In vivo dosimetry in UK external beam radiotherapy: current and future usage

PubMed Central

Graveling, Michael; Hansen, Vibeke N; Brownsword, Kevin; Morgan, Andrew

2017-01-01

Objective: Towards Safer Radiotherapy recommended that radiotherapy (RT) centres should have protocols in place for in vivo dosimetry (IVD) monitoring at the beginning of patient treatment courses (Donaldson S. Towards safer radiotherapy. R Coll Radiol 2008). This report determines IVD implementation in the UK in 2014, the methods used and makes recommendations on future use. Methods: Evidence from peer-reviewed journals was used in conjunction with the first survey of UK RT centre IVD practice since the publication of Towards Safer Radiotherapy. In March 2014, profession-specific questionnaires were sent to radiographer, clinical oncologist and physics staff groups in each of the 66 UK RT centres. Results: Response rates from each group were 74%, 45% and 74%, respectively. 73% of RT centres indicated that they performed IVD. Diodes are the most popular IVD device. Thermoluminescent dosimeter (TLD) is still in use in a number of centres but not as a sole modality, being used in conjunction with diodes and/or electronic portal imaging device (EPID). The use of EPID dosimetry is increasing and is considered of most potential value for both geometric and dosimetric verification. Conclusion: Owing to technological advances, such as electronic data transfer, independent monitor unit checking and daily image-guided radiotherapy, the overall risk of adverse treatment events in RT has been substantially reduced. However, the use of IVD may prevent a serious radiation incident. Point dose IVD is not considered suited to the requirements of verifying advanced RT techniques, leaving EPID dosimetry as the current modality likely to be developed as a future standard. Advances in knowledge: An updated perspective on UK IVD use and provision of professional guidelines for future implementation. PMID:28205452

Developing a computer-controlled simulated digestion system to predict the concentration of metabolizable energy of feedstuffs for rooster.

PubMed

Zhao, F; Ren, L Q; Mi, B M; Tan, H Z; Zhao, J T; Li, H; Zhang, H F; Zhang, Z Y

2014-04-01

Four experiments were conducted to evaluate the effectiveness of a computer-controlled simulated digestion system (CCSDS) for predicting apparent metabolizable energy (AME) and true metabolizable energy (TME) using in vitro digestible energy (IVDE) content of feeds for roosters. In Exp. 1, the repeatability of the IVDE assay was tested in corn, wheat, rapeseed meal, and cottonseed meal with 3 assays of each sample and each with 5 replicates of the same sample. In Exp. 2, the additivity of IVDE concentration in corn, soybean meal, and cottonseed meal was tested by comparing determined IVDE values of the complete diet with values predicted from measurements on individual ingredients. In Exp. 3, linear models to predict AME and TME based on IVDE were developed with 16 calibration samples. In Exp. 4, the accuracy of prediction models was tested by the differences between predicted and determined values for AME or TME of 6 ingredients and 4 diets. In Exp. 1, the mean CV of IVDE was 0.88% (range = 0.20 to 2.14%) for corn, wheat, rapeseed meal, and cottonseed meal. No difference in IVDE was observed between 3 assays of an ingredient, indicating that the IVDE assay is repeatable under these conditions. In Exp. 2, minimal differences (<21 kcal/kg) were observed between determined and calculated IVDE of 3 complete diets formulated with corn, soybean meal, and cottonseed meal, demonstrating that the IVDE values are additive in a complete diet. In Exp. 3, linear relationships between AME and IVDE and between TME and IVDE were observed in 16 calibration samples: AME = 1.062 × IVDE - 530 (R(2) = 0.97, residual standard deviation [RSD] = 146 kcal/kg, P < 0.001) and TME = 1.050 × IVDE - 16 (R(2) = 0.97, RSD = 148 kcal/kg, P < 0.001). Differences of less than 100 kcal/kg were observed between determined and predicted values in 10 and 9 of the 16 calibration samples for AME and TME, respectively. In Exp. 4, differences of less than 100 kcal/kg between determined and predicted values were observed in 3 and 4 of the 6 ingredient samples for AME and TME, respectively, and all 4 diets showed the differences of less than 25 kcal/kg between determined and predicted AME or TME. Our results indicate that the CCSDS is repeatable and additive. This system accurately predicted AME or TME on 17 of the 26 samples and may be a promising method to predict the energetic values of feed for poultry.

Are proteomic technologies ready for IVDs

USDA-ARS?s Scientific Manuscript database

During the last decade as a result of the unparalleled advancements in mass spectrometry-based methods in protein analysis, biomarker research has escalated to new heights in the academic, government and industrial research laboratories. Translation of biomarker research to in vitro diagnostics (IVD...

Expression and regulation of neurotrophins in the nondegenerate and degenerate human intervertebral disc

PubMed Central

Purmessur, Devina; Freemont, Anthony J; Hoyland, Judith A

2008-01-01

Introduction The neurotrophins nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) have been identified in the human intervertebral disc (IVD) and have been implicated in the mechanisms associated with nerve ingrowth and nociception in degeneration of the IVD. The aim of the current study was to investigate an association between neurotrophin expression in the IVD and the severity of disc degeneration, including the effect of disc-related proinflammatory cytokines on neurotrophin and neuropeptide expression in cells derived from the human IVD. Methods Immunohistochemical analysis was performed to examine the expression of NGF, BDNF and their high-affinity receptors Trk-A and Trk-B in human IVD samples, divided into three categories: non-degenerate, moderate degeneration and severe degeneration. In order to study the effect of disc-related cytokines on neurotrophin/neuropeptide gene expression, nucleus pulposus cells derived from non-degenerate and degenerate IVD samples were seeded in alginate and were stimulated with either IL-1β or TNFα for 48 hours. RNA was extracted, cDNA was synthesised and quantitative real-time PCR was performed to examine the expression of NGF, BDNF and substance P. Results Immunohistochemistry showed expression of NGF and BDNF in the native chondrocyte-like cells in all regions of the IVD and in all grades of degeneration. Interestingly only BDNF significantly increased with the severity of degeneration (P < 0.05). Similar expression was observed for Trk-A and Trk-B, although no association with disease severity was demonstrated. In cultured human nucleus pulposus cells, stimulation with IL-1β led to significant increases in NGF and BDNF gene expression (P < 0.05). Treatment with TNFα was associated with an upregulation of substance P expression only. Conclusion Our findings show that both the annulus fibrosus and nucleus pulposus cells of the IVD express the neurotrophins NGF and BDNF, factors that may influence and enhance innervation and pain in the degenerate IVD. Expression of Trk-A and Trk-B by cells of the nondegenerate and degenerate IVD suggests an autocrine role for neurotrophins in regulation of disc cell biology. Furthermore, modulation of neurotrophin expression by IL-1β and modulation of substance P expression by TNFα, coupled with their increased expression in the degenerate IVD, highlights novel roles for these cytokines in regulating nerve ingrowth in the degenerate IVD and associated back pain. PMID:18727839

Special Advanced Studies for Pollution Prevention Delivery Order 0065: The Monitor - Winter 2001

DTIC Science & Technology

2001-04-01

were selected based on previous efforts. These alternatives included Alodine 2000, NCS Rainseal, Sanchem Full Process, and trivalent chromium . CTC’s IVD...12 Alternatives for Chromium Electroplating: ElectroSpark Deposition .................. 13...Requirements ............ 15 Aluminum Substitution for Cadmium/ Chromium ............................................... 16 Review of Cadmium Alternatives

Catabolic cytokine expression in degenerate and herniated human intervertebral discs: IL-1β and TNFα expression profile

PubMed Central

Le Maitre, Christine Lyn; Hoyland, Judith Alison; Freemont, Anthony J

2007-01-01

Low back pain is a common and debilitating disorder. Current evidence implicates intervertebral disc (IVD) degeneration and herniation as major causes, although the pathogenesis is poorly understood. While several cytokines have been implicated in the process of IVD degeneration and herniation, investigations have predominately focused on Interleukin 1 (IL-1) and tumor necrosis factor alpha (TNFα). However, to date no studies have investigated the expression of these cytokines simultaneously in IVD degeneration or herniation, or determined which may be the predominant cytokine associated with these disease states. Using quantitative real time PCR and immunohistochemistry we investigated gene and protein expression for IL-1β, TNFα and their receptors in non-degenerate, degenerate and herniated human IVDs. IL-1β gene expression was observed in a greater proportion of IVDs than TNFα (79% versus 59%). Degenerate and herniated IVDs displayed higher levels of both cytokines than non-degenerate IVDs, although in degenerate IVDs higher levels of IL-1β gene expression (1,300 copies/100 ng cDNA) were observed compared to those of TNFα (250 copies of TNFα/100 ng cDNA). Degenerate IVDs showed ten-fold higher IL-1 receptor gene expression compared to non-degenerate IVDs. In addition, 80% of degenerate IVD cells displayed IL-1 receptor immunopositivity compared to only 30% of cells in non-degenerate IVDs. However, no increase in TNF receptor I gene or protein expression was observed in degenerate or herniated IVDs compared to non-degenerate IVDs. We have demonstrated that although both cytokines are produced by human IVD cells, IL-1β is expressed at higher levels and in more IVDs, particularly in more degenerate IVDs (grades 4 to 12). Importantly, this study has highlighted an increase in gene and protein production for the IL-1 receptor type I but not the TNF receptor type I in degenerate IVDs. The data thus suggest that although both cytokines may be involved in the pathogenesis of IVD degeneration, IL-1 may have a more significant role than TNFα, and thus may be a better target for therapeutic intervention. PMID:17688691

In vivo dosimetry in UK external beam radiotherapy: current and future usage.

PubMed

MacDougall, Niall D; Graveling, Michael; Hansen, Vibeke N; Brownsword, Kevin; Morgan, Andrew

2017-04-01

Towards Safer Radiotherapy recommended that radiotherapy (RT) centres should have protocols in place for in vivo dosimetry (IVD) monitoring at the beginning of patient treatment courses (Donaldson S. Towards safer radiotherapy. R Coll Radiol 2008). This report determines IVD implementation in the UK in 2014, the methods used and makes recommendations on future use. Evidence from peer-reviewed journals was used in conjunction with the first survey of UK RT centre IVD practice since the publication of Towards Safer Radiotherapy. In March 2014, profession-specific questionnaires were sent to radiographer, clinical oncologist and physics staff groups in each of the 66 UK RT centres. Response rates from each group were 74%, 45% and 74%, respectively. 73% of RT centres indicated that they performed IVD. Diodes are the most popular IVD device. Thermoluminescent dosimeter (TLD) is still in use in a number of centres but not as a sole modality, being used in conjunction with diodes and/or electronic portal imaging device (EPID). The use of EPID dosimetry is increasing and is considered of most potential value for both geometric and dosimetric verification. Owing to technological advances, such as electronic data transfer, independent monitor unit checking and daily image-guided radiotherapy, the overall risk of adverse treatment events in RT has been substantially reduced. However, the use of IVD may prevent a serious radiation incident. Point dose IVD is not considered suited to the requirements of verifying advanced RT techniques, leaving EPID dosimetry as the current modality likely to be developed as a future standard. Advances in knowledge: An updated perspective on UK IVD use and provision of professional guidelines for future implementation.

Low Intensity Pulsed Ultrasound (LIPUS) for the treatment of intervertebral disc degeneration

NASA Astrophysics Data System (ADS)

Horne, Devante; Jones, Peter; Salgaonkar, Vasant; Adams, Matt; Ozilgen, B. Arda; Zahos, Peter; Tang, Xinyan; Liebenberg, Ellen; Coughlin, Dezba; Lotz, Jeffrey; Diederich, Chris

2017-02-01

Discogenic back pain presents a major public health issue, with current therapeutic interventions limited to short-term symptom relief without providing regenerative remedies for diseased intervertebral discs (IVD). Many of these interventions are invasive and can diminish the biomechanical integrity of the IVDs. Low intensity pulsed ultrasound (LIPUS) is a potential treatment option that is both non-invasive and regenerative. LIPUS has been shown to be a clinically effective method for the enhancement of wound and fracture healing. Recent in vitro studies have shown that LIPUS stimulation induces an upregulation functional matrix proteins and downregulation of inflammatory factors in cultured IVD cells. However, we do not know the effects of LIPUS on an in vivo model for intervertebral disc degeneration. The objective of this study was to show technical feasibility of building a LIPUS system that can target the rat tail IVD and apply this setup to a model for acute IVD degeneration. A LIPUS exposimetry system was built using a 1.0 MHz planar transducer and custom housing. Ex vivo intensity measurements demonstrated LIPUS delivery to the center of the rat tail IVD. Using an established stab-incision model for disc degeneration, LIPUS was applied for 20 minutes daily for five days. For rats that displayed a significant injury response, LIPUS treatment caused significant upregulation of Collagen II and downregulation of Tumor Necrosis Factor - α gene expression. Our preliminary studies indicate technical feasibility of targeted delivery of ultrasound to a rat tail IVD for studies of LIPUS biological effects.

Application of the polynomial chaos expansion to approximate the homogenised response of the intervertebral disc.

PubMed

Karajan, N; Otto, D; Oladyshkin, S; Ehlers, W

2014-10-01

A possibility to simulate the mechanical behaviour of the human spine is given by modelling the stiffer structures, i.e. the vertebrae, as a discrete multi-body system (MBS), whereas the softer connecting tissue, i.e. the softer intervertebral discs (IVD), is represented in a continuum-mechanical sense using the finite-element method (FEM). From a modelling point of view, the mechanical behaviour of the IVD can be included into the MBS in two different ways. They can either be computed online in a so-called co-simulation of a MBS and a FEM or offline in a pre-computation step, where a representation of the discrete mechanical response of the IVD needs to be defined in terms of the applied degrees of freedom (DOF) of the MBS. For both methods, an appropriate homogenisation step needs to be applied to obtain the discrete mechanical response of the IVD, i.e. the resulting forces and moments. The goal of this paper was to present an efficient method to approximate the mechanical response of an IVD in an offline computation. In a previous paper (Karajan et al. in Biomech Model Mechanobiol 12(3):453-466, 2012), it was proven that a cubic polynomial for the homogenised forces and moments of the FE model is a suitable choice to approximate the purely elastic response as a coupled function of the DOF of the MBS. In this contribution, the polynomial chaos expansion (PCE) is applied to generate these high-dimensional polynomials. Following this, the main challenge is to determine suitable deformation states of the IVD for pre-computation, such that the polynomials can be constructed with high accuracy and low numerical cost. For the sake of a simple verification, the coupling method and the PCE are applied to the same simplified motion segment of the spine as was used in the previous paper, i.e. two cylindrical vertebrae and a cylindrical IVD in between. In a next step, the loading rates are included as variables in the polynomial response functions to account for a more realistic response of the overall viscoelastic intervertebral disc. Herein, an additive split into elastic and inelastic contributions to the homogenised forces and moments is applied.

Development of novel IVD assays: a manufacturer's perspective.

PubMed

Metcalfe, Thomas A

2010-01-01

IVD manufacturers are heavily reliant on novel IVD assays to fuel their growth and drive innovation within the industry. They represent a key part of the IVD industry's value proposition to customers and the healthcare industry in general, driving product differentiation, helping to create demand for new systems and generating incremental revenue. However, the discovery of novel biomarkers and their qualification for a specific clinical purpose is a high risk undertaking and the large, risky investments associated with doing this on a large scale are incompatible with IVD manufacturer's business models. This article describes the sources of novel IVD assays, the processes for discovering and qualifying novel assays and the reliance of IVD manufacturers on collaborations and in-licensing to source new IVD assays for their platforms.

Lumbar annulus fibrosus biomechanical characterization in healthy children by ultrasound shear wave elastography.

PubMed

Vergari, Claudio; Dubois, Guillaume; Vialle, Raphael; Gennisson, Jean-Luc; Tanter, Mickael; Dubousset, Jean; Rouch, Philippe; Skalli, Wafa

2016-04-01

Intervertebral disc (IVD) is key to spine biomechanics, and it is often involved in the cascade leading to spinal deformities such as idiopathic scoliosis, especially during the growth spurt. Recent progress in elastography techniques allows access to non-invasive measurement of cervical IVD in adults; the aim of this study was to determine the feasibility and reliability of shear wave elastography in healthy children lumbar IVD. Elastography measurements were performed in 31 healthy children (6-17 years old), in the annulus fibrosus and in the transverse plane of L5-S1 or L4-L5 IVD. Reliability was determined by three experienced operators repeating measurements. Average shear wave speed in IVD was 2.9 ± 0.5 m/s; no significant correlations were observed with sex, age or body morphology. Intra-operator repeatability was 5.0 % while inter-operator reproducibility was 6.2 %. Intraclass correlation coefficient was higher than 0.9 for each operator. Feasibility and reliability of IVD shear wave elastography were demonstrated. The measurement protocol is compatible with clinical routine and the results show the method's potential to give an insight into spine deformity progression and early detection. • Intervertebral disc mechanical properties are key to spine biomechanics • Feasibility of shear wave elastography in children lumbar disc was assessed • Measurement was fast and reliable • Elastography could represent a novel biomarker for spine pathologies.

Field safety notices released by manufacturers in cases of failure of products for infection testing: analysis of cases reported to the BfArM between 2005 and 2007

PubMed Central

2010-01-01

The European Directive 98/79/EC for in vitro diagnostic medical devices (IVD) regulates marketing and post marketing surveillance of IVD in the European Economic Area. Manufacturers have to inform the responsible Competent Authorities (CA) about incidents and field safety corrective actions (FSCA) related to IVD. In Germany, the Federal Institute for Drugs and Medical Devices (BfArM) is the responsible CA for most IVD, only few IVD as specified in Annex II of the Directive are under the responsibility of the Paul Ehrlich Institute (PEI). In case of a FSCA manufacturers have to inform customers by means of a Field Safety Notice (FSN) which should be sent to BfArM prior to release and is published on the BfArM home-page. Between beginning of 2005 and end of 2007 the BfArM received a total of 1025 reports regarding IVD. From these, 38 related to tests, reagents, calibrators, and control materials for infection testing, 13 related to analysers and general consumables (n = 8 and n = 5, respectively) based on culture techniques, and 7 related to analysers and general consumables (n = 5 and n = 2, respectively) based on molecular biological methods. FSCA were performed in Germany in 32 (84.2%) of all notifications related to tests reagents, calibrators, and control materials as well as in 13 (100%) and 7 (100%) of notifications related to analysers and consumables based on culture techniques and molecular biological methods, respectively. A number of relevant deficiencies regarding the quality of the FSN were separately demonstrated for FSN in German and English language. In brief, manufacturers often sent their FSN to the BfArM with delay. Additionally, a subset of FSN provided insufficient information on the product related risks or the measures to be performed by the customer to mitigate product related risks. Furthermore, customer confirmation forms often were missing in the FSN sent to the BfArM. Our data suggest that for IVD for infection testing FSCA and FSN are frequently performed. For better vigilance performance, manufacturers could shorten the time until release and improve the contents of FSN to ensure the safety of IVD in cases of product related corrective actions. PMID:21147647

Field safety notices released by manufacturers in cases of failure of products for infection testing: analysis of cases reported to the BfArM between 2005 and 2007.

PubMed

Siekmeier, R; Lisson, K; Wetzel, D

2010-11-04

The European Directive 98/79/EC for in vitro diagnostic medical devices (IVD) regulates marketing and post marketing surveillance of IVD in the European Economic Area. Manufacturers have to inform the responsible Competent Authorities (CA) about incidents and field safety corrective actions (FSCA) related to IVD. In Germany, the Federal Institute for Drugs and Medical Devices (BfArM) is the responsible CA for most IVD, only few IVD as specified in Annex II of the Directive are under the responsibility of the Paul Ehrlich Institute (PEI). In case of a FSCA manufacturers have to inform customers by means of a Field Safety Notice (FSN) which should be sent to BfArM prior to release and is published on the BfArM homepage. Between beginning of 2005 and end of 2007 the BfArM received a total of 1025 reports regarding IVD. From these, 38 related to tests, reagents, calibrators, and control materials for infection testing, 13 related to analysers and general consumables (n = 8 and n = 5, respectively) based on culture techniques, and 7 related to analysers and general consumables (n = 5 and n = 2, respectively) based on molecular biological methods. FSCA were performed in Germany in 32 (84.2%) of all notifications related to tests reagents, calibrators, and control materials as well as in 13 (100%) and 7 (100%) of notifications related to analysers and consumables based on culture techniques and molecular biological methods, respectively. A number of relevant deficiencies regarding the quality of the FSN were separately demonstrated for FSN in German and English language. In brief, manufacturers often sent their FSN to the BfArM with delay. Additionally, a subset of FSN provided insufficient information on the product related risks or the measures to be performed by the customer to mitigate product related risks. Furthermore, customer confirmation forms often were missing in the FSN sent to the BfArM. Our data suggest that for IVD for infection testing FSCA and FSN are frequently performed. For better vigilance performance, manufacturers could shorten the time until release and improve the contents of FSN to ensure the safety of IVD in cases of product related corrective actions.

Intervertebral disc segmentation in MR images with 3D convolutional networks

NASA Astrophysics Data System (ADS)

Korez, Robert; Ibragimov, Bulat; Likar, Boštjan; Pernuš, Franjo; Vrtovec, Tomaž

2017-02-01

The vertebral column is a complex anatomical construct, composed of vertebrae and intervertebral discs (IVDs) supported by ligaments and muscles. During life, all components undergo degenerative changes, which may in some cases cause severe, chronic and debilitating low back pain. The main diagnostic challenge is to locate the pain generator, and degenerated IVDs have been identified to act as such. Accurate and robust segmentation of IVDs is therefore a prerequisite for computer-aided diagnosis and quantification of IVD degeneration, and can be also used for computer-assisted planning and simulation in spinal surgery. In this paper, we present a novel fully automated framework for supervised segmentation of IVDs from three-dimensional (3D) magnetic resonance (MR) spine images. By considering global intensity appearance and local shape information, a landmark-based approach is first used for the detection of IVDs in the observed image, which then initializes the segmentation of IVDs by coupling deformable models with convolutional networks (ConvNets). For this purpose, a 3D ConvNet architecture was designed that learns rich high-level appearance representations from a training repository of IVDs, and then generates spatial IVD probability maps that guide deformable models towards IVD boundaries. By applying the proposed framework to 15 3D MR spine images containing 105 IVDs, quantitative comparison of the obtained against reference IVD segmentations yielded an overall mean Dice coefficient of 92.8%, mean symmetric surface distance of 0.4 mm and Hausdorff surface distance of 3.7 mm.

Running exercise strengthens the intervertebral disc

PubMed Central

Belavý, Daniel L.; Quittner, Matthew J.; Ridgers, Nicola; Ling, Yuan; Connell, David; Rantalainen, Timo

2017-01-01

There is currently no evidence that the intervertebral discs (IVDs) can respond positively to exercise in humans. Some authors have argued that IVD metabolism in humans is too slow to respond anabolically to exercise within the human lifespan. Here we show that chronic running exercise in men and women is associated with better IVD composition (hydration and proteoglycan content) and with IVD hypertrophy. Via quantitative assessment of physical activity we further find that accelerations at fast walking and slow running (2 m/s), but not high-impact tasks, lower intensity walking or static positions, correlated to positive IVD characteristics. These findings represent the first evidence in humans that exercise can be beneficial for the IVD and provide support for the notion that specific exercise protocols may improve IVD material properties in the spine. We anticipate that our findings will be a starting point to better define exercise protocols and physical activity profiles for IVD anabolism in humans. PMID:28422125

“If I Were Nick”: Men’s Responses to an Interactive Video Drama Series to Support Smoking Cessation

PubMed Central

2015-01-01

Background Men continue to smoke in greater numbers than women; however, few interventions have been developed and tested to support men’s cessation. Men tend to rely on quitting strategies associated with stereotypical manliness, such as willpower, stoicism, and independence, but they may lack the self-efficacy skills required to sustain a quit. In this paper, we describe the development of and reception to an interactive video drama (IVD) series, composed of 7 brief scenarios, to support and strengthen men’s smoking cessation efforts. The value of IVD in health promotion is predicated on the evidence that viewers engage with the material when they are presented characters with whom they can personally identify. The video dramatizes the challenges unfolding in the life of the main character, Nick, on the first day of his quit and models the skills necessary to embark upon a sustainable quit. Objective The objective was to describe men’s responses to the If I were Nick IVD series as part of a study of QuitNow Men, an innovative smoking cessation website designed for men. Specific objectives were to explore the resonance of the main character of the IVD series with end-users and explore men’s perceptions of the effectiveness of the IVD series for supporting their quit self-management. Methods Seven brief IVD scenarios were developed, filmed with a professional actor, and uploaded to a new online smoking cessation website, QuitNow Men. A sample of 117 men who smoked were recruited into the study and provided baseline data prior to access to the QuitNow Men website for a 6-month period. During this time, 47 men chose to view the IVDs. Their responses to questions about the IVDs were collected in online surveys at 3-month and 6-month time points and analyzed using descriptive statistics. Results The majority of participants indicated they related to the main character, Nick. Participants who “strongly agreed” they could relate to Nick perceived significantly higher levels of support from the IVDs than the “neutral” and “disagree” groups (P<.001, d=2.0, P<.001, d=3.1). The “agree” and “neutral” groups were significantly higher on rated support from the videos than the “disagree” (P<.001, d=2.2, P=.01, d=1.5). Participants’ perception of the main character was independent of participant age, education attainment, or previous quit attempts. Conclusions The findings suggest that IVD interventions may be an important addition to men’s smoking cessation programs. Given that the use of IVD scenarios in health promotion is in its infancy, the positive outcomes from this study signal the potential for IVD and warrant ongoing evaluation in smoking cessation and, more generally, men’s health promotion. PMID:26265410

The Substitution of Ion Vapor Deposited (IVD) Aluminum for Cadmium

DTIC Science & Technology

1990-05-25

Coating Spray 1 0.0030 15 nin a 250TF Epoxy Powder Coating Spray 1 0.0030 15 nln @ 250OF Zinc Phosphate Tank Immersion 1 0.0002 Not Required Whitford: P...1candidate protection systems. MCAIR includ ed barrier-type coatings in the corrosion inhibitors ý;iich wMY be controlled by a~r-to-ic regulations that are

Review of the results of the in vivo dosimetry during total skin electron beam therapy

PubMed Central

Guidi, Gabriele; Gottardi, Giovanni; Ceroni, Paola; Costi, Tiziana

2013-01-01

This work reviews results of in vivo dosimetry (IVD) for total skin electron beam (TSEB) therapy, focusing on new methods, data emerged within 2012. All quoted data are based on a careful review of the literature reporting IVD results for patients treated by means of TSEB therapy. Many of the reviewed papers refer mainly to now old studies and/or old guidelines and recommendations (by IAEA, AAPM and EORTC), because (due to intrinsic rareness of TSEB-treated pathologies) only a limited number of works and reports with a large set of numerical data and proper statistical analysis is up-to-day available in scientific literature. Nonetheless, a general summary of the results obtained by the now numerous IVD techniques available is reported; innovative devices and methods, together with areas of possible further and possibly multicenter investigations for TSEB therapies are highlighted. PMID:24936333

45 CFR 309.150 - What start-up costs are allowable for Tribal IV-D programs carried out under § 309.65(b) of this...

Code of Federal Regulations, 2010 CFR

2010-10-01

... 45 Public Welfare 2 2010-10-01 2010-10-01 false What start-up costs are allowable for Tribal IV-D... ENFORCEMENT (IV-D) PROGRAM Tribal IV-D Program Funding § 309.150 What start-up costs are allowable for Tribal IV-D programs carried out under § 309.65(b) of this part? Federal funds are available for costs of...

Qualitative grading of disc degeneration by magnetic resonance in the lumbar and cervical spine: lack of correlation with histology in surgical cases.

PubMed

Davies, B M; Atkinson, R A; Ludwinski, F; Freemont, A J; Hoyland, J A; Gnanalingham, K K

2016-08-01

Clinically, magnetic resonance (MR) imaging is the most effective non-invasive tool for assessing IVD degeneration. Histological examination of the IVD provides a more detailed assessment of the pathological changes at a tissue level. However, very few reports have studied the relationship between these techniques. Identifying a relationship may allow more detailed staging of IVD degeneration, of importance in targeting future regenerative therapies. To investigate the relationship between MR and histological grading of IVD degeneration in the cervical and lumbar spine in patients undergoing discectomy. Lumbar (N = 99) and cervical (N = 106) IVD samples were obtained from adult patients undergoing discectomy surgery for symptomatic IVD herniation and graded to ascertain a histological grade of degeneration. The pre-operative MR images from these patients were graded for the degree of IVD (MR grade) and vertebral end-plate degeneration (Modic Changes, MC). The relationship between histological and MR grades of degeneration were studied. In lumbar and cervical IVD the majority of samples (93%) exhibited moderate levels of degeneration (ie MR grades 3-4) on pre-operative MR scans. Histologically, most specimens displayed moderate to severe grades of degeneration in lumbar (99%) and cervical spine (93%). MR grade was weakly correlated with patient age in lumbar and cervical study groups. MR and histological grades of IVD degeneration did not correlate in lumbar or cervical study groups. MC were more common in the lumbar than cervical spine (e.g. 39 versus 20% grade 2 changes; p < 0.05), but failed to correlate with MR or histological grades for degeneration. In this surgical series, the resected IVD tissue displayed moderate to severe degeneration, but there is no correlation between MR and histological grades using a qualitative classification system. There remains a need for a quantitative, non-invasive, pre-clinical measure of IVD degeneration that correlates with histological changes seen in the IVD.

Detrimental effects of discectomy on intervertebral disc biology can be decelerated by growth factor treatment during surgery: a large animal organ culture model.

PubMed

Illien-Jünger, Svenja; Lu, Young; Purmessur, Devina; Mayer, Jillian E; Walter, Benjamin A; Roughley, Peter J; Qureshi, Sheeraz A; Hecht, Andrew C; Iatridis, James C

2014-11-01

Lumbar discectomies are common surgical interventions that treat radiculopathy by removing herniated and loose intervertebral disc (IVD) tissues. However, remaining IVD tissue can continue to degenerate resulting in long-term clinical problems. Little information is available on the effects of discectomy on IVD biology. Currently, no treatments exist that can suspend or reverse the degeneration of the remaining IVD. To improve the knowledge on how discectomy procedures influence IVD physiology and to assess the potential of growth factor treatment as an augmentation during surgery. To determine effects of discectomy on IVDs with and without transforming growth factor beta 3 (TGFβ3) augmentation using bovine IVD organ culture. This study determined effects of discectomy with and without TGFβ3 injection using 1-, 6-, and 19-day organ culture experiments. Treated IVDs were injected with 0.2 μg TGFβ3 in 20 μL phosphate-buffered saline+bovine serum albumin into several locations of the discectomy site. Cell viability, gene expression, nitric oxide (NO) release, IVD height, aggrecan degradation, and proteoglycan content were determined. Discectomy significantly increased cell death, aggrecan degradation, and NO release in healthy IVDs. Transforming growth factor beta 3 injection treatment prevented or mitigated these effects for the 19-day culture period. Discectomy procedures induced cell death, catabolism, and NO production in healthy IVDs, and we conclude that post-discectomy degeneration is likely to be associated with cell death and matrix degradation. Transforming growth factor beta 3 injection augmented discectomy procedures by acting to protect IVD tissues by maintaining cell viability, limiting matrix degradation, and suppressing NO. We conclude that discectomy procedures can be improved with injectable therapies at the time of surgery although further in vivo and human studies are required. Copyright © 2014 Elsevier Inc. All rights reserved.

Evaluation of Lumbar Intervertebral Disc Degeneration Using T1ρ and T2 Magnetic Resonance Imaging in a Rabbit Disc Injury Model.

PubMed

Ishikawa, Tetsuhiro; Watanabe, Atsuya; Kamoda, Hiroto; Miyagi, Masayuki; Inoue, Gen; Takahashi, Kazuhisa; Ohtori, Seiji

2018-04-01

An in vivo histologic and magnetic resonance imaging (MRI) study of lumbar intervertebral disc (IVD) degeneration was conducted. To clarify the sensitivity and efficacy of T1ρ/T2 mapping for IVD degeneration, the correlation between T1ρ/T2 mapping and degenerative grades and histological findings in the lumbar IVD were investigated. The early signs of IVD degeneration are proteoglycan loss, dehydration, and collagen degradation. Recently, several quantitative MRI techniques have been developed; T2 mapping can be used to evaluate hydration and collagen fiber integrity within cartilaginous tissue, and T1ρ mapping can be used to evaluate hydration and proteoglycan content. Using New Zealand White rabbits, annular punctures of the IVD were made 10 times at L2/3, 5 times at L3/4, and one time at L4/5 using an 18-gauge needle (n=6) or a 21-gauge needle (n=6). At 4 and 8 weeks post-surgery, MRI was performed including T1ρ and T2 mapping. The degree of IVD degeneration was macroscopically assessed using the Thompson grading system. All specimens were cut for hematoxylin and eosin, safranin-O, and toluidine blue staining. Disc degeneration became more severe as the number of punctures increased and when the larger needle was used. T1ρ and T2 values were significantly different between grade 1 and grade 3 IVDs, grade 1 and grade 4 IVDs, grade 2 and grade 3 IVDs, and grade 2 and grade 4 IVDs ( p <0.05). There was a significant difference between grade 1 and grade 2 IVDs only in terms of T1ρ values ( p <0.05). T1ρ and T2 quantitative MRI could detect these small differences. Our results suggest that T1ρ and T2 mapping are sensitive to degenerative changes of lumbar IVDs and that T1ρ mapping can be used as a clinical tool to identify early IVD degeneration.

Anti-inflammatory Chitosan/Poly-γ-glutamic acid nanoparticles control inflammation while remodeling extracellular matrix in degenerated intervertebral disc.

PubMed

Teixeira, Graciosa Q; Leite Pereira, Catarina; Castro, Flávia; Ferreira, Joana R; Gomez-Lazaro, Maria; Aguiar, Paulo; Barbosa, Mário A; Neidlinger-Wilke, Cornelia; Goncalves, Raquel M

2016-09-15

Intervertebral disc (IVD) degeneration is one of the most common causes of low back pain (LBP), the leading disorder in terms of years lived with disability. Inflammation can play a role in LPB, while impairs IVD regeneration. In spite of this, different inflammatory targets have been purposed in the context of IVD regeneration. Anti-inflammatory nanoparticles (NPs) of Chitosan and Poly-(γ-glutamic acid) with a non-steroidal anti-inflammatory drug, diclofenac (Df), were previously shown to counteract a pro-inflammatory response of human macrophages. Here, the effect of intradiscal injection of Df-NPs in degenerated IVD was evaluated. For that, Df-NPs were injected in a bovine IVD organ culture in pro-inflammatory/degenerative conditions, upon stimulation with needle-puncture and interleukin (IL)-1β. Df-NPs were internalized by IVD cells, down-regulating IL-6, IL-8, MMP1 and MMP3, and decreasing PGE2 production, compared with IL-1β-stimulated IVD punches. Interestingly, at the same time, Df-NPs promoted an up-regulation of extracellular matrix (ECM) proteins, namely collagen type II and aggrecan. Allover, this study suggests that IVD treatment with Df-NPs not only reduces inflammation, but also delays and/or decreases ECM degradation, opening perspectives to new intradiscal therapies for IVD degeneration, based on the modulation of inflammation. Degeneration of the IVD is an age-related progressive process considered to be the major cause of spine disorders. The pro-inflammatory environment and biomechanics of the degenerated IVD is a challenge for regenerative therapies. The novelty of this work is the intradiscal injection of an anti-inflammatory therapy based on Chitosan (Ch)/Poly-(γ-glutamic acid) (γ-PGA) nanoparticles (NPs) with an anti-inflammatory drug (diclofenac, Df), previously developed by us. This drug delivery system was tested in a pro-inflammatory/degenerative intervertebral disc ex vivo model. The main findings support the success of an anti-inflammatory therapy for degenerated IVD that not only reduces inflammation but also promotes native IVD matrix production. Copyright © 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

SU-E-T-484: In Vivo Dosimetry Tolerances in External Beam Fast Neutron Therapy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Young, L; Gopan, O

Purpose: Optical stimulated luminescence (OSL) dosimetry with Landauer Al2O3:C nanodots was developed at our institution as a passive in vivo dosimetry (IVD) system for patients treated with fast neutron therapy. The purpose of this study was to establish clinically relevant tolerance limits for detecting treatment errors requiring further investigation. Methods: Tolerance levels were estimated by conducting a series of IVD expected dose calculations for square field sizes ranging between 2.8 and 28.8 cm. For each field size evaluated, doses were calculated for open and internal wedged fields with angles of 30°, 45°, or 60°. Theoretical errors were computed for variationsmore » of incorrect beam configurations. Dose errors, defined as the percent difference from the expected dose calculation, were measured with groups of three nanodots placed in a 30 x 30 cm solid water phantom, at beam isocenter (150 cm SAD, 1.7 cm Dmax). The tolerances were applied to IVD patient measurements. Results: The overall accuracy of the nanodot measurements is 2–3% for open fields. Measurement errors agreed with calculated errors to within 3%. Theoretical estimates of dosimetric errors showed that IVD measurements with OSL nanodots will detect the absence of an internal wedge or a wrong wedge angle. Incorrect nanodot placement on a wedged field is more likely to be caught if the offset is in the direction of the “toe” of the wedge where the dose difference in percentage is about 12%. Errors caused by an incorrect flattening filter size produced a 2% measurement error that is not detectable by IVD measurement alone. Conclusion: IVD with nanodots will detect treatment errors associated with the incorrect implementation of the internal wedge. The results of this study will streamline the physicists’ investigations in determining the root cause of an IVD reading that is out of normally accepted tolerances.« less

Assessment of the knowledge of graphical symbols labelled on malaria rapid diagnostic tests in four international settings

PubMed Central

2011-01-01

Background Graphical symbols on in vitro diagnostics (IVD symbols) replace the need for text in different languages and are used on malaria rapid diagnostic tests (RDTs) marketed worldwide. The present study assessed the comprehension of IVD symbols labelled on malaria RDT kits among laboratory staff in four different countries. Methods Participants (n = 293) in Belgium (n = 96), the Democratic Republic of the Congo (DRC, n = 87), Cambodia (n = 59) and Cuba (n = 51) were presented with an anonymous questionnaire with IVD symbols extracted from ISO 15223 and EN 980 presented as stand-alone symbols (n = 18) and in context (affixed on RDT packages, n = 16). Responses were open-ended and scored for correctness by local professionals. Results Presented as stand-alone, three and five IVD symbols were correctly scored for comprehension by 67% and 50% of participants; when contextually presented, five and seven symbols reached the 67% and 50% correct score respectively. 'Batch code' scored best (correctly scored by 71.3% of participants when presented as stand-alone), 'Authorized representative in the European Community' scored worst (1.4% correct). Another six IVD symbols were scored correctly by less than 10% of participants: 'Do not reuse', 'In vitro diagnostic medical device', 'Sufficient for', 'Date of manufacture', 'Authorised representative in EC', and 'Do not use if package is damaged'. Participants in Belgium and Cuba both scored six symbols above the 67% criterion, participants from DRC and Cambodia scored only two and one symbols above this criterion. Low correct scores were observed for safety-related IVD symbols, such as for 'Biological Risk' (42.7%) and 'Do not reuse' (10.9%). Conclusion Comprehension of IVD symbols on RDTs among laboratory staff in four international settings was unsatisfactory. Administrative and outreach procedures should be undertaken to assure their acquaintance by end-users. PMID:22047089

45 CFR 310.15 - What are the safeguards and processes that comprehensive Tribal IV-D agencies must have in place...

Code of Federal Regulations, 2010 CFR

2010-10-01

... IV-D Systems and Office Automation? 310.15 Section 310.15 Public Welfare Regulations Relating to... AND OFFICE AUTOMATION Requirements for Computerized Tribal IV-D Systems and Office Automation § 310.15... ensure the security and privacy of Computerized Tribal IV-D Systems and Office Automation? (a...

45 CFR 310.40 - What requirements apply for accessing systems and records for monitoring Computerized Tribal IV-D...

Code of Federal Regulations, 2010 CFR

2010-10-01

... records for monitoring Computerized Tribal IV-D Systems and Office Automation? 310.40 Section 310.40... COMPUTERIZED TRIBAL IV-D SYSTEMS AND OFFICE AUTOMATION Accountability and Monitoring Procedures for... monitoring Computerized Tribal IV-D Systems and Office Automation? In accordance with Part 95 of this title...

45 CFR 310.25 - What conditions apply to acquisitions of Computerized Tribal IV-D Systems?

Code of Federal Regulations, 2014 CFR

2014-10-01

... acquisitions of Computerized Tribal IV-D Systems? (a) APD Approval. A comprehensive Tribal IV-D agency must... include a clause that provides that the comprehensive Tribal IV-D agency will have all ownership rights to... use and to authorize others to use for Federal Government purposes, such software, modifications and...

45 CFR 310.25 - What conditions apply to acquisitions of Computerized Tribal IV-D Systems?

Code of Federal Regulations, 2011 CFR

2011-10-01

... acquisitions of Computerized Tribal IV-D Systems? (a) APD Approval. A comprehensive Tribal IV-D agency must... include a clause that provides that the comprehensive Tribal IV-D agency will have all ownership rights to... use and to authorize others to use for Federal Government purposes, such software, modifications and...

