bluuebunny/crossref_metadata_2025_split huggingface dataset:
Optimization of the Expansion and Differentiation of Rabbit Chondrocytes In Vitro
Authors: Mounts T. , Ginley N. , Schluchter M. , Dennis J.E. 
Published: February 2012

Objective: To develop a tissue culture expansion method for rabbit chondrocytes that promotes robust expansion while preserving chondrogenic potential. Design: Rabbit chondrocytes isolated from articular or auricular chondrocytes were assessed for chondrogenic differentiation potential versus population doubling using different expansion and differentiation conditions. Expansion conditions included serum alone, serum plus basic fibroblast growth factor 2 (FGF-2), and serum plus insulin-like growth factor 1 (IGF-1) and FGF-2. Differentiation conditions consisted of defined medium with and without bone morphogenetic protein 2 (BMP-2). Results: Nonsupplemented chondrocytes showed limited expandability, whereas supplementation with FGF-2 allowed articular chondrocytes to be expanded past 10 population doublings (PDs) and allowed auricular chondrocytes to expand past 15 population doublings. Differentiation, as measured by glycosaminoglycan production in aggregate cultures, was minimal in articular chondrocytes without BMP-2 supplementation and diminished to less than 50% maximal in auricular chondrocytes by PD 20. However, when FGF-2 was used during expansion and BMP-2 used during differentiation, both articular and auricular chondrocytes retained greater than 50% maximal differentiation for more than 25 PDs. The addition of IGF-1 to FGF-2 during expansion decreased chondrogenicity of auricular chondrocytes exposed to BMP-2, whereas for articular chondrocytes, chondrogenic expression increased. Conclusion: These results demonstrate that FGF-2, for expansion, and BMP-2, for differentiation, dramatically increase the functional expansion of auricular and articular chondrocytes and provide a methodology to expand sufficient numbers of chondrocytes for tissue engineering applications.
Evaluating Osteoarthritic Chondrocytes through a Novel 3-Dimensional In Vitro System for Cartilage Tissue Engineering and Regeneration
Authors: Li Hanwei , Davison Noel , Moroni Lorenzo , Feng Felicia , Crist Joshua , Salter Erin , Bingham Clifton O. , Elisseeff Jennifer 
Published: February 2012

Objective: To characterize and evaluate osteoarthritic (OA) chondrocytes, in comparison to normal chondrocytes, through a novel 3-dimensional (3-D) culture system, poly(ethylene-glycol) diacrylate (PEGDA). The cytokine interleukin 1β (IL-1β) was also used to simulate an in vitro OA model. Methods: Normal and OA chondrocytes were cultured in monolayer and analyzed for changes in cartilage-specific gene expressions due to passage number. Then, cells were encapsulated in PEGDA to evaluate phenotype and matrix production capabilities through the in vitro culture system. Characterization was conducted with polymerase chain reaction (PCR), biochemical analyses, and histological staining. 3-D encapsulated chondrocytes (human and bovine) were also treated with IL-1β to characterize how the cytokine affects gene transcription and extracellular matrix (ECM) content. Results: In 2-dimensional monolayer, anabolic genes were down-regulated significantly in both normal and OA chondrocytes. In 3-D culture, OA chondrocytes demonstrated significantly higher expressions of catabolic genes when compared to normal cells. Differentiation medium resulted in significantly more matrix production than growth medium from OA chondrocytes, indicated through histological staining. In addition, normal chondrocytes responded more significantly to exogenous administration of IL-1β than OA chondrocytes. Temporary initial stimulation of IL-1β to OA chondrocytes resulted in comparable gene expressions to untreated cells after 3 weeks of in vitro culture. Conclusions: Our findings demonstrate the use of OA chondrocytes in tissue engineering and their significance for potential future cartilage regeneration research through their matrix production capabilities and the use of a hydrogel culture system.
Involvement of a periodontal pathogen, Porphyromonas gingivalis on the pathogenesis of non-alcoholic fatty liver disease
Authors: Yoneda Masato , Naka Shuhei , Nakano Kazuhiko , Wada Koichiro , Endo Hiroki , Mawatari Hironori , Imajo Kento , Nomura Ryota , Hokamura Kazuya , Ono Masafumi , Murata Shogo , Tohnai Iwai , Sumida Yoshio , Shima Toshihide , Kuboniwa Masae , Umemura Kazuo , Kamisaki Yoshinori , Amano Atsuo , Okanoue Takeshi , Ooshima Takashi , Nakajima Atsushi 
Published: February 2012

Abstract

Background
Non-alcoholic fatty liver disease (NAFLD) is a hepatic manifestation of metabolic syndrome that is closely associated with multiple factors such as obesity, hyperlipidemia and type 2 diabetes mellitus. However, other risk factors for the development of NAFLD are unclear. With the association between periodontal disease and the development of systemic diseases receiving increasing attention recently, we conducted this study to investigate the relationship between NAFLD and infection with Porphyromonas gingivalis (P. gingivalis), a major causative agent of periodontitis.

