck junction than the posterior approach for patients undergoing hip resurfacing. This may be reflected subsequently in a lower incidence of fracture of the femoral neck and avascular necrosis.
Does bone quality predict loosening of cemented total hip replacements?
Authors: Nixon M. , Taylor G. , Sheldon P. , Iqbal S. J. , Harper W.
Published: October 2007

We matched 78 patients with a loose cemented Charnley Elite Plus total hip replacement (THR) by age, gender, race, prosthesis and time from surgery with 49 patients with a well-fixed stable hip replacement, to determine if poor bone quality predisposes to loosening. Clinical, radiological, biomechanical and bone mineral density indicators of bone quality were assessed.

Patients with loose replacements had more pain, were more likely to have presented with atrophic arthritis and to have a history of fragility fracture, narrower femoral cortices and lower peri-prosthetic or lumbar spine bone mineral density (all t-test, p < 0.01). They also tended to be smokers (chi-squared test, p = 0.08). Vitamin-D deficiency was common, but not significantly different between the two groups (t-test, p = 0.31)

In this series of cemented hip replacements performed between 1994 and 1998, aseptic loosening was associated with poor bone quality. Patients with a THR should be screened for osteoporosis and have regular radiological surveillance.
Successful treatment of refractory tibial nonunion using calcium sulphate and bone marrow stromal cell implantation
Authors: Bajada S. , Harrison P. E. , Ashton B. A. , Cassar-Pullicino V. N. , Ashammakhi N. , Richardson J. B.
Published: October 2007

Successful healing of a nine-year tibial nonunion resistant to six previous surgical procedures was achieved by tissue engineering. We used autologous bone marrow stromal cells (BMSCs) expanded to 5 × 106 cells after three weeks’ tissue culture. Calcium sulphate (CaSO4) in pellet form was combined with these cells at operation. The nonunion was clinically and radiologically healed two months after implantation.

This is the description of on healing of a long-standing tibial nonunion by tissue engineering. The successful combination of BMSCs and CaSO4 has not to our knowledge been reported in a clinical setting.
The risks and benefits of radiotherapy with massive endoprosthetic replacement
Authors: Jeys L. M. , Luscombe J. S. , Grimer R. J. , Abudu A. , Tillman R. M. , Carter S. R.
Published: October 2007

Between 1966 and 2001, 1254 patients underwent excision of a bone tumour with endoprosthetic replacement. All patients who had radiotherapy were identified. Their clinical details were retrieved from their records.

A total of 63 patients (5%) had received adjunctive radiotherapy, 29 pre-operatively and 34 post-operatively. The mean post-operative Musculoskeletal Tumor Society scores of irradiated patients were significantly lower (log-rank test, p = 0.009). The infection rate in the group who had not been irradiated was 9.8% (117 of 1191), compared with 20.7% (6 of 29) in those who had pre-operative radiotherapy and 35.3% (12 of 34) in those who radiotherapy post-operatively. The infection-free survival rate at ten years was 85.5% for patients without radiotherapy, 74.1% for those who had pre-operative radiotherapy and 44.8% for those who had post-operative radiotherapy (log-rank test, p < 0.001). The ten-year limb salvage rate was 89% for those who did not have radiotherapy and 76% for those who did (log-rank test, p = 0.02).

Radiotherapy increased the risk of revision (log-rank test, p = 0.015). A total of ten amputations were necessary to control infection, of which nine were successful. Radiotherapy may be necessary for the treatment of a bone sarcoma but increases the risk of deep infection for which amputation may be the only solution.
Incidence of deep-vein thrombosis in patients with fractures of the ankle treated in a plaster cast
Authors: Patil S. , Gandhi J. , Curzon I. , Hui A. C. W.
Published: October 2007

Stable fractures of the ankle can be successfully treated non-operatively by a below-knee plaster cast. In some centres, patients with this injury are routinely administered low-molecular-weight heparin, to reduce the risk of deep-vein thrombosis (DVT). We have assessed the incidence of DVT in 100 patients in the absence of any thromboprophylaxis. A colour Doppler duplex ultrasound scan was done at the time of the removal of the cast.