45 CFR 310.25 - What conditions apply to acquisitions of Computerized Tribal IV-D Systems?

Code of Federal Regulations, 2013 CFR

2013-10-01

... acquisitions of Computerized Tribal IV-D Systems? (a) APD Approval. A comprehensive Tribal IV-D agency must... include a clause that provides that the comprehensive Tribal IV-D agency will have all ownership rights to... use and to authorize others to use for Federal Government purposes, such software, modifications and...

In vivo bioavailability and in vitro bioaccessibility of perfluorooctanoic acid (PFOA) in food matrices: correlation analysis and method development.

PubMed

Li, Kan; Li, Chao; Yu, Nan-Yang; Juhasz, Albert L; Cui, Xin-Yi; Ma, Lena Q

2015-01-06

Food is a major source of human exposure to perfluorooctanoic acid (PFOA), however, PFOA bioavailability in food has not been studied. An in vivo mouse model and three in vitro methods (unified BARGE method, UBM; physiologically based extraction test, PBET; and in vitro digestion method, IVD) were used to determine the relative bioavailability and bioaccessibility of PFOA in the presence of 17 foods. PFOA was mixed with foods of different nutritional compositions and fed to mice over a 7-d period. PFOA relative bioavailability was determined by comparing PFOA accumulation in the liver following PFOA exposure via food to that in water. PFOA bioavailability relative to water ranged from 4.30 ± 0.80 to 69.0 ± 11.9% and was negatively correlated with lipid content (r = 0.76). This was possibly due to competitive sorption of free fatty acids with PFOA onto transporters on intestine epithelial cells. Besides, cations in the gastrointestinal tract, such as Ca(2+) and Mg(2+), are capable of complexing PFOA and partitioning to the lipid phase. On the other hand, when assessed using in vitro assays, PFOA bioaccessibility varied with methods, being 8.7-73% (UBM), 9.8-99% (PBET), and 21-114% (IVD). PFOA bioaccessibility was negatively correlated with lipid content when assessed using UBM (r = 0.82); however, a poor correlation with food composition was observed for PBET and IVD (r = 0.01-0.50). When in vivo and in vitro data were compared, a strong correlation was observed for UBM (r = 0.79), but poor relationships were observed for PBET and IVD (r = 0.11-0.22). This was probably because the higher lipolysis ability and presence of Ca(2+) and Mg(2+) in the gastrointestinal fluid of UBM resulted in a lower potential to form stable micelles compared to PBET and IVD. These results indicated that PFOA relative bioavailability was mainly affected by lipid content in foods, and UBM has the potential to determine PFOA bioaccessibility in food samples.

Moderate-intensity running causes intervertebral disc compression in young adults.

PubMed

Kingsley, Michael Ian; D'Silva, Lindsay Antonio; Jennings, Cameron; Humphries, Brendan; Dalbo, Vincent James; Scanlan, Aaron Terrance

2012-11-01

Decreased intervertebral disc (IVD) volume can result in diminished load-carrying capacity of the spinal region. Although moderate-intensity running is generally advocated for apparently healthy adults, running causes a loss in stature that is thought to reflect IVD compression. The aim of this investigation was to use magnetic resonance imaging (MRI) to quantify the influence of moderate-intensity treadmill running on IVD height and volume in the thoracic and lumbar regions of the vertebral column. A clinic-based repeated-measures design was used in eight healthy young asymptomatic adults. After preliminary measurements and familiarization (day 1), participants reported to the clinic on two further occasions. MRI scans and stature measurements were completed at baseline (day 2), preexercise (day 3), and after 30 min of moderate-intensity treadmill running (postexercise, day 3). Mean height and volume were derived for all thoracic and lumbar IVDs from digitized MRIs, and stature was determined with a stadiometer. Moderate-intensity running resulted in 6.3% ± 0.9% reduction in mean IVD height and 6.9% ± 1.0% reduction in calculated IVD volume. The day-to-day variation in mean IVD height and volume were 0.6% ± 0.6% and 0.4% ± 0.6%, respectively. This is the first study to quantify the influence of moderate-intensity running on IVD height and volume. Changes in IVD height and volume were observed throughout the thoracic and lumbar vertebral regions. These findings suggest that future studies evaluating the influence of various loading activities and recovery techniques on IVD structure should consider thoracic as well as lumbar regions of the spine.

Beam related response of in vivo diode detectors for external radiotherapy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Baci, Syrja, E-mail: sbarci2013@gmail.com; Telhaj, Ervis; Malkaj, Partizan

2016-03-25

In Vivo Dosimetry (IVD) is a set of methods used in cancer treatment clinics to determine the real dose of radiation absorbed by target volume in a patient’s body. IVD has been widely implemented in radiotherapy treatment centers and is now recommended part of Quality Assurance program by many International health and radiation organizations. Because of cost and lack of specialized personnel, IVD has not been practiced as yet, in Albanian radiotherapy clinics. At Hygeia Hospital Tirana, patients are irradiated with high energy photons generated by Elekta Synergy Accelerators. We have recently started experimenting with the purpose of establishing anmore » IVD practice at this hospital. The first set of experiments was aimed at calibration of diodes that are going to be used for IVD. PMMA, phantoms by PTW were used to calibrate p – type Si, semiconductor diode dosimeters, made by PTW Freiburg for entrance dose. Response of the detectors is affected by energy of the beam, accumulated radiation dose, dose rate, temperature, angle against the beam axis, etc. Here we present the work done for calculating calibration factor and correction factors of source to surface distance, field size, and beam incidence for the entrance dose for both 6 MV photon beam and 18 MV photon beam. Dependence of dosimeter response was found to be more pronounced with source to surface distance as compared to other variables investigated.« less

45 CFR 310.15 - What are the safeguards and processes that comprehensive Tribal IV-D agencies must have in place...

Code of Federal Regulations, 2014 CFR

2014-10-01

... comprehensive Tribal IV-D agencies must have in place to ensure the security and privacy of Computerized Tribal... ensure the security and privacy of Computerized Tribal IV-D Systems and Office Automation? (a..., accuracy, completeness, access to, and use of data in the Computerized Tribal IV-D System and Office...

45 CFR 309.135 - What requirements apply to funding, obligating and liquidating Federal title IV-D grant funds?

Code of Federal Regulations, 2011 CFR

2011-10-01

... 45 Public Welfare 2 2011-10-01 2011-10-01 false What requirements apply to funding, obligating and... (IV-D) PROGRAM Tribal IV-D Program Funding § 309.135 What requirements apply to funding, obligating and liquidating Federal title IV-D grant funds? (a) Funding period—(1) Ongoing funding. Federal title...

45 CFR 309.135 - What requirements apply to funding, obligating and liquidating Federal title IV-D grant funds?

Code of Federal Regulations, 2012 CFR

2012-10-01

... 45 Public Welfare 2 2012-10-01 2012-10-01 false What requirements apply to funding, obligating and... (IV-D) PROGRAM Tribal IV-D Program Funding § 309.135 What requirements apply to funding, obligating and liquidating Federal title IV-D grant funds? (a) Funding period—(1) Ongoing funding. Federal title...

45 CFR 309.135 - What requirements apply to funding, obligating and liquidating Federal title IV-D grant funds?

Code of Federal Regulations, 2014 CFR

2014-10-01

... 45 Public Welfare 2 2014-10-01 2012-10-01 true What requirements apply to funding, obligating and... (IV-D) PROGRAM Tribal IV-D Program Funding § 309.135 What requirements apply to funding, obligating and liquidating Federal title IV-D grant funds? (a) Funding period—(1) Ongoing funding. Federal title...

45 CFR 309.135 - What requirements apply to funding, obligating and liquidating Federal title IV-D grant funds?

Code of Federal Regulations, 2013 CFR

2013-10-01

... 45 Public Welfare 2 2013-10-01 2012-10-01 true What requirements apply to funding, obligating and... (IV-D) PROGRAM Tribal IV-D Program Funding § 309.135 What requirements apply to funding, obligating and liquidating Federal title IV-D grant funds? (a) Funding period—(1) Ongoing funding. Federal title...

45 CFR 309.135 - What requirements apply to funding, obligating and liquidating Federal title IV-D grant funds?

Code of Federal Regulations, 2010 CFR

2010-10-01

... 45 Public Welfare 2 2010-10-01 2010-10-01 false What requirements apply to funding, obligating and... (IV-D) PROGRAM Tribal IV-D Program Funding § 309.135 What requirements apply to funding, obligating and liquidating Federal title IV-D grant funds? (a) Funding period—(1) Ongoing funding. Federal title...

45 CFR 309.35 - What are the procedures for review of a Tribal IV-D program application, plan or plan amendment?

Code of Federal Regulations, 2010 CFR

2010-10-01

...-D program application, plan or plan amendment? 309.35 Section 309.35 Public Welfare Regulations... ENFORCEMENT (IV-D) PROGRAM Tribal IV-D Program Application Procedures § 309.35 What are the procedures for review of a Tribal IV-D program application, plan or plan amendment? (a) The Secretary will promptly...

Can Exercise Positively Influence the Intervertebral Disc?

PubMed

Belavý, Daniel L; Albracht, Kirsten; Bruggemann, Gert-Peter; Vergroesen, Pieter-Paul A; van Dieën, Jaap H

2016-04-01

To better understand what kinds of sports and exercise could be beneficial for the intervertebral disc (IVD), we performed a review to synthesise the literature on IVD adaptation with loading and exercise. The state of the literature did not permit a systematic review; therefore, we performed a narrative review. The majority of the available data come from cell or whole-disc loading models and animal exercise models. However, some studies have examined the impact of specific sports on IVD degeneration in humans and acute exercise on disc size. Based on the data available in the literature, loading types that are likely beneficial to the IVD are dynamic, axial, at slow to moderate movement speeds, and of a magnitude experienced in walking and jogging. Static loading, torsional loading, flexion with compression, rapid loading, high-impact loading and explosive tasks are likely detrimental for the IVD. Reduced physical activity and disuse appear to be detrimental for the IVD. We also consider the impact of genetics and the likelihood of a 'critical period' for the effect of exercise in IVD development. The current review summarises the literature to increase awareness amongst exercise, rehabilitation and ergonomic professionals regarding IVD health and provides recommendations on future directions in research.

Direct bacterial identification in positive blood cultures by use of two commercial matrix-assisted laser desorption ionization-time of flight mass spectrometry systems.

PubMed

Chen, Jonathan H K; Ho, Pak-Leung; Kwan, Grace S W; She, Kevin K K; Siu, Gilman K H; Cheng, Vincent C C; Yuen, Kwok-Yung; Yam, Wing-Cheong

2013-06-01

Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for the identification of bacteria and fungi was recently introduced in microbiology laboratories. This technology could greatly improve the clinical management of patients and guidance for chemotherapy. In this study, we used a commercial MALDI Sepsityper extraction method to evaluate the performance of two commercial MALDI-TOF MS systems, the Vitek MS IVD (bioMérieux) and the Microflex LT Biotyper (Bruker Daltonics) for direct bacterial identification in positive blood cultures. In 181 monomicrobial cultures, both systems generated genus to species level identifications for >90% of the specimens (Biotyper, 177/181 [97.8%]; Vitek MS IVD, 167/181 [92.3%]). Overall, the Biotyper system generated significantly more accurate identifications than the Vitek MS IVD system (P = 0.016; 177 versus 167 out of 181 specimens). The Biotyper system identified the minority species among polymicrobial blood cultures. We also compared the performance of an in-house extraction method with that of the Sepsityper on both MALDI-TOF MS systems. The in-house method generated more correct identifications at the genus level than the Sepsityper (96.7% versus 93.5%) on the Biotyper system, whereas the two methods exhibited the same performance level (88.0% versus 88.0%) on the Vitek MS IVD system. Our study confirmed the practical advantages of MALDI-TOF MS, and our in-house extraction method reduced the reagent cost to $1 per specimen, with a shorter turnaround time of 3 h, which is highly cost-effective for a diagnostic microbiology service.

How Long and Low Can You Go? Effect of Conformation on the Risk of Thoracolumbar Intervertebral Disc Extrusion in Domestic Dogs

PubMed Central

Packer, Rowena M. A.; Hendricks, Anke; Volk, Holger A.; Shihab, Nadia K.; Burn, Charlotte C.

2013-01-01

Intervertebral disc extrusion (IVDE) is a common neurological disorder in certain dog breeds, resulting in spinal cord compression and injury that can cause pain and neurological deficits. Most disc extrusions are reported in chondrodystrophic breeds (e.g. Dachshunds, Basset Hounds, Pekingese), where selection for ‘long and low’ morphologies is linked with intervertebral discs abnormalities that predispose dogs to IVDE. The aim of this study was to quantify the relationship between relative thoracolumbar vertebral column length and IVDE risk in diverse breeds. A 14 month cross-sectional study of dogs entering a UK small animal referral hospital for diverse disorders including IVDE was carried out. Dogs were measured on breed-defining morphometrics, including back length (BL) and height at the withers (HW). Of 700 dogs recruited from this referral population, measured and clinically examined, 79 were diagnosed with thoracolumbar IVDE following diagnostic imaging ± surgery. The BL:HW ratio was positively associated with IVDE risk, indicating that relatively longer dogs were at increased risk, e.g. the probability of IVDE was 0.30 for Miniature Dachshunds when BL:HW ratio equalled 1.1, compared to 0.68 when BL:HW ratio equalled 1.5. Additionally, both being overweight and skeletally smaller significantly increased IVDE risk. Therefore, selection for longer backs and miniaturisation should be discouraged in high-risk breeds to reduce IVDE risk. In higher risk individuals, maintaining a lean body shape is particularly important to reduce the risk of IVDE. Results are reported as probabilities to aid decision-making regarding breed standards and screening programmes reflecting the degree of risk acceptable to stakeholders. PMID:23894518

How long and low can you go? Effect of conformation on the risk of thoracolumbar intervertebral disc extrusion in domestic dogs.

PubMed

Packer, Rowena M A; Hendricks, Anke; Volk, Holger A; Shihab, Nadia K; Burn, Charlotte C

2013-01-01

Intervertebral disc extrusion (IVDE) is a common neurological disorder in certain dog breeds, resulting in spinal cord compression and injury that can cause pain and neurological deficits. Most disc extrusions are reported in chondrodystrophic breeds (e.g. Dachshunds, Basset Hounds, Pekingese), where selection for 'long and low' morphologies is linked with intervertebral discs abnormalities that predispose dogs to IVDE. The aim of this study was to quantify the relationship between relative thoracolumbar vertebral column length and IVDE risk in diverse breeds. A 14 month cross-sectional study of dogs entering a UK small animal referral hospital for diverse disorders including IVDE was carried out. Dogs were measured on breed-defining morphometrics, including back length (BL) and height at the withers (HW). Of 700 dogs recruited from this referral population, measured and clinically examined, 79 were diagnosed with thoracolumbar IVDE following diagnostic imaging ± surgery. The BL:HW ratio was positively associated with IVDE risk, indicating that relatively longer dogs were at increased risk, e.g. the probability of IVDE was 0.30 for Miniature Dachshunds when BL:HW ratio equalled 1.1, compared to 0.68 when BL:HW ratio equalled 1.5. Additionally, both being overweight and skeletally smaller significantly increased IVDE risk. Therefore, selection for longer backs and miniaturisation should be discouraged in high-risk breeds to reduce IVDE risk. In higher risk individuals, maintaining a lean body shape is particularly important to reduce the risk of IVDE. Results are reported as probabilities to aid decision-making regarding breed standards and screening programmes reflecting the degree of risk acceptable to stakeholders.

Histological and reference system for the analysis of mouse intervertebral disc.

PubMed

Tam, Vivian; Chan, Wilson C W; Leung, Victor Y L; Cheah, Kathryn S E; Cheung, Kenneth M C; Sakai, Daisuke; McCann, Matthew R; Bedore, Jake; Séguin, Cheryle A; Chan, Danny

2018-01-01

A new scoring system based on histo-morphology of mouse intervertebral disc (IVD) was established to assess changes in different mouse models of IVD degeneration and repair. IVDs from mouse strains of different ages, transgenic mice, or models of artificially induced IVD degeneration were assessed. Morphological features consistently observed in normal, and early/later stages of degeneration were categorized into a scoring system focused on nucleus pulposus (NP) and annulus fibrosus (AF) changes. "Normal NP" exhibited a highly cellularized cell mass that decreased with natural ageing and in disc degeneration. "Normal AF" consisted of distinct concentric lamellar structures, which was disrupted in severe degeneration. NP/AF clefts indicated more severe changes. Consistent scores were obtained between experienced and new users. Altogether, our scoring system effectively differentiated IVD changes in various strains of wild-type and genetically modified mice and in induced models of IVD degeneration, and is applicable from the post-natal stage to the aged mouse. This scoring tool and reference resource addresses a pressing need in the field for studying IVD changes and cross-study comparisons in mice, and facilitates a means to normalize mouse IVD assessment between different laboratories. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:233-243, 2018. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

3D multi-scale FCN with random modality voxel dropout learning for Intervertebral Disc Localization and Segmentation from Multi-modality MR Images.

PubMed

Li, Xiaomeng; Dou, Qi; Chen, Hao; Fu, Chi-Wing; Qi, Xiaojuan; Belavý, Daniel L; Armbrecht, Gabriele; Felsenberg, Dieter; Zheng, Guoyan; Heng, Pheng-Ann

2018-04-01

Intervertebral discs (IVDs) are small joints that lie between adjacent vertebrae. The localization and segmentation of IVDs are important for spine disease diagnosis and measurement quantification. However, manual annotation is time-consuming and error-prone with limited reproducibility, particularly for volumetric data. In this work, our goal is to develop an automatic and accurate method based on fully convolutional networks (FCN) for the localization and segmentation of IVDs from multi-modality 3D MR data. Compared with single modality data, multi-modality MR images provide complementary contextual information, which contributes to better recognition performance. However, how to effectively integrate such multi-modality information to generate accurate segmentation results remains to be further explored. In this paper, we present a novel multi-scale and modality dropout learning framework to locate and segment IVDs from four-modality MR images. First, we design a 3D multi-scale context fully convolutional network, which processes the input data in multiple scales of context and then merges the high-level features to enhance the representation capability of the network for handling the scale variation of anatomical structures. Second, to harness the complementary information from different modalities, we present a random modality voxel dropout strategy which alleviates the co-adaption issue and increases the discriminative capability of the network. Our method achieved the 1st place in the MICCAI challenge on automatic localization and segmentation of IVDs from multi-modality MR images, with a mean segmentation Dice coefficient of 91.2% and a mean localization error of 0.62 mm. We further conduct extensive experiments on the extended dataset to validate our method. We demonstrate that the proposed modality dropout strategy with multi-modality images as contextual information improved the segmentation accuracy significantly. Furthermore, experiments conducted on extended data collected from two different time points demonstrate the efficacy of our method on tracking the morphological changes in a longitudinal study. Copyright © 2018 Elsevier B.V. All rights reserved.

Computation models simulating notochordal cell extinction during early ageing of an intervertebral disc.

PubMed

Louman-Gardiner, K M; Coombe, D; Hunter, C J

2011-12-01

Lower back pain due to intervertebral disc (IVD) degeneration is a prevalent problem which drastically affects the quality of life of millions of sufferers. Healthy IVDs begin with high populations of notochordal cells in the nucleus pulposus, while by the second stage of degeneration, these cells will be replaced by chondrocyte-like cells. Because the IVD is avascular, these cells rely on passive diffusion of nutrients to survive. It is thought that this transition in cell phenotype causes the shift of the IVD's physical properties, which impede the flow of nutrients. Our computational model of the IVD illustrates its ability to simulate the evolving chemical and mechanical environments occurring during the early ageing process. We demonstrate that, due to the insufficient nutrient supply and accompanying changes in physical properties of the IVD, there was a resultant exponential decay in the number of notochordal cells over time.

Combined Dynamic Contrast Enhanced Liver MRI and MRA Using Interleaved Variable Density Sampling

PubMed Central

Rahimi, Mahdi Salmani; Korosec, Frank R.; Wang, Kang; Holmes, James H.; Motosugi, Utaroh; Bannas, Peter; Reeder, Scott B.

2014-01-01

Purpose To develop and evaluate a method for volumetric contrast-enhanced MR imaging of the liver, with high spatial and temporal resolutions, for combined dynamic imaging and MR angiography using a single injection of contrast. Methods An interleaved variable density (IVD) undersampling pattern was implemented in combination with a real-time-triggered, time-resolved, dual-echo 3D spoiled gradient echo sequence. Parallel imaging autocalibration lines were acquired only once during the first time-frame. Imaging was performed in ten subjects with focal nodular hyperplasia (FNH) and compared with their clinical MRI. The angiographic phase of the proposed method was compared to a dedicated MR angiogram acquired during a second injection of contrast. Results A total of 21 FNH, 3 cavernous hemangiomas, and 109 arterial segments were visualized in 10 subjects. The temporally-resolved images depicted the characteristic arterial enhancement pattern of the lesions with a 4 s update rate. Images were graded as having significantly higher quality compared to the clinical MRI. Angiograms produced from the IVD method provided non-inferior diagnostic assessment compared to the dedicated MRA. Conclusion Using an undersampled IVD imaging method, we have demonstrated the feasibility of obtaining high spatial and temporal resolution dynamic contrast-enhanced imaging and simultaneous MRA of the liver. PMID:24639130

CRISPR Epigenome Editing of AKAP150 in DRG Neurons Abolishes Degenerative IVD-Induced Neuronal Activation.

PubMed

Stover, Joshua D; Farhang, Niloofar; Berrett, Kristofer C; Gertz, Jason; Lawrence, Brandon; Bowles, Robby D

2017-09-06

Back pain is a major contributor to disability and has significant socioeconomic impacts worldwide. The degenerative intervertebral disc (IVD) has been hypothesized to contribute to back pain, but a better understanding of the interactions between the degenerative IVD and nociceptive neurons innervating the disc and treatment strategies that directly target these interactions is needed to improve our understanding and treatment of back pain. We investigated degenerative IVD-induced changes to dorsal root ganglion (DRG) neuron activity and utilized CRISPR epigenome editing as a neuromodulation strategy. By exposing DRG neurons to degenerative IVD-conditioned media under both normal and pathological IVD pH levels, we demonstrate that degenerative IVDs trigger interleukin (IL)-6-induced increases in neuron activity to thermal stimuli, which is directly mediated by AKAP and enhanced by acidic pH. Utilizing this novel information on AKAP-mediated increases in nociceptive neuron activity, we developed lentiviral CRISPR epigenome editing vectors that modulate endogenous expression of AKAP150 by targeted promoter histone methylation. When delivered to DRG neurons, these epigenome-modifying vectors abolished degenerative IVD-induced DRG-elevated neuron activity while preserving non-pathologic neuron activity. This work elucidates the potential for CRISPR epigenome editing as a targeted gene-based pain neuromodulation strategy. Copyright © 2017 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.

Notochordal cell conditioned medium stimulates mesenchymal stem cell differentiation toward a young nucleus pulposus phenotype

PubMed Central

2010-01-01

Introduction Mesenchymal stem cells (MSCs) offer promise for intervertebral disc (IVD) repair and regeneration because they are easily isolated and expanded, and can differentiate into several mesenchymal tissues. Notochordal (NC) cells contribute to IVD development, incorporate into the nucleus pulposus (NP), and stimulate mature disc cells. However, there have been no studies investigating the effects of NC cells on adult stem cell differentiation. The premise of this study is that IVD regeneration is more similar to IVD development than to IVD maintenance, and we hypothesize that soluble factors from NC cells differentiate MSCs to a phenotype characteristic of nucleus pulposus (NP) cells during development. The eventual clinical goal would be to isolate or chemically/recombinantly produce the active agent to induce the therapeutic effects, and to use it as either an injectable therapy for early intervention on disc disease, or in developing appropriately pre-differentiated MSC cells in a tissue engineered NP construct. Methods Human MSCs from bone marrow were expanded and pelleted to form high-density cultures. MSC pellets were exposed to either control medium (CM), chondrogenic medium (CM with dexamethasone and transforming growth factor, (TGF)-β3) or notochordal cell conditioned medium (NCCM). NCCM was prepared from NC cells maintained in serum free medium for four days. After seven days culture, MSC pellets were analyzed for appearance, biochemical composition (glycosaminoglycans and DNA), and gene expression profile (sox-9, collagen types-II and III, laminin-β1 and TIMP1(tissue inhibitor of metalloproteinases-1)). Results Significantly higher glycosaminoglycan accumulation was seen in NCCM treated pellets than in CM or TGFβ groups. With NCCM treatment, increased gene expression of collagen III, and a trend of increasing expression of laminin-β1 and decreased expression of sox-9 and collagen II relative to the TGFβ group was observed. Conclusions Together, results suggest NCCM stimulates mesenchymal stem cell differentiation toward a potentially NP-like phenotype with some characteristics of the developing IVD. PMID:20565707

Poster — Thur Eve — 25: Sensitivity to inhomogeneities for an in-vivo EPID dosimetry method

DOE Office of Scientific and Technical Information (OSTI.GOV)

Peca, Stefano; Brown, Derek; Department of Physics and Astronomy, University of Calgary, Calgary, AB

2014-08-15

Introduction: The electronic portal imaging device (EPID) has the potential to be used for in vivo dosimetry during radiotherapy as an additional dose delivery check. We recently proposed a simple method of using the EPID for 2D-IVD based on correlation ratios. In this work we have investigated the sensitivity of our EPID-IVD to inhomogeneities. Methods: We used slab phantoms that simulate water, bone, and lung, arranged in various geometries. To simulate body contours non-orthogonal to the field, we used a water wedge. CT data of these phantoms was imported into MATLAB, in conjunction with EPID images acquired during irradiation, tomore » calculate dose inside the phantom in isocenter plane. Each phantom was irradiated using a linear accelerator while images were acquired with the EPID (cine mode). Comparisons between EPID-calculated and TPS dose maps were: pixel-by-pixel dose difference, and 3%,3mm gamma evaluation. Results: In the homogeneous case, CAX dose difference was <1%, and 3%,3mm gamma analysis yielded 99% of points with gamma<1. For the inhomogeneous phantoms, agreement decreased with increasing inhomogeneity reaching up to 10% CAX dose difference with 10cm of lung. Results from the water wedge phantom suggest that the EPID-calculated dose can account for surface irregularities of approximately ±3cm. Conclusions: The EPID-based IVD investigated has limitations in the presence of large inhomogeneities. Nonetheless, CAX doses never differed by >15% from the TPS. This suggests that this EPID-IVD is capable of detecting gross dose delivery errors even in the presence of inhomogeneities, supporting its utility as an additional patient safety device.« less

MRI signal distribution within the intervertebral disc as a biomarker of adolescent idiopathic scoliosis and spondylolisthesis.

PubMed

Gervais, Julien; Périé, Delphine; Parent, Stefan; Labelle, Hubert; Aubin, Carl-Eric

2012-12-03

Early stages of scoliosis and spondylolisthesis entail changes in the intervertebral disc (IVD) structure and biochemistry. The current clinical use of MR T2-weighted images is limited to visual inspection. Our hypothesis is that the distribution of the MRI signal intensity within the IVD in T2-weighted images depends on the spinal pathology and on its severity. Therefore, this study aims to develop the AMRSID (analysis of MR signal intensity distribution) method to analyze the 3D distribution of the MR signal intensity within the IVD and to evaluate their sensitivity to scoliosis and spondylolisthesis and their severities. This study was realized on 79 adolescents who underwent a MRI acquisition (sagittal T2-weighted images) before their orthopedic or surgical treatment. Five groups were considered: low severity scoliosis (Cobb angle ≤50°), high severity scoliosis (Cobb angles >50°), low severity spondylolisthesis (Meyerding grades I and II), high severity spondylolisthesis (Meyerding grades III, IV and V) and control. The distribution of the MRI signal intensity within the IVD was analyzed using the descriptive statistics of histograms normalized by either cerebrospinal fluid or bone signal intensity, weighted centers and volume ratios. Differences between pathology and severity groups were assessed using one- and two-way ANOVAs. There were significant (p < 0.05) variations of indices between scoliosis, spondylolithesis and control groups and between low and high severity groups. The cerebrospinal fluid normalization was able to detect differences between healthy and pathologic IVDs whereas the bone normalization, which reflects both bone and IVD health, detected more differences between the severities of these pathologies. This study proves for the first time that changes in the intervertebral disc, non visible to the naked eye on sagittal T2-weighted MR images of the spine, can be detected from specific indices describing the distribution of the MR signal intensity. Moreover, these indices are able to discriminate between scoliosis and spondylolisthesis and their severities, and provide essential information on the composition and structure of the discs whatever the pathology considered. The AMRSID method may have the potential to complement the current diagnostic tools available in clinics to improve the diagnostic with earlier biomarkers, the prognosis of evolution and the treatment options of scoliosis and spondylolisthesis.

Interactive Videodisc in Vocational Education. ERIC Digest No. 105.

ERIC Educational Resources Information Center

Kerka, Sandra

Interactive videodisc (IVD) offers a combination of media with practical applications in vocational education. IVD is superior to videotapes and other media in quality, applicability, and effectiveness. IVD can be used in different settings and for a variety of instructional applications. Although not appropriate for every learning situation, IVD…

45 CFR 303.7 - Provision of services in intergovernmental IV-D cases.

Code of Federal Regulations, 2011 CFR

2011-10-01

... ENFORCEMENT (CHILD SUPPORT ENFORCEMENT PROGRAM), ADMINISTRATION FOR CHILDREN AND FAMILIES, DEPARTMENT OF HEALTH AND HUMAN SERVICES STANDARDS FOR PROGRAM OPERATIONS § 303.7 Provision of services in intergovernmental IV-D cases. (a) General responsibilities. A State IV-D agency must: (1) Establish and use...

45 CFR 309.155 - What uses of Tribal IV-D program funds are not allowable?

Code of Federal Regulations, 2011 CFR

2011-10-01

... SUPPORT ENFORCEMENT (CHILD SUPPORT ENFORCEMENT PROGRAM), ADMINISTRATION FOR CHILDREN AND FAMILIES, DEPARTMENT OF HEALTH AND HUMAN SERVICES TRIBAL CHILD SUPPORT ENFORCEMENT (IV-D) PROGRAM Tribal IV-D Program... used for: (a) Activities related to administering other programs, including those under the Social...

45 CFR 310.25 - What conditions apply to acquisitions of Computerized Tribal IV-D Systems?

Code of Federal Regulations, 2010 CFR

2010-10-01

... FAMILIES, DEPARTMENT OF HEALTH AND HUMAN SERVICES COMPUTERIZED TRIBAL IV-D SYSTEMS AND OFFICE AUTOMATION... Acquisition Threshold; (c) Software and ownership rights. (1) All procurement and contract instruments must... Computerized Tribal IV-D System software or enhancements thereof and all associated documentation designed...

A comparison between porcine, ovine, and bovine intervertebral disc anatomy and single lamella annulus fibrosus tensile properties.

PubMed

Monaco, Lauren A; DeWitte-Orr, Stephanie J; Gregory, Diane E

2016-02-01

This project aimed to compare gross anatomical measures and biomechanical properties of single lamellae from the annulus fibrosus of ovine and porcine lumbar vertebrae, and bovine tail vertebrae. The morphology of the vertebrae of these species differ significantly both from each other and from human, yet how these differences alter biomechanical properties is unknown. Geometric parameters measured in this study included: 1) absolute and relative intervertebral (IVD) and vertebral body height and 2) absolute and relative intervertebral disc (IVD) anterior-posterior (AP) and medial-lateral (ML) widths. Single lamella tensile properties included toe-region stress and stretch ratio, stiffness, and tensile strength. As expected, the bovine tail IVD revealed a more circular shape compared with both the ovine and porcine lumbar IVD. The bovine tail also had the largest IVD to vertebral body height ratio (due to having the highest absolute IVD height). Bovine tail lamellae were also found to be strongest and stiffest (in tension) while ovine lumbar lamellae were weakest and most compliant. Histological analysis revealed the greatest proportion of collagen in the bovine corroborating findings of increased strength and stiffness. The observed differences in anatomical shape, connective tissue composition, and tensile properties need to be considered when choosing an appropriate model for IVD research. © 2015 Wiley Periodicals, Inc.

3D characterization of morphological changes in the intervertebral disc and endplate during aging: A propagation phase contrast synchrotron micro-tomography study

PubMed Central

Cao, Yong; Liao, Shenghui; Zeng, Hao; Ni, Shuangfei; Tintani, Francis; Hao, Yongqiang; Wang, Lei; Wu, Tianding; Lu, Hongbin; Duan, Chunyue; Hu, Jianzhong

2017-01-01

A better understanding of functional changes in the intervertebral disc (IVD) and interaction with endplate is essential to elucidate the pathogenesis of IVD degeneration disease (IDDD). To date, the simultaneous depiction of 3D micro-architectural changes of endplate with aging and interaction with IVD remains a technical challenge. We aim to characterize the 3D morphology changes of endplate and IVD during aging using PPCST. The lumbar vertebral level 4/5 IVDs harvested from 15-day-, 4- and 24-month-old mice were initially evaluated by PPCST with histological sections subsequently analyzed to confirm the imaging efficiency. Quantitative assessments of age-related trends after aging, including mean diameter, volume fraction and connectivity of the canals, and endplate porosity and thickness, reached a peak at 4 months and significantly decreased at 24 months. The IVD volume consistently exhibited same trend of variation with the endplate after aging. In this study, PPCST simultaneously provided comprehensive details of 3D morphological changes of the IVD and canal network in the endplate and the interaction after aging. The results suggest that PPCST has the potential to provide a new platform for attaining a deeper insight into the pathogenesis of IDDD, providing potential therapeutic targets. PMID:28266560

Quantitative Chemical Exchange Saturation Transfer MRI of Intervertebral Disc in a Porcine Model

PubMed Central

Zhou, Zhengwei; Bez, Maxim; Tawackoli, Wafa; Giaconi, Joseph; Sheyn, Dmitriy; de Mel, Sandra; Maya, Marcel M.; Pressman, Barry D.; Gazit, Zulma; Pelled, Gadi; Gazit, Dan; Li, Debiao

2017-01-01

Purpose Previous studies have associated low pH in interver-tebral discs (IVDs) with discogenic back pain. The purpose of this study was to determine whether quantitative CEST (qCEST) MRI can be used to detect pH changes in IVDs in vivo. Methods The exchange rate ksw between glycosaminoglycan (GAG) protons and water protons was determined from qCEST analysis. Its dependence on pH value was investigated in GAG phantoms with varying pH and concentrations. The relationship between ksw and pH was studied further in vivo in a porcine model on a 3T MR scanner and validated using a pH meter. Sodium lactate was injected into the IVDs to induce various pH values within the discs ranging from 5 to 7. Results Phantom and animal results revealed that ksw measured using qCEST MRI is highly correlated with pH level. In the animal studies, the relationship can be described as ksw =9.2 × 106 × 10−pH + 196.9, R2 = 0.7883. Conclusion The exchange rate between GAG and water protons determined from qCEST MRI is closely correlated with pH value. This technique has the potential to noninvasively measure pH in the IVDs of patients with discogenic pain. PMID:27670140

Organ culture bioreactors--platforms to study human intervertebral disc degeneration and regenerative therapy.

PubMed

Gantenbein, Benjamin; Illien-Jünger, Svenja; Chan, Samantha C W; Walser, Jochen; Haglund, Lisbet; Ferguson, Stephen J; Iatridis, James C; Grad, Sibylle

2015-01-01

In recent decades the application of bioreactors has revolutionized the concept of culturing tissues and organs that require mechanical loading. In intervertebral disc (IVD) research, collaborative efforts of biomedical engineering, biology and mechatronics have led to the innovation of new loading devices that can maintain viable IVD organ explants from large animals and human cadavers in precisely defined nutritional and mechanical environments over extended culture periods. Particularly in spine and IVD research, these organ culture models offer appealing alternatives, as large bipedal animal models with naturally occurring IVD degeneration and a genetic background similar to the human condition do not exist. Latest research has demonstrated important concepts including the potential of homing of mesenchymal stem cells to nutritionally or mechanically stressed IVDs, and the regenerative potential of "smart" biomaterials for nucleus pulposus or annulus fibrosus repair. In this review, we summarize the current knowledge about cell therapy, injection of cytokines and short peptides to rescue the degenerating IVD. We further stress that most bioreactor systems simplify the real in vivo conditions providing a useful proof of concept. Limitations are that certain aspects of the immune host response and pain assessments cannot be addressed with ex vivo systems. Coccygeal animal disc models are commonly used because of their availability and similarity to human IVDs. Although in vitro loading environments are not identical to the human in vivo situation, 3D ex vivo organ culture models of large animal coccygeal and human lumbar IVDs should be seen as valid alternatives for screening and feasibility testing to augment existing small animal, large animal, and human clinical trial experiments.

Vessel architecture in human knee cartilage in children: an in vivo susceptibility-weighted imaging study at 7 T.

PubMed

Kolb, Alexander; Robinson, Simon; Stelzeneder, David; Schreiner, Markus; Chiari, Catharina; Windhager, Reinhard; Trattnig, Siegfried; Bohndorf, Klaus

2018-02-26

To evaluate the clinical feasibility of ultrahigh field 7-T SWI to visualize vessels and assess their density in the immature epiphyseal cartilage of human knee joints. 7-T SWI of 12 knees (six healthy volunteers, six patients with osteochondral abnormalities; mean age 10.7 years; 3 female, 9 male) were analysed by two readers, classifying intracartilaginous vessel densities (IVD) in three grades (no vessels, low IVD and high IVD) in defined femoral, tibial and patellar zones. Differences between patients and volunteers, IVDs in different anatomic locations, differences between cartilage overlying osteochondral abnormalities and corresponding normal zones, and differences in age groups were analysed. Interrater reliability showed moderate agreement between the two readers (κ = 0.58, p < 0.001). The comparison of IVDs between patients and volunteers revealed no significant difference (p = 0.706). The difference between zones in the cartilage overlying osteochondral abnormalities to corresponding normal zones showed no significant difference (p = 0.564). IVDs were related to anatomic location, with decreased IVDs in loading areas (p = 0.003). IVD was age dependent, with more vessels present in the younger participants (p = 0.001). The use of SWI in conjunction with ultrahigh field MRI makes the in vivo visualization of vessels in the growing cartilage of humans feasible, providing insights into the role of the vessel network in acquired disturbances. • SWI facilitates in vivo visualization of vessels in the growing human cartilage. • Interrater reliability of the intracartilaginous vessel grading was moderate. • Intracartilaginous vessel densities are dependent on anatomical location and age.

45 CFR 310.5 - What options are available for Computerized Tribal IV-D Systems and office automation?

Code of Federal Regulations, 2010 CFR

2010-10-01

... to conduct automated data processing and recordkeeping activities through Office Automation... IV-D Systems and office automation? 310.5 Section 310.5 Public Welfare Regulations Relating to Public... AUTOMATION Requirements for Computerized Tribal IV-D Systems and Office Automation § 310.5 What options are...

45 CFR 309.170 - What statistical and narrative reporting requirements apply to Tribal IV-D programs?

Code of Federal Regulations, 2011 CFR

2011-10-01

... 45 Public Welfare 2 2011-10-01 2011-10-01 false What statistical and narrative reporting... (IV-D) PROGRAM Statistical and Narrative Reporting Requirements § 309.170 What statistical and... organizations must submit the following information and statistics for Tribal IV-D program activity and caseload...

45 CFR 309.170 - What statistical and narrative reporting requirements apply to Tribal IV-D programs?

Code of Federal Regulations, 2010 CFR

2010-10-01

... 45 Public Welfare 2 2010-10-01 2010-10-01 false What statistical and narrative reporting... (IV-D) PROGRAM Statistical and Narrative Reporting Requirements § 309.170 What statistical and... organizations must submit the following information and statistics for Tribal IV-D program activity and caseload...

"If I Were Nick": Men's Responses to an Interactive Video Drama Series to Support Smoking Cessation.

PubMed

Bottorff, Joan L; Sarbit, Gayl; Oliffe, John L; Kelly, Mary T; Lohan, Maria; Stolp, Sean; Sharp, Paul

2015-08-10

Men continue to smoke in greater numbers than women; however, few interventions have been developed and tested to support men's cessation. Men tend to rely on quitting strategies associated with stereotypical manliness, such as willpower, stoicism, and independence, but they may lack the self-efficacy skills required to sustain a quit. In this paper, we describe the development of and reception to an interactive video drama (IVD) series, composed of 7 brief scenarios, to support and strengthen men's smoking cessation efforts. The value of IVD in health promotion is predicated on the evidence that viewers engage with the material when they are presented characters with whom they can personally identify. The video dramatizes the challenges unfolding in the life of the main character, Nick, on the first day of his quit and models the skills necessary to embark upon a sustainable quit. The objective was to describe men's responses to the If I were Nick IVD series as part of a study of QuitNow Men, an innovative smoking cessation website designed for men. Specific objectives were to explore the resonance of the main character of the IVD series with end-users and explore men's perceptions of the effectiveness of the IVD series for supporting their quit self-management. Seven brief IVD scenarios were developed, filmed with a professional actor, and uploaded to a new online smoking cessation website, QuitNow Men. A sample of 117 men who smoked were recruited into the study and provided baseline data prior to access to the QuitNow Men website for a 6-month period. During this time, 47 men chose to view the IVDs. Their responses to questions about the IVDs were collected in online surveys at 3-month and 6-month time points and analyzed using descriptive statistics. The majority of participants indicated they related to the main character, Nick. Participants who "strongly agreed" they could relate to Nick perceived significantly higher levels of support from the IVDs than the "neutral" and "disagree" groups (P<.001, d=2.0, P<.001, d=3.1). The "agree" and "neutral" groups were significantly higher on rated support from the videos than the "disagree" (P<.001, d=2.2, P=.01, d=1.5). Participants' perception of the main character was independent of participant age, education attainment, or previous quit attempts. The findings suggest that IVD interventions may be an important addition to men's smoking cessation programs. Given that the use of IVD scenarios in health promotion is in its infancy, the positive outcomes from this study signal the potential for IVD and warrant ongoing evaluation in smoking cessation and, more generally, men's health promotion.