Methods
The detection frequencies of periodontal bacteria in oral samples collected from 150 biopsy-proven NAFLD patients (102 with non-alcoholic steatohepatitis (NASH) and 48 with non-alcoholic fatty liver (NAFL) patients) and 60 non-NAFLD control subjects were determined. Detection of P. gingivalis and other periodontopathic bacteria were detected by PCR assay. In addition, effect of P. gingivalis-infection on mouse NAFLD model was investigated. To clarify the exact contribution of P. gingivalis-induced periodontitis, non-surgical periodontal treatments were also undertaken for 3 months in 10 NAFLD patients with periodontitis.

Results
The detection frequency of P. gingivalis in NAFLD patients was significantly higher than that in the non-NAFLD control subjects (46.7% vs. 21.7%, odds ratio: 3.16). In addition, the detection frequency of P. gingivalis in NASH patients was markedly higher than that in the non-NAFLD subjects (52.0%, odds ratio: 3.91). Most of the P. gingivalis fimbria detected in the NAFLD patients was of invasive genotypes, especially type II (50.0%). Infection of type II P. gingivalis on NAFLD model of mice accelerated the NAFLD progression. The non-surgical periodontal treatments on NAFLD patients carried out for 3 months ameliorated the liver function parameters, such as the serum levels of AST and ALT.

Conclusions
Infection with high-virulence P. gingivalis might be an additional risk factor for the development/progression of NAFLD/NASH.
A phase I study evaluating the pharmacokinetics, safety and tolerability of an antibody-based tissue factor antagonist in subjects with acute lung injury or acute respiratory distress syndrome
Authors: Morris Peter E , Steingrub Jay S , Huang Bee Y , Tang Shamay , Liu Patrick M , Rhode Peter R , Wong Hing C 
Published: February 2012

Abstract

Background
The tissue factor (TF)-dependent extrinsic pathway has been suggested to be a central mechanism by which the coagulation cascade is locally activated in the lungs of patients with acute lung injury and acute respiratory distress syndrome (ALI/ARDS) and thus represents an attractive target for therapeutic intervention. This study was designed to determine the pharmacokinetic and safety profiles of ALT-836, an anti-TF antibody, in patients with ALI/ARDS.

Methods
This was a prospective, randomized, placebo-controlled, dose-escalation Phase I clinical trial in adult patients who had suspected or proven infection, were receiving mechanical ventilation and had ALI/ARDS (PaO2/FiO2 ≤ 300 mm). Eighteen patients (6 per cohort) were randomized in a 5:1 ratio to receive ALT-836 or placebo, and were treated within 48 hours after meeting screening criteria. Cohorts of patients were administered a single intravenously dose of 0.06, 0.08 or 0.1 mg/kg ALT-836 or placebo. Blood samples were taken for pharmacokinetic and immunogenicity measurements. Safety was assessed by adverse events, vital signs, ECGs, laboratory, coagulation and pulmonary function parameters.

Results
Pharmacokinetic analysis showed a dose dependent exposure to ALT-836 across the infusion range of 0.06 to 0.1 mg/kg. No anti-ALT-836 antibody response was observed in the study population during the trial. No major bleeding episodes were reported in the ALT-836 treated patients. The most frequent adverse events were anemia, observed in both placebo and ALT-836 treated patients, and ALT-836 dose dependent, self-resolved hematuria, which suggested 0.08 mg/kg as an acceptable dose level of ALT-836 in this patient population.

Conclusions
Overall, this study showed that ALT-836 could be safely administered to patients with sepsis-induced ALI/ARDS.

Trial registration
ClinicalTrials.gov: NCT01438853
Unsupervised machine-learning method for improving the performance of ambulatory fall-detection systems
Authors: Yuwono Mitchell , Moulton Bruce D , Su Steven W , Celler Branko G , Nguyen Hung T 
Published: February 2012

Abstract

Background
Falls can cause trauma, disability and death among older people. Ambulatory accelerometer devices are currently capable of detecting falls in a controlled environment. However, research suggests that most current approaches can tend to have insufficient sensitivity and specificity in non-laboratory environments, in part because impacts can be experienced as part of ordinary daily living activities.