Five patients did develop DVT, though none had clinical signs suggestive of it. One case involved the femoral and another the popliteal vein. No patient developed pulmonary embolism. As the incidence of DVT after ankle fractures is low, we do not recommend routine thromboprophylaxis.
A biomechanical analysis of multistrand repairs with the Silfverskiöld peripheral cross-stitch
Authors: Hirpara K. M. , Sullivan P. J. , Raheem O. , O’Sullivan M. E.
Published: October 2007

We compared the bulking and tensile strength of the Pennington modified Kessler, Cruciate and the Savage repairs in an ex vivo model. A total of 60 porcine tendons were randomised to three groups, half repaired using a core suture alone and the remainder employing a core and peripheral technique. The tendons were distracted to failure. The force required to produce a 3 mm gap, the ultimate strength, the mode of failure and bulking for each repair were assessed. We found that there was a significant increase in strength without an increase in bulk as the number of strands increased. The Cruciate repair was significantly more likely to fail by suture pullout than the Pennington modified Kessler or Savage repairs. We advise the use of the Savage repair, especially in the thumb, and a Cruciate when a Savage is not possible. The Pennington modified Kessler repair should be reserved for multiple tendon injuries.
The evolution of the compartment syndrome since 1948 as recorded in the JBJS (B)
Authors: Klenerman L.
Published: October 2007

This paper describes how we came to understand the pathophysiology of Volkmann’s ischaemic contracture with references to relevant papers in this Journal, and the investigation and management of acute compartment syndrome is briefly discussed.
Treatment With Once-Weekly Alendronate 70 mg Compared With Once-Weekly Risedronate 35 mg in Women With Postmenopausal Osteoporosis: A Randomized Double-Blind Study
Authors: Rosen Clifford J , Hochberg Marc C , Bonnick Sydney L , McClung Michael , Miller Paul , Broy Susan , Kagan Risa , Chen Erluo , Petruschke Richard A , Thompson Desmond E , de Papp Anne E
Published: October 2007

Once-weekly alendronate 70 mg and once-weekly risedronate 35 mg are indicated for the treatment of postmenopausal osteoporosis. These two agents were compared in a 12-month head-to-head trial. Greater gains in BMD and greater reductions in markers of bone turnover were seen with alendronate compared with risedronate with similar tolerability.

Introduction: The nitrogen-containing bisphosphonates, alendronate and risedronate, are available in once-weekly (OW) formulations for the treatment of postmenopausal osteoporosis. A 12-month, head-to-head study was performed to compare these agents in the treatment of postmenopausal women with low BMD.

Materials and Methods: A total of 1053 patients from 78 U.S. sites were randomized to OW alendronate 70 mg (N = 520) or risedronate 35 mg (N = 533), taken in the morning after fasting. Endpoints included BMD changes over 6 and 12 months at the hip trochanter, total hip, femoral neck, and lumbar spine (LS); percent of patients with predefined levels of change in trochanter and LS BMD at 12 months; and change in biochemical markers of bone turnover at 3, 6, and 12 months. Tolerability was evaluated by adverse experience (AE) reporting.

Results: Significantly greater increases in hip trochanter BMD were seen with alendronate (3.4%) than risedronate (2.1%) at 12 months (treatment difference, 1.4%; p < 0.001) as well as 6 months (treatment difference, 1.3%; p < 0.001). Significantly greater gains in BMD were seen with alendronate at all BMD sites measured (12-month difference: total hip, 1.0%; femoral neck, 0.7%; LS, 1.2%). Significant differences were seen as early as 6 months at all sites. A greater percentage of patients had ≥0% (p < 0.001) and ≥3% (p < 0.01) gain in trochanter and spine BMD at 12 months with alendronate than risedronate. Significantly greater (p < 0.001) reductions in all biochemical markers of bone turnover occurred with alendronate compared with risedronate by 3 months. No significant differences were seen between treatment groups in the incidence of upper gastrointestinal AEs or AEs causing discontinuation.

Conclusions: In this 12-month, head-to-head trial of alendronate and risedronate, given in accordance with the approved OW regimens for treatment of osteoporosis in postmenopausal women, alendronate produced greater gains in BMD and greater reductions in markers of bone turnover than risedronate. The greater antiresorptive effect of alendronate was seen as early as 3 months, and the tolerability profiles were similar.
Femoral Neck Trabecular Microstructure in Ovariectomized Ewes Treated With Calcitonin: MRI Microscopic Evaluation
Authors: Jiang Yebin , Zhao Jenny , Geusens Piet , Liao Er-Yuan , Adriaensens Peter , Gelan Jan , Azria Moïse , Boonen Steven , Caulin Francine , Lynch John A , Ouyang Xiaolong , Genant Harry K
Published: October 2007

Ovariectomy induces deterioration of the trabecular structure in the femoral neck of ewes, as depicted by MR microscopic imaging. This structural deterioration is prevented by salmon calcitonin treatment.

Introduction: This study evaluated the trabecular (Tb) microarchitecture of an ovariectomy (OVX)-induced osteoporotic model in ewes and determined the effects of salmon calcitonin (sCT), an osteoclast inhibitor, on the Tb structure. This is the first report of OVX-induced changes in the Tb structure in the femoral neck in the ewes and effect of sCT on the microarchitecture.