Poly(γ-glutamic acid) and poly(γ-glutamic acid)-based nanocomplexes enhance type II collagen production in intervertebral disc.

PubMed

Antunes, Joana C; Pereira, Catarina Leite; Teixeira, Graciosa Q; Silva, Ricardo V; Caldeira, Joana; Grad, Sibylle; Gonçalves, Raquel M; Barbosa, Mário A

2017-01-01

Intervertebral disc (IVD) degeneration often leads to low back pain, which is one of the major causes of disability worldwide, affecting more than 80% of the population. Although available treatments for degenerated IVD decrease symptoms' progression, they fail to address the underlying causes and to restore native IVD properties. Poly(γ-glutamic acid) (γ-PGA) has recently been shown to support the production of chondrogenic matrix by mesenchymal stem/stromal cells. γ-PGA/chitosan (Ch) nanocomplexes (NCs) have been proposed for several biomedical applications, showing advantages compared with either polymer alone. Hence, this study explores the potential of γ-PGA and γ-PGA/Ch NCs for IVD regeneration. Nucleotomised bovine IVDs were cultured ex vivo upon injection of γ-PGA (pH 7.4) and γ-PGA/Ch NCs (pH 5.0 and pH 7.4). Tissue metabolic activity and nucleus pulposus DNA content were significantly reduced when NCs were injected in acidic-buffered solution (pH 5.0). However, at pH 7.4, both γ-PGA and NCs promoted sulphated glycosaminoglycan production and significant type II collagen synthesis, as determined at the protein level. This study is a first proof of concept that γ-PGA and γ-PGA/Ch NCs promote recovery of IVD native matrix, opening new perspectives on the development of alternative therapeutic approaches for IVD degeneration.

Prevalence of Age-Related Changes in Ovine Lumbar Intervertebral Discs during Computed Tomography and Magnetic Resonance Imaging

PubMed Central

Nisolle, Jean-François; Bihin, Benoît; Kirschvink, Nathalie; Neveu, Fabienne; Clegg, Peter; Dugdale, Alexandra; Wang, Xiaoqing; Vandeweerd, Jean-Michel

2016-01-01

Ovine models are used to study intervertebral disc (IVD) degeneration. The objective of the current study was to assess the naturally occurring age-related changes of the IVD that can be diagnosed by CT and MRI in the lumbar spine of sheep. We used CT and T2-weighted MR images to score the IVD (L6S1 to L1L2) in 41 sheep (age, 6 mo to 11 y) that were euthanized for reasons not related to musculoskeletal disease. T2 mapping and measurement of T2 time of L6S1 to L2L3 were performed in 22 of the sheep. Degenerative changes manifested as early as 2 y of age and occurred at every IVD level. Discs were more severely damaged in older sheep. The age effect of the L6S1 IVD was larger than the average age effect for the other IVD. The current study provides evidence that lesions similar to those encountered in humans can be identified by CT and MRI in lumbar spine of sheep. Ideally, research animals should be assessed at the initiation of preclinical trials to determine the extent of prevalent degenerative changes. The ovine lumbosacral disc seems particularly prone to degeneration and might be a favorable anatomic site for studying IVD degeneration. PMID:27538861

Evaluation of VITEK mass spectrometry (MS), a matrix-assisted laser desorption ionization time-of-flight MS system for identification of anaerobic bacteria.

PubMed

Lee, Wonmok; Kim, Myungsook; Yong, Dongeun; Jeong, Seok Hoon; Lee, Kyungwon; Chong, Yunsop

2015-01-01

By conventional methods, the identification of anaerobic bacteria is more time consuming and requires more expertise than the identification of aerobic bacteria. Although the matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) systems are relatively less studied, they have been reported to be a promising method for the identification of anaerobes. We evaluated the performance of the VITEK MS in vitro diagnostic (IVD; 1.1 database; bioMérieux, France) in the identification of anaerobes. We used 274 anaerobic bacteria isolated from various clinical specimens. The results for the identification of the bacteria by VITEK MS were compared to those obtained by phenotypic methods and 16S rRNA gene sequencing. Among the 249 isolates included in the IVD database, the VITEK MS correctly identified 209 (83.9%) isolates to the species level and an additional 18 (7.2%) at the genus level. In particular, the VITEK MS correctly identified clinically relevant and frequently isolated anaerobic bacteria to the species level. The remaining 22 isolates (8.8%) were either not identified or misidentified. The VITEK MS could not identify the 25 isolates absent from the IVD database to the species level. The VITEK MS showed reliable identifications for clinically relevant anaerobic bacteria.

45 CFR 310.20 - What are the conditions for funding the installation, operation, maintenance and enhancement of...

Code of Federal Regulations, 2010 CFR

2010-10-01

... installation, operation, maintenance and enhancement of Computerized Tribal IV-D Systems and Office Automation... HEALTH AND HUMAN SERVICES COMPUTERIZED TRIBAL IV-D SYSTEMS AND OFFICE AUTOMATION Funding for Computerized Tribal IV-D Systems and Office Automation § 310.20 What are the conditions for funding the installation...

45 CFR 310.35 - Under what circumstances would emergency FFP be available for Computerized Tribal IV-D Systems?

Code of Federal Regulations, 2010 CFR

2010-10-01

... 45 Public Welfare 2 2010-10-01 2010-10-01 false Under what circumstances would emergency FFP be... AND OFFICE AUTOMATION Funding for Computerized Tribal IV-D Systems and Office Automation § 310.35 Under what circumstances would emergency FFP be available for Computerized Tribal IV-D Systems? (a...

Disc cell clusters in pathological human intervertebral discs are associated with increased stress protein immunostaining.

PubMed

Sharp, Christopher A; Roberts, Sally; Evans, Helena; Brown, Sharon J

2009-11-01

Intervertebral disc (IVD) cells within the annulus fibrosus (AF) and nucleus pulposus (NP) maintain distinct functional extracellular matrices and operate within a potentially noxious and stressful environment. How disc cells respond to stress and whether stress is responsible for triggering degeneration is unknown. Disc cell proliferation and cluster formation are most marked in degenerate IVDs, possibly indicating attempts at matrix repair. In other tissues, stress proteins increase rapidly after stress protecting cell function and, although implicated in degeneration of articular cartilage, have received little attention in degenerative IVD pathologies. We have compared the distribution of stress protein immunolocalization in pathological and control IVDs. Disc tissues were obtained at surgery from 43 patients with degenerative disc disease (DDD) and herniation, and 12 controls at postmortem. Tissues were immunostained with a polyclonal antibody for heat shock factor 1 (HSF-1) and monoclonal antibodies for the heat shock proteins, Hsp27 and Hsp72, using an indirect immunoperoxidase method. Positively stained cells were expressed as a percentage of the total. Cell cluster formation was also assessed. The proportion of cells in clusters was similar in the AF (both 2%) and NP (8 and 9%) of control and DDD samples, whereas in herniated tissues this was increased (AF 12%, NP 14%). Stress antigen staining tended to be more frequent in clustered rather than in single/doublet cells, and this was significant (P < 0.005) in both the AF and NP of herniated discs. Clustered cells, which are most common in herniated discs, may be mounting a protective response to abnormal environmental factors associated with disc degeneration. A better understanding of the stress response in IVD cells may allow its utilization in disc cell therapies.

A 10-Gene Classifier for Indeterminate Thyroid Nodules: Development and Multicenter Accuracy Study

PubMed Central

González, Hernán E.; Martínez, José R.; Vargas-Salas, Sergio; Solar, Antonieta; Veliz, Loreto; Cruz, Francisco; Arias, Tatiana; Loyola, Soledad; Horvath, Eleonora; Tala, Hernán; Traipe, Eufrosina; Meneses, Manuel; Marín, Luis; Wohllk, Nelson; Diaz, René E.; Véliz, Jesús; Pineda, Pedro; Arroyo, Patricia; Mena, Natalia; Bracamonte, Milagros; Miranda, Giovanna; Bruce, Elsa

2017-01-01

Background: In most of the world, diagnostic surgery remains the most frequent approach for indeterminate thyroid cytology. Although several molecular tests are available for testing in centralized commercial laboratories in the United States, there are no available kits for local laboratory testing. The aim of this study was to develop a prototype in vitro diagnostic (IVD) gene classifier for the further characterization of nodules with an indeterminate thyroid cytology. Methods: In a first stage, the expression of 18 genes was determined by quantitative polymerase chain reaction (qPCR) in a broad histopathological spectrum of 114 fresh-tissue biopsies. Expression data were used to train several classifiers by supervised machine learning approaches. Classifiers were tested in an independent set of 139 samples. In a second stage, the best classifier was chosen as a model to develop a multiplexed-qPCR IVD prototype assay, which was tested in a prospective multicenter cohort of fine-needle aspiration biopsies. Results: In tissue biopsies, the best classifier, using only 10 genes, reached an optimal and consistent performance in the ninefold cross-validated testing set (sensitivity 93% and specificity 81%). In the multicenter cohort of fine-needle aspiration biopsy samples, the 10-gene signature, built into a multiplexed-qPCR IVD prototype, showed an area under the curve of 0.97, a positive predictive value of 78%, and a negative predictive value of 98%. By Bayes' theorem, the IVD prototype is expected to achieve a positive predictive value of 64–82% and a negative predictive value of 97–99% in patients with a cancer prevalence range of 20–40%. Conclusions: A new multiplexed-qPCR IVD prototype is reported that accurately classifies thyroid nodules and may provide a future solution suitable for local reference laboratory testing. PMID:28521616

Organ Culture Bioreactors – Platforms to Study Human Intervertebral Disc Degeneration and Regenerative Therapy

PubMed Central

Gantenbein, Benjamin; Illien-Jünger, Svenja; Chan, Samantha CW; Walser, Jochen; Haglund, Lisbet; Ferguson, Stephen J; Iatridis, James C; Grad, Sibylle

2015-01-01

In recent decades the application of bioreactors has revolutionized the concept of culturing tissues and organs that require mechanical loading. In intervertebral disc (IVD) research, collaborative efforts of biomedical engineering, biology and mechatronics have led to the innovation of new loading devices that can maintain viable IVD organ explants from large animals and human cadavers in precisely defined nutritional and mechanical environments over extended culture periods. Particularly in spine and IVD research, these organ culture models offer appealing alternatives, as large bipedal animal models with naturally occurring IVD degeneration and a genetic background similar to the human condition do not exist. Latest research has demonstrated important concepts including the potential of homing of mesenchymal stem cells to nutritionally or mechanically stressed IVDs, and the regenerative potential of “smart” biomaterials for nucleus pulposus or annulus fibrosus repair. In this review, we summarize the current knowledge about cell therapy, injection of cytokines and short peptides to rescue the degenerating IVD. We further stress that most bioreactor systems simplify the real in vivo conditions providing a useful proof of concept. Limitations are that certain aspects of the immune host response and pain assessments cannot be addressed with ex vivo systems. Coccygeal animal disc models are commonly used because of their availability and similarity to human IVDs. Although in vitro loading environments are not identical to the human in vivo situation, 3D ex vivo organ culture models of large animal coccygeal and human lumbar IVDs should be seen as valid alternatives for screening and feasibility testing to augment existing small animal, large animal, and human clinical trial experiments. PMID:25764196

Impaired intervertebral disc development and premature disc degeneration in mice with notochord-specific deletion of CCN2.

PubMed

Bedore, Jake; Sha, Wei; McCann, Matthew R; Liu, Shangxi; Leask, Andrew; Séguin, Cheryle A

2013-10-01

Currently, our ability to treat intervertebral disc (IVD) degeneration is hampered by an incomplete understanding of disc development and aging. The specific function of matricellular proteins, including CCN2, during these processes remains an enigma. The aim of this study was to determine the tissue-specific localization of CCN proteins and to characterize their role in IVD tissues during embryonic development and age-related degeneration by using a mouse model of notochord-specific CCN2 deletion. Expression of CCN proteins was assessed in IVD tissues from wild-type mice beginning on embryonic day 15.5 to 17 months of age. Given the enrichment of CCN2 in notochord-derived tissues, we generated notochord-specific CCN2-null mice to assess the impact on the IVD structure and extracellular matrix composition. Using a combination of histologic evaluation and magnetic resonance imaging (MRI), IVD health was assessed. Loss of the CCN2 gene in notochord-derived cells disrupted the formation of IVDs in embryonic and newborn mice, resulting in decreased levels of aggrecan and type II collagen and concomitantly increased levels of type I collagen within the nucleus pulposus. CCN2-knockout mice also had altered expression of CCN1 (Cyr61) and CCN3 (Nov). Mirroring its role during early development, notochord-specific CCN2 deletion accelerated age-associated degeneration of IVDs. Using a notochord-specific gene targeting strategy, this study demonstrates that CCN2 expression by nucleus pulposus cells is essential to the regulation of IVD development and age-associated tissue maintenance. The ability of CCN2 to regulate the composition of the intervertebral disc suggests that it may represent an intriguing clinical target for the treatment of disc degeneration. Copyright © 2013 by the American College of Rheumatology.

Biologic canine and human intervertebral disc repair by notochordal cell-derived matrix: from bench towards bedside.

PubMed

Bach, Frances C; Tellegen, Anna R; Beukers, Martijn; Miranda-Bedate, Alberto; Teunissen, Michelle; de Jong, Willem A M; de Vries, Stefan A H; Creemers, Laura B; Benz, Karin; Meij, Björn P; Ito, Keita; Tryfonidou, Marianna A

2018-05-29

The socioeconomic burden of chronic back pain related to intervertebral disc (IVD) disease is high and current treatments are only symptomatic. Minimally invasive strategies that promote biological IVD repair should address this unmet need. Notochordal cells (NCs) are replaced by chondrocyte-like cells (CLCs) during IVD maturation and degeneration. The regenerative potential of NC-secreted substances on CLCs and mesenchymal stromal cells (MSCs) has already been demonstrated. However, identification of these substances remains elusive. Innovatively, this study exploits the regenerative NC potential by using healthy porcine NC-derived matrix (NCM) and employs the dog as a clinically relevant translational model. NCM increased the glycosaminoglycan and DNA content of human and canine CLC aggregates and facilitated chondrogenic differentiation of canine MSCs in vitro . Based on these results, NCM, MSCs and NCM+MSCs were injected in mildly (spontaneously) and moderately (induced) degenerated canine IVDs in vivo and, after six months of treatment, were analyzed. NCM injected in moderately (induced) degenerated canine IVDs exerted beneficial effects at the macroscopic and MRI level, induced collagen type II-rich extracellular matrix production, improved the disc height, and ameliorated local inflammation. MSCs exerted no (additive) effects. In conclusion, NCM induced in vivo regenerative effects on degenerated canine IVDs. NCM may, comparable to demineralized bone matrix in bone regeneration, serve as 'instructive matrix', by locally releasing growth factors and facilitating tissue repair. Therefore, intradiscal NCM injection could be a promising regenerative treatment for IVD disease, circumventing the cumbersome identification of bioactive NC-secreted substances.

Biological intervertebral disc replacement: an in vivo model and comparison of two surgical techniques to approach the rat caudal disc

PubMed Central

Gebhard, Harry; James, Andrew R.; Bowles, Robby D.; Dyke, Jonathan P.; Saleh, Tatianna; Doty, Stephen P.; Bonassar, Lawrence J.; Härtl, Roger

2011-01-01

Study design: Prospective randomized animal study. Objective: To determine a surgical technique for reproducible and functional intervertebral disc replacement in an orthotopic animal model. Methods: The caudal 3/4 intervertebral disc (IVD) of the rat tail was approached by two surgical techniques: blunt dissection, stripping and retracting (Technique 1) or incising and repairing (Technique 2) the dorsal longitudinal tendons. The intervertebral disc was dissected and removed, and then either discarded or reinserted. Outcome measures were perioperative complications, spontaneous tail movement, 7T MRI (T1- and T2-sequences for measurement of disc space height (DSH) and disc hydration). Microcomputed tomographic imaging (micro CT) was additionally performed postmortem. Results: No vascular injuries occurred and no systemic or local infections were observed over the course of 1 month. Tail movements were maintained. With tendon retraction (Technique 1) gross loss of DSH occurred with both discectomy and reinsertion. Tendon division (Technique 2) maintained DSH with IVD reinsertion but not without. The DSH was demonstrated on MRI measurement. A new scoring system to assess IVD appearances was described. Conclusions: The rat tail model, with a tendon dividing surgical technique, can function as an orthotopic animal model for IVD research. Mechanical stimulation is maintained by preserved tail movements. 7T MRI is a feasible modality for longitudinal monitoring for the rat caudal disc. PMID:22956934

Conditioned Medium Derived from Notochordal Cell-Rich Nucleus Pulposus Tissue Stimulates Matrix Production by Canine Nucleus Pulposus Cells and Bone Marrow-Derived Stromal Cells

PubMed Central

de Vries, Stefan A.H.; Potier, Esther; van Doeselaar, Marina; Meij, Björn P.; Tryfonidou, Marianna A.

2015-01-01

Objectives: Conditioned medium derived from notochordal cell-rich nucleus pulposus tissue (NCCM) was previously shown to have a stimulatory effect on bone marrow stromal cells (BMSCs) and nucleus pulposus cells (NPCs) individually, in mixed species in vitro cell models. The objective of the current study was to assess the stimulatory effect of NCCM on NPCs in a homologous canine in vitro model and to investigate whether combined stimulation with NCCM and addition of BMSCs provides a synergistic stimulatory effect. Methods: BMSCs and NPCs were harvested from chondrodystrophic dogs with confirmed early intervertebral disc (IVD) degeneration. NCCM was produced from NP tissue of nonchondrodystrophic dogs with healthy IVDs. BMSCs or NPCs alone (3×106 cells/mL) and NPCs+BMSCs (6×106 cells/mL; mixed 1:1) were cultured for 4 weeks in 1.2% alginate beads under base medium (BM), NCCM, or with addition of 10 ng/mL transforming growth factor-β1 (TGF-β1) as a positive control. Beads were assessed for glycosaminoglycan (GAG) and DNA contents by biochemical assays, GAG deposition by Alcian blue staining, and gene expression (aggrecan, versican, collagen 1 and 2, SOX9, A disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5), and matrix metalloproteinase 13 [MMP13]) with real-time quantitative RT-PCR. Results: NCCM increased NPC proliferation, proteoglycan production, and expression of genes associated with a healthy NP-like phenotype. BMSCs also showed increased proteoglycan production under NCCM, but these effects were not observed at the gene level. Combined stimulation of NPCs with NCCM and coculturing with BMSCs did not result in increased proteoglycan content compared to stimulation with NCCM alone. Discussion: NCCM stimulates matrix production by both NPCs and BMSCs and directs NPCs toward a healthier phenotype. NCCM is therefore promising for IVD regeneration and identification of the bioactive components will be helpful to further develop this approach. In the current study, no synergistic effect of adding BMSCs was observed. PMID:25370929

Rectal diazepam solution is as good as rectal administration of intravenous diazepam in the first-aid cessation of seizures in children with intractable epilepsy.

PubMed

Chiang, Lin-Mei; Wang, Huei-Shyong; Shen, Hsin-Hsien; Deng, Shin-Tang; Tseng, Chi-Hao; Chen, Yu-In; Chou, Ming-Liang; Hung, Po-Cheng; Lin, Kuang-Lin

2011-02-01

Acute seizures are readily recognizable episodes requiring urgent treatment. This study was conducted to compare the efficacy and safety of suppository use of rectal diazepam solution [Stesolid rectal tube (SRT), Alpharma, Inc., Lierskogen, Norway] with those of intravenous diazepam (IVD), Li Ta Pharma Co, Ltd., Taichung, Taiwan for control of acute seizures in children with intractable epilepsy. Subjects were patients, aged 1-18 years, with intractable epilepsy under at least three kinds of antiepileptic treatments. Caregivers were trained to rectally administer SRT or IVD (dosage varying from 0.2 to 0.5mg per kilogram of body weight) and to monitor respiration condition, seizure severity, and adverse drug effects. Among the 24 subjects, 9 males and 15 females, treated for a period of 3 months, the ages ranged from 2 to 18 years, with a mean of 9.1 years. Seizure types were generalized tonic and/or clonic. Seizure frequency varied from once per week to 20 times per day. Twenty-one (87.5%) of them had mental retardation and/or developmental delay, and 103 of the 127 (81.1%) IVD administrations and 90 of the 103 (87.3%) SRT administrations resulted in rapid cessation of seizures within 10 minutes. Each first dose failed to control seizures in 24 and 13 episodes, respectively. A second dose of IVD achieved cessation of seizure in 21 of the 24 episodes and a second dose of SRT in 12 of the 13 episodes within another 10 minutes. Four episodes (3 with rectal IVD and 1 with SRT) of prolonged seizure beyond 20 minutes needed IVD injection at our emergency room. Sedation occurred in 17% of patients, which was attributed to IVD in 8% and SRT in 9% of patients. No respiratory depression was attributable to IVD or SRT. There was no significant statistical difference in efficacy and safety between these two forms of diazepam. Rectal diazepam solution, administered by capable caregivers, is as effective and safe as rectal administration of IVD for children with intractable epilepsy. Copyright © 2011. Published by Elsevier B.V.

Angle-ply biomaterial scaffold for annulus fibrosus repair replicates native tissue mechanical properties, restores spinal kinematics, and supports cell viability.

PubMed

Borem, Ryan; Madeline, Allison; Walters, Joshua; Mayo, Henry; Gill, Sanjitpal; Mercuri, Jeremy

2017-08-01

Annulus fibrosus (AF) damage commonly occurs due to intervertebral disc (IVD) degeneration/herniation. The dynamic mechanical role of the AF is essential for proper IVD function and thus it is imperative that biomaterials developed to repair the AF withstand the mechanical rigors of the native tissue. Furthermore, these biomaterials must resist accelerated degradation within the proteolytic environment of degenerate IVDs while supporting integration with host tissue. We have previously reported a novel approach for developing collagen-based, multi-laminate AF repair patches (AFRPs) that mimic the angle-ply architecture and basic tensile properties of the human AF. Herein, we further evaluate AFRPs for their: tensile fatigue and impact burst strength, IVD attachment strength, and contribution to functional spinal unit (FSU) kinematics following IVD repair. Additionally, AFRP resistance to collagenase degradation and cytocompatibility were assessed following chemical crosslinking. In summary, AFRPs demonstrated enhanced durability at high applied stress amplitudes compared to human AF and withstood radially-directed biaxial stresses commonly borne by the native tissue prior to failure/detachment from IVDs. Moreover, FSUs repaired with AFRPs and nucleus pulposus (NP) surrogates had their axial kinematic parameters restored to intact levels. Finally, carbodiimide crosslinked AFRPs resisted accelerated collagenase digestion without detrimentally effecting AFRP tensile properties or cytocompatibility. Taken together, AFRPs demonstrate the mechanical robustness and enzymatic stability required for implantation into the damaged/degenerate IVD while supporting AF cell infiltration and viability. The quality of life for millions of individuals globally is detrimentally impacted by IVD degeneration and herniation. These pathologies often result in the structural demise of IVD tissue, particularly the annulus fibrosus (AF). Biomaterials developed for AF repair have yet to demonstrate the mechanical strength and durability required for utilization in the spine. Herein, we demonstrate the development of an angle-ply AF repair patch (AFRP) that can resist the application of physiologically relevant stresses without failure and which contributes to the restoration of functional spinal unit axial kinematics following repair. Furthermore, we show that this biomaterial can resist accelerated degradation in a simulated degenerate environment and supports AF cell viability. Copyright © 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

The viscoelastic standard nonlinear solid model: predicting the response of the lumbar intervertebral disk to low-frequency vibrations.

PubMed

Groth, Kevin M; Granata, Kevin P

2008-06-01

Due to the mathematical complexity of current musculoskeletal spine models, there is a need for computationally efficient models of the intervertebral disk (IVD). The aim of this study is to develop a mathematical model that will adequately describe the motion of the IVD under axial cyclic loading as well as maintain computational efficiency for use in future musculoskeletal spine models. Several studies have successfully modeled the creep characteristics of the IVD using the three-parameter viscoelastic standard linear solid (SLS) model. However, when the SLS model is subjected to cyclic loading, it underestimates the load relaxation, the cyclic modulus, and the hysteresis of the human lumbar IVD. A viscoelastic standard nonlinear solid (SNS) model was used to predict the response of the human lumbar IVD subjected to low-frequency vibration. Nonlinear behavior of the SNS model was simulated by a strain-dependent elastic modulus on the SLS model. Parameters of the SNS model were estimated from experimental load deformation and stress-relaxation curves obtained from the literature. The SNS model was able to predict the cyclic modulus of the IVD at frequencies of 0.01 Hz, 0.1 Hz, and 1 Hz. Furthermore, the SNS model was able to quantitatively predict the load relaxation at a frequency of 0.01 Hz. However, model performance was unsatisfactory when predicting load relaxation and hysteresis at higher frequencies (0.1 Hz and 1 Hz). The SLS model of the lumbar IVD may require strain-dependent elastic and viscous behavior to represent the dynamic response to compressive strain.

Setup in a clinical workflow and impact on radiotherapy routine of an in vivo dosimetry procedure with an electronic portal imaging device

PubMed Central

Piermattei, Angelo; Kang, Shengwei; Xiao, Mingyong; Tang, Bin; Liao, Xiongfei; Xin, Xin; Grusio, Mattia

2018-01-01

High conformal techniques such as intensity-modulated radiation therapy and volumetric-modulated arc therapy are widely used in overloaded radiotherapy departments. In vivo dosimetric screening is essential in this environment to avoid important dosimetric errors. This work examines the feasibility of introducing in vivo dosimetry (IVD) checks in a radiotherapy routine. The causes of dosimetric disagreements between delivered and planned treatments were identified and corrected during the course of treatment. The efficiency of the corrections performed and the added workload needed for the entire procedure were evaluated. The IVD procedure was based on an electronic portal imaging device. A total of 3682 IVD tests were performed for 147 patients who underwent head and neck, abdomen, pelvis, breast, and thorax radiotherapy treatments. Two types of indices were evaluated and used to determine if the IVD tests were within tolerance levels: the ratio R between the reconstructed and planned isocentre doses and a transit dosimetry based on the γ-analysis of the electronic portal images. The causes of test outside tolerance level were investigated and corrected and IVD test was repeated during subsequent fraction. The time needed for each step of the IVD procedure was registered. Pelvis, abdomen, and head and neck treatments had 10% of tests out of tolerance whereas breast and thorax treatments accounted for up to 25%. The patient setup was the main cause of 90% of the IVD tests out of tolerance and the remaining 10% was due to patient morphological changes. An average time of 42 min per day was sufficient to monitor a daily workload of 60 patients in treatment. This work shows that IVD performed with an electronic portal imaging device is feasible in an overloaded department and enables the timely realignment of the treatment quality indices in order to achieve a patient’s final treatment compliant with the one prescribed. PMID:29432473

Annulus Fibrosus Repair Using High-Density Collagen Gel: An In Vivo Ovine Model.

PubMed

Pennicooke, Brenton; Hussain, Ibrahim; Berlin, Connor; Sloan, Stephen R; Borde, Brandon; Moriguchi, Yu; Lang, Gernot; Navarro-Ramirez, Rodrigo; Cheetham, Jonathan; Bonassar, Lawrence J; Härtl, Roger

2018-02-15

Ovine in vivo study. To perform lateral approach lumbar surgery in an ovine model to administer an injectable riboflavin cross-linked high-density collagen (HDC) gel and to assess its ability to mitigate intervertebral disc (IVD) degeneration after induced annulus fibrosus (AF) injury. Biological-based injectable gels have shown efficacy in restoring biomechanical, radiographic, and histological parameters in IVD-injured animal models. Riboflavin cross-linked HDC gel has previously demonstrated retention of nucleus pulposus (NP) tissue, reduced loss of disc height, and prevention of terminal cellular degenerative changes in rat-tail spines. However, this biological therapy has never been tested in large animal models. Forty lumbar IVDs were accessed from eight sheep via lateral approach surgery. IVDs were randomly assigned to healthy control, injury and HDC treatment, or negative control with injury and no treatment. IVD injury was carried out using a drill-bit through the AF followed by needle puncture of the NP. Sheep were followed for 16 weeks and underwent qualitative/quantitative magnetic resonance imaging, x-ray, and histological analyses of collagen and proteoglycan content. The lateral approach to the ovine lumbar spine to deliver HDC gel proved to be safe and reproducible. IVDs treated with the HDC gel revealed less degenerative changes at the microscopic level based on AF and NP histology. However, mean Pfirrmann grade, T2 relaxation time, NP voxel size, and disc height index were not significantly different between the two injury groups. Injectable HDC gel can be administered safely via lateral approach surgery in an ovine AF injury model. IVDs treated with HDC gel demonstrated less degeneration at the microscopic level though radiographic changes were slight when comparing treated to untreated IVDs. Future studies will need to elucidate the role of injury technique and time frame for follow-up in correlating histological and radiographical outcomes. N /A.

Metameric pattern of intervertebral disc/vertebral body is generated independently of Mesp2/Ripply-mediated rostro-caudal patterning of somites in the mouse embryo.

PubMed

Takahashi, Yu; Yasuhiko, Yukuto; Takahashi, Jun; Takada, Shinji; Johnson, Randy L; Saga, Yumiko; Kanno, Jun

2013-08-15

The vertebrae are derived from the sclerotome of somites. Formation of the vertebral body involves a process called resegmentation, by which the caudal half of a sclerotome is combined with the rostral half of the next sclerotome. To elucidate the relationship between resegmentation and rostro-caudal patterning of somite, we used the Uncx4.1-LacZ transgene to characterize the resegmentation process. Our observations suggested that in the thoracic and lumbar vertebrae, the Uncx4.1-expressing caudal sclerotome gave rise to the intervertebral disc (IVD) and rostral portion of the vertebral body (VB). In the cervical vertebrae, the Uncx4.1-expressing caudal sclerotome appeared to contribute to the IVD and both caudal and rostral ends of the VB. This finding suggests that the rostro-caudal gene expression boundary does not necessarily coincide with the resegmentation boundary. This conclusion was supported by analyses of Mesp2 KO and Ripply1/2 double KO embryos lacking rostral and caudal properties, respectively. Resegmentation was not observed in Mesp2 KO embryos, but both the IVD and whole VB were formed from the caudalized sclerotome. Expression analysis of IVD marker genes including Pax1 in the wild-type, Mesp2 KO, and Ripply1/2 DKO embryos also supported the idea that a metameric pattern of IVD/VB is generated independently of Mesp2/Ripply-mediated rostro-caudal patterning of somite. However, in the lumbar region, IVD differentiation appeared to be stimulated by the caudal property and suppressed by the rostral property. Therefore, we propose that rostro-caudal patterning of somites is not a prerequisite for metameric patterning of the IVD and VB, but instead required to stimulate IVD differentiation in the caudal half of the sclerotome. Copyright © 2013 Elsevier Inc. All rights reserved.

The Substitution of IVD (Ion Vapor Deposition) Aluminum for Cadmium

DTIC Science & Technology

1989-08-01

environment, it can finu its A in( the water suHpl o- fooo chain. Also, with electroplated cadmium processing, tnere are auditional nazaros associated win...no , - r e CUr r in ( polut io n- r elIt e costs. Tne po IIut ion controlI costs for all caG;iul n processing , nclIUd ino, clpitl treatm’ient , col...H T P, H VacuumCadmium None T. O. H T. P. H Key T - Toxic materials 0 - OSHA standards imposed P - Polution control required H - Hazardous waste

Improved display of cervical intervertebral discs on water (iodine) images: incidental findings from single-source dual-energy CT angiography of head and neck arteries.

PubMed

Wu, Qingxia; Shi, Dapeng; Cheng, Tianming; Liu, Hongming; Hu, Niuniu; Chang, Xiaowan; Guo, Ying; Wang, Meiyun

2018-06-19

To (a) assess the diagnostic performance of material decomposition (MD) water (iodine) images for the evaluation of cervical intervertebral discs (IVDs) in patients who underwent dual-energy head and neck CT angiography (HNCTA) compared with 70-keV images and (b) to explore the correlation of water concentration with the T2 relaxation time of IVDs. Twenty-four consecutive patients who underwent dual-energy HNCTA and cervical spine MRI were studied. The diagnostic performance of water (iodine), 70-keV and MR images for IVD bulge and herniation was assessed. A subjective image score for each image set was recorded. The signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) of IVDs to the cervical spinal cord were compared between water (iodine) and 70-keV images. Disc water concentration as measured on water (iodine) images was correlated with T2 relaxation time. IVD evaluations for bulge and herniation did not differ significantly among the three image sets (pairwise comparisons; all p > 0.05). SNR and CNR were significantly improved on water (iodine) images compared with those on 70-keV images (p < 0.001). Although water (iodine) images showed higher image quality scores when evaluating IVDs compared with 70-keV images, the difference is not significant (all adjusted p > 0.05). IVD water concentration exhibited no correlation with relative T2 relaxation time (all p > 0.05). Water (iodine) images facilitated analysis of cervical IVDs by providing higher SNR and CNR compared with 70-keV images. The disc water concentration measured on water (iodine) images exhibited no correlation with relative T2 relaxation time. • There was no significant difference in cervical IVD evaluations for bulge and herniation among water (iodine) images, 70-keV images and MR images. • Water (iodine) images provided higher objective and subjective image quality than 70-keV images, though the difference of subjective evaluation was not statistically significant. • The disc water concentration exhibited no correlation with relative T2 relaxation time, which reflects the inferiority of the water (iodine) images in evaluating disc water content compared with T2 maps.

Automated detection, 3D segmentation and analysis of high resolution spine MR images using statistical shape models

NASA Astrophysics Data System (ADS)

Neubert, A.; Fripp, J.; Engstrom, C.; Schwarz, R.; Lauer, L.; Salvado, O.; Crozier, S.

2012-12-01

Recent advances in high resolution magnetic resonance (MR) imaging of the spine provide a basis for the automated assessment of intervertebral disc (IVD) and vertebral body (VB) anatomy. High resolution three-dimensional (3D) morphological information contained in these images may be useful for early detection and monitoring of common spine disorders, such as disc degeneration. This work proposes an automated approach to extract the 3D segmentations of lumbar and thoracic IVDs and VBs from MR images using statistical shape analysis and registration of grey level intensity profiles. The algorithm was validated on a dataset of volumetric scans of the thoracolumbar spine of asymptomatic volunteers obtained on a 3T scanner using the relatively new 3D T2-weighted SPACE pulse sequence. Manual segmentations and expert radiological findings of early signs of disc degeneration were used in the validation. There was good agreement between manual and automated segmentation of the IVD and VB volumes with the mean Dice scores of 0.89 ± 0.04 and 0.91 ± 0.02 and mean absolute surface distances of 0.55 ± 0.18 mm and 0.67 ± 0.17 mm respectively. The method compares favourably to existing 3D MR segmentation techniques for VBs. This is the first time IVDs have been automatically segmented from 3D volumetric scans and shape parameters obtained were used in preliminary analyses to accurately classify (100% sensitivity, 98.3% specificity) disc abnormalities associated with early degenerative changes.

Human mesenchymal stem cell differentiation to NP-like cells in chitosan-glycerophosphate hydrogels.

PubMed

Richardson, Stephen M; Hughes, Nesta; Hunt, John A; Freemont, Anthony J; Hoyland, Judith A

2008-01-01

Intervertebral disc (IVD) degeneration is one of the major causes of low back pain. As current clinical treatments are aimed at restoring biomechanical function and providing symptomatic relief, interest in methods focused on biological repair has increased. Several tissue engineering approaches using different cell types and hydrogels/scaffolds have been proposed. Owing to the unsuitable nature of degenerate cells for tissue engineering attention has focused on the use of mesenchymal stem cells (MSCs). Additionally, while rigid scaffolds have been demonstrated to allow MSC differentiation to the chondrocyte-like cells of the IVD, hydrogels are being increasingly studied as they allow minimally invasive implantation without extensive damage to the IVD. Here, we have studied the temperature-sensitive hydrogel chitosan-glycerophosphate (C/Gp), seeded with human MSCs and cultured for 4 weeks in standard medium. We have analysed the gene and protein expression profile of the MSCs and compared it to that of both nucleus pulposus (NP) cells and articular chondrocytes cultured in C/Gp. Gene expression analysis for chondrocytic-cell marker genes demonstrated differentiation of MSCs to a phenotype which showed similarities to both articular chondrocytes and NP cells. Conventional PCR demonstrated a lack of expression of osteogenic marker genes and the hypertrophic marker gene type X collagen. MSCs also secreted both proteoglycans and collagens in a ratio, which more closely resembled that of NP cells than articular chondrocytes. These results therefore suggest that MSC-seeded C/Gp gels could be used clinically for the regeneration of the degenerate human IVD.

Intervertebral disc-derived stem cells: implications for regenerative medicine and neural repair.

PubMed

Erwin, W Mark; Islam, Diana; Eftekarpour, Eftekhar; Inman, Robert D; Karim, Muhammad Zia; Fehlings, Michael G

2013-02-01

An in vitro and in vivo evaluation of intervertebral disc (IVD)-derived stem/progenitor cells. To determine the chondrogenic, adipogenic, osteogenic, and neurogenic differentiation capacity of disc-derived stem/progenitor cells in vitro and neurogenic differentiation in vivo. Tissue repair strategies require a source of appropriate cells that could be used to replace dead or damaged cells and tissues such as stem cells. Here we examined the potential use of IVD-derived stem cells in regenerative medicine approaches and neural repair. Nonchondrodystrophic canine IVD nucleus pulposus (NP) cells were used to generate stem/progenitor cells (NP progenitor cells [NPPCs]) and the NPPCs were differentiated in vitro into chondrogenic, adipogenic, and neurogenic lineages and in vivo into the neurogenic lineage. NPPCs were compared with bone marrow-derived mesenchymal (stromal) stem cells in terms of the expression of stemness genes. The expression of the neural crest marker protein 0 and the Brachyury gene were evaluated in NP cells and NPPCs. NPPCs contain stem/progenitor cells and express "stemness" genes such as Sox2, Oct3/4, Nanog, CD133, Nestin, and neural cell adhesion molecule but differ from mesenchymal (stromal) stem cells in the higher expression of the Nanog gene by NPPCs. NPPCs do not express protein 0 or the Brachyury gene both of which are expressed by the totality of IVD NP cells. The percentage of NPPCs within the IVD is 1% of the total as derived by colony-forming assay. NPPCs are capable of differentiating along chondrogenic, adipogenic, and neurogenic lineages in vitro and into oligodendrocyte, neuron, and astroglial specific precursor cells in vivo within the compact myelin-deficient shiverer mouse. We propose that the IVD NP represents a regenerative niche suggesting that the IVD could represent a readily accessible source of precursor cells for neural repair and regeneration.

Kinetic and Spectral Properties of Isovaleryl-CoA Dehydrogenase and Interaction with Ligands

PubMed Central

Mohsen, Al-Walid A.; Vockley, Jerry

2014-01-01

Isovaleryl-CoA dehydrogenase (IVD) catalyzes the conversion of isovaleryl-CoA to 3-methylcrotonyl-CoA and the transfer of electrons to the electron transfer flavoprotein (ETF). Recombinant human IVD purifies with bound CoA-persulfide. A modified purification protocol was developed to isolate IVD without bound CoA-persulfide and to protect the protein thiols from oxidation. The CoA-persulfide-free IVD specific activity was 112.5 µmol porcine ETF•min−1•mg−1, which was ~20-fold higher than that of its CoA-persulfide bound form. The Km and catalytic efficiency (kcat/Km) for isovaleryl-CoA were 1.0 µM and 4.3 × 106•M−1•sec−1 per monomer, respectively, and its Km for ETF was 2.0 µM. Anaerobic titration of isovaleryl-CoA into an IVD solution resulted in a stable blue complex with increased absorbance at 310 nm, decreased absorbance at 373 and 447 nm, and the appearance of the charge transfer complex band at 584 nm. The apparent dissociation constant (KD app) determined spectrally for isovaleryl-CoA was 0.54 µM. Isovaleryl-CoA, acetoacetyl-CoA, methylenecyclopropylacetyl-CoA, and ETF induced CD spectral changes at the 250–500 nm region while isobutyryl-CoA did not, suggesting conformational changes occur at the flavin ring that are ligand specific. Replacement of the IVD Trp166 with a Phe did not block IVD interaction with ETF, indicating that its indole ring is not essential for electron transfer to ETF. A twelve amino acid synthetic peptide that matches the sequence of the ETF docking peptide competitively inhibited the enzyme reaction when ETF was used as the electron acceptor with a Ki of 1.5 mM. PMID:25450250

Assessment of the knowledge of graphical symbols labelled on malaria rapid diagnostic tests in four international settings.