Method
We used a waist-worn wireless tri-axial accelerometer combined with digital signal processing, clustering and neural network classifiers. The method includes the application of Discrete Wavelet Transform, Regrouping Particle Swarm Optimization, Gaussian Distribution of Clustered Knowledge and an ensemble of classifiers including a multilayer perceptron and Augmented Radial Basis Function (ARBF) neural networks.

Results
Preliminary testing with 8 healthy individuals in a home environment yields 98.6% sensitivity to falls and 99.6% specificity for routine Activities of Daily Living (ADL) data. Single ARB and MLP classifiers were compared with a combined classifier. The combined classifier offers the greatest sensitivity, with a slight reduction in specificity for routine ADL and an increased specificity for exercise activities. In preliminary tests, the approach achieves 100% sensitivity on in-group falls, 97.65% on out-group falls, 99.33% specificity on routine ADL, and 96.59% specificity on exercise ADL.

Conclusion
The pre-processing and feature-extraction steps appear to simplify the signal while successfully extracting the essential features that are required to characterize a fall. The results suggest this combination of classifiers can perform better than MLP alone. Preliminary testing suggests these methods may be useful for researchers who are attempting to improve the performance of ambulatory fall-detection systems.
Reducing widespread pipe sharing and risky sex among crystal methamphetamine smokers in Toronto: do safer smoking kits have a potential role to play?
Authors: Hunter Charlotte , Strike Carol , Barnaby Lorraine , Busch Adam , Marshall Chantel , Shepherd Susan , Hopkins Shaun 
Published: February 2012

Abstract

Background
Crystal methamphetamine smoking is associated with many negative health consequences, including the potential for transmission of hepatitis. We examined whether or not a kit for crystal methamphetamine smoking might have some potential to reduce the negative health effects of crystal methamphetamine smoking.

Methods
Five focus groups were conducted with crystal methamphetamine smokers recruited by community health agencies and youth shelters in Toronto, Canada. Target groups included homeless/street-involved youth, sex workers, men who have sex with men, and youth in the party scene. Participants (n = 32) were asked questions about motivations for crystal methamphetamine use, the process of smoking, health problems experienced, sharing behaviour, risky sexual practices, and the ideal contents of a harm reduction kit.

Results
Pipe sharing was widespread among participants and was deemed integral to the social experience of smoking crystal methamphetamine. Heated pipes were unlikely to cause direct injuries, but participants mentioned having dry, cracked lips, which may be a vector for disease transmission. Many reported having sex with multiple partners and being less likely to use condoms while on the drug. Demand for harm reduction kits was mixed.

Conclusions
Changing pipe sharing behaviours may be difficult because many participants considered sharing to be integral to the social experience of smoking crystal methamphetamine. Within the context of a broader health promotion and prevention program, pilot testing of safer smoking kits to initiate discussion and education on the risks associated with sharing pipes and unprotected sex for some communities (e.g., homeless/street-involved youth) is worth pursuing.
Cyclin D1 overexpression and poor clinical outcomes in Taiwanese oral cavity squamous cell carcinoma
Authors: Huang Shiang-Fu , Cheng Sou-De , Chuang Wen-Yu , Chen I-How , Liao Chun-Ta , Wang Hung-Ming , Hsieh Ling-Ling 
Published: February 2012

Abstract

Background
Cyclin D1 gene regulates cell cycle and plays an important role in the tumorigenesis of human cancers. The association between cyclin D1, clinicopathologic parameters and prognosis in oral cavity squamous cell carcinoma (OSCC) is inconclusive.

Methods
A total of 264 male OSCCs were examined for cyclin D1 protein expression using immunohistochemistry (IHC). The expression levels of cyclin D1 were defined as overexpression when more than 10% of tumor cells displayed nuclear staining with moderate to strong intensity.