Materials and Methods: Ewes (5–8 years old, n = 28) were equally allocated into sham (Sham), OVX injected with vehicle, or OVX injected with sCT at 50 or 100 IU, three injections per week. They were killed 6 months after OVX. The femoral neck was examined with an MR imager at 9.4 T in axial, coronal, and sagittal planes. An internal calibration procedure as a means of standardizing image analysis was used to adjust the segmentation threshold. Data from all three planes were averaged.

Results and Conclusions: Compared with Sham, OVX induced significant changes (p < 0.0125) in the MRI-derived femoral neck Tb structure: Tb bone volume fraction (BV/TV), −18%; Tb number, −20%; Tb separation, +23%; number of free ends, +28%; number of nodes, −39%; number of Tb branches, −23%; mean length of Tb branches, −19%. Compared with OVX, treatment of sCT at 100 IU significantly improved all the Tb structural parameters to the Sham level (p < 0.0001 ∼ p = 0.0281), whereas 50 IU significantly increased the Tb number and the mean length of the Tb branches. BV/TV explained 74% of the variation of compressive stress of the trabecular cylinder cores of the femoral neck. Combining all structural parameters in a multivariate regression analysis significantly improved the explanation to 84%, and adding BMD further improved the predictive ability of the model to 92%. We conclude that OVX induces deterioration of the MRI-derived Tb microstructure in the femoral neck of ewes. sCT treatment prevents OVX-induced changes. The femoral neck microarchitecture significantly correlates with its biomechanical properties. Combining microstructural parameters with BMD further improves the prediction of bone biomechanical properties. The effects of sCT on OVX ewes may help explain reduced fracture risk in postmenopausal osteoporotic women treated with sCT.
Fluid Shear-Induced ATP Secretion Mediates Prostaglandin Release in MC3T3-E1 Osteoblasts
Authors: Genetos Damian C , Geist Derik J , Liu Dawei , Donahue Henry J , Duncan Randall L
Published: October 2007

ATP is rapidly released from osteoblasts in response to mechanical load. We examined the mechanisms involved in this release and established that shear-induced ATP release was mediated through vesicular fusion and was dependent on Ca2+ entry into the cell through L-type voltage-sensitive Ca2+ channels. Degradation of secreted ATP by apyrase prevented shear-induced PGE2 release.

Introduction Fluid shear induces a rapid rise in intracellular calcium ([Ca2+]i) in osteoblasts that mediates many of the cellular responses associated with mechanotransduction in bone. A potential mechanism for this increase in [Ca2+]i is the activation of purinergic (P2) receptors resulting from shear-induced extracellular release of ATP. This study was designed to determine the effects of fluid shear on ATP release and the possible mechanisms associated with this release.

Materials and Methods MC3T3-E1 preosteoblasts were plated on type I collagen, allowed to proliferate to 90% confluency, and subjected to 12 dynes/cm2 laminar fluid flow using a parallel plate flow chamber. ATP release into the flow media was measured using a luciferin/luciferase assay. Inhibitors of channels, gap junctional intercellular communication (GJIC), and vesicular formation were added before shear and maintained in the flow medium for the duration of the experiment.

Results and Conclusions Fluid shear produced a transient increase in ATP release compared with static MC3T3-E1 cells (59.8 ± 15.7 versus 6.2 ± 1.8 nM, respectively), peaking within 1 minute of onset. Inhibition of calcium entry through the L-type voltage-sensitive Ca2+ channel (L-VSCC) with nifedipine or verapamil significantly attenuated shear-induced ATP release. Channel inhibition had no effect on basal ATP release in static cells. Ca2+-dependent ATP release in response to shear seemed to result from vesicular release and not through gap hemichannels. Vesicle disruption with N-ethylmaleimide, brefeldin A, or monensin prevented increases in flow-induced ATP release, whereas inhibition of gap hemichannels with either 18α-glycyrrhetinic acid or 18β-glycyrrhetinic acid did not. Degradation of extracellular ATP with apyrase prevented shear-induced increases in prostaglandin E2 (PGE2) release. These data suggest a time line of mechanotransduction wherein fluid shear activates L-VSCCs to promote Ca2+ entry that, in turn, stimulates vesicular ATP release. Furthermore, these data suggest that P2 receptor activation by secreted ATP mediates flow-induced prostaglandin release.
Long-Term Disuse Osteoporosis Seems Less Sensitive to Bisphosphonate Treatment Than Other Osteoporosis
Authors: Li Chao Yang , Price Christopher , Delisser Kemesha , Nasser Philip , Laudier Damien , Clement Mariza , Jepsen Karl J , Schaffler Mitchell B
Published: October 2007

We sought to determine whether risedronate can preserve cortical bone mass and mechanical properties during long-term disuse in dogs, assessed by histomorphometry and biomechanics on metacarpal diaphyses. Risedronate slowed cortical thinning and partially preserved mechanical properties, but it was unable to suppress bone loss to the degree seen in other osteoporoses.