PubMed

Hermans, Veerle; Monzote, Lianet; Van den Sande, Björn; Mukadi, Pierre; Sopheak, Thai; Gillet, Philippe; Jacobs, Jan

2011-11-02

Graphical symbols on in vitro diagnostics (IVD symbols) replace the need for text in different languages and are used on malaria rapid diagnostic tests (RDTs) marketed worldwide. The present study assessed the comprehension of IVD symbols labelled on malaria RDT kits among laboratory staff in four different countries. Participants (n = 293) in Belgium (n = 96), the Democratic Republic of the Congo (DRC, n = 87), Cambodia (n = 59) and Cuba (n = 51) were presented with an anonymous questionnaire with IVD symbols extracted from ISO 15223 and EN 980 presented as stand-alone symbols (n = 18) and in context (affixed on RDT packages, n = 16). Responses were open-ended and scored for correctness by local professionals. Presented as stand-alone, three and five IVD symbols were correctly scored for comprehension by 67% and 50% of participants; when contextually presented, five and seven symbols reached the 67% and 50% correct score respectively. 'Batch code' scored best (correctly scored by 71.3% of participants when presented as stand-alone), 'Authorized representative in the European Community' scored worst (1.4% correct). Another six IVD symbols were scored correctly by less than 10% of participants: 'Do not reuse', 'In vitro diagnostic medical device', 'Sufficient for', 'Date of manufacture', 'Authorised representative in EC', and 'Do not use if package is damaged'. Participants in Belgium and Cuba both scored six symbols above the 67% criterion, participants from DRC and Cambodia scored only two and one symbols above this criterion. Low correct scores were observed for safety-related IVD symbols, such as for 'Biological Risk' (42.7%) and 'Do not reuse' (10.9%). Comprehension of IVD symbols on RDTs among laboratory staff in four international settings was unsatisfactory. Administrative and outreach procedures should be undertaken to assure their acquaintance by end-users.

Acidic pH promotes intervertebral disc degeneration: Acid-sensing ion channel -3 as a potential therapeutic target.

PubMed

Gilbert, Hamish T J; Hodson, Nathan; Baird, Pauline; Richardson, Stephen M; Hoyland, Judith A

2016-11-17

The aetiology of intervertebral disc (IVD) degeneration remains poorly understood. Painful IVD degeneration is associated with an acidic intradiscal pH but the response of NP cells to this aberrant microenvironmental factor remains to be fully characterised. The aim here was to address the hypothesis that acidic pH, similar to that found in degenerate IVDs, leads to the altered cell/functional phenotype observed during IVD degeneration, and to investigate the involvement of acid-sensing ion channel (ASIC) -3 in the response. Human NP cells were treated with a range of pH, from that of a non-degenerate (pH 7.4 and 7.1) through to mildly degenerate (pH 6.8) and severely degenerate IVD (pH 6.5 and 6.2). Increasing acidity of pH caused a decrease in cell proliferation and viability, a shift towards matrix catabolism and increased expression of proinflammatory cytokines and pain-related factors. Acidic pH resulted in an increase in ASIC-3 expression. Importantly, inhibition of ASIC-3 prevented the acidic pH induced proinflammatory and pain-related phenotype in NP cells. Acidic pH causes a catabolic and degenerate phenotype in NP cells which is inhibited by blocking ASIC-3 activity, suggesting that this may be a useful therapeutic target for treatment of IVD degeneration.

[Analysis of the results of total cervical disc arthroplasty using a M6-C prosthesis: a multicenter study].

PubMed

Byval'tsev, V A; Kalinin, A A; Stepanov, I A; Pestryakov, Yu Ya; Shepelev, V V

Cervical spondylosis and intervertebral disc (IVD) degeneration are the most common cause for compression of the spinal cord and/or its roots. Total IVD arthroplasty, as a modern alternative to surgical treatment of IVD degeneration, is gaining popularity in many neurosurgical clinics around the world. Aim - the study aim was to conduct a multicenter analysis of cervical spine arthroplasty with an IVD prosthesis M6-C ('Spinal Kinetics', USA). The study included 112 patients (77 males and 35 females). All patients underwent single-level discectomy with implantation of the artificial IVD prosthesis M6-C. The follow-up period was up to 36 months. Dynamic assessment of the prosthesis was based on clinical parameters (pain intensity in the cervical spine and upper extremities (visual analog scale - VAS); quality of life (Neck Disability Index - NDI)); and subjective satisfaction with the results of surgical treatment (Macnab scale) and instrumental data (range of motion in the operated spinal motion segment, degree of heterotopic ossification (McAfee-Suchomel classification), and time course of degenerative changes in the adjacent segments).

aSi EPIDs for the in-vivo dosimetry of static and dynamic beams

NASA Astrophysics Data System (ADS)

Piermattei, A.; Cilla, S.; Azario, L.; Greco, F.; Russo, M.; Grusio, M.; Orlandini, L.; Fidanzio, A.

2015-10-01

Portal imaging by amorphous silicon (aSi) photodiode is currently the most applied technology for in-vivo dosimetry (IVD) of static and dynamic radiotherapy beams. The strategy, adopted in this work to perform the IVD procedure by aSi EPID, is based on: in patient reconstruction of the isocenter dose and day to day comparison between 2D-portal images to verify the reproducibility of treatment delivery. About 20.000 tests have been carried out in this last 3 years in 8 radiotherapy centers using the SOFTDISO program. The IVD results show that: (i) the procedure can be implemented for linacs of different manufacturer, (ii) the IVD analysis can be obtained on a computer screen, in quasi real time (about 2 min after the treatment delivery) and (iii) once the causes of the discrepancies were eliminated, all the global IVD tests for single patient were within the acceptance criteria defined by: ±5% for the isocenter dose, and Pγ<1≥90% of the checked points for the 2D portal image γ-analysis. This work is the result of a project supported by the Istituto Nazionale di Fisica Nucleare (INFN) and Università Cattolica del S.Cuore (UCSC).

Trueness verification of actual creatinine assays in the European market demonstrates a disappointing variability that needs substantial improvement. An international study in the framework of the EC4 creatinine standardization working group.

PubMed

Delanghe, Joris R; Cobbaert, Christa; Galteau, Marie-Madeleine; Harmoinen, Aimo; Jansen, Rob; Kruse, Rolf; Laitinen, Päivi; Thienpont, Linda M; Wuyts, Birgitte; Weykamp, Cas; Panteghini, Mauro

2008-01-01

The European In Vitro Diagnostics (IVD) directive requires traceability to reference methods and materials of analytes. It is a task of the profession to verify the trueness of results and IVD compatibility. The results of a trueness verification study by the European Communities Confederation of Clinical Chemistry (EC4) working group on creatinine standardization are described, in which 189 European laboratories analyzed serum creatinine in a commutable serum-based material, using analytical systems from seven companies. Values were targeted using isotope dilution gas chromatography/mass spectrometry. Results were tested on their compliance to a set of three criteria: trueness, i.e., no significant bias relative to the target value, between-laboratory variation and within-laboratory variation relative to the maximum allowable error. For the lower and intermediate level, values differed significantly from the target value in the Jaffe and the dry chemistry methods. At the high level, dry chemistry yielded higher results. Between-laboratory coefficients of variation ranged from 4.37% to 8.74%. Total error budget was mainly consumed by the bias. Non-compensated Jaffe methods largely exceeded the total error budget. Best results were obtained for the enzymatic method. The dry chemistry method consumed a large part of its error budget due to calibration bias. Despite the European IVD directive and the growing needs for creatinine standardization, an unacceptable inter-laboratory variation was observed, which was mainly due to calibration differences. The calibration variation has major clinical consequences, in particular in pediatrics, where reference ranges for serum and plasma creatinine are low, and in the estimation of glomerular filtration rate.

Improving the Process of Adjusting the Parameters of Finite Element Models of Healthy Human Intervertebral Discs by the Multi-Response Surface Method.

PubMed

Gómez, Fátima Somovilla; Lorza, Rubén Lostado; Bobadilla, Marina Corral; García, Rubén Escribano

2017-09-21

The kinematic behavior of models that are based on the finite element method (FEM) for modeling the human body depends greatly on an accurate estimate of the parameters that define such models. This task is complex, and any small difference between the actual biomaterial model and the simulation model based on FEM can be amplified enormously in the presence of nonlinearities. The current paper attempts to demonstrate how a combination of the FEM and the MRS methods with desirability functions can be used to obtain the material parameters that are most appropriate for use in defining the behavior of Finite Element (FE) models of the healthy human lumbar intervertebral disc (IVD). The FE model parameters were adjusted on the basis of experimental data from selected standard tests (compression, flexion, extension, shear, lateral bending, and torsion) and were developed as follows: First, three-dimensional parameterized FE models were generated on the basis of the mentioned standard tests. Then, 11 parameters were selected to define the proposed parameterized FE models. For each of the standard tests, regression models were generated using MRS to model the six stiffness and nine bulges of the healthy IVD models that were created by changing the parameters of the FE models. The optimal combination of the 11 parameters was based on three different adjustment criteria. The latter, in turn, were based on the combination of stiffness and bulges that were obtained from the standard test FE simulations. The first adjustment criteria considered stiffness and bulges to be equally important in the adjustment of FE model parameters. The second adjustment criteria considered stiffness as most important, whereas the third considered the bulges to be most important. The proposed adjustment methods were applied to a medium-sized human IVD that corresponded to the L3-L4 lumbar level with standard dimensions of width = 50 mm, depth = 35 mm, and height = 10 mm. Agreement between the kinematic behavior that was obtained with the optimized parameters and that obtained from the literature demonstrated that the proposed method is a powerful tool with which to adjust healthy IVD FE models when there are many parameters, stiffnesses, and bulges to which the models must adjust.

Improving the Process of Adjusting the Parameters of Finite Element Models of Healthy Human Intervertebral Discs by the Multi-Response Surface Method

PubMed Central

Somovilla Gómez, Fátima

2017-01-01

The kinematic behavior of models that are based on the finite element method (FEM) for modeling the human body depends greatly on an accurate estimate of the parameters that define such models. This task is complex, and any small difference between the actual biomaterial model and the simulation model based on FEM can be amplified enormously in the presence of nonlinearities. The current paper attempts to demonstrate how a combination of the FEM and the MRS methods with desirability functions can be used to obtain the material parameters that are most appropriate for use in defining the behavior of Finite Element (FE) models of the healthy human lumbar intervertebral disc (IVD). The FE model parameters were adjusted on the basis of experimental data from selected standard tests (compression, flexion, extension, shear, lateral bending, and torsion) and were developed as follows: First, three-dimensional parameterized FE models were generated on the basis of the mentioned standard tests. Then, 11 parameters were selected to define the proposed parameterized FE models. For each of the standard tests, regression models were generated using MRS to model the six stiffness and nine bulges of the healthy IVD models that were created by changing the parameters of the FE models. The optimal combination of the 11 parameters was based on three different adjustment criteria. The latter, in turn, were based on the combination of stiffness and bulges that were obtained from the standard test FE simulations. The first adjustment criteria considered stiffness and bulges to be equally important in the adjustment of FE model parameters. The second adjustment criteria considered stiffness as most important, whereas the third considered the bulges to be most important. The proposed adjustment methods were applied to a medium-sized human IVD that corresponded to the L3–L4 lumbar level with standard dimensions of width = 50 mm, depth = 35 mm, and height = 10 mm. Agreement between the kinematic behavior that was obtained with the optimized parameters and that obtained from the literature demonstrated that the proposed method is a powerful tool with which to adjust healthy IVD FE models when there are many parameters, stiffnesses, and bulges to which the models must adjust. PMID:28934161

Continuous in vitro evolution of a ribozyme ligase: a model experiment for the evolution of a biomolecule.

PubMed

Ledbetter, Michael P; Hwang, Tony W; Stovall, Gwendolyn M; Ellington, Andrew D

2013-01-01

Evolution is a defining criterion of life and is central to understanding biological systems. However, the timescale of evolutionary shifts in phenotype limits most classroom evolution experiments to simple probability simulations. In vitro directed evolution (IVDE) frequently serves as a model system for the study of Darwinian evolution but produces noticeable phenotypic shifts in a matter of hours. An IVDE demonstration lab would serve to both directly demonstrate how Darwinian selection can act on a pool of variants and introduce students to an essential method of modern molecular biology. To produce an IVDE demonstration lab, continuous IVDE of a T500 ribozyme ligase population has been paired with a fluorescent strand displacement reporter system to visualize the selection of improved catalytic function. A ribozyme population is taken through rounds of isothermal amplification dependent on the self-ligation of a T7 promoter. As the population is selectively enriched with better ligase activity, the strand displacement system allows for the monitoring of the population's ligation rate. The strand displacement reporter system permits the detection of ligated ribozyme. Once ligated with the T7 promoter, the 5' end of the ribozyme displaces paired fluorophore-quencher oligonucleotides, in turn, generating visible signal upon UV light excitation. As the ligation rate of the population increases, due to the selection for faster ligating species, the fluorescent signal develops more rapidly. The pairing of the continuous isothermal system with the fluorescent reporting scheme allows any user, provided with minimal materials, to model the continuous directed evolution of a biomolecule. Copyright © 2013 Wiley-Liss, Inc.

Audio-frequency analysis of inductive voltage dividers based on structural models

DOE Office of Scientific and Technical Information (OSTI.GOV)

Avramov, S.; Oldham, N.M.; Koffman, A.D.

1994-12-31

A Binary Inductive Voltage Divider (BIVD) is compared with a Decade Inductive Voltage Divider (DIVD) in an automatic IVD bridge. New detection and injection circuitry was designed and used to evaluate the IVDs with either the input or output tied to ground potential. In the audio frequency range the DIVD and BIVD error patterns are characterized for both in-phase and quadrature components. Differences between results obtained using a new error decomposition scheme based on structural modeling, and measurements using conventional IVD standards are reported.

Evaluation of Water Content in Lumbar Intervertebral Discs and Facet Joints Before and After Physiological Loading Using T2 Mapping MRI.

PubMed

Yamabe, Daisuke; Murakami, Hideki; Chokan, Kou; Endo, Hirooki; Oikawa, Ryosuke; Sawamura, Shoitsu; Doita, Minoru

2017-12-15

T2 mapping was used to quantify the water content of lumbar spine intervertebral discs (IVDs) and facet joints before and after physiological loading. The aim of this study was to clarify the interaction between lumbar spine IVD and facet joints as load-bearing structures by measuring the water content of their matrix after physiological loading using T2 mapping magnetic resonance imaging (MRI). To date, few reports have functionally evaluated lumbar spine IVD and facet joints, and their interaction in vivo. T2 mapping may help detect changes in the water content of IVD and articular cartilage of facet joints before and after physiological loading, thereby enabling the evaluation of changes in interacted water retention between IVD and facet joints. Twenty asymptomatic volunteers (10 female and 10 male volunteers; mean age, 19.3 years; age range, 19-20 years) underwent MRI before and after physiological loading such as lumbar flexion, extension, and rotation. Each IVD from L1/2 to L5/S1 was sliced at center of the disc space, and the T2 value was measured at the nucleus pulposus (NP), anterior annulus fibrosus (AF), posterior AF, and bilateral facet joints. In the NP, T2 values significantly decreased after exercise at every lumbar spinal level. In the anterior AF, there were no significant differences in T2 values at any level. In the posterior AF, T2 values significantly increased only at L4/5. In the bilateral facet joints, T2 values significantly decreased after exercise at every level. There was a significant decrease in the water content of facet joints and the NP at every lumbar spinal level after dynamic loading by physical lumbar exercise. These changes appear to play an important and interactional role in the maintenance of the interstitial matrix in the IVD NP and cartilage in the facet joint. 3.

Superovulation, in vitro fertilization (IVF) and in vitro development (IVD) protocols for inbred BALB/cJ mice in comparison with outbred NMRI mice.

PubMed

Golkar-Narenji, Afsaneh; Gourabi, Hamid; Eimani, Hussein; Barekati, Zeinab; Akhlaghi, Aliasghar

2012-10-01

To study assisted reproductive technology (ART) protocols including superovulation, in vitro fertilization (IVF) and in vitro development (IVD) for BALB/cJ mice in comparison with a common ART protocol for NMRI mice. Adult NMRI and BALB/cJ mice were superovulated using a 48 h G-interval. In order to find a more suitable G-interval for the BALB/cJ strain, G-intervals including 44, 46 and 50 h were also examined. Superovulation rates were recorded in all groups. IVF rate of BALB/c oocytes in T6 and mHTF media were compared. IVD rates of BALB/cJ zygotes in mHTF, T6 and G1V 5 /G2V 5 media were compared. In addition, IVF and IVD rates of BALB/cJ and NMRI oocytes were compared in T6 medium during IVF-IVD procedures. In BALB/cJ mice the highest superovulation rates were observed with 44-46 h G-intervals. However, with a 48 h G-interval, superovulation rates were significantly lower in BALB/cJ compared to NMRI mice ( p < 0.05). mHTF medium significantly increased in vitro fertilization of BALB/cJ oocytes compared to T6 medium ( p < 0.05). Fertilization rate of NMRI oocytes was significantly higher than BALB/cJ oocytes in T6 medium ( p < 0.05). The BALB/cJ embryo IVD was significantly higher in G1/G2 medium compared to mHTF and T6 media ( p < 0.01). Superovulation with 48 h G-interval and using T6 during all in vitro procedures produces embryos more efficiently for NMRI mice than for BALB/cJ mice. For BALB/cJ mice, a protocol including superovulation with a 44-46 h G-interval, using mHTF during IVF and G1V 5 /G2V 5 medium during IVD, may improve in vitro embryo production.

Loss of HIF-1α in the notochord results in cell death and complete disappearance of the nucleus pulposus.

PubMed

Merceron, Christophe; Mangiavini, Laura; Robling, Alexander; Wilson, Tremika LeShan; Giaccia, Amato J; Shapiro, Irving M; Schipani, Ernestina; Risbud, Makarand V

2014-01-01

The intervertebral disc (IVD) is one of the largest avascular organs in vertebrates. The nucleus pulposus (NP), a highly hydrated and proteoglycan-enriched tissue, forms the inner portion of the IVD. The NP is surrounded by a multi-lamellar fibrocartilaginous structure, the annulus fibrosus (AF). This structure is covered superior and inferior side by cartilaginous endplates (CEP). The NP is a unique tissue within the IVD as it results from the differentiation of notochordal cells, whereas, AF and CEP derive from the sclerotome. The hypoxia inducible factor-1α (HIF-1α) is expressed in NP cells but its function in NP development and homeostasis is largely unknown. We thus conditionally deleted HIF-1α in notochordal cells and investigated how loss of this transcription factor impacts NP formation and homeostasis at E15.5, birth, 1 and 4 months of age, respectively. Histological analysis, cell lineage studies, and TUNEL assay were performed. Morphologic changes of the mutant NP cells were identified as early as E15.5, followed, postnatally, by the progressive disappearance and replacement of the NP with a novel tissue that resembles fibrocartilage. Notably, lineage studies and TUNEL assay unequivocally proved that NP cells did not transdifferentiate into chondrocyte-like cells but they rather underwent massive cell death, and were completely replaced by a cell population belonging to a lineage distinct from the notochordal one. Finally, to evaluate the functional consequences of HIF-1α deletion in the NP, biomechanical testing of mutant IVD was performed. Loss of the NP in mutant mice significantly reduced the IVD biomechanical properties by decreasing its ability to absorb mechanical stress. These findings are similar to the changes usually observed during human IVD degeneration. Our study thus demonstrates that HIF-1α is essential for NP development and homeostasis, and it raises the intriguing possibility that this transcription factor could be involved in IVD degeneration in humans.

Kinetic and spectral properties of isovaleryl-CoA dehydrogenase and interaction with ligands.

PubMed

Mohsen, Al-Walid A; Vockley, Jerry

2015-01-01

Isovaleryl-CoA dehydrogenase (IVD) catalyzes the conversion of isovaleryl-CoA to 3-methylcrotonyl-CoA and the transfer of electrons to the electron transfer flavoprotein (ETF). Recombinant human IVD purifies with bound CoA-persulfide. A modified purification protocol was developed to isolate IVD without bound CoA-persulfide and to protect the protein thiols from oxidation. The CoA-persulfide-free IVD specific activity was 112.5 μmol porcine ETF min(-)(1) mg(-)(1), which was ∼20-fold higher than that of its CoA-persulfide bound form. The Km and catalytic efficiency (kcat/Km) for isovaleryl-CoA were 1.0 μM and 4.3 × 10(6) M(-1) s(-1) per monomer, respectively, and its Km for ETF was 2.0 μM. Anaerobic titration of isovaleryl-CoA into an IVD solution resulted in a stable blue complex with increased absorbance at 310 nm, decreased absorbance at 373 and 447 nm, and the appearance of the charge transfer complex band at 584 nm. The apparent dissociation constant (KDapp) determined spectrally for isovaleryl-CoA was 0.54 μM. Isovaleryl-CoA, acetoacetyl-CoA, methylenecyclopropyl-acetyl-CoA, and ETF induced CD spectral changes at the 250-500 nm region while isobutyryl-CoA did not, suggesting conformational changes occur at the flavin ring that are ligand specific. Replacement of the IVD Trp166 with a Phe did not block IVD interaction with ETF, indicating that its indole ring is not essential for electron transfer to ETF. A twelve amino acid synthetic peptide that matches the sequence of the ETF docking peptide competitively inhibited the enzyme reaction when ETF was used as the electron acceptor with a Ki of 1.5 mM. Copyright © 2014 Elsevier B.V. and Société française de biochimie et biologie Moléculaire (SFBBM). All rights reserved.

APOC3 Loss-of-Function Mutations, Remnant Cholesterol, Low-Density Lipoprotein Cholesterol, and Cardiovascular Risk: Mediation- and Meta-Analyses of 137 895 Individuals.

PubMed

Wulff, Anders B; Nordestgaard, Børge G; Tybjærg-Hansen, Anne

2018-03-01

Loss-of-function mutations in APOC3 associate with low remnant cholesterol levels and low risk of ischemic vascular disease (IVD). Because some studies show an additional association with low levels of low-density lipoprotein cholesterol (LDL-C), low LDL-C may explain the low risk of IVD in APOC3 loss-of-function heterozygotes. We tested to what extent the low risk of IVD in APOC3 loss-of-function heterozygotes is mediated by low plasma remnant cholesterol and LDL-C. In APOC3 loss-of-function heterozygotes versus noncarriers, we first determined remnant cholesterol and LDL-C levels in meta-analyses of 137 895 individuals. Second, we determined whether the association with LDL-C was masked by lipid-lowering therapy. Finally, using mediation analysis, we determined the fraction of the low risk of IVD and ischemic heart disease mediated by remnant cholesterol and LDL-C. In meta-analyses, remnant cholesterol was 43% lower (95% confidence interval, 40%-47%), and LDL-C was 4% lower (1%-6%) in loss-of-function heterozygotes (n=776) versus noncarriers. In the general population, LDL-C was 3% lower in loss-of-function heterozygotes versus noncarriers, 4% lower when correcting for lipid-lowering therapy, and 3% lower in untreated individuals ( P values, 0.06-0.008). Remnant cholesterol mediated 37% of the observed 41% lower risk of IVD and 54% of the observed 36% lower risk of ischemic heart disease; corresponding values mediated by LDL-C were 1% and 2%. The low risk of IVD observed in APOC3 loss-of-function heterozygotes is mainly mediated by the associated low remnant cholesterol and not by low LDL-C. Furthermore, the contribution of LDL-C to IVD risk was not masked by lipid-lowering therapy. This suggests APOC3 and remnant cholesterol as important new targets for reducing cardiovascular risk. © 2018 American Heart Association, Inc.

Integrin – Dependent Mechanotransduction in Mechanically Stimulated Human Annulus Fibrosus Cells: Evidence for an Alternative Mechanotransduction Pathway Operating with Degeneration

PubMed Central

Gilbert, Hamish T. J.; Nagra, Navraj S.; Freemont, Anthony J.; Millward-Sadler, Sarah J.; Hoyland, Judith A.

2013-01-01

Intervertebral disc (IVD) cells derived from degenerate tissue respond aberrantly to mechanical stimuli, potentially due to altered mechanotransduction pathways. Elucidation of the altered, or alternative, mechanotransduction pathways operating with degeneration could yield novel targets for the treatment of IVD disease. Our aim here was to investigate the involvement of RGD-recognising integrins and associated signalling molecules in the response to cyclic tensile strain (CTS) of human annulus fibrosus (AF) cells derived from non-degenerate and degenerate IVDs. AF cells from non-degenerate and degenerate human IVDs were cyclically strained with and without function blocking RGD – peptides with 10% strain, 1.0 Hz for 20 minutes using a Flexercell® strain device. QRT-PCR and Western blotting were performed to analyse gene expression of type I collagen and ADAMTS -4, and phosphorylation of focal adhesion kinase (FAK), respectively. The response to 1.0 Hz CTS differed between the two groups of AF cells, with decreased ADAMTS -4 gene expression and decreased type I collagen gene expression post load in AF cells derived from non-degenerate and degenerate IVDs, respectively. Pre-treatment of non-degenerate AF cells with RGD peptides prevented the CTS-induced decrease in ADAMTS -4 gene expression, but caused an increase in expression at 24 hours, a response not observed in degenerate AF cells where RGD pre-treatment failed to inhibit the mechano-response. In addition, FAK phosphorylation increased in CTS stimulated AF cells derived from non-degenerate, but not degenerate IVDs, with RGD pre-treatment inhibiting the CTS – dependent increase in phosphorylated FAK. Our findings suggest that RGD -integrins are involved in the 1.0 Hz CTS – induced mechano-response observed in AF cells derived from non-degenerate, but not degenerate IVDs. This data supports our previous work, suggesting an alternative mechanotransduction pathway may be operating in degenerate AF cells. PMID:24039840

45 CFR 308.1 - Self-assessment implementation methodology.

Code of Federal Regulations, 2010 CFR

2010-10-01

... selects statistically valid samples of cases from the IV-D program universe of cases; and (3) The State establishes a procedure for the design of samples and assures that no portions of the IV-D case universe are...

45 CFR 308.1 - Self-assessment implementation methodology.

Code of Federal Regulations, 2012 CFR

2012-10-01

... selects statistically valid samples of cases from the IV-D program universe of cases; and (3) The State establishes a procedure for the design of samples and assures that no portions of the IV-D case universe are...

45 CFR 308.1 - Self-assessment implementation methodology.

Code of Federal Regulations, 2011 CFR

2011-10-01

... selects statistically valid samples of cases from the IV-D program universe of cases; and (3) The State establishes a procedure for the design of samples and assures that no portions of the IV-D case universe are...

45 CFR 308.1 - Self-assessment implementation methodology.

Code of Federal Regulations, 2013 CFR

2013-10-01

... selects statistically valid samples of cases from the IV-D program universe of cases; and (3) The State establishes a procedure for the design of samples and assures that no portions of the IV-D case universe are...

45 CFR 308.1 - Self-assessment implementation methodology.

Code of Federal Regulations, 2014 CFR

2014-10-01

... selects statistically valid samples of cases from the IV-D program universe of cases; and (3) The State establishes a procedure for the design of samples and assures that no portions of the IV-D case universe are...

Preconditioning of mesenchymal stromal cells toward nucleus pulposus-like cells by microcryogels-based 3D cell culture and syringe-based pressure loading system.

PubMed

Zeng, Yang; Feng, Siyu; Liu, Wei; Fu, Qinyouen; Li, Yaqian; Li, Xiaokang; Chen, Chun; Huang, Chenyu; Ge, Zigang; Du, Yanan

2017-04-01

To precondition mesenchymal stromal/stem cells (MSCs) with mechanical stimulation may enhance cell survival and functions following implantation in load bearing environment such as nucleus pulposus (NP) in intervertebral disc (IVD). In this study, preconditioning of MSCs toward NP-like cells was achieved in previously developed poly (ethylene glycol) diacrylate (PEGDA) microcryogels (PMs) within a syringe-based three-dimensional (3D) culture system which provided a facile and cost-effective pressure loading approach. PMs loaded with alginate and MSCs could be incubated in a sealable syringe which could be air-compressed to apply pressure loading through a programmable syringe pump. Expression levels of chondrogenic marker genes SOX9, COL II, and ACAN were significantly upregulated in MSCs when pressure loading of 0.2 MPa or 0.8 MPa was implemented. Expression levels of COL I and COL X were downregulated when pressure loading was applied. In a nude mouse model, MSCs loaded in PMs mechanically stimulated for three days were subcutaneously injected using the same culture syringe. Three weeks postinjection, more proteoglycans (PGs) were deposited and more SOX9 and COL II but less COL I and COL X were stained in 0.2 MPa group. Furthermore, injectable MSCs-loaded PMs were utilized in an ex vivo rabbit IVD organ culture model that demonstrated the leak-proof function and enhanced cell retention of PMs assisted cell delivery to a load bearing environment for potential NP regeneration. This microcryogels-based 3D cell culture and syringe-based pressure loading system represents a novel method for 3D cell culture with mechanical stimulation for better function. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 507-520, 2017. © 2015 Wiley Periodicals, Inc.

2D segmentation of intervertebral discs and its degree of degeneration from T2-weighted magnetic resonance images

NASA Astrophysics Data System (ADS)

Castro-Mateos, Isaac; Pozo, José Maria; Lazary, Aron; Frangi, Alejandro F.

2014-03-01

Low back pain (LBP) is a disorder suffered by a large population around the world. A key factor causing this illness is Intervertebral Disc (IVD) degeneration, whose early diagnosis could help in preventing this widespread condition. Clinicians base their diagnosis on visual inspection of 2D slices of Magnetic Resonance (MR) images, which is subject to large interobserver variability. In this work, an automatic classification method is presented, which provides the Pfirrmann degree of degeneration from a mid-sagittal MR slice. The proposed method utilizes Active Contour Models, with a new geometrical energy, to achieve an initial segmentation, which is further improved using fuzzy C-means. Then, IVDs are classified according to their degree of degeneration. This classification is attained by employing Adaboost on five specific features: the mean and the variance of the probability map of the nucleus using two different approaches and the eccentricity of the fitting ellipse to the contour of the IVD. The classification method was evaluated using a cohort of 150 intervertebral discs assessed by three experts, resulting in a mean specificity (93%) and sensitivity (83%) similar to the one provided by every expert with respect to the most voted value. The segmentation accuracy was evaluated using the Dice Similarity Index (DSI) and Root Mean Square Error (RMSE) of the point-to-contour distance. The mean DSI ± 2 standard deviation was 91:7% ±5:6%, the mean RMSE was 0:82mm and the 95 percentile was 1:36mm. These results were found accurate when compared to the state-of-the-art.

An Anisotropic Multiphysics Model for Intervertebral Disk

PubMed Central

Gao, Xin; Zhu, Qiaoqiao; Gu, Weiyong

2016-01-01

Intervertebral disk (IVD) is the largest avascular structure in human body, consisting of three types of charged hydrated soft tissues. Its mechanical behavior is nonlinear and anisotropic, due mainly to nonlinear interactions among different constituents within tissues. In this study, a more realistic anisotropic multiphysics model was developed based on the continuum mixture theory and employed to characterize the couplings of multiple physical fields in the IVD. Numerical simulations demonstrate that this model is capable of systematically predicting the mechanical and electrochemical signals within the disk under various loading conditions, which is essential in understanding the mechanobiology of IVD. PMID:27099402

Wnt signaling activates Shh signaling in early postnatal intervertebral discs, and re-activates Shh signaling in old discs in the mouse.

PubMed

Winkler, Tamara; Mahoney, Eric J; Sinner, Debora; Wylie, Christopher C; Dahia, Chitra Lekha

2014-01-01

Intervertebral discs (IVDs) are strong fibrocartilaginous joints that connect adjacent vertebrae of the spine. As discs age they become prone to failure, with neurological consequences that are often severe. Surgical repair of discs treats the result of the disease, which affects as many as one in seven people, rather than its cause. An ideal solution would be to repair degenerating discs using the mechanisms of their normal differentiation. However, these mechanisms are poorly understood. Using the mouse as a model, we previously showed that Shh signaling produced by nucleus pulposus cells activates the expression of differentiation markers, and cell proliferation, in the postnatal IVD. In the present study, we show that canonical Wnt signaling is required for the expression of Shh signaling targets in the IVD. We also show that Shh and canonical Wnt signaling pathways are down-regulated in adult IVDs. Furthermore, this down-regulation is reversible, since re-activation of the Wnt or Shh pathways in older discs can re-activate molecular markers of the IVD that are lost with age. These data suggest that biological treatments targeting Wnt and Shh signaling pathways may be feasible as a therapeutic for degenerative disc disease.

Lumbar spine paraspinal muscle and intervertebral disc height changes in astronauts after long-duration spaceflight on the International Space Station

PubMed Central

Chang, DG; Healey, RM; Snyder, AJ; Sayson, JV; Macias, BR; Coughlin, DG; Bailey, JF; Parazynski, SE; Lotz, JC; Hargens, AR

2017-01-01

Study Design Prospective case series Objective Evaluate lumbar paraspinal muscle (PSM) cross-sectional area and intervertebral disc (IVD) height changes induced by a 6-month space mission on the International Space Station (ISS). The long-term objective of this project is to promote spine health and prevent spinal injury during space missions as well as here on Earth. Summary of Background NASA crewmembers have a 4.3 times higher risk of herniated IVDs, compared to the general and military aviator populations. The highest risk occurs during the first year after a mission. Microgravity exposure during long-duration spaceflights results in ~5cm lengthening of body height, spinal pain, and skeletal deconditioning. How the PSMs and IVDs respond during spaceflight is not well described. Methods Six NASA crewmembers were imaged supine with a 3T MRI. Imaging was conducted pre-flight, immediately post-flight and then 33 to 67 days after landing. Functional cross-sectional area (FCSA) measurements of the PSMs were performed at the L3-4 level. FCSA was measured by grayscale thresholding within the posterior lumbar extensors to isolate lean muscle on T2-weighted scans. IVD heights were measured at the anterior, middle and posterior sections of all lumbar levels. Repeated measures ANOVA was used to determine significance at p<0.05, followed by post-hoc testing. Results Paraspinal lean muscle mass, as indicated by the FCSA, decreased from 86% of the total PSM cross-sectional area down to 72%, immediately after the mission. Recovery of 68% of the post-flight loss occurred over the next 6 weeks, still leaving a significantly lower lean muscle fractional content compared to pre-flight values. In contrast, lumbar IVD heights were not appreciably different at any time point. Conclusions The data reveal lumbar spine PSM atrophy after long-duration spaceflight. Some FCSA recovery was seen with 46 days post-flight in a terrestrial environment, but it remained incomplete compared to pre-flight levels. PMID:27779600

A Finite Element Model to Simulate Formation of the Inverted-V Deformity

PubMed Central

Tjoa, Tjoson; Manuel, Cyrus T.; Leary, Ryan P.; Harb, Rani; Protsenko, Dmitriy E.; Wong, Brian J. F.

2018-01-01

IMPORTANCE Computational modeling can be used to mimic the forces acting on the nasal framework that lead to the inverted-V deformity (IVD) after surgery and potentially determine long-range outcomes. OBJECTIVE To demonstrate the use of the finite element method (FEM) to predict the formation of the IVD after separation of the upper lateral cartilages (ULCs) from the nasal septum. DESIGN, SETTING, AND PARTICIPANTS A computer model of a nose was derived from human computed tomographic data. The septum and upper and lower lateral cartilages were designed to fit within the soft-tissue envelope using computer-aided design software. Mechanical properties were obtained from the literature. The 3 simulations created included (1) partial fusion of the ULCs to the septum, (2) separation of the ULCs from the septum, and (3) a fully connected model to serve as a control. Forces caused by wound healing were prescribed at the junction of the disarticulated ULCs and septum. Using FEM software, equilibrium stress and strain were calculated. Displacement of the soft tissue along the nasal dorsum was measured and evaluated for evidence of morphologic change consistent with the IVD. MAIN OUTCOME AND MEASURES Morphologic changes on the computer models in response to each simulation. RESULTS When a posteroinferior force vector was applied along the nasal dorsum, the areas of highest stress were along the medial edge of the ULCs and at the junction of the ULCs and the nasal bones. With full detachment of ULCs and the dorsal septum, the characteristic IVD was observed. Both separation FEMs produced a peak depression of 0.3 mm along the nasal dorsum. CONCLUSIONS AND RELEVANCE The FEM can be used to simulate the long-term structural complications of a surgical maneuver in rhinoplasty, such as the IVD. When applied to other rhinoplasty maneuvers, the use of FEMs may be useful to simulate the long-term outcomes, particularly when long-term clinical results are not available. In the future, use of FEMs may simulate rhinoplasty results beyond simply morphing the outer contours of the nose and allow estimation of potentially long-term clinical outcomes that may not be readily apparent. LEVEL OF EVIDENCE NA. PMID:26720757

An investigation of the feasibility of interactive videodisc as a training mode for VDOT.

DOT National Transportation Integrated Search

1991-01-01

This report addressed the potential application of interactive videodisc (IVD) technology in the Virginia Department of Transportation (VDOT). The research revealed that IVD is a growing force as a training vehicle in several industries and instituti...

Lumbar intervertebral disc allograft transplantation: long-term mobility and impact on the adjacent segments.

PubMed

Huang, Yong-Can; Xiao, Jun; Lu, William W; Leung, Victor Y L; Hu, Yong; Luk, Keith D K

2017-03-01

Fresh-frozen intervertebral disc (IVD) allograft transplantation has been successfully performed in the human cervical spine. Whether this non-fusion technology could truly decrease adjacent segment disease is still unknown. This study evaluated the long-term mobility of the IVD-transplanted segment and the impact on the adjacent spinal segments in a goat model. Twelve goats were used. IVD allograft transplantation was performed at lumbar L4/L5 in 5 goats; the other 7 goats were used as the untreated control (5) and for the supply of allografts (2). Post-operation lateral radiographs of the lumbar spine in the neutral, full-flexion and full-extension positions were taken at 1, 3, 6, 9 and 12 months. Disc height (DH) of the allograft and the adjacent levels was calculated and range of motion (ROM) was measured using the Cobb's method. The anatomy of the adjacent discs was observed histologically. DH of the transplanted segment was decreased significantly after 3 months but no further reduction was recorded until the final follow-up. No obvious alteration was seen in the ROM of the transplanted segment at different time points with the ROM at 12 months being comparable to that of the untreated control. The DH and ROM in the adjacent segments were well maintained during the whole observation period. At post-operative 12 months, the ROM of the adjacent levels was similar to that of the untreated control and the anatomical morphology was well preserved. Lumbar IVD allograft transplantation in goats could restore the segmental mobility and did not negatively affect the adjacent segments after 12 months.