Results
Overexpression of cyclin D1 was found in 97 (36.7%) OSCCs. Cyclin D1 protein overexpression was significantly associated with lymph node metastasis (P = 0.002), tumor cell differentiation (P = 0.031) and tumor stage (P = 0.051), but not associated with age onset, cigarette smoking, alcohol drinking, or areca quid chewing. Overexpression of cyclin D1 was also significantly associated with poor clinical outcomes in terms of disease-free survival (DFS, P = 0.002) and overall survival (OS, P < 0.001). The effects of cyclin D1 protein overexpression on DFS (hazard ratio (HR) = 1.540; 95% confidence interval (CI), 1.068 - 2.222) and OS (HR = 1.702; 95% CI, 1.168 - 2.480) were still existed after adjusting for clinicopathological paremeters (such as age, primary tumor status, tumor cell differentiation, and lymph node metastasis) using logistic multivariate analysis.

Conclusion
Cyclin D1 protein worked as an independent prognostic factor and can be as a biomarker for the aggressiveness of OSCC.
Developing the dielectric mechanisms of polyetherimide/multiwalled carbon nanotube/(Ba0.8Sr0.2)(Ti0.9Zr0.1)O3 composites
Authors: Su Chean-Cheng , Wu Chia-Ching , Yang Cheng-Fu 
Published: February 2012

Abstract
Various amounts of multiwalled carbon nanotubes [MWNTs] were embedded into polyetherimide [PEI] to form PEI/MWNT composites, and their dielectric properties were measured at 1 MHz. The Lichtenecker mixing rule was used to find a reasonable dielectric constant for the MWNTs used in this study. The dielectric constants of the developed composites were significantly increased, and the loss tangents were significantly decreased as 2.0 wt.% (Ba0.8Sr0.2)(Ti0.9Zr0.1)O3 ceramic powder [BSTZ] was added to the PEI/MWNTs to form PEI/MWNT/BSTZ composites. The Lichtenecker and Yamada mixing rules were used to predict the dielectric constants of the PEI/MWNT and PEI/MWNT/BSTZ composites. Equivalent electrical conduction models of both composites were established using the two mixing rules. In addition, the theoretical bases of the two mixing rules were used to explain the measured results for the PEI/MWNT and PEI/BSTZ/MWNT composites.
Naloxone inhibits immune cell function by suppressing superoxide production through a direct interaction with gp91 phox subunit of NADPH oxidase
Authors: Wang Qingshan , Zhou Hui , Gao Huiming , Chen Shih-Heng , Chu Chun-Hsien , Wilson Belinda , Hong Jau-Shyong 
Published: February 2012

AbstractBackgroundBoth (-) and (+)-naloxone attenuate inflammation-mediated neurodegeneration by inhibition of microglial activation through superoxide reduction in an opioid receptor-independent manner. Multiple lines of evidence have documented a pivotal role of overactivated NADPH oxidase (NOX2) in inflammation-mediated neurodegeneration. We hypothesized that NOX2 might be a novel action site of naloxone to mediate its anti-inflammatory actions.MethodsInhibition of NOX-2-derived superoxide by (-) and (+)-naloxone was measured in lipopolysaccharide (LPS)-treated midbrain neuron-glia cultures and phorbol myristate acetate (PMA)-stimulated neutrophil membranes by measuring the superoxide dismutase (SOD)-inhibitable reduction of tetrazolium salt (WST-1) or ferricytochrome c. Further, various ligand (3H-naloxone) binding assays were performed in wild type and gp91phox-/-neutrophils and transfected COS-7 and HEK293 cells. The translocation of cytosolic subunit p47phoxto plasma membrane was assessed by western blot.ResultsBoth (-) and (+)-naloxone equally inhibited LPS- and PMA-induced superoxide production with an IC50 of 1.96 and 2.52 μM, respectively. Competitive binding of3H-naloxone with cold (-) and (+)-naloxone in microglia showed equal potency with an IC50 of 2.73 and 1.57 μM, respectively.3H-Naloxone binding was elevated in COS-7 and HEK293 cells transfected with gp91phox; in contrast, reduced3H-naloxone binding was found in neutrophils deficient in gp91phoxor in the presence of a NOX2 inhibitor. The specificity and an increase in binding capacity of3H-naloxone were further demonstrated by 1) an immunoprecipitation study using gp91phoxantibody, and 2) activation of NOX2 by PMA. Finally, western blot studies showed that naloxone suppressed translocation of the cytosolic subunit p47phoxto the membrane, leading to NOX2 inactivation.ConclusionsStrong evidence is provided indicating that NOX2 is a non-opioid novel binding site for naloxone, which is critical in mediating its inhibitory effect on microglia overactivation and superoxide production.
Receptor chimeras demonstrate that the C-terminal domain of the human cytomegalovirus US27 gene product is necessary and sufficient for intracellular receptor localization
Authors: Stapleton Lance K , Arnolds Kathleen L , Lares Angela P , Devito Tori M , Spencer Juliet V 
Published: February 2012

Abstract

Background
Human cytomegalovirus (HCMV) is ubiquitous in the population but generally causes only mild or asymptomatic infection except in immune suppressed individuals. HCMV employs numerous strategies for manipulating infected cells, including mimicry of G-protein coupled receptors (GPCRs). The HCMV US27 gene product is a putative GPCR, yet no ligand or signaling has been identified for this receptor. In the present study, immunofluorescence microscopy was used to examine the cellular distribution of wild type US27, as well as US27 deletion mutants and chimeric receptors.