Introduction: Disuse induces dramatic bone loss resulting from greatly elevated osteoclastic resorption. Targeting osteoclasts with antiresorptive agents, such as bisphosphonates, should be an effective countermeasure for preventing disuse osteoporosis.

Materials and Methods: Single forelimbs from beagles (5–7 years old, n = 28) were immobilized (IM) for 12 months. Age-matched, non-IM dogs served as controls. One-half the animals received either risedronate (RIS, 1 mg/kg) or vehicle daily. Histomorphometry was performed on second metacarpal mid-diaphyses. Cortical mechanical properties were determined by testing third metacarpal diaphyses in four-point bending.

Results: IM caused marked reduction in cortical area (−42%) and cortical thinning (−40%) through endocortical resorption, extensive intracortical tunneling, and periosteal resorption; both bone resorption and formation were significantly elevated over control levels on all envelopes. IM also decreased maximum load and stiffness by ∼80% compared with controls. RIS reduced both periosteal bone loss and marrow cavity expansion; however, cortical area remained significantly lower in RIS-treated IM animals than in untreated non-IM controls (−16%). RIS also increased resorption indices in all envelopes compared with nontreated IM, indicating that RIS suppressed osteoclast activity but not osteoclast recruitment. RIS did not affect bone formation. RIS treatment conserved some whole bone mechanical properties, but they were still significantly lower than in controls. There were no significant differences in tissue level material properties among the groups.

Conclusion: RIS treatment reduces cortical bone loss at periosteal and endocortical surfaces caused by long-term immobilization, thus partially conserving tissue mechanical properties. This modest effect contrasts with more dramatic actions of the bisphosphonate in other osteoporoses. Our results suggest that risedronate impairs osteoclastic function but cannot completely overcome the intense stimulus for osteoclast recruitment during prolonged disuse.
Hyaluronan Increases RANKL Expression in Bone Marrow Stromal Cells Through CD44
Authors: Cao Jay J , Singleton Patrick A , Majumdar Sharmila , Boudignon Benjamin , Burghardt Andrew , Kurimoto Pam , Wronski Thomas J , Bourguignon Lilly YW , Halloran Bernard P
Published: October 2007

HA activates CD44 to stimulate RANKL expression in bone marrow stromal cells. HA stimulation of RANKL is blocked by anti-CD44 antibody and is absent in cells from CD44−/− mice. CD44−/− mice exhibit thicker cortical bone and a smaller medullary cavity, but indices of bone resorption are not affected.

Introduction: Hyaluronan (HA), the major nonprotein glycosaminoglycan component of the extracellular matrix in mammalian bone marrow, functions in part through its receptor, CD44, to stimulate a series of intracellular signaling events that lead to cell migration, adhesion, and activation. To determine whether HA activation of CD44 influences RANKL and osteoprotegerin (OPG) expression and whether CD44 is functionally important in bone metabolism, we studied whole bone and bone marrow stromal cells (BMSCs) from wildtype and CD44−/− mice.

Materials and Methods: BMSCs from wildtype and CD44−/− mice at 7 weeks of age were cultured and treated with either HA or anti-CD44 antibody. The levels of mRNA of RANKL, OPG, CD44, alkaline phosphatase (ALP), osteocalcin (OC), and αI collagen (COLL) were determined by quantitative real-time RT-PCR. Levels of RANKL and CD44 protein were measured by immunoblotting, and expression of CD44 in whole bone was determined by immunohistochemical staining. Double immunofluorescence staining and confocal microscopy were used to study colocalization of Cbfa1, CD44, and HA. Tibias were imaged using μCT, and cancellous and cortical parameters were measured. Osteoblast and osteoclast surface in the distal femoral metaphysis and osteoclast on the endocortical surface at the tibio-fibular junction were measured using quantitative histomorphometry. Differences were analyzed using ANOVA and the Newman-Keuls test.

Results: Addition of HA dose-dependently increased RANKL mRNA (3.6-fold) and protein (3-fold) levels in BMSCs. Stimulation of RANKL by HA could be blocked with anti-CD44 antibody. Treatment of cells with HA or anti-CD44 antibody had no significant effect on OPG mRNA levels. Both CD44 and HA localized on the plasma membrane in cells expressing Cbfa1. HA localization on the cell membrane disappeared when cells were preincubated with anti-CD44 antibody. Compared with control mice, cortical bone of CD44−/− was thicker, and medullary area was smaller at both 7 and 17 weeks, but at 7 weeks, indices of bone resorption were normal. At 17 weeks of age, tibial mass of CD44−/− mice was higher than control mice. CD44−/− animals expressed less RANKL in whole bone (−30%) and in BMSCs (−50%). Cells from CD44−/− animals failed to respond to either HA or CD44 antibody treatment.

Conclusions: HA can increase RANKL expression in BMSCs through CD44.