Increased expression of matrix metalloproteinase-10, nerve growth factor and substance P in the painful degenerate intervertebral disc

PubMed Central

Richardson, Stephen M; Doyle, Paul; Minogue, Ben M; Gnanalingham, Kanna; Hoyland, Judith A

2009-01-01

Introduction Matrix metalloproteinases (MMPs) are known to be involved in the degradation of the nucleus pulposus (NP) during intervertebral disc (IVD) degeneration. This study investigated MMP-10 (stromelysin-2) expression in the NP during IVD degeneration and correlated its expression with pro-inflammatory cytokines and molecules involved in innervation and nociception during degeneration which results in low back pain (LBP). Methods Human NP tissue was obtained at postmortem (PM) from patients without a history of back pain and graded as histologically normal or degenerate. Symptomatic degenerate NP samples were also obtained at surgery for LBP. Expression of MMP-10 mRNA and protein was analysed using real-time polymerase chain reaction and immunohistochemistry. Gene expression for pro-inflammatory cytokines interleukin-1 (IL-1) and tumour necrosis factor-alpha (TNF-α), nerve growth factor (NGF) and the pain-associated neuropeptide substance P were also analysed. Correlations between MMP-10 and IL-1, TNF-α and NGF were assessed along with NGF with substance P. Results MMP-10 mRNA was significantly increased in surgical degenerate NP when compared to PM normal and PM degenerate samples. MMP-10 protein was also significantly higher in degenerate surgical NP samples compared to PM normal. IL-1 and MMP-10 mRNA demonstrated a significant correlation in surgical degenerate samples, while TNF-α was not correlated with MMP-10 mRNA. NGF was significantly correlated with both MMP-10 and substance P mRNA in surgical degenerate NP samples. Conclusions MMP-10 expression is increased in the symptomatic degenerate IVD, where it may contribute to matrix degradation and initiation of nociception. Importantly, this study suggests differences in the pathways involved in matrix degradation between painful and pain-free IVD degeneration. PMID:19695094

Effects of Tobacco Smoking on the Degeneration of the Intervertebral Disc: A Finite Element Study

PubMed Central

Elmasry, Shady; Asfour, Shihab; de Rivero Vaccari, Juan Pablo; Travascio, Francesco

2015-01-01

Tobacco smoking is associated with numerous pathological conditions. Compelling experimental evidence associates smoking to the degeneration of the intervertebral disc (IVD). In particular, it has been shown that nicotine down-regulates both the proliferation rate and glycosaminoglycan (GAG) biosynthesis of disc cells. Moreover, tobacco smoking causes the constriction of the vascular network surrounding the IVD, thus reducing the exchange of nutrients and anabolic agents from the blood vessels to the disc. It has been hypothesized that both nicotine presence in the IVD and the reduced solute exchange are responsible for the degeneration of the disc due to tobacco smoking, but their effects on tissue homeostasis have never been quantified. In this study, a previously presented computational model describing the homeostasis of the IVD was deployed to investigate the effects of impaired solute supply and nicotine-mediated down-regulation of cell proliferation and biosynthetic activity on the health of the disc. We found that the nicotine-mediated down-regulation of cell anabolism mostly affected the GAG concentration at the cartilage endplate, reducing it up to 65% of the value attained in normal physiological conditions. In contrast, the reduction of solutes exchange between blood vessels and disc tissue mostly affected the nucleus pulposus, whose cell density and GAG levels were reduced up to 50% of their normal physiological levels. The effectiveness of quitting smoking on the regeneration of a degenerated IVD was also investigated, and showed to have limited benefit on the health of the disc. A cell-based therapy in conjunction with smoke cessation provided significant improvements in disc health, suggesting that, besides quitting smoking, additional treatments should be implemented in the attempt to recover the health of an IVD degenerated by tobacco smoking. PMID:26301590

On the Relative Relevance of Subject-Specific Geometries and Degeneration-Specific Mechanical Properties for the Study of Cell Death in Human Intervertebral Disk Models

PubMed Central

Malandrino, Andrea; Pozo, José M.; Castro-Mateos, Isaac; Frangi, Alejandro F.; van Rijsbergen, Marc M.; Ito, Keita; Wilke, Hans-Joachim; Dao, Tien Tuan; Ho Ba Tho, Marie-Christine; Noailly, Jérôme

2015-01-01

Capturing patient- or condition-specific intervertebral disk (IVD) properties in finite element models is outmost important in order to explore how biomechanical and biophysical processes may interact in spine diseases. However, disk degenerative changes are often modeled through equations similar to those employed for healthy organs, which might not be valid. As for the simulated effects of degenerative changes, they likely depend on specific disk geometries. Accordingly, we explored the ability of continuum tissue models to simulate disk degenerative changes. We further used the results in order to assess the interplay between these simulated changes and particular IVD morphologies, in relation to disk cell nutrition, a potentially important factor in disk tissue regulation. A protocol to derive patient-specific computational models from clinical images was applied to different spine specimens. In vitro, IVD creep tests were used to optimize poro-hyperelastic input material parameters in these models, in function of the IVD degeneration grade. The use of condition-specific tissue model parameters in the specimen-specific geometrical models was validated against independent kinematic measurements in vitro. Then, models were coupled to a transport-cell viability model in order to assess the respective effects of tissue degeneration and disk geometry on cell viability. While classic disk poro-mechanical models failed in representing known degenerative changes, additional simulation of tissue damage allowed model validation and gave degeneration-dependent material properties related to osmotic pressure and water loss, and to increased fibrosis. Surprisingly, nutrition-induced cell death was independent of the grade-dependent material properties, but was favored by increased diffusion distances in large IVDs. Our results suggest that in situ geometrical screening of IVD morphology might help to anticipate particular mechanisms of disk degeneration. PMID:25717471

Detection of low back pain using pH level-dependent imaging of the intervertebral disc using the ratio of R1ρ dispersion and -OH chemical exchange saturation transfer (RROC).

PubMed

Liu, Qi; Tawackoli, Wafa; Pelled, Gadi; Fan, Zhaoyang; Jin, Ning; Natsuaki, Yutaka; Bi, Xiaoming; Gart, Avrom; Bae, Hyun; Gazit, Dan; Li, Debiao

2015-03-01

Low pH is associated with intervertebral disc (IVD)-generated low back pain (LBP). The purpose of this work was to develop an in vivo pH level-dependent magnetic resonance imaging (MRI) method for detecting discogenic LBP, without using exogenous contrast agents. The ratio of R1ρ dispersion and chemical exchange saturation transfer (CEST) (RROC) was used for pH-level dependent imaging of the IVD while eliminating the effect of labile proton concentration. The technique was validated by numerical simulations and studies on phantoms and ex vivo porcine spines. Four male (ages 42.8 ± 18.3) and two female patients (ages 55.5 ± 2.1) with LBP and scheduled for discography were examined with the method on a 3.0 Tesla MR scanner. RROC measurements were compared with discography outcomes using paired t-test. Simulation and phantom results indicated RROC is a concentration independent and pH level-dependent technique. Porcine spine study results found higher RROC value was related to lower pH level. Painful discs based on discography had significant higher RROC values than those with negative diagnosis (P < 0.05). RROC imaging is a promising pH level dependent MRI technique that has the potential to be a noninvasive imaging tool to detect painful IVDs in vivo. © 2014 Wiley Periodicals, Inc.

Digital Audio: A Sound Design Element.

ERIC Educational Resources Information Center

Barron, Ann; Varnadoe, Susan

1992-01-01

Discussion of incorporating audio into videodiscs for multimedia educational applications highlights a project developed for the Navy that used digital audio in an interactive video delivery system (IVDS) for training sonar operators. Storage constraints with videodiscs are explained, design requirements for the IVDS are described, and production…

Perception of parents as demonstrating the inherent merit of their values: relations with self-congruence and subjective well-being.

PubMed

Yu, Shi; Assor, Avi; Liu, Xiangping

2015-02-01

This study focuses on the parenting practice of inherent value demonstration (IVD), involving parents' tendency to express their values in behaviours and appear satisfied and vital while doing so. Data from Chinese college students (n = 89) confirmed the hypothesis that offspring's perception of their parents as engaged in IVD predicts offspring's subjective well-being (SWB) through sense of self-congruence. Importantly, these relations emerged also when controlling for fundamental autonomy-supportive (FAS) parenting practices such as taking children's perspective, minimising control and allowing choice. These findings are consistent with the view that parents concerned with their children's sense of autonomy may do well to engage in IVD in addition to more fundamental autonomy-supportive practices. Future research may examine the role of IVD in promoting authentic values that serve as an internal compass that guides children to act in ways that feel self-congruent. © 2014 International Union of Psychological Science.

Disruption of biomineralization pathways in spinal tissues of a mouse model of diffuse idiopathic skeletal hyperostosis.

PubMed

Ii, Hisataka; Warraich, Sumeeta; Tenn, Neil; Quinonez, Diana; Holdsworth, David W; Hammond, James R; Dixon, S Jeffrey; Séguin, Cheryle A

2016-09-01

Equilibrative nucleoside transporter 1 (ENT1) mediates passage of adenosine across the plasma membrane. We reported previously that mice lacking ENT1 (ENT1(-/-)) exhibit progressive ectopic mineralization of spinal tissues resembling diffuse idiopathic skeletal hyperostosis (DISH) in humans. Here, we investigated mechanisms underlying aberrant mineralization in ENT1(-/-) mice. Micro-CT revealed ectopic mineralization of spinal tissues in both male and female ENT1(-/-) mice, involving the annulus fibrosus of the intervertebral discs (IVDs) of older mice. IVDs were isolated from wild-type and ENT1(-/-) mice at 2months of age (prior to disc mineralization), 4, and 6months of age (disc mineralization present) and processed for real-time PCR, cell isolation, or histology. Relative to the expression of ENTs in other tissues, ENT1 was the primary nucleoside transporter expressed in wild-type IVDs and mediated the functional uptake of [(3)H]2-chloroadenosine by annulus fibrosus cells. No differences in candidate gene expression were detected in IVDs from ENT1(-/-) and wild-type mice at 2 or 4months of age. However, at 6months of age, expression of genes that inhibit biomineralization Mgp, Enpp1, Ank, and Spp1 were reduced in IVDs from ENT1(-/-) mice. To assess whether changes detected in ENT1(-/-) mice were cell autonomous, annulus fibrosus cell cultures were established. Compared to wild-type cells, cells isolated from ENT1(-/-) IVDs at 2 or 6months of age demonstrated greater activity of alkaline phosphatase, a promoter of biomineralization. Cells from 2-month-old ENT1(-/-) mice also showed greater mineralization than wild-type. Interestingly, altered localization of alkaline phosphatase activity was detected in the inner annulus fibrosus of ENT1(-/-) mice in vivo. Alkaline phosphatase activity, together with the marked reduction in mineralization inhibitors, is consistent with the mineralization of IVDs seen in ENT1(-/-) mice at older ages. These findings establish that both cell-autonomous and systemic mechanisms contribute to ectopic mineralization in ENT1(-/-) mice. Copyright © 2016 Elsevier Inc. All rights reserved.

75 FR 38611 - Child Support Enforcement Program; Intergovernmental Child Support

Federal Register 2010, 2011, 2012, 2013, 2014

2010-07-02

...This rule revises Federal requirements for establishing and enforcing intergovernmental support obligations in Child Support Enforcement (IV-D) program cases receiving services under title IV-D of the Social Security Act (the Act). This final rule revises previous interstate requirements to apply to case processing in all intergovernmental cases; requires the responding State IV-D agency to pay the cost of genetic testing; clarifies responsibility for determining in which State tribunal a controlling order determination is made where multiple support orders exist; recognizes and incorporates electronic communication advancements; and makes conforming changes to the Federal substantial compliance audit and State self-assessment requirements.

Early onset of disc degeneration in SM/J mice is associated with changes in ion transport systems and fibrotic events.

PubMed

Zhang, Ying; Xiong, Chi; Kudelko, Mateusz; Li, Yan; Wang, Cheng; Wong, Yuk Lun; Tam, Vivian; Rai, Muhammad Farooq; Cheverud, James; Lawson, Heather A; Sandell, Linda; Chan, Wilson C W; Cheah, Kathryn S E; Sham, Pak C; Chan, Danny

2018-04-09

Intervertebral disc degeneration (IDD) causes back pain and sciatica, affecting quality of life and resulting in high economic/social burden. The etiology of IDD is not well understood. Along with aging and environmental factors, genetic factors also influence the onset, progression and severity of IDD. Genetic studies of risk factors for IDD using human cohorts are limited by small sample size and low statistical power. Animal models amenable to genetic and functional studies of IDD provide desirable alternatives. Despite differences in size and cellular content as compared to human intervertebral discs (IVDs), the mouse is a powerful model for genetics and assessment of cellular changes relevant to human biology. Here, we provide evidence for early onset disc degeneration in SM/J relative to LG/J mice with poor and good tissue healing capacity respectively. In the first few months of life, LG/J mice maintain a relatively constant pool of notochordal-like cells in the nucleus pulposus (NP) of the IVD. In contrast, chondrogenic events are observed in SM/J mice beginning as early as one-week-old, with progressive fibrotic changes. Further, the extracellular matrix changes in the NP are consistent with IVD degeneration. Leveraging on the genomic data of two parental and two recombinant inbred lines, we assessed the genetic contribution to the NP changes and identified processes linked to the regulation of ion transport systems. Significantly, "transport" system is also in the top three gene ontology (GO) terms from a comparative proteomic analysis of the mouse NP. These findings support the potential of the SM/J, LG/J and their recombinant inbred lines for future genetic and biological analysis in mice and validation of candidate genes and biological relevance in human cohort studies. The proteomic data has been deposited to the ProteomeXchange Consortium via the PRIDE [1] partner repository with the dataset identifier PXD008784. Copyright © 2017. Published by Elsevier B.V.

Fibrin-genipin annulus fibrosus sealant as a delivery system for anti-TNFα drug.

PubMed

Likhitpanichkul, Morakot; Kim, Yesul; Torre, Olivia M; See, Eugene; Kazezian, Zepur; Pandit, Abhay; Hecht, Andrew C; Iatridis, James C

2015-09-01

Intervertebral discs (IVDs) are attractive targets for local drug delivery because they are avascular structures with limited transport. Painful IVDs are in a chronic inflammatory state. Although anti-inflammatories show poor performance in clinical trials, their efficacy treating IVD cells suggests that sustained, local drug delivery directly to painful IVDs may be beneficial. The purpose of this study was to determine if genipin cross-linked fibrin (FibGen) with collagen Type I hollow spheres (CHS) can serve as a drug-delivery carrier for infliximab, the anti-tumor necrosis factor α (TNFα) drug. Infliximab was chosen as a model drug because of the known role of TNFα in increasing downstream production of several pro-inflammatory cytokines and pain mediators. Genipin cross-linked fibrin was used as drug carrier because it is adhesive, injectable, and slowly degrading hydrogel with the potential to seal annulus fibrosus (AF) defects. CHS allow simple and nondamaging drug loading and could act as a drug reservoir to improve sustained delivery. This is a study of biomaterials and human AF cell culture to determine drug release kinetics and efficacy. Infliximab was delivered at low and high concentrations using FibGen with and without CHS. Gels were analyzed for structure, drug release kinetics, and efficacy treating human AF cells after release. Fibrin showed rapid infliximab drug release but degraded quickly. CHS alone showed a sustained release profile, but the small spheres may not remain in a degenerated IVD with fissures. Genipin cross-linked fibrin showed steady and low levels of infliximab release that was increased when loaded with higher drug concentrations. Infliximab was bound in CHS when delivered within FibGen and was only released after enzymatic degradation. The infliximab released over 20 days retained its bioactivity as confirmed by the sustained reduction of interleukin (IL)-1β, IL-6, IL-8, and TNFα concentrations produced by AF cells. Direct mixing of infliximab into FibGen was the simplest drug-loading protocol capable of sustained release. Results show feasibility of using drug-loaded FibGen for delivery of infliximab and, in the context with the literature, show potential to seal AF defects and partially restore IVD biomechanics. Future investigations are required to determine if drug-loaded FibGen can effectively deliver drugs, seal AF defects, and promote IVD repair or prevent further IVD degeneration in vivo. Copyright © 2015 Elsevier Inc. All rights reserved.

In vivo dosimetry in external beam radiotherapy.

PubMed

Mijnheer, Ben; Beddar, Sam; Izewska, Joanna; Reft, Chester

2013-07-01

In vivo dosimetry (IVD) is in use in external beam radiotherapy (EBRT) to detect major errors, to assess clinically relevant differences between planned and delivered dose, to record dose received by individual patients, and to fulfill legal requirements. After discussing briefly the main characteristics of the most commonly applied IVD systems, the clinical experience of IVD during EBRT will be summarized. Advancement of the traditional aspects of in vivo dosimetry as well as the development of currently available and newly emerging noninterventional technologies are required for large-scale implementation of IVD in EBRT. These new technologies include the development of electronic portal imaging devices for 2D and 3D patient dosimetry during advanced treatment techniques, such as IMRT and VMAT, and the use of IVD in proton and ion radiotherapy by measuring the decay of radiation-induced radionuclides. In the final analysis, we will show in this Vision 20∕20 paper that in addition to regulatory compliance and reimbursement issues, the rationale for in vivo measurements is to provide an accurate and independent verification of the overall treatment procedure. It will enable the identification of potential errors in dose calculation, data transfer, dose delivery, patient setup, and changes in patient anatomy. It is the authors' opinion that all treatments with curative intent should be verified through in vivo dose measurements in combination with pretreatment checks.

Shh signaling from the nucleus pulposus is required for the postnatal growth and differentiation of the mouse intervertebral disc.

PubMed

Dahia, Chitra Lekha; Mahoney, Eric; Wylie, Christopher

2012-01-01

Intervertebral discs (IVD) are essential components of the vertebral column. They maintain separation, and provide shock absorbing buffers, between adjacent vertebrae, while also allowing movements between them. Each IVD consists of a central semi-liquid nucleus pulposus (NP) surrounded by a multi-layered fibrocartilagenous annulus fibrosus (AF). Although the IVDs grow and differentiate after birth along with the vertebral column, little is known about the mechanism of this. Understanding the signals that control normal IVD growth and differentiation would also provide potential therapies for degenerative disc disease, which is the major cause of lower back pain and affects a large proportion of the population. In this work, we show that during postnatal growth of the mouse, Sonic hedgehog (Shh) signaling from the NP cells controls many aspects of growth and differentiation of both the NP cells themselves and of the surrounding AF, and that it acts, at least partly, by regulating other signaling pathways in the NP and AF. Recent studies have shown that the NP cells arise from the embryonic notochord, which acts as a major signaling center in the embryo. This work shows that this notochord-derived tissue continues to carry out a major signaling function in the postnatal body and that the IVDs are signaling centers, in addition to their already known functions in the mechanics of vertebral column function.

45 CFR 310.0 - What does this part cover?

Code of Federal Regulations, 2010 CFR

2010-10-01

... COMPUTERIZED TRIBAL IV-D SYSTEMS AND OFFICE AUTOMATION General Provisions § 310.0 What does this part cover... and Office Automation including: (a) The automated systems options for comprehensive Tribal IV-D... and Office Automation in § 310.15 of this part; (d) The conditions for funding the installation...

Multimedia Football Viewing: Embedded Rules, Practice, and Video Context in IVD Procedural Learning.

ERIC Educational Resources Information Center

Kim, Eunsoon; Young, Michael F.

This study investigated the effects of interactive video (IVD) instruction with embedded rules (production system rules) and practice with feedback on learners' academic achievement and perceived self efficacy in the domain of procedural knowledge for watching professional football. Subjects were 71 female volunteers from undergraduate education…

76 FR 20356 - Submission for OMB Review; Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2011-04-12

... DEPARTMENT OF HEALTH AND HUMAN SERVICES Administration for Children and Families Submission for OMB Review; Comment Request Title: State Plan for Child Support under IV-D of the Social Security Act. OMB No.: 0970-0017. Description: The Office of Child Support Enforcement has approved a IV-D State...

45 CFR 309.01 - What does this part cover?

Code of Federal Regulations, 2011 CFR

2011-10-01

... Public Welfare Regulations Relating to Public Welfare OFFICE OF CHILD SUPPORT ENFORCEMENT (CHILD SUPPORT... CHILD SUPPORT ENFORCEMENT (IV-D) PROGRAM Tribal IV-D Program: General Provisions § 309.01 What does this... Social Security Act. Section 455(f) of the Act authorizes direct grants to Indian Tribes and Tribal...

77 FR 7591 - Request for Notification From Consumer Organizations Interested in Participating in the Selection...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-02-13

.... Assurance--Knowledgeable in clinical practice, research specialization, or professional work that has a significant focus on mammography. Certain Panels of the Medical Devices Advisory Committee Clinical Chemistry... chemistry and toxicology in vitro diagnostic devices (IVDs); clinical use of related IVDs in laboratories...

Senecavirus A: overview of experimental studies

USDA-ARS?s Scientific Manuscript database

Introduction Senecavirus A (SVA) is a picornavirus in the same family as Foot and Mouth Disease virus (FMDV) and swine vesicular disease virus (SVDV). SVA has been associated with rare cases of idiopathic vesicular disease (IVD) in the past. A "mini-epidemic" of IVD in US finisher and sow farms beg...

45 CFR 305.64 - Audit procedures and State comments.

Code of Federal Regulations, 2010 CFR

2010-10-01

... Audit procedures and State comments. (a) Prior to the start of the actual audit, Federal auditors will hold an audit entrance conference with the IV-D agency. At that conference, the auditors will explain... fieldwork, Federal auditors will afford the State IV-D agency an opportunity for an audit exit conference at...

Demonstration of Senecavirus A protective immunity in a pig model

USDA-ARS?s Scientific Manuscript database

Although idiopathic vesicular disease (IVD) is rarely diagnosed in US swine, in 2015 there were over 100 cases identified throughout the country. Frequently, Senecavirus A (SVA) has been associated with IVD and is presumed to be the etiologic agent. In recent studies we have shown SVA to induce a v...

45 CFR 305.61 - Penalty for failure to meet IV-D requirements.

Code of Federal Regulations, 2010 CFR

2010-10-01

... HEALTH AND HUMAN SERVICES PROGRAM PERFORMANCE MEASURES, STANDARDS, FINANCIAL INCENTIVES, AND PENALTIES § 305.61 Penalty for failure to meet IV-D requirements. (a) A State will be subject to a financial... order establishment and current collections performance measures as set forth in § 305.40 of this part...

45 CFR 302.80 - Medical support enforcement.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 45 Public Welfare 2 2014-10-01 2012-10-01 true Medical support enforcement. 302.80 Section 302.80... PLAN REQUIREMENTS § 302.80 Medical support enforcement. (a) The State plan may provide that the IV-D agency will secure and enforce medical support obligations under a cooperative agreement between the IV-D...

Vesicular disease in 9-week-old pigs experimentally infected with Senecavirus A

USDA-ARS?s Scientific Manuscript database

Introduction: Senecavirus A (SVA), a picornavirus, has been infrequently associated with cases of idiopathic vesicular disease (IVD) in pigs in the US and Canada since 1988. In 2014 and 2015 there was surge of IVD cases in Brazil and US, respectively. SVA was identified in serum, vesicular fluid, an...

45 CFR 302.80 - Medical support enforcement.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 45 Public Welfare 2 2011-10-01 2011-10-01 false Medical support enforcement. 302.80 Section 302.80... PLAN REQUIREMENTS § 302.80 Medical support enforcement. (a) The State plan may provide that the IV-D agency will secure and enforce medical support obligations under a cooperative agreement between the IV-D...

45 CFR 305.64 - Audit procedures and State comments.

Code of Federal Regulations, 2011 CFR

2011-10-01

... Audit procedures and State comments. (a) Prior to the start of the actual audit, Federal auditors will hold an audit entrance conference with the IV-D agency. At that conference, the auditors will explain... fieldwork, Federal auditors will afford the State IV-D agency an opportunity for an audit exit conference at...

45 CFR 302.20 - Separation of cash handling and accounting functions.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 45 Public Welfare 2 2010-10-01 2010-10-01 false Separation of cash handling and accounting... accounting functions. The State plan shall provide that the following requirements and criteria to separate the cash handling and accounting functions are in effect. (a) IV-D responsibility. The IV-D agency...

Comparison of Saramis 4.12 and IVD 3.0 Vitek MS Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for Identification of Mycobacteria from Solid and Liquid Culture Media

PubMed Central

Leyer, Caroline; Gregorowicz, Guillaume; Mougari, Faiza; Raskine, Laurent; Cambau, Emmanuelle

2017-01-01

ABSTRACT During the last decade, many investigators have studied matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) for identification of mycobacteria. Diverse and contradictory results indicated that optimal level for routine testing has not been reached yet. This work aimed to assess Vitek MS through two distinct versions, Saramis v4.12 RUO and the IVD v3.0, under conditions close to routine laboratory practice. Overall, 111 mycobacterial isolates were subjected to protein extraction and same spectra were matched against both databases. The IVD v3.0 database proved to be superior to Saramis v4.12 and its identification rates remarkably increased, from 67% to 94% for isolates grown on Middlebrook 7H10 solid medium and from 62% to 91% for isolates grown on mycobacterial growth indicator tube (MGIT) liquid medium. With this new version, IVD v3.0, MALDI-TOF MS might be integrated into routine clinical diagnostics, although molecular techniques remain mandatory in some cases. PMID:28424252

Foxa1 and Foxa2 are required for formation of the intervertebral discs.

PubMed

Maier, Jennifer A; Lo, YinTing; Harfe, Brian D

2013-01-01

The intervertebral disc (IVD) is composed of 3 main structures, the collagenous annulus fibrosus (AF), which surrounds the gel-like nucleus pulposus (NP), and hyaline cartilage endplates, which are attached to the vertebral bodies. An IVD is located between each vertebral body. Degeneration of the IVD is thought to be a major cause of back pain, a potentially chronic condition for which there exist few effective treatments. The NP forms from the embryonic notochord. Foxa1 and Foxa2, transcription factors in the forkhead box family, are expressed early during notochord development. However, embryonic lethality and the absence of the notochord in Foxa2 null mice have precluded the study of potential roles these genes may play during IVD formation. Using a conditional Foxa2 allele in conjunction with a tamoxifen-inducible Cre allele (ShhcreER(T2)), we removed Foxa2 from the notochord of E7.5 mice null for Foxa1. Foxa1(-/-);Foxa2(c/c);ShhcreER(T2) double mutant animals had a severely deformed nucleus pulposus, an increase in cell death in the tail, decreased hedgehog signaling, defects in the notochord sheath, and aberrant dorsal-ventral patterning of the neural tube. Embryos lacking only Foxa1 or Foxa2 from the notochord were indistinguishable from control animals, demonstrating a functional redundancy for these genes in IVD formation. In addition, we provide in vivo genetic evidence that Foxa genes are required for activation of Shh in the notochord.

Foxa1 and Foxa2 Are Required for Formation of the Intervertebral Discs

PubMed Central

Maier, Jennifer A.; Lo, YinTing; Harfe, Brian D.

2013-01-01

The intervertebral disc (IVD) is composed of 3 main structures, the collagenous annulus fibrosus (AF), which surrounds the gel-like nucleus pulposus (NP), and hyaline cartilage endplates, which are attached to the vertebral bodies. An IVD is located between each vertebral body. Degeneration of the IVD is thought to be a major cause of back pain, a potentially chronic condition for which there exist few effective treatments. The NP forms from the embryonic notochord. Foxa1 and Foxa2, transcription factors in the forkhead box family, are expressed early during notochord development. However, embryonic lethality and the absence of the notochord in Foxa2 null mice have precluded the study of potential roles these genes may play during IVD formation. Using a conditional Foxa2 allele in conjunction with a tamoxifen-inducible Cre allele (ShhcreERT2), we removed Foxa2 from the notochord of E7.5 mice null for Foxa1. Foxa1−/−;Foxa2c/c;ShhcreERT2 double mutant animals had a severely deformed nucleus pulposus, an increase in cell death in the tail, decreased hedgehog signaling, defects in the notochord sheath, and aberrant dorsal-ventral patterning of the neural tube. Embryos lacking only Foxa1 or Foxa2 from the notochord were indistinguishable from control animals, demonstrating a functional redundancy for these genes in IVD formation. In addition, we provide in vivo genetic evidence that Foxa genes are required for activation of Shh in the notochord. PMID:23383217

Plasma vitamin D and osteo-cartilaginous markers in Italian males affected by intervertebral disc degeneration: Focus on seasonal and pathological trend of type II collagen degradation.

PubMed

Brayda-Bruno, Marco; Viganò, Marco; Cauci, Sabina; Vitale, Jacopo A; de Girolamo, Laura; De Luca, Paola; Lombardi, Giovanni; Banfi, Giuseppe; Colombini, Alessandra

2017-08-01

To evaluate plasma vitamin D and cross-linked C-telopeptides of type I (CTx-I) and type II (CTx-II) collagen concentrations in males with lumbar intervertebral disc degeneration (IVD) compared to healthy controls. Improved knowledge might suggest to optimize the vitamin D status of IVD patients and contribute to clarify mechanisms of cartilage degradation. 79 Italian males with lumbar IVD assessed by Magnetic Resonance Imaging (MRI) and 79 age, sex and BMI-matched healthy controls were enrolled. Plasma 25hydroxyvitamin D (25(OH)D), CTx-I and CTx-II were measured by immunoassays. Circannual seasonality, correlation between biomarkers concentrations and clinical variables were assessed. Overall subjects 25(OH)D and CTx-II showed month rhythmicity with acrophase in August/September and October/November, and nadir in February/March and April/May, respectively. An inverse correlation between 25(OH)D and CTx-I, and a direct correlation between CTx-II and CTx-I were observed. IVD patients, particularly with osteochondrosis, showed higher CTx-II than healthy controls. Month of sampling may affect plasma 25(OH)D and CTx-II concentrations. The correlation between CTx-I and CTx-II suggests an interplay between the osteo-cartilaginous endplate and the fibro-cartilaginous disc. The results of this study highlighted that osteochondrosis associates with increased cartilaginous catabolism. Vitamin D supplementation seems more necessary in winter for lumbar IVD patients. Copyright © 2017 Elsevier B.V. All rights reserved.

A comprehensive training approach for biomedical engineers in biochemistry and in vitro diagnostics technology.

PubMed

Spyropoulos, Basile; Tzavaras, Aris

2007-01-01

The purpose of this paper is to review 20 years (1987-2007) of experience in training young Biomedical Engineers in Biochemistry and in vitro Diagnostics (IVD) Technology. This encountering has resulted in the gradual formation of a comprehensive training package that includes lectures and laboratory practicals, supported by both, traditional and on-line digital means, such as lecture-notes, slides, videos, demos and equipment simulations. Further, this course is maintained up to date by several research and development activities that offer partially feed back to the course and enrich its contents with custom developed devices, methods and application software. In this paper are presented, first, the structure and the components of this course, and second, the most important custom developed novelties, which have been integrated in the IVD Technology laboratory-practicals.

Intradiscal injection of simvastatin results in radiologic, histologic, and genetic evidence of disc regeneration in a rat model of degenerative disc disease

PubMed Central

Than, Khoi D.; Rahman, Shayan U.; Wang, Lin; Khan, Adam; Kyere, Kwaku A.; Than, Tracey T.; Miyata, Yoshinari; Park, Yoon-Shin; La Marca, Frank; Kim, Hyungjin M.; Zhang, Huina; Park, Paul; Lin, Chia-Ying

2014-01-01

BACKGROUND CONTEXT A large percentage of back pain can be attributed to degeneration of the intervertebral disc (IVD). Bone morphogenetic protein-2 (BMP-2) is known to play an important role in chondrogenesis of the IVD. Simvastatin is known to up-regulate expression of BMP-2. Thus, we hypothesized that intradiscal injection of simvastatin in a rat model of degenerative disc disease (DDD) would result in retardation of DDD. PURPOSE To develop a novel conservative treatment for DDD and related discogenic back pain. STUDY DESIGN/SETTING Laboratory investigation. METHODS Disc injury was induced in 272 rats via 21-gauge needle puncture. After 6 weeks, injured discs were treated with simvastatin in a saline or hydrogel carrier. Rats were sacrificed at predetermined time points. Outcome measures assessed were radiologic, histologic, and genetic. Radiologically, the MRI index (number of pixels multiplied by corresponding image densities) was determined. Histologically, disc spaces were read by 3 blinded scorers employing a previously described histological grading scale. Genetically, nuclei pulposi were harvested and polymerase chain reaction was run to determine relative levels of aggrecan, collagen type II, and BMP-2 gene expression. This project was supported by Grant No. R01 AR056649 from NIAMS/NIH. There are no other financial conflicts of interest to report. RESULTS Radiologically, discs treated with 5 mg/mL simvastatin in hydrogel or saline demonstrated MRI indices that were normal through 8 weeks post-treatment, although this was more sustained when delivered in hydrogel. Histologically, discs treated with 5 mg/mL simvastatin in hydrogel demonstrated improved grades in comparison to discs treated at higher doses. Genetically, discs treated with 5 mg/mL of simvastatin in hydrogel demonstrated higher gene expression of aggrecan and collagen type II than control. CONCLUSIONS Degenerate discs treated with 5 mg/mL simvastatin in a hydrogel carrier demonstrated radiographic and histologic features resembling normal, non-injured IVDs. In addition, gene expression of aggrecan and collagen type II (important constituents of the IVD extracellular matrix) was up-regulated in treated discs. Injection of simvastatin into degenerate IVDs may result in retardation of disc degeneration and represents a promising investigational therapy for conservative treatment of DDD. PMID:24291703

Back pain in space and post-flight spine injury: Mechanisms and countermeasure development

NASA Astrophysics Data System (ADS)

Sayson, Jojo V.; Lotz, Jeffrey; Parazynski, Scott; Hargens, Alan R.

2013-05-01

During spaceflight many astronauts experience moderate to severe lumbar pain and deconditioning of paraspinal muscles. There is also a significant incidence of herniated nucleus pulposus (HNP) in astronauts post-flight being most prevalent in cervical discs. Relief of in-flight lumbar back pain is facilitated by assuming a knee-to-chest position. The pathogenesis of lumbar back pain during spaceflight is most likely discogenic and somatic referred (from the sinuvertebral nerves) due to supra-physiologic swelling of the lumbar intervertebral discs (IVDs) due to removal of gravitational compressive loads in microgravity. The knee-to-chest position may reduce lumbar back pain by redistributing stresses through compressive loading to the IVDs, possibly reducing disc volume by fluid outflow across IVD endplates. IVD stress redistribution may reduce Type IV mechanoreceptor nerve impulse propagation in the annulus fibrosus and vertebral endplate resulting in centrally mediated pain inhibition during spinal flexion. Countermeasures for lumbar back pain may include in-flight use of: (1) an axial compression harness to prevent excessive IVD expansion and spinal column elongation; (2) the use of an adjustable pulley exercise developed to prevent atrophy of spine muscle stabilisers; and (3) other exercises that provide Earth-like annular stress with low-load repetitive active spine rotation movements. The overall objective of these countermeasures is to promote IVD health and to prevent degenerative changes that may lead to HNPs post-flight. In response to "NASA's Critical Path Roadmap Risks and Questions" regarding disc injury and higher incidence of HNPs after space flight (Integrated Research Plan Gap-B4), future studies will incorporate pre- and post-flight imaging of International Space Station long-duration crew members to investigate mechanisms of lumbar back pain as well as degeneration and damage to spinal structures. Quantitative results on morphological, biochemical, metabolic, and kinematic spinal changes in the lumbar spine may aid further development of countermeasures to prevent lumbar back pain in microgravity and reduce the incidence of HNPs post-flight.

The change in the diffusion of water in normal and degenerative lumbar intervertebral discs following joint mobilization compared to prone lying.

PubMed

Beattie, Paul F; Donley, Jonathan W; Arnot, Cathy F; Miller, Ronald

2009-01-01

Prospective, repeated measures obtained under treatment and control conditions. The purposes of this study were to provide preliminary evidence regarding the immediate change in the diffusion of water in the nuclear region of normal and degenerative lumbar intervertebral discs (IVDs) following a single session of lumbar joint mobilization, and to compare these findings to the immediate change in the diffusion of water following a 10-minute session of prone lying. There is conflicting evidence regarding the effectiveness and efficacy of lumbar joint mobilization. Increased knowledge of the physiologic effects of lumbar joint mobilization can lead to refinement of its clinical application. A total of 24 people (15 males and 9 females), ranging in age from 22 to 58 years, participated in this study. All subjects had a history of activity-limiting low back pain. Diffusion-weighted magnetic resonance images (DW-MRIs) were obtained immediately before and after a 10-minute session of lumbar joint mobilization. At least 1 month later, a second session was performed in which DW-MRIs were obtained immediately before and after a 10-minute session of prone lying. Following lumbar joint mobilization, a significant increase (P = .002) in the mean values for diffusion of water was observed within degenerative IVDs at L5-S1 (22.2% increase; effect size, 0.97). Degenerative IVDs at L1-2 to L4-5 and normal IVDs at L1-2 to L5-S1 did not demonstrate a change in diffusion following joint mobilization. Prone lying was not associated with a change in diffusion for normal or degenerative IVDs. The stimulus provided by lumbar joint mobilization may influence the diffusion of water in degenerative IVDs at L5-S1; however, these are preliminary findings and the relationship of these findings to pain and function needs further investigation.

The role of interleukin-1 in the pathogenesis of human Intervertebral disc degeneration

PubMed Central

Le Maitre, Christine Lyn; Freemont, Anthony J; Hoyland, Judith Alison

2005-01-01

In this study, we investigated the hypotheses that in human intervertebral disc (IVD) degeneration there is local production of the cytokine IL-1, and that this locally produced cytokine can induce the cellular and matrix changes of IVD degeneration. Immunohistochemistry was used to localize five members of the IL-1 family (IL-1α, IL-1β, IL-1Ra (IL-1 receptor antagonist), IL-1RI (IL-1 receptor, type I), and ICE (IL-1β-converting enzyme)) in non-degenerate and degenerate human IVDs. In addition, cells derived from non-degenerate and degenerate human IVDs were challenged with IL-1 agonists and the response was investigated using real-time PCR for a number of matrix-degrading enzymes, matrix proteins, and members of the IL-1 family. This study has shown that native disc cells from non-degenerate and degenerate discs produced the IL-1 agonists, antagonist, the active receptor, and IL-1β-converting enzyme. In addition, immunopositivity for these proteins, with the exception of IL-1Ra, increased with severity of degeneration. We have also shown that IL-1 treatment of human IVD cells resulted in increased gene expression for the matrix-degrading enzymes (MMP 3 (matrix metalloproteinase 3), MMP 13 (matrix metalloproteinase 13), and ADAMTS-4 (a disintegrin and metalloproteinase with thrombospondin motifs)) and a decrease in the gene expression for matrix genes (aggrecan, collagen II, collagen I, and SOX6). In conclusion we have shown that IL-1 is produced in the degenerate IVD. It is synthesized by native disc cells, and treatment of human disc cells with IL-1 induces an imbalance between catabolic and anabolic events, responses that represent the changes seen during disc degeneration. Therefore, inhibiting IL-1 could be an important therapeutic target for preventing and reversing disc degeneration. PMID:15987475

Correlation study between facet joint cartilage and intervertebral discs in early lumbar vertebral degeneration using T2, T2* and T1ρ mapping

PubMed Central

Zhang, Yi; Hu, Jianzhong; Duan, Chunyue; Hu, Ping; Lu, Hongbin; Peng, Xianjing

2017-01-01

Recent advancements in magnetic resonance imaging have allowed for the early detection of biochemical changes in intervertebral discs and articular cartilage. Here, we assessed the feasibility of axial T2, T2* and T1ρ mapping of the lumbar facet joints (LFJs) to determine correlations between cartilage and intervertebral discs (IVDs) in early lumbar vertebral degeneration. We recruited 22 volunteers and examined 202 LFJs and 101 IVDs with morphological (sagittal and axial FSE T2-weighted imaging) and axial biochemical (T2, T2* and T1ρ mapping) sequences using a 3.0T MRI scanner. IVDs were graded using the Pfirrmann system. Mapping values of LFJs were recorded according to the degeneration grades of IVDs at the same level. The feasibility of T2, T2* and T1ρ in IVDs and LFJs were analyzed by comparing these mapping values across subjects with different rates of degeneration using Kruskal-Wallis tests. A Pearson’s correlation analysis was used to compare T2, T2* and T1ρ values of discs and LFJs. We found excellent reproducibility in the T2, T2* and T1ρ values for the nucleus pulposus (NP), anterior and posterior annulus fibrosus (PAF), and LFJ cartilage (intraclass correlation coefficients 0.806–0.955). T2, T2* and T1ρ mapping (all P<0.01) had good Pfirrmann grade performances in the NP with IVD degeneration. LFJ T2* values were significantly different between grades I and IV (PL = 0.032, PR = 0.026), as were T1ρ values between grades II and III (PL = 0.002, PR = 0.006) and grades III and IV (PL = 0.006, PR = 0.001). Correlations were moderately negative for T1ρ values between LFJ cartilage and NP (rL = −0.574, rR = −0.551), and between LFJ cartilage and PAF (rL = −0.551, rR = −0.499). T1ρ values of LFJ cartilage was weakly correlated with T2 (r = 0.007) and T2* (r = −0.158) values. Overall, we show that axial T1ρ effectively assesses early LFJ cartilage degeneration. Using T1ρ analysis, we propose a link between LFJ degeneration and IVD NP or PAF changes. PMID:28570641

Biocompatibility and intradiscal application of a thermoreversible celecoxib-loaded poly-N-isopropylacrylamide MgFe-layered double hydroxide hydrogel in a canine model.

PubMed

Willems, Nicole; Yang, Hsiao-Yin; Langelaan, Marloes L P; Tellegen, Anna R; Grinwis, Guy C M; Kranenburg, Hendrik-Jan C; Riemers, Frank M; Plomp, Saskia G M; Craenmehr, Eric G M; Dhert, Wouter J A; Papen-Botterhuis, Nicole E; Meij, Björn P; Creemers, Laura B; Tryfonidou, Marianna A

2015-08-20

Chronic low back pain due to intervertebral disc (IVD) degeneration is associated with increased levels of inflammatory mediators. Current medical treatment consists of oral anti-inflammatory drugs to alleviate pain. In this study, the efficacy and safety of a novel thermoreversible poly-N-isopropylacrylamide MgFe-layered double hydroxide (pNIPAAM MgFe-LDH) hydrogel was evaluated for intradiscal controlled delivery of the selective cyclooxygenase (COX) 2 inhibitor and anti-inflammatory drug celecoxib (CXB). Degradation, release behavior, and the ability of a CXB-loaded pNIPAAM MgFe-LDH hydrogel to suppress prostaglandin E2 (PGE2) levels in a controlled manner in the presence of a proinflammatory stimulus (TNF-α) were evaluated in vitro. Biocompatibility was evaluated histologically after subcutaneous injection in mice. Safety of intradiscal application of the loaded and unloaded hydrogels was studied in a canine model of spontaneous mild IVD degeneration by histological, biomolecular, and biochemical evaluation. After the hydrogel was shown to be biocompatible and safe, an in vivo dose-response study was performed in order to determine safety and efficacy of the pNIPAAM MgFe-LDH hydrogel for intradiscal controlled delivery of CXB. CXB release correlated to hydrogel degradation in vitro. Furthermore, controlled release from CXB-loaded hydrogels was demonstrated to suppress PGE2 levels in the presence of TNF-α. The hydrogel was shown to exhibit a good biocompatibility upon subcutaneous injection in mice. Upon intradiscal injection in a canine model, the hydrogel exhibited excellent biocompatibility based on histological evaluation of the treated IVDs. Gene expression and biochemical analyses supported the finding that no substantial negative effects of the hydrogel were observed. Safety of application was further confirmed by the absence of clinical symptoms, IVD herniation or progression of degeneration. Controlled release of CXB resulted in a nonsignificant maximal inhibition (approximately 35 %) of PGE2 levels in the mildly degenerated canine IVDs. In conclusion, this study showed biocompatibility and safe intradiscal application of an MgFe LDH-pNIPAAM hydrogel. Controlled release of CXB resulted in only limited inhibition of PGE2 in this model with mild IVD degeneration, and further studies should concentrate on application of controlled release from this type of hydrogel in animal models with more severe IVD degeneration.