Results
In transiently transfected cells, wild type US27 was found primarily in intracellular compartments, in striking contrast to the cell surface distribution seen for the human cellular chemokine receptor CXCR3. When the N-terminal extracellular domains of the two receptors were swapped, no change in protein localization was observed. However, swapping of the C-terminal intracellular domains resulted in a significant change in receptor distribution. A chimera that contained US27 fused to the C-terminal intracellular tail of CXCR3 exhibited surface distribution similar to that of wild-type CXCR3. When the C-terminal domain of US27 was fused to CXCR3, this chimeric receptor (CXCR3/US27-CT) was found in the same intracellular pattern as wild-type US27. In addition, a US27 mutant lacking the C-terminus (US27ΔCT) failed to accumulate inside the cell and exhibited cell surface distribution. Co-localization with organelle-specific markers revealed that wild-type US27 was found predominantly in the Golgi apparatus and in endosomal compartments, whereas the US27/CXCR3-CT chimera, US27ΔCT and US27Δ348 mutants were not localized to endosomal compartments.

Conclusions
The results indicate that the C-terminal domain of the HCMV US27 protein, which contains a di-leucine endocytic sorting motif, is both necessary and sufficient for intracellular localization, which may also help explain why no cellular ligands have yet been identified for this viral receptor.
Biology and interactions of two distinct monopartite begomoviruses and betasatellites associated with radish leaf curl disease in India
Authors: Singh AK , Chattopadhyay B , Chakraborty S 
Published: February 2012

Abstract

Background
Emerging whitefly transmitted begomoviruses are major pathogens of vegetable and fibre crops throughout the world, particularly in tropical and sub-tropical regions. Mutation, pseudorecombination and recombination are driving forces for the emergence and evolution of new crop-infecting begomoviruses. Leaf curl disease of field grown radish plants was noticed in Varanasi and Pataudi region of northern India. We have identified and characterized two distinct monopartite begomoviruses and associated beta satellite DNA causing leaf curl disease of radish (Raphanus sativus) in India.

Results
We demonstrate that RaLCD is caused by a complex of two Old World begomoviruses and their associated betasatellites. Radish leaf curl virus-Varanasi is identified as a new recombinant species, Radish leaf curl virus (RaLCV) sharing maximum nucleotide identity of 87.7% with Tomato leaf curl Bangladesh virus-[Bangladesh:2] (Accession number AF188481) while the virus causing radish leaf curl disease-Pataudi is an isolate of Croton yellow vein mosaic virus-[India] (CYVMV-IN) (Accession number AJ507777) sharing 95.8% nucleotide identity. Further, RDP analysis revealed that the RaLCV has a hybrid genome, a putative recombinant between Euphorbia leaf curl virus and Papaya leaf curl virus. Cloned DNA of either RaLCV or CYVMV induced mild leaf curl symptoms in radish plants. However, when these clones (RaLCV or CYVMV) were individually co-inoculated with their associated cloned DNA betasatellite, symptom severity and viral DNA levels were increased in radish plants and induced typical RaLCD symptoms. To further extend these studies, we carried out an investigation of the interaction of these radish-infecting begomoviruses and their associated satellite, with two tomato infecting begomoviruses (Tomato leaf curl Gujarat virus and Tomato leaf curl New Delhi virus). Both of the tomato-infecting begomoviruses showed a contrasting and differential interaction with DNA satellites, not only in the capacity to interact with these molecules but also in the modulation of symptom phenotypes by the satellites.

Conclusion
This is the first report and experimental demonstration of Koch's postulate for begomoviruses associated with radish leaf curl disease. Further observations also provide direct evidence of lateral movement of weed infecting begomovirus in the cultivated crops and the present study also suggests that the exchange of betasatellites with other begomoviruses would create a new disease complex posing a serious threat to crop production.