Mechanical behavior of the human lumbar intervertebral disc with polymeric hydrogel nucleus implant: An experimental and finite element study

NASA Astrophysics Data System (ADS)

Joshi, Abhijeet Bhaskar

The origin of the lower back pain is often the degenerated lumbar intervertebral disc (IVD). We are proposing replacement of the degenerated nucleus by a PVA/PVP polymeric hydrogel implant. We hypothesize that a polymeric hydrogel nucleus implant can restore the normal biomechanics of the denucleated IVD by mimicking the natural load transfer phenomenon as in case of the intact IVD. Lumbar IVDs (n = 15) were harvested from human cadavers. In the first part, specimens were tested in four different conditions for compression: Intact, bone in plug, denucleated and Implanted. Hydrogel nucleus implants were chosen to have line-to-line fit in the created nuclear cavity. In the second part, nucleus implant material (modulus) and geometric (height and diameter) parameters were varied and specimens (n = 9) were tested. Nucleus implants with line-to-line fit significantly restored (88%) the compressive stiffness of the denucleated IVD. The synergistic effect between the implant and the intact annulus resulted in the nonlinear increase in implanted IVD stiffness, where Poisson effect of the hydrogel played major role. Nucleus implant parameters were observed to have a significant effect on the compressive stiffness. All implants with modulus in the tested range restored the compressive stiffness. The undersize implants resulted in incomplete restoration while oversize implants resulted in complete restoration compared to the BI condition. Finite element models (FEM) were developed to simulate the actual test conditions and validated against the experimental results for all conditions. The annulus (defined as hyperelastic, isotropic) mainly determined the nonlinear response of the IVD. Validated FEMs predicted 120--3000 kPa as a feasible range for nucleus implant modulus. FEMs also predicted that overdiameter implant would be more effective than overheight implant in terms of stiffness restoration. Underdiameter implants, initially allowed inward deformation of the annulus and hence were less effective compared to underheight implants. This research successfully proved the feasibility of PVA/PVP polymeric hydrogel as a replacement for degenerated nucleus. This approach may reduce the abnormal stresses on the annulus and thus, prevent/postpone the degeneration of the annulus. A validated FEM can be used as a design tool for optimization of hydrogel nucleus implants design and related feasibility studies.

45 CFR 309.80 - What safeguarding procedures must a Tribe or Tribal organization include in a Tribal IV-D plan?

Code of Federal Regulations, 2010 CFR

2010-10-01

... Relating to Public Welfare OFFICE OF CHILD SUPPORT ENFORCEMENT (CHILD SUPPORT ENFORCEMENT PROGRAM), ADMINISTRATION FOR CHILDREN AND FAMILIES, DEPARTMENT OF HEALTH AND HUMAN SERVICES TRIBAL CHILD SUPPORT... Tribal IV-D agency and that are designed to protect the privacy rights of the parties, including: (1...

45 CFR 309.145 - What costs are allowable for Tribal IV-D programs carried out under § 309.65(a) of this part?

Code of Federal Regulations, 2010 CFR

2010-10-01

.... (h) Automated data processing computer systems, including: (1) Planning efforts in the identification, evaluation, and selection of an automated data processing computer system solution meeting the program... existing automated data processing computer system to support Tribal IV-D program operations, and...

45 CFR 309.145 - What costs are allowable for Tribal IV-D programs carried out under § 309.65(a) of this part?

Code of Federal Regulations, 2013 CFR

2013-10-01

.... (h) Automated data processing computer systems, including: (1) Planning efforts in the identification, evaluation, and selection of an automated data processing computer system solution meeting the program... existing automated data processing computer system to support Tribal IV-D program operations, and...

45 CFR 309.145 - What costs are allowable for Tribal IV-D programs carried out under § 309.65(a) of this part?

Code of Federal Regulations, 2014 CFR

2014-10-01

.... (h) Automated data processing computer systems, including: (1) Planning efforts in the identification, evaluation, and selection of an automated data processing computer system solution meeting the program... existing automated data processing computer system to support Tribal IV-D program operations, and...

45 CFR 309.145 - What costs are allowable for Tribal IV-D programs carried out under § 309.65(a) of this part?

Code of Federal Regulations, 2012 CFR

2012-10-01

.... (h) Automated data processing computer systems, including: (1) Planning efforts in the identification, evaluation, and selection of an automated data processing computer system solution meeting the program... existing automated data processing computer system to support Tribal IV-D program operations, and...

45 CFR 309.145 - What costs are allowable for Tribal IV-D programs carried out under § 309.65(a) of this part?

Code of Federal Regulations, 2011 CFR

2011-10-01

.... (h) Automated data processing computer systems, including: (1) Planning efforts in the identification, evaluation, and selection of an automated data processing computer system solution meeting the program... existing automated data processing computer system to support Tribal IV-D program operations, and...

75 FR 36425 - Guidance for Industry and Food and Drug Administration Staff; In Vitro Diagnostic Studies...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-06-25

..., International, and Consumer Assistance, Center for Devices and Radiological Health (CDRH), Food and Drug...-addressed adhesive label to assist that office in processing your request, or fax your request to CDRH at... IVD studies. CDRH and CBER both have regulatory oversight of IVD devices. Information in this guidance...

Analgesic Activity of Some 1,2,4-Triazole Heterocycles Clubbed with Pyrazole, Tetrazole, Isoxazole and Pyrimidine

PubMed Central

Gajanan Khanage, Shantaram; Raju, Appala; Baban Mohite, Popat; Bhanudas Pandhare, Ramdas

2013-01-01

Purpose: In the present study in vivo analgesic activity of some previously synthesized 1,2,4-triazole derivatives containing pyrazole, tetrazole, isoxazole and pyrimidine ring have been evaluated. Methods: Acetic acid induced writhing method and Hot plate method has been described to study analgesic activity of some 1,2,4-triazole derivatives containing pyrazole, tetrazole, isoxazole and pyrimidine as a pharmacological active lead. Results: Thirty six different derivatives containing 1,2,4-triazole ring were subjected to study their in vivo analgesic activity. Chloro, nitro and methoxy, hydroxy and bromo substituted derivatives showed excellent analgesic activity and dimethylamino, furan and phenyl substituted derivatives showed moderate analgesic activity in both of the methods. Compounds IIIa, IIId, IIIf, IIIi, IIIj, IVa, IVb, IVd, IVf, IVh, IVj IV3a and IIj were found to be superior analgesic agents after screening by Acetic acid induced writhing method. Compounds IIIb, IIId, IIIf, IIIh, IIIj, IVa, IVb, IVd, IVf, IVh, IVi, IV3c, IV3e and IIj were showed analgesic potential after screening of Hot plate method. Conclusion: All tested compounds containing 1,2,4-triazole were found to be promising analgesic agents, for this activity pyrazole, tetrazole, isoxazole and pyrimidine leads might be supported. PMID:24312806

MALDI-TOF for the rapid detection of carbapenemase-producing Enterobacteriaceae: comparison of the commercialized MBT STAR®-Carba IVD Kit with two in-house MALDI-TOF techniques and the RAPIDEC® CARBA NP.

PubMed

Dortet, Laurent; Tandé, Didier; de Briel, Dominique; Bernabeu, Sandrine; Lasserre, Camille; Gregorowicz, Guillaume; Jousset, Agnès B; Naas, Thierry

2018-06-11

There is an urgent need for accurate and fast diagnostic tests to identify carbapenemase-producing bacteria. Here, we have evaluated three MALDI-TOF-based techniques to detect carbapenemase-producing Enterobacteriaceae (CPE) from cultured colonies. The performance of three MALDI-TOF-based techniques, including the commercialized MBT STAR®-Carba IVD Kit (Bruker Daltonics) and two in-house protocols performed on the Microflex LT Biotyper (Bruker Daltonics) and the VITEK® MS Plus (bioMérieux), were compared with those of the RAPIDEC® CARBA NP (bioMérieux). A collection of 175 isolates including 120 carbapenemase producers and 55 non-carbapenemase producers was tested. Samples were tested blind in the three participating centres. The repeatability of the MBT STAR®-Carba IVD Kit was also evaluated. The three MALDI-TOF techniques possess sensitivities ranging from 95% to 100% and specificities from 98.2% to 100% compared with 99.2% and 100%, respectively, for the RAPIDEC® CARBA NP. The MBT STAR®-Carba IVD Kit gave highly reproducible results and is the only technique able to provide a concomitant identification of the bacterial isolate. The three MALDI-TOF techniques possess a fast turnaround time (less than 1.5 h). Overall, MALDI-TOF is a reliable technique for the rapid detection of CPE from cultured colonies. MBT STAR®-Carba IVD Kit, the only commercially available assay, could easily be implemented in a clinical microbiology laboratory if it is already equipped with a Microflex LT Biotyper mass spectrometer.

An experimental and finite element poroelastic creep response analysis of an intervertebral hydrogel disc model in axial compression.

PubMed

Silva, P; Crozier, S; Veidt, M; Pearcy, M J

2005-07-01

A hydrogel intervertebral disc (IVD) model consisting of an inner nucleus core and an outer anulus ring was manufactured from 30 and 35% by weight Poly(vinyl alcohol) hydrogel (PVA-H) concentrations and subjected to axial compression in between saturated porous endplates at 200 N for 11 h, 30 min. Repeat experiments (n=4) on different samples (N=2) show good reproducibility of fluid loss and axial deformation. An axisymmetric nonlinear poroelastic finite element model with variable permeability was developed using commercial finite element software to compare axial deformation and predicted fluid loss with experimental data. The FE predictions indicate differential fluid loss similar to that of biological IVDs, with the nucleus losing more water than the anulus, and there is overall good agreement between experimental and finite element predicted fluid loss. The stress distribution pattern indicates important similarities with the biological IVD that includes stress transference from the nucleus to the anulus upon sustained loading and renders it suitable as a model that can be used in future studies to better understand the role of fluid and stress in biological IVDs.

The effects of dynamic loading on the intervertebral disc.

PubMed

Chan, Samantha C W; Ferguson, Stephen J; Gantenbein-Ritter, Benjamin

2011-11-01

Loading is important to maintain the balance of matrix turnover in the intervertebral disc (IVD). Daily cyclic diurnal assists in the transport of large soluble factors across the IVD and its surrounding circulation and applies direct and indirect stimulus to disc cells. Acute mechanical injury and accumulated overloading, however, could induce disc degeneration. Recently, there is more information available on how cyclic loading, especially axial compression and hydrostatic pressure, affects IVD cell biology. This review summarises recent studies on the response of the IVD and stem cells to applied cyclic compression and hydrostatic pressure. These studies investigate the possible role of loading in the initiation and progression of disc degeneration as well as quantifying a physiological loading condition for the study of disc degeneration biological therapy. Subsequently, a possible physiological/beneficial loading range is proposed. This physiological/beneficial loading could provide insight into how to design loading regimes in specific system for the testing of various biological therapies such as cell therapy, chemical therapy or tissue engineering constructs to achieve a better final outcome. In addition, the parameter space of 'physiological' loading may also be an important factor for the differentiation of stem cells towards most ideally 'discogenic' cells for tissue engineering purpose.

Intercellular signaling pathways active during intervertebral disc growth, differentiation, and aging.

PubMed

Dahia, Chitra Lekha; Mahoney, Eric J; Durrani, Atiq A; Wylie, Christopher

2009-03-01

Intervertebral discs at different postnatal ages were assessed for active intercellular signaling pathways. To generate a spatial and temporal map of the signaling pathways active in the postnatal intervertebral disc (IVD). The postnatal IVD is a complex structure, consisting of 3 histologically distinct components, the nucleus pulposus, fibrous anulus fibrosus, and endplate. These differentiate and grow during the first 9 weeks of age in the mouse. Identification of the major signaling pathways active during and after the growth and differentiation period will allow functional analysis using mouse genetics and identify targets for therapy for individual components of the disc. Antibodies specific for individual cell signaling pathways were used on cryostat sections of IVD at different postnatal ages to identify which components of the IVD were responding to major classes of intercellular signal, including sonic hedgehog, Wnt, TGFbeta, FGF, and BMPs. We present a spatial/temporal map of these signaling pathways during growth, differentiation, and aging of the disc. During growth and differentiation of the disc, its different components respond at different times to different intercellular signaling ligands. Most of these are dramatically downregulated at the end of disc growth.

Tissue engineering strategies applied in the regeneration of the human intervertebral disk.

PubMed

Silva-Correia, Joana; Correia, Sandra I; Oliveira, Joaquim M; Reis, Rui L

2013-12-01

Low back pain (LBP) is one of the most common painful conditions that lead to work absenteeism, medical visits, and hospitalization. The majority of cases showing signs of LBP are due to age-related degenerative changes in the intervertebral disk (IVD), which are, in fact, associated with multiple spine pathologies. Traditional and more conservative procedures/clinical approaches only treat the symptoms of disease and not the underlying pathology, thus limiting their long-term efficiency. In the last few years, research and development of new approaches aiming to substitute the nucleus pulposus and annulus fibrosus tissue and stimulate its regeneration has been conducted. Regeneration of the damaged IVD using tissue engineering strategies appears particularly promising in pre-clinical studies. Meanwhile, surgical techniques must be adapted to this new approach in order to be as minimally invasive as possible, reducing recovering time and side effects associated to traditional surgeries. In this review, the current knowledge on IVD, its associated pathologies and current surgical procedures are summarized. Furthermore, it also provides a succinct and up-to-date overview on regenerative medicine research, especially on the newest tissue engineering strategies for IVD regeneration. © 2013.

Energy metabolism of intervertebral disc under mechanical loading.

PubMed

Wang, Chong; Gonzales, Silvia; Levene, Howard; Gu, Weiyong; Huang, Chun-Yuh Charles

2013-11-01

Intervertebral disc (IVD) degeneration is closely associated with low back pain (LBP), which is a major health concern in the U.S. Cellular biosynthesis of extracellular matrix (ECM), which is important for maintaining tissue integrity and preventing tissue degeneration, is an energy demanding process. Due to impaired nutrient support in avascular IVD, adenosine triphosphate (ATP) supply could be a limiting factor for maintaining normal ECM synthesis. Therefore, the objective of this study was to investigate the energy metabolism in the annulus fibrosus (AF) and nucleus pulposus (NP) of porcine IVD under static and dynamic compressions. Under compression, pH decreased and the contents of lactate and ATP increased significantly in both AF and NP regions, suggesting that compression can promote ATP production via glycolysis and reduce pH by increasing lactate accumulation. A high level of extracellular ATP content was detected in the NP region and regulated by compressive loading. Since ATP can serve not only as an intra-cellular energy currency, but also as a regulator of a variety of cellular activities extracellularly through the purinergic signaling pathway, our findings suggest that compression-mediated ATP metabolism could be a novel mechanobiological pathway for regulating IVD metabolism. © 2013 Orthopaedic Research Society.

Comparison of Saramis 4.12 and IVD 3.0 Vitek MS Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry for Identification of Mycobacteria from Solid and Liquid Culture Media.

PubMed

Leyer, Caroline; Gregorowicz, Guillaume; Mougari, Faiza; Raskine, Laurent; Cambau, Emmanuelle; de Briel, Dominique

2017-07-01

During the last decade, many investigators have studied matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for identification of mycobacteria. Diverse and contradictory results indicated that optimal level for routine testing has not been reached yet. This work aimed to assess Vitek MS through two distinct versions, Saramis v4.12 RUO and the IVD v3.0, under conditions close to routine laboratory practice. Overall, 111 mycobacterial isolates were subjected to protein extraction and same spectra were matched against both databases. The IVD v3.0 database proved to be superior to Saramis v4.12 and its identification rates remarkably increased, from 67% to 94% for isolates grown on Middlebrook 7H10 solid medium and from 62% to 91% for isolates grown on mycobacterial growth indicator tube (MGIT) liquid medium. With this new version, IVD v3.0, MALDI-TOF MS might be integrated into routine clinical diagnostics, although molecular techniques remain mandatory in some cases. Copyright © 2017 American Society for Microbiology.

Mobile diagnostics: next-generation technologies for in vitro diagnostics.

PubMed

Shin, Joonchul; Chakravarty, Sudesna; Choi, Wooseok; Lee, Kyungyeon; Han, Dongsik; Hwang, Hyundoo; Choi, Jaekyu; Jung, Hyo-Il

2018-03-26

The emergence of a wide range of applications of smartphones along with advances in 'liquid biopsy' has significantly propelled medical research particularly in the field of in vitro diagnostics (IVD). Herein, we have presented a detailed analysis of IVD, its associated critical concerns and probable solutions. It also demonstrates the transition in terms of analytes from minimally invasive (blood) to non-invasive (urine, saliva and sweat) and depicts how the different features of a smartphone can be integrated for specific diagnostic purposes. This review basically highlights recent advances in the applications of smartphone-based biosensors in IVD taking into account the following factors: accuracy and portability; quantitative and qualitative analysis; and centralization and decentralization tests. Furthermore, the critical concerns and future direction of diagnostics based on smartphones are also discussed.

An HF coaxial bridge for measuring impedance ratios up to 1 MHz

NASA Astrophysics Data System (ADS)

Kucera, J.; Sedlacek, R.; Bohacek, J.

2012-08-01

A four-terminal pair coaxial ac bridge developed for calibrating both resistance and capacitance ratios and working in the frequency range from 100 kHz up to 1 MHz is described. A reference inductive voltage divider (IVD) makes it possible to calibrate ratios 1:1 and 10:1 with uncertainty of a few parts in 105. The IVD is calibrated by means of a series-parallel capacitance device (SPCD). Use of the same ac bridge with minimal changes for calibrating the SPCD, IVD and unknown impedances simplifies the whole calibration process. The bridge balance conditions are fulfilled with simple capacitance and resistance decades and by injecting voltage supplied from the auxiliary direct digital synthesizer. Bridge performance was checked on the basis of resistance ratio measurements and also capacitance ratio measurements.

Assessment of mechanical properties of isolated bovine intervertebral discs from multi-parametric magnetic resonance imaging.

PubMed

Recuerda, Maximilien; Périé, Delphine; Gilbert, Guillaume; Beaudoin, Gilles

2012-10-12

The treatment planning of spine pathologies requires information on the rigidity and permeability of the intervertebral discs (IVDs). Magnetic resonance imaging (MRI) offers great potential as a sensitive and non-invasive technique for describing the mechanical properties of IVDs. However, the literature reported small correlation coefficients between mechanical properties and MRI parameters. Our hypothesis is that the compressive modulus and the permeability of the IVD can be predicted by a linear combination of MRI parameters. Sixty IVDs were harvested from bovine tails, and randomly separated in four groups (in-situ, digested-6h, digested-18h, digested-24h). Multi-parametric MRI acquisitions were used to quantify the relaxation times T1 and T2, the magnetization transfer ratio MTR, the apparent diffusion coefficient ADC and the fractional anisotropy FA. Unconfined compression, confined compression and direct permeability measurements were performed to quantify the compressive moduli and the hydraulic permeabilities. Differences between groups were evaluated from a one way ANOVA. Multi linear regressions were performed between dependent mechanical properties and independent MRI parameters to verify our hypothesis. A principal component analysis was used to convert the set of possibly correlated variables into a set of linearly uncorrelated variables. Agglomerative Hierarchical Clustering was performed on the 3 principal components. Multilinear regressions showed that 45 to 80% of the Young's modulus E, the aggregate modulus in absence of deformation HA0, the radial permeability kr and the axial permeability in absence of deformation k0 can be explained by the MRI parameters within both the nucleus pulposus and the annulus pulposus. The principal component analysis reduced our variables to two principal components with a cumulative variability of 52-65%, which increased to 70-82% when considering the third principal component. The dendograms showed a natural division into four clusters for the nucleus pulposus and into three or four clusters for the annulus fibrosus. The compressive moduli and the permeabilities of isolated IVDs can be assessed mostly by MT and diffusion sequences. However, the relationships have to be improved with the inclusion of MRI parameters more sensitive to IVD degeneration. Before the use of this technique to quantify the mechanical properties of IVDs in vivo on patients suffering from various diseases, the relationships have to be defined for each degeneration state of the tissue that mimics the pathology. Our MRI protocol associated to principal component analysis and agglomerative hierarchical clustering are promising tools to classify the degenerated intervertebral discs and further find biomarkers and predictive factors of the evolution of the pathologies.

Market surveillance of in vitro diagnostics by the BfArM until end 2010: how safe are products for tumor diagnostics?

PubMed

Siekmeier, R; Wetzel, D

2013-01-01

The European Directive 98/79/EC on in vitro diagnostic medical devices (IVD) regulates the marketing and post market surveillance of IVD in the European Economic Area. In cases of incidents and field corrective actions the manufacturers have to inform the responsible Competent Authorities (CA). In Germany, the Federal Institute for Drugs and Medical Devices (BfArM) is the responsible CA for most IVD. In this study all notifications regarding IVD (tests, calibrators, kits, and control materials, except laboratory analyzers) for tumor diagnostics received by the BfArM between begin 1999 until end of 2010 were analyzed. All notifications were analyzed in respect to the type of product, the source of notification, the underlying product defects and the corrective actions performed. In the observation period, a total of 2,851 notifications were received of which 84 were related to IVD for tumor diagnostics included in this study (clinical chemistry - 63, histology - 6, molecular biology - 3, rapid tests - 12). Reports were received from manufacturers (68 cases), CA (8 cases), users (4 cases) and other sources (4 cases). In the group of IVD based on clinical chemistry means, the affected products were mostly those for the measurement of prostate specific antigen (PSA, 14 cases), human chorion gonadotropine (13 cases), carcino embryonic antigen (6 cases), CA 19-9 (6 cases), α(1)-fetoprotein (6 cases) and CA 125 (5 cases), whereas in test strips 9 out of the 12 notifications were related to PSA. Investigations of the manufacturers were able to identify the underlying root causes of product failures in 66 cases (78.6%). In 10 cases (11.9%) the root cause remained unclear and in 6 cases and 2 cases (7.1% and 2.4%) a product failure was excluded or a user error was the underlying cause. Most common root causes of product failures were material defects (24 cases) and manufacturing errors (15 cases). Corrective actions were performed by the manufacturers in 64 cases (76.2%) and were predominantly (multiple entries possible) customer information (62 cases, mandatory in case of a recall), recalls (45 cases), modifications in production or quality management (45 cases) and design changes (14 cases). The obtained results suggest that the system for post marketing surveillance of IVD is an established tool to enhance product safety and provides valuable information on product specific problems serving for improvement of product safety.

45 CFR 309.35 - What are the procedures for review of a Tribal IV-D program application, plan or plan amendment?

Code of Federal Regulations, 2014 CFR

2014-10-01

...) The effective date of a plan or plan amendment may not be earlier than the first day of the fiscal... program application, plan or plan amendment? 309.35 Section 309.35 Public Welfare Regulations Relating to... of a Tribal IV-D program application, plan or plan amendment? (a) The Secretary will promptly review...

45 CFR 310.30 - Under what circumstances would FFP be suspended or disallowed in the costs of Computerized Tribal...

Code of Federal Regulations, 2010 CFR

2010-10-01

... 45 Public Welfare 2 2010-10-01 2010-10-01 false Under what circumstances would FFP be suspended or... SYSTEMS AND OFFICE AUTOMATION Funding for Computerized Tribal IV-D Systems and Office Automation § 310.30 Under what circumstances would FFP be suspended or disallowed in the costs of Computerized Tribal IV-D...

45 CFR 309.130 - How will Tribal IV-D programs be funded and what forms are required?

Code of Federal Regulations, 2012 CFR

2012-10-01

...-D Program Funding § 309.130 How will Tribal IV-D programs be funded and what forms are required? (a... grants of less than $1 million per 12-month funding period will receive a single annual award. Tribes and Tribal organizations eligible for grants of $1 million or more per 12-month funding period will receive...

45 CFR 309.130 - How will Tribal IV-D programs be funded and what forms are required?

Code of Federal Regulations, 2014 CFR

2014-10-01

...-D Program Funding § 309.130 How will Tribal IV-D programs be funded and what forms are required? (a... grants of less than $1 million per 12-month funding period will receive a single annual award. Tribes and Tribal organizations eligible for grants of $1 million or more per 12-month funding period will receive...

45 CFR 309.130 - How will Tribal IV-D programs be funded and what forms are required?

Code of Federal Regulations, 2010 CFR

2010-10-01

...-D Program Funding § 309.130 How will Tribal IV-D programs be funded and what forms are required? (a... grants of less than $1 million per 12-month funding period will receive a single annual award. Tribes and Tribal organizations eligible for grants of $1 million or more per 12-month funding period will receive...

45 CFR 309.130 - How will Tribal IV-D programs be funded and what forms are required?

Code of Federal Regulations, 2011 CFR

2011-10-01

...-D Program Funding § 309.130 How will Tribal IV-D programs be funded and what forms are required? (a... grants of less than $1 million per 12-month funding period will receive a single annual award. Tribes and Tribal organizations eligible for grants of $1 million or more per 12-month funding period will receive...

45 CFR 309.130 - How will Tribal IV-D programs be funded and what forms are required?

Code of Federal Regulations, 2013 CFR

2013-10-01

...-D Program Funding § 309.130 How will Tribal IV-D programs be funded and what forms are required? (a... grants of less than $1 million per 12-month funding period will receive a single annual award. Tribes and Tribal organizations eligible for grants of $1 million or more per 12-month funding period will receive...

45 CFR 309.75 - What administrative and management procedures must a Tribe or Tribal organization include in a...

Code of Federal Regulations, 2010 CFR

2010-10-01

... application fee must be uniformly applied by the Tribe or Tribal organization and must be: (i) A flat amount... a Tribe or Tribal organization include in a Tribal IV-D plan? 309.75 Section 309.75 Public Welfare... procedures must a Tribe or Tribal organization include in a Tribal IV-D plan? A Tribe or Tribal organization...

Effect of Load Carriage on Lumbar Spine Kinematics

DTIC Science & Technology

2013-01-01

reference frame and lordosis was reduced during all tasks with load. Superior levels became more lordotic, whereas inferior levels became more... lordosis , and IVD compressibility have been measured in both young 17 and adult 13 populations using upright MRI. However, these data cannot be...the kinematic behavior of the overall lumbar spine and func- tional spinal units. We hypothesized that IVD compression and lumbar lordosis increased

Quantitative evaluation of lumbar intervertebral disc degeneration by axial T2* mapping.

PubMed

Huang, Leitao; Liu, Yuan; Ding, Yi; Wu, Xia; Zhang, Ning; Lai, Qi; Zeng, Xianjun; Wan, Zongmiao; Dai, Min; Zhang, Bin

2017-12-01

To quantitatively evaluate the clinical value and demonstrate the potential benefits of biochemical axial T2* mapping-based grading of early stages of degenerative disc disease (DDD) using 3.0-T magnetic resonance imaging (MRI) in a clinical setting.Fifty patients with low back pain and 20 healthy volunteers (control) underwent standard MRI protocols including axial T2* mapping. All the intervertebral discs (IVDs) were classified morphologically. Lumbar IVDs were graded using Pfirrmann score (I to IV). The T2* values of the anterior annulus fibrosus (AF), posterior AF, and nucleus pulposus (NP) of each lumbar IVD were measured. The differences between groups were analyzed regarding specific T2* pattern at different regions of interest.The T2* values of the NP and posterior AF in the patient group were significantly lower than those in the control group (P < .01). The T2* value of the anterior AF was not significantly different between the patients and the controls (P > .05). The mean T2*values of the lumbar IVD in the patient group were significantly lower, especially the posterior AF, followed by the NP, and finally, the anterior AF. In the anterior AF, comparison of grade I with grade III and grade I with grade IV showed statistically significant differences (P = .07 and P = .08, respectively). Similarly, in the NP, comparison of grade I with grade III, grade I with grade IV, grade II with grade III, and grade II with grade IV showed statistically significant differences (P < .001). In the posterior AF, comparison of grade II with grade IV showed a statistically significant difference (P = .032). T2 values decreased linearly with increasing degeneration based on the Pfirrmann scoring system (ρ < -0.5, P < .001).Changes in the T2* value can signify early degenerative IVD diseases. Hence, T2* mapping can be used as a diagnostic tool for quantitative assessment of IVD degeneration. Copyright © 2017 The Authors. Published by Wolters Kluwer Health, Inc. All rights reserved.

Quantitative evaluation of lumbar intervertebral disc degeneration by axial T2∗ mapping

PubMed Central

Huang, Leitao; Liu, Yuan; Ding, Yi; Wu, Xia; Zhang, Ning; Lai, Qi; Zeng, Xianjun; Wan, Zongmiao; Dai, Min; Zhang, Bin

2017-01-01

Abstract To quantitatively evaluate the clinical value and demonstrate the potential benefits of biochemical axial T2∗ mapping-based grading of early stages of degenerative disc disease (DDD) using 3.0-T magnetic resonance imaging (MRI) in a clinical setting. Fifty patients with low back pain and 20 healthy volunteers (control) underwent standard MRI protocols including axial T2∗ mapping. All the intervertebral discs (IVDs) were classified morphologically. Lumbar IVDs were graded using Pfirrmann score (I to IV). The T2∗ values of the anterior annulus fibrosus (AF), posterior AF, and nucleus pulposus (NP) of each lumbar IVD were measured. The differences between groups were analyzed regarding specific T2∗ pattern at different regions of interest. The T2∗ values of the NP and posterior AF in the patient group were significantly lower than those in the control group (P < .01). The T2∗ value of the anterior AF was not significantly different between the patients and the controls (P > .05). The mean T2∗values of the lumbar IVD in the patient group were significantly lower, especially the posterior AF, followed by the NP, and finally, the anterior AF. In the anterior AF, comparison of grade I with grade III and grade I with grade IV showed statistically significant differences (P = .07 and P = .08, respectively). Similarly, in the NP, comparison of grade I with grade III, grade I with grade IV, grade II with grade III, and grade II with grade IV showed statistically significant differences (P < .001). In the posterior AF, comparison of grade II with grade IV showed a statistically significant difference (P = .032). T2∗ values decreased linearly with increasing degeneration based on the Pfirrmann scoring system (ρ < −0.5, P < .001). Changes in the T2∗ value can signify early degenerative IVD diseases. Hence, T2∗ mapping can be used as a diagnostic tool for quantitative assessment of IVD degeneration. PMID:29390547

Bone Marrow Stem Cells in Response to Intervertebral Disc-Like Matrix Acidity and Oxygen Concentration: Implications for Cell-based Regenerative Therapy.

PubMed

Naqvi, Syeda M; Buckley, Conor T

2016-05-01

In vitro culture of porcine bone marrow stem cells (BMSCs) in varying pH microenvironments in a three-dimensional hydrogel system. To characterize the response of BMSCs to varying pH environments (blood [pH 7.4], healthy intervertebral disc (IVD) (pH 7.1), mildly degenerated IVD (pH 6.8), and severely degenerated IVD (pH 6.5) in three-dimensional culture under normoxic (20%) and hypoxic (5%) conditions. The IVD is an avascular organ relying on diffusion of essential nutrients through the cartilaginous endplates (CEPs) thereby creating a challenging microenvironment. Within a degenerated IVD, oxygen and glucose concentrations decrease further (<5% oxygen, <5 mmol/L glucose) and matrix acidity (

Whole-body vibration of mice induces progressive degeneration of intervertebral discs associated with increased expression of Il-1β and multiple matrix degrading enzymes.

PubMed

McCann, Matthew R; Veras, Matthew A; Yeung, Cynthia; Lalli, Gurkeet; Patel, Priya; Leitch, Kristyn M; Holdsworth, David W; Dixon, S Jeffrey; Séguin, Cheryle A

2017-05-01

Whole-body vibration (WBV) is a popular fitness trend based on claims of increased muscle mass, weight loss and reduced joint pain. Following its original implementation as a treatment to increase bone mass in patients with osteoporosis, WBV has been incorporated into clinical practice for musculoskeletal disorders, including back pain. However, our recent studies revealed damaging effects of WBV on joint health in a murine model. In this report, we examined potential mechanisms underlying disc degeneration following exposure of mice to WBV. Ten-week-old male mice were exposed to WBV (45 Hz, 0.3 g peak acceleration, 30 min/day, 5 days/week) for 4 weeks, 8 weeks, or 4 weeks WBV followed by 4 weeks recovery. Micro-computed tomography (micro-CT), histological, and gene expression analyses were used to assess the effects of WBV on spinal tissues. Exposure of mice to 4 or 8 weeks of WBV did not alter total body composition or induce significant changes in vertebral bone density. On the other hand, WBV-induced intervertebral disc (IVD) degeneration, associated with decreased disc height and degenerative changes in the annulus fibrosus (AF) that did not recover within 4 weeks after cessation of WBV. Gene expression analysis showed that WBV for 8 weeks induced expression of Mmp3, Mmp13, and Adamts5 in IVD tissues, changes preceded by increased expression of Il-1β. Progressive IVD degeneration induced by WBV was associated with increased expression of Il-1β within the IVD that preceded Mmp and Adamts gene induction. Moreover, WBV-induced IVD degeneration is not reversed following cessation of vibration. Copyright © 2017 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.

Effects of radial compression on a novel simulated intervertebral disc-like assembly using bone marrow-derived mesenchymal stem cell cell-sheets for annulus fibrosus regeneration.

PubMed

See, Eugene Yong-Shun; Toh, Siew Lok; Goh, James Cho-Hong

2011-10-01

The aim of this study was to develop a tissue engineering approach in regenerating the annulus fibrosus (AF) as part of an overall strategy to produce a tissue-engineered intervertebral disc (IVD) replacement. To determine whether a rehabilitative simulation regime on bone marrow–derived mesenchymal stem cell cell-sheet is able to aid the regeneration of the AF. No previous study has used bone marrow–derived mesenchymal stem cell cell-sheets simulated by a rehabilitative regime to regenerate the AF. The approach was to use bone marrow–derived stem cells to form cell-sheets and incorporating them onto silk scaffolds to simulate the native lamellae of the AF. The in vitro experimental model used to study the efficacy of such a system was made up of the tissue engineering AF construct wrapped around a silicone disc to form a simulated IVD-like assembly. The assembly was cultured within a custom-designed bioreactor that provided a compressive mechanical stimulation onto the silicone disc. The silicone nucleus pulposus would bulge radially and compress the simulated AF to mimic the physiological conditions. The simulated IVD-like assembly was compressed using a rehabilitative regime that lasted for 4 weeks at 0.25 Hz, for 15 minutes each day. With the rehabilitative regime, the cell-sheets remained viable but showed a decrease in cell numbers and viability. Gene expression analysis showed significant upregulation of IVD-related genes and there was an increased ratio of collagen type II to collagen type I found within the extracellular matrix. The results suggested that a rehabilitative regime caused extensive remodeling to take place within the simulated IVD-like assembly, producing extracellular matrix similar to that found in the inner AF.

Accelerated cellular senescence in degenerate intervertebral discs: a possible role in the pathogenesis of intervertebral disc degeneration

PubMed Central

Le Maitre, Christine Lyn; Freemont, Anthony John; Hoyland, Judith Alison

2007-01-01

Current evidence implicates intervertebral disc degeneration as a major cause of low back pain, although its pathogenesis is poorly understood. Numerous characteristic features of disc degeneration mimic those seen during ageing but appear to occur at an accelerated rate. We hypothesised that this is due to accelerated cellular senescence, which causes fundamental changes in the ability of disc cells to maintain the intervertebral disc (IVD) matrix, thus leading to IVD degeneration. Cells isolated from non-degenerate and degenerate human tissue were assessed for mean telomere length, senescence-associated β-galactosidase (SA-β-gal), and replicative potential. Expression of P16INK4A (increased in cellular senescence) was also investigated in IVD tissue by means of immunohistochemistry. RNA from tissue and cultured cells was used for real-time polymerase chain reaction analysis for matrix metalloproteinase-13, ADAMTS 5 (a disintegrin and metalloprotease with thrombospondin motifs 5), and P16INK4A. Mean telomere length decreased with age in cells from non-degenerate tissue and also decreased with progressive stages of degeneration. In non-degenerate discs, there was an age-related increase in cellular expression of P16INK4A. Cells from degenerate discs (even from young patients) exhibited increased expression of P16INK4A, increased SA-β-gal staining, and a decrease in replicative potential. Importantly, there was a positive correlation between P16INK4A and matrix-degrading enzyme gene expression. Our findings indicate that disc cell senescence occurs in vivo and is accelerated in IVD degeneration. Furthermore, the senescent phenotype is associated with increased catabolism, implicating cellular senescence in the pathogenesis of IVD degeneration. PMID:17498290

Fibrin-Genipin Adhesive Hydrogel for Annulus Fibrosus Repair: Performance Evaluation with Large Animal Organ Culture, In Situ Biomechanics, and In Vivo Degradation Tests

PubMed Central

Likhitpanichkul, M.; Dreischarf, M.; Illien-Junger, S.; Walter, B. A.; Nukaga, T.; Long, R. G; Sakai, D.; Hecht, A. C.; Iatridis, J. C.

2015-01-01

Annulus fibrosus (AF) defects from annular tears, herniation, and discectomy procedures are associated with painful conditions and accelerated intervertebral disc (IVD) degeneration. Currently, no effective treatments exist to repair AF damage, restore IVD biomechanics and promote tissue regeneration. An injectable fibrin-genipin adhesive hydrogel (Fib-Gen) was evaluated for its performance repairing large AF defects in a bovine caudal IVD model using ex vivo organ culture and biomechanical testing of motion segments, and for its in vivo longevity and biocompatibility in a rat model by subcutaneous implantation. Fib-Gen sealed AF defects, prevented IVD height loss, and remained well-integrated with native AF tissue following approximately 14,000 cycles of compression in 6-day organ culture experiments. Fib-Gen repair also retained high viability of native AF cells near the repair site, reduced nitric oxide released to the media, and showed evidence of AF cell migration into the gel. Biomechanically, Fib-Gen fully restored compressive stiffness to intact levels validating organ culture findings. However, only partial restoration of tensile and torsional stiffness was obtained, suggesting opportunities to enhance this formulation. Subcutaneous implantation results, when compared with the literature, suggested Fib-Gen exhibited similar biocompatibility behaviour to fibrin alone but degraded much more slowly. We conclude that injectable Fib-Gen successfully sealed large AF defects, promoted functional restoration with improved motion segment biomechanics, and served as a biocompatible adhesive biomaterial that had greatly enhanced in vivo longevity compared to fibrin. Fib-Gen offers promise for AF repairs that may prevent painful conditions and accelerated degeneration of the IVD, and warrants further material development and evaluation. PMID:25036053

Fibrin-genipin adhesive hydrogel for annulus fibrosus repair: performance evaluation with large animal organ culture, in situ biomechanics, and in vivo degradation tests.

PubMed

Likhitpanichkul, M; Dreischarf, M; Illien-Junger, S; Walter, B A; Nukaga, T; Long, R G; Sakai, D; Hecht, A C; Iatridis, J C

2014-07-18

Annulus fibrosus (AF) defects from annular tears, herniation, and discectomy procedures are associated with painful conditions and accelerated intervertebral disc (IVD) degeneration. Currently, no effective treatments exist to repair AF damage, restore IVD biomechanics and promote tissue regeneration. An injectable fibrin-genipin adhesive hydrogel (Fib-Gen) was evaluated for its performance repairing large AF defects in a bovine caudal IVD model using ex vivo organ culture and biomechanical testing of motion segments, and for its in vivo longevity and biocompatibility in a rat model by subcutaneous implantation. Fib-Gen sealed AF defects, prevented IVD height loss, and remained well-integrated with native AF tissue following approximately 14,000 cycles of compression in 6-day organ culture experiments. Fib-Gen repair also retained high viability of native AF cells near the repair site, reduced nitric oxide released to the media, and showed evidence of AF cell migration into the gel. Biomechanically, Fib-Gen fully restored compressive stiffness to intact levels validating organ culture findings. However, only partial restoration of tensile and torsional stiffness was obtained, suggesting opportunities to enhance this formulation. Subcutaneous implantation results, when compared with the literature, suggested Fib-Gen exhibited similar biocompatibility behaviour to fibrin alone but degraded much more slowly. We conclude that injectable Fib-Gen successfully sealed large AF defects, promoted functional restoration with improved motion segment biomechanics, and served as a biocompatible adhesive biomaterial that had greatly enhanced in vivo longevity compared to fibrin. Fib-Gen offers promise for AF repairs that may prevent painful conditions and accelerated degeneration of the IVD, and warrants further material development and evaluation.

Low level light therapy modulates inflammatory mediators secreted by human annulus fibrosus cells during intervertebral disc degeneration in vitro.

PubMed

Hwang, Min Ho; Shin, Jae Hee; Kim, Kyoung Soo; Yoo, Chang Min; Jo, Ga Eun; Kim, Joo Han; Choi, Hyuk

2015-01-01

Intervertebral disc degeneration (IVD) is one of the important causes of low back pain and is associated with inflammation induced by interaction between macrophages and the human annulus fibrosus (AF) cells. Low-level light therapy (LLLT) has been widely known to regulate inflammatory reaction. However, the effect of LLLT on macrophage-mediated inflammation in the AF cells has not been studied till date. The aim of this study is to mimic the inflammatory microenvironment and to investigate the anti-inflammatory effect of LLLT at a range of wavelengths (405, 532 and 650 nm) on the AF treated with macrophage-like THP-1 cells conditioned medium (MCM) containing proinflammatory cytokines and chemokines (interleukin-1beta, tumor necrosis factor-alpha, interleukin-6 and 8). We observed that AF cells exposed to MCM secrete significantly higher concentrations of IL-6, IL-8, IL-1β and TNF-α. LLLT markedly inhibited secretion of IL-6 at 405 nm in a time-dependent manner. Level of IL-8 was significantly decreased at all wavelengths in a time-dependent manner. We showed that MCM can induce the inflammatory microenvironment in AF cells and LLLT selectively suppressed IL-6 and 8 levels. The results indicate that LLLT is a potential method of IVD treatment and provide insights into further investigation of its anti-inflammation effect on IVD. © 2015 The American Society of Photobiology.

A minimally invasive in-fiber Bragg grating sensor for intervertebral disc pressure measurements

NASA Astrophysics Data System (ADS)

Dennison, Christopher R.; Wild, Peter M.; Wilson, David R.; Cripton, Peter A.

2008-08-01

We present an in-fiber Bragg grating (FBG) based intervertebral disc (IVD) pressure sensor that has pressure sensitivity seven times greater than that of a bare fiber, and a major diameter and sensing area of only 400 µm and 0.03 mm2, respectively. This is the only optical, the smallest and the most mechanically compliant disc pressure sensor reported in the literature. This is also an improvement over other FBG pressure sensors that achieve increased sensitivity through mechanical amplification schemes, usually resulting in major diameters and sensing lengths of many millimeters. Sensor sensitivity is predicted using numerical models, and the predicted sensitivity is verified through experimental calibrations. The sensor is validated by conducting IVD pressure measurements in porcine discs and comparing the FBG measurements to those obtained using the current standard sensor for IVD pressure. The predicted sensitivity of the FBG sensor matched with that measured experimentally. IVD pressure measurements showed excellent repeatability and agreement with those obtained from the standard sensor. Unlike the current larger sensors, the FBG sensor could be used in discs with small disc height (i.e. cervical or degenerated discs). Therefore, there is potential to conduct new measurements that could lead to new understanding of the biomechanics.

Hyperosmotically induced volume change and calcium signaling in intervertebral disk cells: the role of the actin cytoskeleton.

PubMed

Pritchard, Scott; Erickson, Geoffrey R; Guilak, Farshid

2002-11-01

Loading of the spine alters the osmotic environment in the intervertebral disk (IVD) as interstitial water is expressed from the tissue. Cells from the three zones of the IVD, the anulus fibrosus (AF), transition zone (TZ), and nucleus pulposus (NP), respond to osmotic stress with altered biosynthesis through a pathway that may involve calcium (Ca(2+)) as a second messenger. We examined the hypothesis that IVD cells respond to hyperosmotic stress by increasing the concentration of intracellular calcium ([Ca(2+)](i)) through a mechanism involving F-actin. In response to hyperosmotic stress, control cells from all zones decreased in volume and cells from the AF and TZ exhibited [Ca(2+)](i) transients, while cells from the NP did not. Extracellular Ca(2+) was necessary to initiate [Ca(2+)](i) transients. Stabilization of F-actin with phalloidin prevented the Ca(2+) response in AF and TZ cells and decreased the rate of volume change in cells from all zones, coupled with an increase in the elastic moduli and apparent viscosity. Conversely, actin breakdown with cytochalasin D facilitated Ca(2+) signaling while decreasing the elastic moduli and apparent viscosity for NP cells. These results suggest that hyperosmotic stress induces volume change in IVD cells and may initiate [Ca(2+)](i) transients through an actin-dependent mechanism.

Age-related carbonylation of fibrocartilage structural proteins drives tissue degenerative modification.

PubMed

Scharf, Brian; Clement, Cristina C; Yodmuang, Supansa; Urbanska, Aleksandra M; Suadicani, Sylvia O; Aphkhazava, David; Thi, Mia M; Perino, Giorgio; Hardin, John A; Cobelli, Neil; Vunjak-Novakovic, Gordana; Santambrogio, Laura

2013-07-25

Aging-related oxidative stress has been linked to degenerative modifications in different organs and tissues. Using redox proteomic analysis and illustrative tandem mass spectrometry mapping, we demonstrate oxidative posttranslational modifications in structural proteins of intervertebral discs (IVDs) isolated from aging mice. Increased protein carbonylation was associated with protein fragmentation and aggregation. Complementing these findings, a significant loss of elasticity and increased stiffness was measured in fibrocartilage from aging mice. Studies using circular dichroism and intrinsic tryptophan fluorescence revealed a significant loss of secondary and tertiary structures of purified collagens following oxidation. Collagen unfolding and oxidation promoted both nonenzymatic and enzymatic degradation. Importantly, induction of oxidative modification in healthy fibrocartilage recapitulated the biochemical and biophysical modifications observed in the aging IVD. Together, these results suggest that protein carbonylation, glycation, and lipoxidation could be early events in promoting IVD degenerative changes. Copyright © 2013 Elsevier Ltd. All rights reserved.

Safety of laboratory analyzers for infection testing - results of the market surveillance by the BfArM until end 2007

PubMed Central

2009-01-01

The European Directive 98/79/EC on in vitro diagnostic medical devices (IVD) stipulates the marketing and post market surveillance of IVD in the European Economic Area. In cases of issues and field corrective actions, the manufacturers have to inform the responsible Competent Authorities (CA). In Germany, the Federal Institute for Drugs and Medical Devices (BfArM) is the responsible CA for most IVD, with a small subset of IVD for immune hematological and infectiological testing as well as tissue typing as specified in Annex II of the Directive, being within the responsibility of the Paul-Ehrlich-Institute (PEI). In this study, all issues regarding laboratory analyzers for infection testing and their consumables, but not reagents, kits and general culture media, reported to the BfArM between begin 1999 and end of 2007 were analyzed in respect to the sources of report, the underlying product failure and the performed corrective actions. Within the observation period a total of 1471 reports for IVD were received of which 73 related to the IVD for infection testing were included in our study. Reports were predominantly received from manufacturers (56) and competent authorities (15). Affected products were most frequently those for immunological analysis (42) whereas those based on culturing techniques (17) and molecular biological techniques (14) played only minor roles. In all these groups, laboratory analyzers (55) were more frequently affected than their consumables (18). Investigations of the manufacturers were able to identify the underlying root causes of product failures in 62 cases (84.9%). In 2 cases (2.7%) the root cause remained unclear and in 9 cases (12.3%) a product failure was excluded or a user error was the underlying cause. Product failures in laboratory analyzers were most frequently caused by software errors (31) and constructional faults (8) whereas the predominant cause of product failure in consumables were errors in production and quality control (8). Manufacturers issued corrective measures in 66 cases (90.4%) from which 49 and 17 were related to laboratory analyzers and their consumables, respectively. Based on the underlying root causes of product failures these were predominantly customer information (48), recalls (40), software-updates (30) and design changes (9) in the product group of laboratory analyzers as well as customer information (16), recalls (12) and modifications of production and quality management (11) in the group of consumables. The results and experiences obtained since 1999 suggest that the system for post marketing surveillance of IVD is an established tool to ensure product safety, even though the current system can be further enhanced. PMID:20156760

Assessment of Intervertebral Disc Degeneration Based on Quantitative MRI Analysis: an in vivo study

PubMed Central

Grunert, Peter; Hudson, Katherine D.; Macielak, Michael R.; Aronowitz, Eric; Borde, Brandon H.; Alimi, Marjan; Njoku, Innocent; Ballon, Douglas; Tsiouris, Apostolos John; Bonassar, Lawrence J.; Härtl, Roger

2015-01-01

Study design Animal experimental study Objective To evaluate a novel quantitative imaging technique for assessing disc degeneration. Summary of Background Data T2-relaxation time (T2-RT) measurements have been used to quantitatively assess disc degeneration. T2 values correlate with the water content of inter vertebral disc tissue and thereby allow for the indirect measurement of nucleus pulposus (NP) hydration. Methods We developed an algorithm to subtract out MRI voxels not representing NP tissue based on T2-RT values. Filtered NP voxels were used to measure nuclear size by their amount and nuclear hydration by their mean T2-RT. This technique was applied to 24 rat-tail intervertebral discs’ (IVDs), which had been punctured with an 18-gauge needle according to different techniques to induce varying degrees of degeneration. NP voxel count and average T2-RT were used as parameters to assess the degeneration process at 1 and 3 months post puncture. NP voxel counts were evaluated against X-ray disc height measurements and qualitative MRI studies based on the Pfirrmann grading system. Tails were collected for histology to correlate NP voxel counts to histological disc degeneration grades and to NP cross-sectional area measurements. Results NP voxel count measurements showed strong correlations to qualitative MRI analyses (R2=0.79, p<0.0001), histological degeneration grades (R2=0.902, p<0.0001) and histological NP cross-sectional area measurements (R2=0.887, p<0.0001). In contrast to NP voxel counts, the mean T2-RT for each punctured group remained constant between months 1 and 3. The mean T2-RTs for the punctured groups did not show a statistically significant difference from those of healthy IVDs (63.55ms ±5.88ms month 1 and 62.61ms ±5.02ms) at either time point. Conclusion The NP voxel count proved to be a valid parameter to quantitatively assess disc degeneration in a needle puncture model. The mean NP T2-RT does not change significantly in needle-puncture induced degenerated IVDs. IVDs can be segmented into different tissue components according to their innate T2-RT. PMID:24384655

A pilot study of nimotuzumab combined with cisplatin and 5-FU in patients with advanced esophageal squamous cell carcinoma

PubMed Central

Ling, Yang; Chen, Jia; Tao, Min; Chu, Xiaoyuan; Zhang, Xizhi

2012-01-01

Objective To observe the short-term effect and adverse reaction of Nimotuzumab in combination with chemotherapy on advanced esophageal squamous cell carcinoma (ESCC). Method 19 patients were treated with the following protocol: Nimotuzumab 400mg/time/week in the 1st week, 200mg/time/week from the 2nd to 8th week, intravenous drip (IVD); Cisplatin 80 mg/m2, IVD, 4 weeks a cycle and repeated again; 5-FU 750 mg/m2, continuous 24-hours pump-in × 5 days, 4 weeks a cycle and repeated again. Result 16 of all 19 patients can be evaluated. After treatment, RP is 42.1% (95% CI, 19.9-64.3%) and DCR is 68.4%; the main side effects include arrest of bone marrow, gastrointestinal reactions, asthenia, etc. Conclusion Nimotuzumab in combination with cisplatin/5-FU regimens in patients with advanced ESCC is safe and effective, which deserves a further expanded sample research. PMID:22295168

Rheological and mechanical properties of acellular and cell-laden methacrylated gellan gum hydrogels.

PubMed

Silva-Correia, Joana; Gloria, Antonio; Oliveira, Mariana B; Mano, João F; Oliveira, Joaquim M; Ambrosio, Luigi; Reis, Rui L

2013-12-01

Tissue engineered hydrogels hold great potential as nucleus pulposus substitutes (NP), as they promote intervertebral disc (IVD) regeneration and re-establish its original function. But, the key to their success in future clinical applications greatly depends on its ability to replicate the native 3D micro-environment and circumvent their limitation in terms of mechanical performance. In the present study, we investigated the rheological/mechanical properties of both ionic- (iGG-MA) and photo-crosslinked methacrylated gellan gum (phGG-MA) hydrogels. Steady shear analysis, injectability and confined compression stress-relaxation tests were carried out. The injectability of the reactive solutions employed for the preparation of iGG-MA and phGG-MA hydrogels was first studied, then the zero-strain compressive modulus and permeability of the acellular hydrogels were evaluated. In addition, human intervertebral disc (hIVD) cells encapsulated in both iGG-MA and phGG-MA hydrogels were cultured in vitro, and its mechanical properties also investigated under dynamic mechanical analysis at 37°C and pH 7.4. After 21 days of culturing, hIVD cells were alive (Calcein AM) and the E' of ionic-crosslinked hydrogels and photo-crosslinked was higher than that observed for acellular hydrogels. Our study suggests that methacrylated gellan gum hydrogels present promising mechanical and biological performance as hIVD cells were producing extracellular matrix. Copyright © 2013 Wiley Periodicals, Inc., a Wiley Company.

Comparative and quantitative proteomic analysis of normal and degenerated human annulus fibrosus cells.

PubMed

Ye, Dongping; Liang, Weiguo; Dai, Libing; Zhou, Longqiang; Yao, Yicun; Zhong, Xin; Chen, Honghui; Xu, Jiake

2015-05-01

Degeneration of the intervertebral disc (IVD) is a major chronic medical condition associated with back pain. To better understand the pathogenesis of IVD degeneration, we performed comparative and quantitative proteomic analyses of normal and degenerated human annulus fibrosus (AF) cells and identified proteins that are differentially expressed between them. Annulus fibrosus cells were isolated and cultured from patients with lumbar disc herniation (the experimental group, degenerated AF cells) and scoliosis patients who underwent orthopaedic surgery (the control group, normal AF cells). Comparative proteomic analyses of normal and degenerated cultured AF cells were carried out using 2-D electrophoresis, mass spectrometric analyses, and database searching. Quantitative analyses of silver-stained 2-D electrophoresis gels of normal and degenerated cultured AF cells identified 10 protein spots that showed the most altered differential expression levels between the two groups. Among these, three proteins were decreased, including heat shock cognate 71-kDa protein, glucose-6-phosphate 1-dehydrogenase, and protocadherin-23, whereas seven proteins were increased, including guanine nucleotide-binding protein G(i) subunit α-2, superoxide dismutase, transmembrane protein 51, adenosine receptor A3, 26S protease regulatory subunit 8, lipid phosphate phosphatase-related protein, and fatty acyl-crotonic acid reductase 1. These differentially expressed proteins might be involved in the pathophysiological process of IVD degeneration and have potential values as biomarkers of the degeneration of IVD. © 2015 Wiley Publishing Asia Pty Ltd.

Anisotropic Multishell Analytical Modeling of an Intervertebral Disk Subjected to Axial Compression.

PubMed

Demers, Sébastien; Nadeau, Sylvie; Bouzid, Abdel-Hakim

2016-04-01

Studies on intervertebral disk (IVD) response to various loads and postures are essential to understand disk's mechanical functions and to suggest preventive and corrective actions in the workplace. The experimental and finite-element (FE) approaches are well-suited for these studies, but validating their findings is difficult, partly due to the lack of alternative methods. Analytical modeling could allow methodological triangulation and help validation of FE models. This paper presents an analytical method based on thin-shell, beam-on-elastic-foundation and composite materials theories to evaluate the stresses in the anulus fibrosus (AF) of an axisymmetric disk composed of multiple thin lamellae. Large deformations of the soft tissues are accounted for using an iterative method and the anisotropic material properties are derived from a published biaxial experiment. The results are compared to those obtained by FE modeling. The results demonstrate the capability of the analytical model to evaluate the stresses at any location of the simplified AF. It also demonstrates that anisotropy reduces stresses in the lamellae. This novel model is a preliminary step in developing valuable analytical models of IVDs, and represents a distinctive groundwork that is able to sustain future refinements. This paper suggests important features that may be included to improve model realism.

26 CFR 1.734-1 - Optional adjustment to basis of undistributed partnership property.

Code of Federal Regulations, 2010 CFR

2010-04-01

... partner. See § 1.460-4(k)(2)(iv)(D) for a rule determining the partnership's basis in a long-term contract accounted for under a long-term contract method of accounting. The provisions of this paragraph (b)(1) are... which the adjustment has been allocated. (e) Recovery of adjustments to basis of partnership property—(1...

Quality Control of Next-generation Sequencing-based In vitro Diagnostic Test for Onco-relevant Mutations Using Multiplex Reference Materials in Plasma.

PubMed

Liu, Donglai; Zhou, Haiwei; Shi, Dawei; Shen, Shu; Tian, Yabin; Wang, Lin; Lou, Jiatao; Cong, Rong; Lu, Juan; Zhang, Henghui; Zhao, Meiru; Zhu, Shida; Cao, Zhisheng; Jin, Ruilin; Wang, Yin; Zhang, Xiaoni; Yang, Guohua; Wang, Youchun; Zhang, Chuntao

2018-01-01

Background: Widespread clinical implementation of next-generation sequencing (NGS)-based cancer in vitro diagnostic tests (IVDs) highlighted the urgency to establish reference materials which could provide full control of the process from nucleic acid extraction to test report generation. The formalin-fixed, paraffin-embedded (FFPE) tissue and blood plasma containing circulating tumor deoxyribonucleic acid (ctDNA) were mostly used for clinically detecting onco-relevant mutations. Methods: We respectively developed multiplex FFPE and plasma reference materials covering three clinically onco-relevant mutations within the epidermal growth factor receptor ( EGFR ) gene at serial allelic frequencies. All reference materials were quantified and validated via droplet digital polymerase chain reaction (ddPCR), and then were distributed to eight domestic manufacturers for the collaborative evaluation of the performance of several domestic NGS-based cancer IVDs covering four major NGS platforms (NextSeq, HiSeq, Ion Proton and BGISEQ). Results: All expected mutations except one at extremely low allelic frequencies were detected, despite some differences in coefficient of variation (CV) which increased with the decrease of allelic frequency (CVs ranging from 18% to 106%). It was worth noting that the CV value seemed to correlate with a particular mutation as well. The repeatability of determination of different mutations was L858R>T790M>19del. Conclusions: The results indicated our reference materials would be pivotal for quality control of NGS-based cancer IVDs and would guide the further development of reference materials covering more onco-relevant mutations.

Expression and functional roles of estrogen receptor GPR30 in human intervertebral disc.

PubMed

Wei, Aiqun; Shen, Bojiang; Williams, Lisa A; Bhargav, Divya; Yan, Feng; Chong, Beng H; Diwan, Ashish D

2016-04-01

Estrogen withdrawal, a characteristic of female aging, is associated with age-related intervertebral disc (IVD) degeneration. The function of estrogen is mediated by two classic nuclear receptors, estrogen receptor (ER)-α and -β, and a membrane bound G-protein-coupled receptor 30 (GPR30). To date, the expression and function of GPR30 in human spine is poorly understood. This study aimed to evaluate GPR30 expression in IVD, and its role in estrogen-related regulation of proliferation and apoptosis of disc nucleus pulposus (NP) cells. GPR30 expression was examined in 30 human adult NP and 9 fetal IVD. Results showed that GPR30 was expressed in NP cells at both mRNA and protein levels. In human fetal IVD, GPR30 protein was expressed in the NP at 12-14 weeks gestation, but was undetectable at 8-11 weeks. The effect of 17β-estradiol (E2) on GPR30-mediated proliferation and interleukin-1β (IL-1β)-induced apoptosis of NP cells was investigated. Cultured NP cells were treated with or without E2, GPR30 antagonist G36, and ER antagonist ICI 182,780. NP cell viability was tested by MTS assay. Apoptosis was determined by flow cytometry using fluorescence labeled annexin-V, TUNEL assay and immumnocytochemical staining of activated caspase-3. E2 enhanced cell proliferation and prevented IL-1β-induced cell death, but the effect was partially blocked by G36 and completely abrogated by a combination of ICI 182,780 and G36. This study demonstrates that GPR30 is expressed in human IVD to transmit signals triggering E2-induced NP cell proliferation and protecting against IL-1β-induced apoptosis. The effects of E2 on NP cells require both GPR30 and classic estrogen receptors. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

Lumbar Spine Paraspinal Muscle and Intervertebral Disc Height Changes in Astronauts After Long-Duration Spaceflight on the International Space Station.

PubMed

Chang, Douglas G; Healey, Robert M; Snyder, Alexander J; Sayson, Jojo V; Macias, Brandon R; Coughlin, Dezba G; Bailey, Jeannie F; Parazynski, Scott E; Lotz, Jeffrey C; Hargens, Alan R

2016-12-15

Prospective case series. Evaluate lumbar paraspinal muscle (PSM) cross-sectional area and intervertebral disc (IVD) height changes induced by a 6-month space mission on the International Space Station. The long-term objective of this project is to promote spine health and prevent spinal injury during space missions and here on Earth. National Aeronautics and Space Administration (NASA) crewmembers have a 4.3 times higher risk of herniated IVDs, compared with the general and military aviator populations. The highest risk occurs during the first year after a mission. Microgravity exposure during long-duration spaceflights results in approximately 5 cm lengthening of body height, spinal pain, and skeletal deconditioning. How the PSMs and IVDs respond during spaceflight is not well described. Six NASA crewmembers were imaged supine with a 3 Tesla magnetic resonance imaging. Imaging was conducted preflight, immediately postflight, and then 33 to 67 days after landing. Functional cross-sectional area (FCSA) measurements of the PSMs were performed at the L3-4 level. FCSA was measured by grayscale thresholding within the posterior lumbar extensors to isolate lean muscle on T2-weighted scans. IVD heights were measured at the anterior, middle, and posterior sections of all lumbar levels. Repeated measures analysis of variance was used to determine significance at P < 0.05, followed by post-hoc testing. Paraspinal lean muscle mass, as indicated by the FCSA, decreased from 86% of the total PSM cross-sectional area down to 72%, immediately after the mission. Recovery of 68% of the postflight loss occurred during the next 6 weeks, still leaving a significantly lower lean muscle fractional content compared with preflight values. In contrast, lumbar IVD heights were not appreciably different at any time point. The data reveal lumbar spine PSM atrophy after long-duration spaceflight. Some FCSA recovery was seen with 46 days postflight in a terrestrial environment, but it remained incomplete compared with preflight levels. 4.

Comparison of Intravitreal Bevacizumab and Intravitreal Diclofenac in the Treatment of Diabetic Macular Edema: a 6-month Follow-up

PubMed Central

FAGHIHI, Hooshang; YAHYAPOUR, Hanif; MAHMOUDZADEH, Raziyeh; FAGHIHI, Shahin

2017-01-01

The aim of this study was to compare the effect of intravitreal diclofenac, a non-steroidal anti-inflammatory drug (NSAID), with that of bevacizumab, a well-known anti-vascular endothelial growth factor (VEGF) drug, in the treatment of diabetic macular edema (DME). Diclofenac was chosen in this study because it has both features of NSAIDs and corticosteroids by inhibiting the cyclooxygenase (COX) and lipoxygenase pathways, respectively. In this non-randomized comparative interventional case series, 64 eyes from 32 patients with bilateral naïve DME were selected and every eye was randomly assigned to intravitreal injection of bevacizumab (IVB) or diclofenac (IVD). After exclusion of some patients because of short follow-up duration or less than two intravitreal injections, finally, 52 eyes from 26 patients were analyzed. Of those, 26 eyes received 500 µg/0.1 mL IVD and 26 eyes received 1.25 mg IVB. After 6 months of follow-up, the results indicated that visual acuity was significantly improved from 0.50 ± 0.13 in IVB and 0.52 ± 0.12 LogMAR in IVD at baseline to 0.2 ± 0.1 and 0.29 ± 0.07, respectively. Central macular thickness (CMT) and macular volume were measured based on spectral-domain optical coherence tomography (OCT) at month 1, 3, and 6. Both groups showed a significant reduction in CMT and macular volume from baseline but there was no significant difference between the IVB and IVD groups. Interestingly, IVD, but not IVB, decreased intraocular pressure (IOP), which is a desirable effect. There was no serious complication due to injections. This study sheds light into the long-term effects of NSAIDs and may support the idea that inflammation suppression by NSAIDs may have the same results as anti-VEGF administration. PMID:29392145

Comparison of Intravitreal Bevacizumab and Intravitreal Diclofenac in the Treatment of Diabetic Macular Edema: a 6-month Follow-up.

PubMed

Faghihi, Hooshang; Yahyapour, Hanif; Mahmoudzadeh, Raziyeh; Faghihi, Shahin

2017-01-01

The aim of this study was to compare the effect of intravitreal diclofenac, a non-steroidal anti-inflammatory drug (NSAID), with that of bevacizumab, a well-known anti-vascular endothelial growth factor (VEGF) drug, in the treatment of diabetic macular edema (DME). Diclofenac was chosen in this study because it has both features of NSAIDs and corticosteroids by inhibiting the cyclooxygenase (COX) and lipoxygenase pathways, respectively. In this non-randomized comparative interventional case series, 64 eyes from 32 patients with bilateral naïve DME were selected and every eye was randomly assigned to intravitreal injection of bevacizumab (IVB) or diclofenac (IVD). After exclusion of some patients because of short follow-up duration or less than two intravitreal injections, finally, 52 eyes from 26 patients were analyzed. Of those, 26 eyes received 500 µg/0.1 mL IVD and 26 eyes received 1.25 mg IVB. After 6 months of follow-up, the results indicated that visual acuity was significantly improved from 0.50 ± 0.13 in IVB and 0.52 ± 0.12 LogMAR in IVD at baseline to 0.2 ± 0.1 and 0.29 ± 0.07, respectively. Central macular thickness (CMT) and macular volume were measured based on spectral-domain optical coherence tomography (OCT) at month 1, 3, and 6. Both groups showed a significant reduction in CMT and macular volume from baseline but there was no significant difference between the IVB and IVD groups. Interestingly, IVD, but not IVB, decreased intraocular pressure (IOP), which is a desirable effect. There was no serious complication due to injections. This study sheds light into the long-term effects of NSAIDs and may support the idea that inflammation suppression by NSAIDs may have the same results as anti-VEGF administration.

Expression of receptors for putative anabolic growth factors in human intervertebral disc: implications for repair and regeneration of the disc.

PubMed

Le Maitre, Christine L; Richardson, Stephen M A; Baird, Pauline; Freemont, Anthony J; Hoyland, Judith A

2005-12-01

Low back pain (LBP) is a common, debilitating and economically important disorder. Current evidence implicates loss of intervertebral disc (IVD) matrix consequent upon 'degeneration' as a major cause of LBP. Degeneration of the IVD involves increases in degradative enzymes and decreases in the extracellular matrix (ECM) component in a process that is controlled by a range of cytokines and growth factors. Studies have suggested using anabolic growth factors to regenerate the normal matrix of the IVD, hence restoring disc height and reversing degenerative disc disease. However, for such therapies to be successful it is vital that the target cells (i.e. the disc cells) express the appropriate receptors. This immunohistochemical study has for the first time investigated the expression and localization of four potentially beneficial growth factor receptors (i.e. TGFbetaRII, BMPRII, FGFR3 and IGFRI) in non-degenerate and degenerate human IVDs. Receptor expression was quantified across regions of the normal and degenerate disc and showed that cells of the nucleus pulposus (NP) and inner annulus fibrosus (IAF) expressed significantly higher levels of the four growth factor receptors investigated. There were no significant differences between the four growth factor expression in non-degenerate and degenerate biopsies. However, expression of TGFbetaRII, FGFR3 and IGFRI, but not BMP RII, were observed in the ingrowing blood vessels that characterize part of the disease aetiology. In conclusion, this study has demonstrated the expression of the four growth factor receptors at similar levels in the chondrocyte-like cells of the NP and IAF in both non-degenerate and degenerate discs, implicating a role in normal disc homeostasis and suggesting that the application of these growth factors to the degenerate human IVD would stimulate matrix production. However, the expression of some of the growth factor receptors on ingrowing blood vessels might be problematic in a therapeutic approach. Copyright 2005 Pathological Society of Great Britain and Ireland.

75 FR 8508 - Computerized Tribal IV-D Systems and Office Automation

Federal Register 2010, 2011, 2012, 2013, 2014

2010-02-25

...This rule enables Tribes and Tribal organizations currently operating comprehensive Tribal Child Support Enforcement programs under Title IV-D of the Social Security Act (the Act) to apply for and receive direct Federal funding for the costs of automated data processing. This rule addresses the Secretary's commitment to provide instructions and guidance to Tribes and Tribal organizations on requirements for applying for, and upon approval, securing Federal Financial Participation (FFP) in the costs of installing, operating, maintaining, and enhancing automated data processing systems.

Histomorphometric comparative study of blood vessels and their pattern in follicular cyst, odontogenic keratocyst, and ameloblastoma.

PubMed

Seifi, Safora; Feizi, Farideh; Khafri, Thoraya; Aram, Mehrdad

2013-03-01

The present study aimed at assessment and histomorphometric analysis of intratumoral and peritumoral (cystic) blood vessels in odontogenic lesions and their pattern on their clinical behavior by immunohistochemistry and morphometry. In a descriptive and analytical cross-sectional study, 45 paraffin blocks of ameloblastoma, odontogenic keratocyst, and follicular cyst were selected and stained immunohistochemically for CD34. In each slide, images of 3 microscopic fields with the highest microvessel density in intratumoral and peritumoral (cystic) areas were captured at 40× magnification with attached camera system. Inner vascular diameter (IVD) and outer vascular diameter (OVD), cross-sectional area (CSA), and the wall thickness (WT) of the vessels were measured with Motic Plus 2 software. The vascular pattern in odontogenic lesions was analyzed. Outer vascular diameter, IVD, and CSA of the vessels in peritumoral (cystic) areas were greater in ameloblastoma than keratocyst (P = 0.001) and follicular cyst (P < 0.001). However, WT of the blood vessels did not show any significant statistical difference among the 3 odontogenic lesions (P = 0.05). The differences in OVD, IVD (P = 0.8), CSA (P = 0.6), and WT (P = 0.4) of the blood vessels in intratumoral (cystic) areas were not statistically significant. The blood vessel pattern was circumferential in ameloblastoma, and it was directional in keratocyst and follicular cyst. Morphometric specifications of blood vessels (IVD, OVD, CSA) and their pattern in peritumoral (cystic) areas may influence the aggressive clinical behavior of ameloblastoma in comparison with keratocyst and follicular cyst.

Human disc degeneration is associated with increased MMP 7 expression.

PubMed

Le Maitre, C L; Freemont, A J; Hoyland, J A

2006-01-01

During intervertebral disc (IVD) degeneration, normal matrix synthesis decreases and degradation of disc matrix increases. A number of proteases that are increased during disc degeneration are thought to be involved in its pathogenesis. Matrix metalloproteinase 7 (MMP 7) (Matrilysin, PUMP-1) is known to cleave the major matrix molecules found within the IVD, i.e., the proteoglycan aggrecan and collagen type II. To date, however, it is not known how its expression changes with degeneration or its exact location. We investigated the localization of MMP 7 in human, histologically graded, nondegenerate, degenerated and prolapsed discs to ascertain whether MMP 7 is up-regulated during disc degeneration. Samples of human IVD tissue were fixed in neutral buffered formalin, embedded in paraffin, and sections stained with hematoxylin and eosin to score the degree of morphological degeneration. Immunohistochemistry was performed to localize MMP 7 in 41 human IVDs with varying degrees of degeneration. We found that the chondrocyte-like cells of the nucleus pulposus and inner annulus fibrosus were MMP 7 immunopositive; little immunopositivity was observed in the outer annulus. Nondegenerate discs showed few immunopositive cells. A significant increase in the proportion of MMP 7 immunopositive cells was seen in the nucleus pulposus of discs classified as showing intermediate levels of degeneration and a further increase was seen in discs with severe degeneration. Prolapsed discs showed more MMP 7 immunopositive cells compared to nondegenerated discs, but fewer than those seen in cases of severe degeneration.

Novel immortal human cell lines reveal subpopulations in the nucleus pulposus

PubMed Central

2014-01-01

Introduction Relatively little is known about cellular subpopulations in the mature nucleus pulposus (NP). Detailed understanding of the ontogenetic, cellular and molecular characteristics of functional intervertebral disc (IVD) cell populations is pivotal to the successful development of cell replacement therapies and IVD regeneration. In this study, we aimed to investigate whether phenotypically distinct clonal cell lines representing different subpopulations in the human NP could be generated using immortalization strategies. Methods Nondegenerate healthy disc material (age range, 8 to 15 years) was obtained as surplus surgical material. Early passage NP monolayer cell cultures were initially characterized using a recently established NP marker set. NP cells were immortalized by simian virus 40 large T antigen (SV40LTag) and human telomerase reverse transcriptase expression. Immortalized cells were clonally expanded and characterized based on collagen type I, collagen type II, α1 (COL2A1), and SRY-box 9 (SOX9) protein expression profiles, as well as on expression of a subset of established in vivo NP cell lineage markers. Results A total of 54 immortal clones were generated. Profiling of a set of novel NP markers (CD24, CA12, PAX1, PTN, FOXF1 and KRT19 mRNA) in a representative set of subclones substantiated successful immortalization of multiple cellular subpopulations from primary isolates and confirmed their NP origin and/or phenotype. We were able to identify two predominant clonal NP subtypes based on their morphological characteristics and their ability to induce SOX9 and COL2A1 under conventional differentiation conditions. In addition, cluster of differentiation 24 (CD24)–negative NP responder clones formed spheroid structures in various culture systems, suggesting the preservation of a more immature phenotype compared to CD24-positive nonresponder clones. Conclusions Here we report the generation of clonal NP cell lines from nondegenerate human IVD tissue and present a detailed characterization of NP cellular subpopulations. Differential cell surface marker expression and divergent responses to differentiation conditions suggest that the NP subtypes may correspond to distinct maturation stages and represent distinct NP cell subpopulations. Hence, we provide evidence that the immortalization strategy that we applied is capable of detecting cell heterogeneity in the NP. Our cell lines yield novel insights into NP biology and provide promising new tools for studies of IVD development, cell function and disease. PMID:24972717

Multifidus Muscle Changes After Back Injury Are Characterized by Structural Remodeling of Muscle, Adipose and Connective Tissue, but Not Muscle Atrophy: Molecular and Morphological Evidence.

PubMed

Hodges, Paul W; James, Gregory; Blomster, Linda; Hall, Leanne; Schmid, Annina; Shu, Cindy; Little, Chris; Melrose, James

2015-07-15

Longitudinal case-controlled animal study. To investigate putative cellular mechanisms to explain structural changes in muscle and adipose and connective tissues of the back muscles after intervertebral disc (IVD) injury. Structural back muscle changes are ubiquitous with back pain/injury and considered relevant for outcome, but their exact nature, time course, and cellular mechanisms remain elusive. We used an animal model that produces phenotypic back muscle changes after IVD injury to study these issues at the cellular/molecular level. Multifidus muscle was harvested from both sides of the spine at L1-L2 and L3-L4 IVDs in 27 castrated male sheep at 3 (n = 10) or 6 (n = 17) months after a surgical anterolateral IVD injury at both levels. Ten control sheep underwent no surgery (3 mo, n = 4; 6 mo, n = 6). Tissue was harvested at L4 for histological analysis of cross-sectional area of muscle and adipose and connective tissue (whole muscle), plus immunohistochemistry to identify proportion and cross-sectional area of individual muscle fiber types in the deepest fascicle. Quantitative polymerase chain reaction measured gene expression of typical cytokines/signaling molecules at L2. Contrary to predictions, there was no multifidus muscle atrophy (whole muscle or individual fiber). There was increased adipose and connective tissue (fibrotic proliferation) cross-sectional area and slow-to-fast muscle fiber transition at 6 but not 3 months. Within the multifidus muscle, increases in the expression of several cytokines (tumor necrosis factor α and interleukin-1β) and molecules that signal trophic/atrophic processes for the 3 tissue types (e.g., growth factor pathway [IGF-1, PI3k, Akt1, mTOR], potent tissue modifiers [calcineurin, PCG-1α, and myostatin]) were present. This study provides cellular evidence that refutes the presence of multifidus muscle atrophy accompanying IVD degeneration at this intermediate time point. Instead, adipose/connective tissue increased in parallel with the expression of the genes that provide putative mechanisms for multifidus structural remodeling. This provides novel targets for pharmacological and physical interventions. N/A.

Human cells derived from degenerate intervertebral discs respond differently to those derived from non-degenerate intervertebral discs following application of dynamic hydrostatic pressure.

PubMed

Le Maitre, Christine Lyn; Frain, Jennie; Fotheringham, Andrew P; Freemont, Anthony J; Hoyland, Judith Alison

2008-01-01

The intervertebral disc (IVD) is one of the body's most important load-bearing structures with the major mechanical force experienced in the nucleus pulposus (NP) being hydrostatic pressure (HP). Physiological levels of HP have an anabolic effect on IVD matrix metabolism in cells derived from non-degenerate animal and herniated IVD while excessive HP has a catabolic effect. However, no studies have investigated the response of non-degenerate and degenerate human disc cells derived from non-herniated discs to HP. Here we investigate the effect of physiological HP on such cells using a novel loading rig. Human IVD cells (both NP and AF) cultured in alginate were subjected to dynamic HP (0.8-1.7 MPa 0.5 Hz) for 2 h. Cell viability was assessed, RNA extracted and qRT-PCR for 18 s, c-fos, Sox-9, collagen type II, aggrecan and MMP-3 performed. Cell viability was unaffected by the loading regime. In non-degenerate NP cells, HP increased c-fos, aggrecan, Sox-9 and collagen type II (significantly so in the case of c-fos and aggrecan), but not MMP-3 gene expression. In contrast, application of HP to AF or degenerate NP cells had no effect on target gene expression. Our data shows that cells obtained from the healthy NP respond to dynamic HP by up-regulating genes indicative of healthy matrix homeostasis. However, responses differed in degenerate NP cells suggesting that an altered mechanotransduction pathway may be operational.

Musculoskeletal Modeling of the Lumbar Spine to Explore Functional Interactions between Back Muscle Loads and Intervertebral Disk Multiphysics

PubMed Central

Toumanidou, Themis; Noailly, Jérôme

2015-01-01

During daily activities, complex biomechanical interactions influence the biophysical regulation of intervertebral disks (IVDs), and transfers of mechanical loads are largely controlled by the stabilizing action of spine muscles. Muscle and other internal forces cannot be easily measured directly in the lumbar spine. Hence, biomechanical models are important tools for the evaluation of the loads in those tissues involved in low-back disorders. Muscle force estimations in most musculoskeletal models mainly rely, however, on inverse calculations and static optimizations that limit the predictive power of the numerical calculations. In order to contribute to the development of predictive systems, we coupled a predictive muscle model with the passive resistance of the spine tissues, in a L3–S1 musculoskeletal finite element model with osmo-poromechanical IVD descriptions. The model included 46 fascicles of the major back muscles that act on the lower spine. The muscle model interacted with activity-related loads imposed to the osteoligamentous structure, as standing position and night rest were simulated through distributed upper body mass and free IVD swelling, respectively. Calculations led to intradiscal pressure values within ranges of values measured in vivo. Disk swelling led to muscle activation and muscle force distributions that seemed particularly appropriate to counterbalance the anterior body mass effect in standing. Our simulations pointed out a likely existence of a functional balance between stretch-induced muscle activation and IVD multiphysics toward improved mechanical stability of the lumbar spine understanding. This balance suggests that proper night rest contributes to mechanically strengthen the spine during day activity. PMID:26301218

Hydrogels in acellular and cellular strategies for intervertebral disc regeneration.

PubMed

Pereira, D R; Silva-Correia, J; Oliveira, J M; Reis, R L

2013-02-01

Low back pain is an extremely common illness syndrome that causes patient suffering and disability and requires urgent solutions to improve the quality of life of these patients. Treatment options aimed to regenerate the intervertebral disc (IVD) are still under development. The cellular complexity of IVD, and consequently its fine regulatory system, makes it a challenge to the scientific community. Biomaterials-based therapies are the most interesting solutions to date, whereby tissue engineering and regenerative medicine (TE&RM) strategies are included. By using such strategies, i.e., combining biomaterials, cells, and biomolecules, the ultimate goal of reaching a complete integration between native and neo-tissue can be achieved. Hydrogels are promising materials for restoring IVD, mainly nucleus pulposus (NP). This study presents an overview of the use of hydrogels in acellular and cellular strategies for intervertebral disc regeneration. To better understand IVD and its functioning, this study will focus on several aspects: anatomy, pathophysiology, cellular and biomolecular performance, intrinsic healing processes, and current therapies. In addition, the application of hydrogels as NP substitutes will be addressed due to their similarities to NP mechanical properties and extracellular matrix. These hydrogels can be used in cellular strategies when combined with cells from different sources, or in acellular strategies by performing the functionalization of the hydrogels with biomolecules. In addition, a brief summary of therapies based on simple injection for primary biological repair will be examined. Finally, special emphasis will focus on reviewing original studies reporting on the use of autologous cells and biomolecules such as platelet-rich plasma and their potential clinical applications. Copyright © 2011 John Wiley & Sons, Ltd.

The deformation behavior of the cervical spine segment

NASA Astrophysics Data System (ADS)

Kolmakova, T. V.; Rikun, Yu. A.

2017-09-01

The paper describes the model of the cervical spine segment (C3-C4) and the calculation results of its deformation behavior at flexion. The segment model was built based on the experimental literature data taking into account the presence of the cortical and cancellous bone tissue of vertebral bodies. Degenerative changes of the intervertebral disk (IVD) were simulated through a reduction of the disc height and an increase of Young's modulus. The construction of the geometric model of the cervical spine segment and the calculations of the stress-strain state were carried out in the ANSYS software complex. The calculation results show that the biggest protrusion of the IVD in bending direction of segment is observed when IVD height is reduced. The disc protrusion is reduced with an increase of Young's modulus. The largest protrusion in the direction of flexion of the segment is the intervertebral disk with height of 4.3 mm and elastic modulus of 2.5 MPa. The results of the study can be useful to specialists in the field of biomechanics, medical materials science and prosthetics.

Randomized control trial follow-up: Online program and waiting period for unmarried parents in Title IV-D Court.

PubMed

Rudd, Brittany N; Poladian, Ani R; Holtzworth-Munroe, Amy; Applegate, Amy G; D'Onofrio, Brian M

2017-04-01

Despite a lack of research on parent programs for separating unmarried parents, many judicial officers mandate participation. Rudd, Holtzworth-Munroe, Reyome, Applegate, and D'Onofrio (2015) conducted the only randomized controlled trial of any online parent program for separating parents, ProudToParent.org (PTP), and related court processes (e.g., having a waiting period between the establishment of paternity and the court hearing regarding child related issues vs. having the hearing the same day). They recruited a unique sample of 182 cases in a Title IV-D Court (i.e., a court for primarily low income parents) (Authorization of Appropriations, 42 U.S.C. § 651, 2013), in which paternity was previously contested but subsequently established via court-ordered genetic testing. Unexpectedly, cases assigned to PTP and a waiting period were the least likely to reach agreement at their court hearing. In the current study, we extend these results to examine the impact of the study conditions on relitigation in the year following the court hearing; only 11.2% of cases filed a motion, and 7.8% had a hearing. The group that was least likely to reach full initial agreement (i.e., assigned to PTP and the waiting period) were the most likely to relitigate. Further, controlling for study conditions, reaching a full agreement in the Title IV-D court decreased the odds of having a court hearing in the following year. Reaching agreements on the specific issues involved in such cases (e.g., custody, child support) reduced the likelihood of both motions and hearings in the year after the Title IV-D hearings. The implications of these findings are discussed. (PsycINFO Database Record (c) 2017 APA, all rights reserved).

Intradiscal application of a PCLA-PEG-PCLA hydrogel loaded with celecoxib for the treatment of back pain in canines: What's in it for humans?

PubMed

Tellegen, Anna R; Willems, Nicole; Beukers, Martijn; Grinwis, Guy C M; Plomp, Saskia G M; Bos, Clemens; van Dijk, Maarten; de Leeuw, Mike; Creemers, Laura B; Tryfonidou, Marianna A; Meij, Björn P

2018-03-01

Chronic low back pain is a common clinical problem in both the human and canine population. Current pharmaceutical treatment often consists of oral anti-inflammatory drugs to alleviate pain. Novel treatments for degenerative disc disease focus on local application of sustained released drug formulations. The aim of this study was to determine safety and feasibility of intradiscal application of a poly(ε-caprolactone-co-lactide)-b-poly(ethylene glycol)-bpoly(ε-caprolactone-co-lactide) PCLA-PEG-PCLA hydrogel releasing celecoxib, a COX-2 inhibitor. Biocompatibility was evaluated after subcutaneous injection in mice, and safety of intradiscal injection of the hydrogel was evaluated in experimental dogs with early spontaneous intervertebral disc (IVD) degeneration. COX-2 expression was increased in IVD samples surgically obtained from canine patients, indicating a role of COX-2 in clinical IVD disease. Ten client-owned dogs with chronic low back pain related to IVD degeneration received an intradiscal injection with the celecoxib-loaded hydrogel. None of the dogs showed adverse reactions after intradiscal injection. The hydrogel did not influence magnetic resonance imaging signal at long-term follow-up. Clinical improvement was achieved by reduction of back pain in 9 of 10 dogs, as was shown by clinical examination and owner questionnaires. In 3 of 10 dogs, back pain recurred after 3 months. This study showed the safety and effectiveness of intradiscal injections in vivo with a thermoresponsive PCLA-PEG-PCLA hydrogel loaded with celecoxib. In this set-up, the dog can be used as a model for the development of novel treatment modalities in both canine and human patients with chronic low back pain. Copyright © 2017 John Wiley & Sons, Ltd.

Isovaleric Acidemia: New Aspects of Genetic and Phenotypic Heterogeneity

PubMed Central

Vockley, Jerry; Ensenauer, Regina

2008-01-01

Isovaleric acidemia (IVA) is an autosomal recessive inborn error of leucine metabolism caused by a deficiency of the mitochondrial enzyme isovaleryl-CoA dehydrogenase (IVD) resulting in the accumulation of derivatives of isovaleryl-CoA. It was the first organic acidemia recognized in humans and can cause significant morbidity and mortality. Early diagnosis and treatment with a protein restricted diet and supplementation with carnitine and glycine are effective in promoting normal development in severely affected individuals. Both intra- and inter-familial variability have been recognized. Initially, two phenotypes with either an acute neonatal or a chronic intermittent presentation were described. More recently, a third group of individuals with mild biochemical abnormalities who can be asymptomatic have been identified through newborn screening of blood spots by tandem mass spectrometry. IVD is a flavoenzyme that catalyzes the conversion of isovaleryl-CoA to 3-methylcrotonyl-CoA and transfers electrons to the electron transfer flavoprotein. Human IVD has been purified from tissue and recombinant sources and its biochemical and physical properties have been extensively studied. Molecular analysis of the IVD gene from patients with IVA has allowed characterization of different types of mutations in this gene. One missense mutation, 932C>T (A282V), is particularly common in patients identified through newborn screening with mild metabolite elevations and who have remained asymptomatic to date. This mutation leads to a partially active enzyme with altered catalytic properties; however, its effects on clinical outcome and the necessity of therapy are still unknown. A better understanding of the heterogeneity of this disease and the relevance of genotype/phenotype correlations to clinical management of patients are among the challenges remaining in the study of this disorder in the coming years. PMID:16602101

Increased expression of matrix metalloproteinase-10, nerve growth factor and substance P in the painful degenerate intervertebral disc.

PubMed

Richardson, Stephen M; Doyle, Paul; Minogue, Ben M; Gnanalingham, Kanna; Hoyland, Judith A

2009-01-01

Matrix metalloproteinases (MMPs) are known to be involved in the degradation of the nucleus pulposus (NP) during intervertebral disc (IVD) degeneration. This study investigated MMP-10 (stromelysin-2) expression in the NP during IVD degeneration and correlated its expression with pro-inflammatory cytokines and molecules involved in innervation and nociception during degeneration which results in low back pain (LBP). Human NP tissue was obtained at postmortem (PM) from patients without a history of back pain and graded as histologically normal or degenerate. Symptomatic degenerate NP samples were also obtained at surgery for LBP. Expression of MMP-10 mRNA and protein was analysed using real-time polymerase chain reaction and immunohistochemistry. Gene expression for pro-inflammatory cytokines interleukin-1 (IL-1) and tumour necrosis factor-alpha (TNF-alpha), nerve growth factor (NGF) and the pain-associated neuropeptide substance P were also analysed. Correlations between MMP-10 and IL-1, TNF-alpha and NGF were assessed along with NGF with substance P. MMP-10 mRNA was significantly increased in surgical degenerate NP when compared to PM normal and PM degenerate samples. MMP-10 protein was also significantly higher in degenerate surgical NP samples compared to PM normal. IL-1 and MMP-10 mRNA demonstrated a significant correlation in surgical degenerate samples, while TNF-alpha was not correlated with MMP-10 mRNA. NGF was significantly correlated with both MMP-10 and substance P mRNA in surgical degenerate NP samples. MMP-10 expression is increased in the symptomatic degenerate IVD, where it may contribute to matrix degradation and initiation of nociception. Importantly, this study suggests differences in the pathways involved in matrix degradation between painful and pain-free IVD degeneration.

Genetic and functional studies of the intervertebral disc: a novel murine intervertebral disc model.

PubMed

Pelle, Dominic W; Peacock, Jacqueline D; Schmidt, Courtney L; Kampfschulte, Kevin; Scholten, Donald J; Russo, Scott S; Easton, Kenneth J; Steensma, Matthew R

2014-01-01

Intervertebral disc (IVD) homeostasis is mediated through a combination of micro-environmental and biomechanical factors, all of which are subject to genetic influences. The aim of this study is to develop and characterize a genetically tractable, ex vivo organ culture model that can be used to further elucidate mechanisms of intervertebral disc disease. Specifically, we demonstrate that IVD disc explants (1) maintain their native phenotype in prolonged culture, (2) are responsive to exogenous stimuli, and (3) that relevant homeostatic regulatory mechanisms can be modulated through ex-vivo genetic recombination. We present a novel technique for isolation of murine IVD explants with demonstration of explant viability (CMFDA/propidium iodide staining), disc anatomy (H&E), maintenance of extracellular matrix (ECM) (Alcian Blue staining), and native expression profile (qRT-PCR) as well as ex vivo genetic recombination (mT/mG reporter mice; AdCre) following 14 days of culture in DMEM media containing 10% fetal bovine serum, 1% L-glutamine, and 1% penicillin/streptomycin. IVD explants maintained their micro-anatomic integrity, ECM proteoglycan content, viability, and gene expression profile consistent with a homeostatic drive in culture. Treatment of genetically engineered explants with cre-expressing adenovirus efficaciously induced ex vivo genetic recombination in a variety of genetically engineered mouse models. Exogenous administration of IL-1ß and TGF-ß3 resulted in predicted catabolic and anabolic responses, respectively. Genetic recombination of TGFBR1fl/fl explants resulted in constitutively active TGF-ß signaling that matched that of exogenously administered TGF-ß3. Our results illustrate the utility of the murine intervertebral disc explant to investigate mechanisms of intervertebral disc degeneration.

Genetic and Functional Studies of the Intervertebral Disc: A Novel Murine Intervertebral Disc Model

PubMed Central

Pelle, Dominic W.; Peacock, Jacqueline D.; Schmidt, Courtney L.; Kampfschulte, Kevin; Scholten, Donald J.; Russo, Scott S.; Easton, Kenneth J.; Steensma, Matthew R.

2014-01-01

Intervertebral disc (IVD) homeostasis is mediated through a combination of micro-environmental and biomechanical factors, all of which are subject to genetic influences. The aim of this study is to develop and characterize a genetically tractable, ex vivo organ culture model that can be used to further elucidate mechanisms of intervertebral disc disease. Specifically, we demonstrate that IVD disc explants (1) maintain their native phenotype in prolonged culture, (2) are responsive to exogenous stimuli, and (3) that relevant homeostatic regulatory mechanisms can be modulated through ex-vivo genetic recombination. We present a novel technique for isolation of murine IVD explants with demonstration of explant viability (CMFDA/propidium iodide staining), disc anatomy (H&E), maintenance of extracellular matrix (ECM) (Alcian Blue staining), and native expression profile (qRT-PCR) as well as ex vivo genetic recombination (mT/mG reporter mice; AdCre) following 14 days of culture in DMEM media containing 10% fetal bovine serum, 1% L-glutamine, and 1% penicillin/streptomycin. IVD explants maintained their micro-anatomic integrity, ECM proteoglycan content, viability, and gene expression profile consistent with a homeostatic drive in culture. Treatment of genetically engineered explants with cre-expressing adenovirus efficaciously induced ex vivo genetic recombination in a variety of genetically engineered mouse models. Exogenous administration of IL-1ß and TGF-ß3 resulted in predicted catabolic and anabolic responses, respectively. Genetic recombination of TGFBR1fl/fl explants resulted in constitutively active TGF-ß signaling that matched that of exogenously administered TGF-ß3. Our results illustrate the utility of the murine intervertebral disc explant to investigate mechanisms of intervertebral disc degeneration. PMID:25474689

The fabrication and characterization of a multi-laminate, angle-ply collagen patch for annulus fibrosus repair.

PubMed

McGuire, Rachel; Borem, Ryan; Mercuri, Jeremy

2017-12-01

One major limitation of intervertebral disc (IVD) repair is that no ideal biomaterial has been developed that effectively mimics the angle-ply collagen architecture and mechanical properties of the native annulus fibrosus (AF). Furthermore, it would be beneficial to devise a simple, scalable process by which to manufacture a biomimetic biomaterial that could function as a mechanical repair patch to be secured over a large defect in the outer AF that will support AF tissue regeneration. Such a biomaterial would: (1) enable the employment of early-stage interventional strategies to treat IVD degeneration (i.e. nucleus pulposus arthroplasty); (2) prevent IVD re-herniation in patients with large AF defects; and (3) serve as a platform to develop full-thickness AF and whole IVD tissue engineering strategies. Due to the innate collagen fibre alignment and mechanical strength of pericardium, a procedure was developed to assemble multi-laminate angle-ply AF patches derived from decellularized pericardial tissue. Patches were subsequently assessed histologically to confirm angle-ply microarchitecture, and mechanically assessed for biaxial burst strength and tensile properties. Additionally, patch cytocompatibility was evaluated following seeding with bovine AF cells. This study demonstrated the effective removal of porcine cell remnants from the pericardium, and the ability to reliably produce multi-laminate patches with angle-ply architecture using a simple assembly technique. Resultant patches demonstrated their inherent ability to resist biaxial burst pressures reminiscent of intradiscal pressures commonly borne by the AF, and exhibited tensile strength and modulus values reported for native human AF. Furthermore, the biomaterial supported AF cell viability, infiltration and proliferation. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

In vivo dosimetry with optically stimulated luminescent dosimeters for conformal and intensity-modulated radiation therapy: A 2-year multicenter cohort study.

PubMed

Riegel, Adam C; Chen, Yu; Kapur, Ajay; Apicello, Laura; Kuruvilla, Abraham; Rea, Anthony J; Jamshidi, Abolghassem; Potters, Louis

Optically stimulated luminescent dosimeters (OSLDs) are utilized for in vivo dosimetry (IVD) of modern radiation therapy techniques such as intensity modulated radiation therapy (IMRT) and volumetric modulated arc therapy (VMAT). Dosimetric precision achieved with conventional techniques may not be attainable. In this work, we measured accuracy and precision for a large sample of clinical OSLD-based IVD measurements. Weekly IVD measurements were collected from 4 linear accelerators for 2 years and were expressed as percent differences from planned doses. After outlier analysis, 10,224 measurements were grouped in the following way: overall, modality (photons, electrons), treatment technique (3-dimensional [3D] conformal, field-in-field intensity modulation, inverse-planned IMRT, and VMAT), placement location (gantry angle, cardinality, and central axis positioning), and anatomical site (prostate, breast, head and neck, pelvis, lung, rectum and anus, brain, abdomen, esophagus, and bladder). Distributions were modeled via a Gaussian function. Fitting was performed with least squares, and goodness-of-fit was assessed with the coefficient of determination. Model means (μ) and standard deviations (σ) were calculated. Sample means and variances were compared for statistical significance by analysis of variance and the Levene tests (α = 0.05). Overall, μ ± σ was 0.3 ± 10.3%. Precision for electron measurements (6.9%) was significantly better than for photons (10.5%). Precision varied significantly among treatment techniques (P < .0001) with field-in-field lowest (σ = 7.2%) and IMRT and VMAT highest (σ = 11.9% and 13.4%, respectively). Treatment site models with goodness-of-fit greater than 0.90 (6 of 10) yielded accuracy within ±3%, except for head and neck (μ = -3.7%). Precision varied with treatment site (range, 7.3%-13.0%), with breast and head and neck yielding the best and worst precision, respectively. Placement on the central axis of cardinal gantry angles yielded more precise results (σ = 8.5%) compared with other locations (range, 10.5%-11.4%). Accuracy of ±3% was achievable. Precision ranged from 6.9% to 13.4% depending on modality, technique, and treatment site. Simple, standardized locations may improve IVD precision. These findings may aid development of patient-specific tolerances for OSLD-based IVD. Copyright © 2016 American Society for Radiation Oncology. Published by Elsevier Inc. All rights reserved.

Measurement uncertainty: Friend or foe?

PubMed

Infusino, Ilenia; Panteghini, Mauro

2018-02-02

The definition and enforcement of a reference measurement system, based on the implementation of metrological traceability of patients' results to higher order reference methods and materials, together with a clinically acceptable level of measurement uncertainty, are fundamental requirements to produce accurate and equivalent laboratory results. The uncertainty associated with each step of the traceability chain should be governed to obtain a final combined uncertainty on clinical samples fulfilling the requested performance specifications. It is important that end-users (i.e., clinical laboratory) may know and verify how in vitro diagnostics (IVD) manufacturers have implemented the traceability of their calibrators and estimated the corresponding uncertainty. However, full information about traceability and combined uncertainty of calibrators is currently very difficult to obtain. Laboratory professionals should investigate the need to reduce the uncertainty of the higher order metrological references and/or to increase the precision of commercial measuring systems. Accordingly, the measurement uncertainty should not be considered a parameter to be calculated by clinical laboratories just to fulfil the accreditation standards, but it must become a key quality indicator to describe both the performance of an IVD measuring system and the laboratory itself. Copyright © 2018 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Total disc replacement using a tissue-engineered intervertebral disc in vivo: new animal model and initial results

PubMed Central

Gebhard, Harry; Bowles, Robby; Dyke, Jonathan; Saleh, Tatianna; Doty, Stephen; Bonassar, Lawrence; Härtl, Roger

2010-01-01

Study type: Basic science Introduction: Chronic back pain due to degenerative disc disease (DDD) is among the most important medical conditions causing morbidity and significant health care costs. Surgical treatment options include disc replacement or fusion surgery, but are associated with significant short- and long-term risks.1 Biological tissue-engineering of human intervertebral discs (IVD) could offer an important alternative.2 Recent in vitro data from our group have shown successful engineering and growth of ovine intervertebral disc composites with circumferentially aligned collagen fibrils in the annulus fibrosus (AF) (Figure 1).3 Figure 1 Tissue-engineered composite disc a Experimental steps to generate composite tissue-engineered IVDs3 b Example of different AF formulations on collagen alignment in the AF. Second harmonic generation and two-photon excited fluorescence images of seeded collagen gels (for AF) of 1 and 2.5 mg/ml over time. At seeding, cells and collagen were homogenously distributed in the gels. Over time, AF cells elongated and collagen aligned parallel to cells. Less contraction and less alignment is noted after 3 days in the 2.5 mg/mL gel. c Imaging-based creation of a virtual disc model that will serve as template for the engineered disc. Total disc dimensions (AF and NP) were retrieved from micro-computer tomography (CT) (left images), and nucleus pulposus (NP) dimensions alone were retrieved from T2-weighted MRI images (right images). Merging of MRI and micro-CT models revealed a composite disc model (middle image)—Software: Microview, GE Healthcare Inc., Princeton, NJ; and slicOmatic v4.3, TomoVision, Montreal, Canada. d Flow chart describing the process for generating multi-lamellar tissue engineered IVDs. IVDs are produced by allowing cell-seeded collagen layers to contract around a cell-seeded alginate core (NP) over time Objective: The next step is to investigate if biological disc implants survive, integrate, and restore function to the spine in vivo. A model will be developed that allows efficient in vivo testing of tissue-engineered discs of various compositions and characteristics. Methods: Athymic rats were anesthetized and a dorsal approach was chosen to perform a microsurgical discectomy in the rat caudal spine (Fig. 2,Fig. 3). Control group I (n = 6) underwent discectomy only, Control group II (n = 6) underwent discectomy, followed by reimplantation of the autologous disc. Two treatment groups (group III, n = 6, 1 month survival; group IV, n = 6, 6 months survival) received a tissue-engineered composite disc implant. The rodents were followed clinically for signs of infection, pain level and wound healing. X-rays and magnetic resonance imaging (MRI) were assessed postoperatively and up to 6 months after surgery (Fig. 6,Fig. 7). A 7 Tesla MRI (Bruker) was implemented for assessment of the operated level as well as the adjacent disc (hydration). T2-weighted sequences were interpreted by a semiquantitative score (0 = no signal, 1 = weak signal, 2 = strong signal and anatomical features of a normal disc). Histology was performed with staining for proteoglycans (Alcian blue) and collagen (Picrosirius red) (Fig. 4,Fig. 5). Figure 2 Disc replacement surgery a Operative situs with native disc that has been disassociated from both adjacent vertebrae b Native disc (left) and tissue-engineered implant (right) c Implant in situ before wound closureAF: Annulus fi brosus, nP: nucleus pulposus, eP: endplate, M: Muscle, T: Tendon, s: skin, art: artery, GP: Growth plate, B: Bone Figure 3 Disc replacement surgery. Anatomy of the rat caudal disc space a Pircrosirius red stained axial cut of native disc space b Saffranin-O stained sagittal cut of native disc space Figure 4 Histologies of three separate motion segments from three different rats. Animal one = native IVD, Animal two = status after discectomy, Animal three = tissue-engineered implant (1 month) a–c H&E (overall tissue staining for light micrsocopy) d–f Alcian blue (proteoglycans) g–i Picrosirius red (collagen I and II) Figure 5 Histology from one motion segment four months after implantation of a bio-engineered disc construct a Picrosirius red staining (collagen) b Polarized light microscopy showing collagen staining and collagen organization in AF region c Increased Safranin-O staining (proteoglycans) in NP region of the disc implant d Higher magnification of figure 5c: Integration between implanted tissue-engineered total disc replacement and vertebral body bone Figure 6 MRI a Disc space height measurements in flash/T1 sequence (top: implant (714.0 micrometer), bottom: native disc (823.5 micrometer) b T2 sequence, red circle surrounding the implant NP Figure 7 7 Tesla MRI imaging of rat tail IVDs showing axial images (preliminary pilot data) a Diffusion tensor imaging (DTI) on two explanted rat tail discs in Formalin b Higher magnification of a, showing directional alignment of collagen fibers (red and green) when compared to the color ball on top which maps fibers' directional alignment (eg, fibers directing from left to right: red, from top to bottom: blue) c Native IVD in vivo (successful imaging of top and bottom of the IVD (red) d Gradient echo sequence (GE) showing differentiation between NP (light grey) and AF (dark margin) e GE of reimplanted tail IVD at the explantation level f T1Rho sequence demonstrating the NP (grey) within the AF (dark margin), containing the yellow marked region of interest for value acquisition (preliminary data are consistent with values reported in the literature). g T2 image of native IVD in vivo for monitoring of hydration (white: NP) Results: The model allowed reproducible and complete discectomies as well as disc implantation in the rat tail spine without any surgical or postoperative complications. Discectomy resulted in immediate collapse of the disc space. Preliminary results indicate that disc space height was maintained after disc implantation in groups II, III and IV over time. MRI revealed high resolution images of normal intervertebral discs in vivo. Eight out of twelve animals (groups III and IV) showed a positive signal in T2-weighted images after 1 month (grade 0 = 4, grade 1 = 4, grade 2 = 4). Positive staining was seen for collagen as well as proteoglycans at the site of disc implantation after 1 month in each of the six animals with engineered implants (group III). Analysis of group IV showed positive T2 signal in five out of six animals and disc-height preservation in all animals after 6 months. Conclusions: This study demonstrates for the first time that tissue-engineered composite IVDs with circumferentially aligned collagen fibrils survive and integrate with surrounding vertebral bodies when placed in the rat spine for up to 6 months. Tissue-engineered composite IVDs restored function to the rat spine as indicated by maintenance of disc height and vertebral alignment. A significant finding was that maintenance of the composite structure in group III was observed, with increased proteoglycan staining in the nucleus pulposus region (Figure 4d–f). Proteoglycan and collagen matrix as well as disc height preservation and positive T2 signals in MRI are promising parameters and indicate functionality of the implants. PMID:23637671

Cytotoxic Effects of the Radiocontrast Agent Iotrolan and Anesthetic Agents Bupivacaine and Lidocaine in Three-Dimensional Cultures of Human Intervertebral Disc Nucleus Pulposus Cells: Identification of the Apoptotic Pathways

PubMed Central

Iwasaki, Koji; Sudo, Hideki; Yamada, Katsuhisa; Ito, Manabu; Iwasaki, Norimasa

2014-01-01

Background Discography and discoblock are imaging procedures used to diagnose discogenic low back pain. Although needle puncture of the intervertebral disc (IVD) itself induces disc degeneration, the agents used in these procedures may also have harmful effects on IVD cells. The purpose of this study was to analyze whether radiocontrast agents and local anesthetic agents have detrimental effects on human nucleus pulposus (NP) cells. Methods Healthy human NP cells were cultured for 7 days in three-dimensional (3D) cell–alginate bead composites, and were then exposed to clinically relevant doses of a radiocontrast agent (iotrolan) or local anesthetic (lidocaine or bupivacaine). Cell viability and apoptosis were measured by confocal microscopy and flow cytometry. On the basis of caspase expression profiles, the apoptotic pathways activated by the agents were identified by Western blot analysis. Results The radiocontrast agent iotrolan did not affect NP cell viability or induce apoptosis. In contrast, both the anesthetic agents significantly decreased cell viability and increased the apoptotic cell number in a time- and dose-dependent manner. After 120 min, 2% lidocaine and 0.5% bupivacaine decreased percent live cells to 13% and 10%, respectively (p<0.05). The number of apoptotic cells was doubled by increasing lidocaine dosage from 1% to 2% (23% and 42%) and bupivacaine from 0.25% to 0.50% (25% and 48%) (p<0.05). Western blot analysis revealed that both anesthetic agents upregulated cleaved caspase-3 and caspase-8, whereas only bupivacaine upregulated cleaved caspase-9. Conclusions/Significance The present study demonstrates that iotrolan does not affect the viability of healthy human NP cells. In contrast, the two anesthetic agents commonly used in discography or discoblock may cause extensive damage to IVDs by inducing apoptotic cell death. PMID:24642945

Novel Immortal Cell Lines Support Cellular Heterogeneity in the Human Annulus Fibrosus

PubMed Central

van den Akker, Guus G. H.; Surtel, Don A. M.; Cremers, Andy; Richardson, Stephen M.; Hoyland, Judith A.; van Rhijn, Lodewijk W.

2016-01-01

Introduction Loss of annulus fibrosus (AF) integrity predisposes to disc herniation and is associated with IVD degeneration. Successful implementation of biomedical intervention therapy requires in-depth knowledge of IVD cell biology. We recently generated unique clonal human nucleus pulposus (NP) cell lines. Recurring functional cellular phenotypes from independent donors provided pivotal evidence for cell heterogeneity in the mature human NP. In this study we aimed to generate and characterize immortal cell lines for the human AF from matched donors. Methods Non-degenerate healthy disc material was obtained as surplus surgical material. AF cells were immortalized by simian virus Large T antigen (SV40LTAg) and human telomerase (hTERT) expression. Early passage cells and immortalized cell clones were characterized based on marker gene expression under standardized culturing and in the presence of Transforming Growth factor β (TGFβ). Results The AF-specific expression signature included COL1A1, COL5A1, COL12A1, SFRP2 and was largely maintained in immortal AF cell lines. Remarkably, TGFβ induced rapid 3D sheet formation in a subgroup of AF clones. This phenotype was associated with inherent differences in Procollagen type I processing and maturation, and correlated with differential mRNA expression of Prolyl 4-hydroxylase alpha polypeptide 1 and 3 (P4HA1,3) and Lysyl oxidase (LOX) between clones and differential P4HA3 protein expression between AF cells in histological sections. Conclusion We report for the first time the generation of representative human AF cell lines. Gene expression profile analysis and functional comparison of AF clones revealed variation between immortalized cells and suggests phenotypic heterogeneity in the human AF. Future characterization of AF cellular (sub-)populations aims to combine identification of additional specific AF marker genes and their biological relevance. Ultimately this knowledge will contribute to clinical application of cell-based technology in IVD repair. PMID:26794306

Molecular signaling in intervertebral disk development.

PubMed

DiPaola, Christian P; Farmer, James C; Manova, Katia; Niswander, Lee A

2005-09-01

The purpose of this investigation is to identify and study the expression pattern of pertinent molecular factors involved in the differentiation of the intervertebral disk (IVD). It is likely that hedgehog genes and the BMP inhibitors are key factors involved in spinal joint formation. Radioactive in situ hybridization with mRNA probes for pax-1, SHH, IHH and Noggin gene was performed on mouse embryo and adult tissue. Immunohistochemistry was performed to localize hedgehog receptor, "patched" (ptc). From 14.5 dpc until birth pax-1 mRNA was expressed in the developing anulus fibrosus (AF). During the same developmental period Noggin mRNA is highly expressed throughout the spine, in the developing AF, while ptc protein and SHH mRNA were expressed in the developing nucleus pulposus (NP). IHH mRNA was expressed by condensing chondrocytes of the vertebral bodies and later becomes confined to the vertebral endplate. We show for the first time that pax-1 is expressed in the adult intervertebral disk. Ptc expression in the NP is an indicator of hedgehog protein signaling in the developing IVD. The expression pattern of the BMP inhibitor Noggin appears to be important for the normal formation of the IVD and may prove to play a role in its segmental pattern formation.

Managing Disruptive Behavior by Patients and Physicians: A Responsibility of the Dialysis Facility Medical Director.

PubMed

Jones, Edward R; Goldman, Richard S

2015-08-07

The Centers for Medicare & Medicaid Services' Conditions for Coverage make the medical director of an ESRD facility responsible for all aspects of care, including high-quality health care delivery (e.g., safe, effective, timely, efficient, and patient centered). Because of the high-pressure environment of the dialysis facility, conflicts are common. Conflict frequently occurs when aberrant behaviors disrupt the dialysis facility. Patients, family members, friends, and, less commonly appreciated, nephrology clinicians (i.e., nephrologists and advanced care practitioners) may manifest disruptive behavior. Disruptive behavior in the dialysis facility impairs the ability to deliver high-quality care. Furthermore, disruptive behavior is the leading cause for involuntary discharge (IVD) or involuntary transfer (IVT) of a patient from a facility. IVD usually results in loss of continuity of care, increased emergency department visits, and increased unscheduled, acute dialysis treatments. A sufficient number of IVDs and IVTs also trigger an extensive review of the facility by the regional ESRD Networks, exposing the facility to possible Medicare-imposed sanctions. Medical directors must be equipped to recognize and correct disruptive behavior. Nephrology-based literature and tools exist to help dialysis facility medical directors successfully address and resolve disruptive behavior before medical directors must involuntarily discharge a patient or terminate an attending clinician. Copyright © 2015 by the American Society of Nephrology.

The effect of creep on human lumbar intervertebral disk impact mechanics.

PubMed

Jamison, David; Marcolongo, Michele S

2014-03-01

The intervertebral disk (IVD) is a highly hydrated tissue, with interstitial fluid making up 80% of the wet weight of the nucleus pulposus (NP), and 70% of the annulus fibrosus (AF). It has often been modeled as a biphasic material, consisting of both a solid and fluid phase. The inherent porosity and osmotic potential of the disk causes an efflux of fluid while under constant load, which leads to a continuous displacement phenomenon known as creep. IVD compressive stiffness increases and NP pressure decreases as a result of creep displacement. Though the effects of creep on disk mechanics have been studied extensively, it has been limited to nonimpact loading conditions. The goal of this study is to better understand the influence of creep and fluid loss on IVD impact mechanics. Twenty-four human lumbar disk samples were divided into six groups according to the length of time they underwent creep (tcreep = 0, 3, 6, 9, 12, 15 h) under a constant compressive load of 400 N. At the end of tcreep, each disk was subjected to a sequence of impact loads of varying durations (timp = 80, 160, 320, 400, 600, 800, 1000 ms). Energy dissipation (ΔE), stiffness in the toe (ktoe) and linear (klin) regions, and neutral zone (NZ) were measured. Analyzing correlations with tcreep, there was a positive correlation with ΔE and NZ, along with a negative correlation with ktoe. There was no strong correlation between tcreep and klin. The data suggest that the IVD mechanical response to impact loading conditions is altered by fluid content and may result in a disk that exhibits less clinical stability and transfers more load to the AF. This could have implications for risk of diskogenic pain as a function of time of day or tissue hydration.

In vivo effects of bupivacaine and gadobutrol on the intervertebral disc following discoblock and discography: a histological analysis.

PubMed

Strube, Patrick; Pfitzner, Berit M; Streitparth, Florian; Hartwig, Tony; Putzier, Michael

2017-01-01

The aim of the present study was to histologically compare chondrotoxicity in surgically harvested intervertebral discs (IVDs) of patients following discoblock, discography, or no preoperative intervention. Thirty patients (IVD degeneration Modic ≥ 2°, Pfirrmann 3° or 4°) at L4/5 or L5/S1 who were planned for anterior lumbar interbody fusion were randomly assigned to three groups (open MRI: group DG - discography with gadobutrol; group DB - discoblock with bupivacaine at 4 weeks prior to surgery; group C - no intervention). The intervertebral discs were histologically evaluated and compared using ANOVA and Bonferroni tests for cell count, apoptosis, and proliferation. A reduced cell count (groups DG vs. DB vs. C: 14.9 ± 7.1, 9.2 ± 3.8, and 16.6 ± 5.2 cells/mm 2 , respectively; p ANOVA  = 0.016), increased apoptosis (groups DG vs. DB vs. C: 34.9 ± 10.2, 47.4 ± 16.3, 32.6 ± 12.2 %, respectively; p ANOVA  = 0.039) and increased cell proliferation (post hoc pDB vs. DG or C p < 0.001; for 3-7 cell monoclonal cell nests: groups DG vs. DB vs. C: 2.4 ± 1, 3.9 ± 1, 2.2 ± 1.1, respectively; p intervention x nest size  = 0.006) were found in the IVDs of patients in group DB. This in vivo study suggests that chondrotoxic effects occur in IVD cells after the intradiscal injection of bupivacaine but not after gadobutrol administration. • Local bupivacaine administration to intervertebral discs leads to cell toxicity and proliferation. • Gadobutrol demonstrated no significant effect on cell count, apoptosis, or cell proliferation. • In vivo cytotoxicity was demonstrated histologically in humans for the first time. • Addition/administration of bupivacaine during discographies must be judged critically.

Differentiation of Mouse Induced Pluripotent Stem Cells (iPSCs) into Nucleus Pulposus-Like Cells In Vitro

PubMed Central

Chen, Jun; Lee, Esther J.; Jing, Liufang; Christoforou, Nicolas; Leong, Kam W.; Setton, Lori A.

2013-01-01

A large percentage of the population may be expected to experience painful symptoms or disability associated with intervertebral disc (IVD) degeneration – a condition characterized by diminished integrity of tissue components. Great interest exists in the use of autologous or allogeneic cells delivered to the degenerated IVD to promote matrix regeneration. Induced pluripotent stem cells (iPSCs), derived from a patient’s own somatic cells, have demonstrated their capacity to differentiate into various cell types although their potential to differentiate into an IVD cell has not yet been demonstrated. The overall objective of this study was to assess the possibility of generating iPSC-derived nucleus pulposus (NP) cells in a mouse model, a cell population that is entirely derived from notochord. This study employed magnetic activated cell sorting (MACS) to isolate a CD24+ iPSC subpopulation. Notochordal cell-related gene expression was analyzed in this CD24+ cell fraction via real time RT-PCR. CD24+ iPSCs were then cultured in a laminin-rich culture system for up to 28 days, and the mouse NP phenotype was assessed by immunostaining. This study also focused on producing a more conducive environment for NP differentiation of mouse iPSCs with addition of low oxygen tension and notochordal cell conditioned medium (NCCM) to the culture platform. iPSCs were evaluated for an ability to adopt an NP-like phenotype through a combination of immunostaining and biochemical assays. Results demonstrated that a CD24+ fraction of mouse iPSCs could be retrieved and differentiated into a population that could synthesize matrix components similar to that in native NP. Likewise, the addition of a hypoxic environment and NCCM induced a similar phenotypic result. In conclusion, this study suggests that mouse iPSCs have the potential to differentiate into NP-like cells and suggests the possibility that they may be used as a novel cell source for cellular therapy in the IVD. PMID:24086564

Finite element based nonlinear normalization of human lumbar intervertebral disc stiffness to account for its morphology.

PubMed

Maquer, Ghislain; Laurent, Marc; Brandejsky, Vaclav; Pretterklieber, Michael L; Zysset, Philippe K

2014-06-01

Disc degeneration, usually associated with low back pain and changes of intervertebral stiffness, represents a major health issue. As the intervertebral disc (IVD) morphology influences its stiffness, the link between mechanical properties and degenerative grade is partially lost without an efficient normalization of the stiffness with respect to the morphology. Moreover, although the behavior of soft tissues is highly nonlinear, only linear normalization protocols have been defined so far for the disc stiffness. Thus, the aim of this work is to propose a nonlinear normalization based on finite elements (FE) simulations and evaluate its impact on the stiffness of human anatomical specimens of lumbar IVD. First, a parameter study involving simulations of biomechanical tests (compression, flexion/extension, bilateral torsion and bending) on 20 FE models of IVDs with various dimensions was carried out to evaluate the effect of the disc's geometry on its compliance and establish stiffness/morphology relations necessary to the nonlinear normalization. The computed stiffness was then normalized by height (H), cross-sectional area (CSA), polar moment of inertia (J) or moments of inertia (Ixx, Iyy) to quantify the effect of both linear and nonlinear normalizations. In the second part of the study, T1-weighted MRI images were acquired to determine H, CSA, J, Ixx and Iyy of 14 human lumbar IVDs. Based on the measured morphology and pre-established relation with stiffness, linear and nonlinear normalization routines were then applied to the compliance of the specimens for each quasi-static biomechanical test. The variability of the stiffness prior to and after normalization was assessed via coefficient of variation (CV). The FE study confirmed that larger and thinner IVDs were stiffer while the normalization strongly attenuated the effect of the disc geometry on its stiffness. Yet, notwithstanding the results of the FE study, the experimental stiffness showed consistently higher CV after normalization. Assuming that geometry and material properties affect the mechanical response, they can also compensate for one another. Therefore, the larger CV after normalization can be interpreted as a strong variability of the material properties, previously hidden by the geometry's own influence. In conclusion, a new normalization protocol for the intervertebral disc stiffness in compression, flexion, extension, bilateral torsion and bending was proposed, with the possible use of MRI and FE to acquire the discs' anatomy and determine the nonlinear relations between stiffness and morphology. Such protocol may be useful to relate the disc's mechanical properties to its degree of degeneration.

Biological performance of cell-encapsulated methacrylated gellan gum-based hydrogels for nucleus pulposus regeneration.

PubMed

Tsaryk, Roman; Silva-Correia, Joana; Oliveira, Joaquim Miguel; Unger, Ronald E; Landes, Constantin; Brochhausen, Christoph; Ghanaati, Shahram; Reis, Rui L; Kirkpatrick, C James

2017-03-01

Limitations of current treatments for intervertebral disc (IVD) degeneration have promoted interest in the development of tissue-engineering approaches. Injectable hydrogels loaded with cells can be used as a substitute material for the inner IVD part, the nucleus pulposus (NP), and provide an opportunity for minimally invasive treatment of IVD degeneration. The NP is populated by chondrocyte-like cells; therefore, chondrocytes and mesenchymal stem cells (MSCs), stimulated to differentiate along the chondrogenic lineage, could be used to promote NP regeneration. In this study, the in vitro and in vivo response of human bone marrow-derived MSCs and nasal chondrocytes (NCs) to modified gellan gum-based hydrogels was investigated. Both ionic- (iGG-MA) and photo-crosslinked (phGG-MA) methacrylated gellan gum hydrogels show no cytotoxicity in extraction assays with MSCs and NCs. Furthermore, the materials do not induce pro-inflammatory responses in endothelial cells. Moreover, MSCs and NCs can be encapsulated into the hydrogels and remain viable for at least 2 weeks, although apoptosis is observed in phGG-MA. Importantly, encapsulated MSCs and NCs show signs of in vivo chondrogenesis in a subcutaneous implantation of iGG-MA. Altogether, the data endorse the potential use of modified gellan gum-based hydrogel as a suitable material in NP tissue engineering. Copyright © 2014 John Wiley & Sons, Ltd. Copyright © 2014 John Wiley & Sons, Ltd.

Intervertebral Disc Cells Produce Interleukins Found in Patients with Back Pain.

PubMed

Zhang, Yejia; Chee, Ana; Shi, Peng; Adams, Sherrill L; Markova, Dessislava Z; Anderson, David Greg; Smith, Harvey E; Deng, Youping; Plastaras, Christopher T; An, Howard S

2016-06-01

To examine the link between cytokines in intervertebral disc (IVD) tissues and axial back pain. In vitro study with human IVD cells cultured from cadaveric donors and annulus fibrosus (AF) tissues from patients. Cultured nucleus pulposus (NP) and AF cells were stimulated with interleukin (IL)-1β. IL-8 and IL-7 gene expression was analyzed using real-time polymerase chain reaction. IL-8 protein was quantified by enzyme-linked immunosorbent assay. After IL-1β stimulation, IL-8 gene expression increased 26,541 fold in NP cells and 22,429 fold in AF cells, whereas protein released by the NP and AF cells increased 2,389- and 1,784-fold, respectively. IL-7 gene expression increased 3.3-fold in NP cells (P < 0.05).Cytokine profiles in AF tissues collected from patients undergoing surgery for back pain (painful group) or scoliosis (controls) were compared by cytokine array. IL-8 protein in the AF tissues from patients with back pain was 1.81-fold of that in controls. IL-7 and IL-10 in AF tissues from the painful group were 6.87 and 4.63 times greater than the corresponding values in controls, respectively (P < 0.05). Inflammatory mediators found in AF tissues from patients with discogenic back pain are likely produced by IVD cells and may play a key role in back pain.

Changes in digestibility and cell-wall constituents of some agricultural by-products due to gamma irradiation and urea treatments

NASA Astrophysics Data System (ADS)

Al-Masri, M. R.; Guenther, K. D.

1999-07-01

The effects of different doses of gamma irradiation (0, 100, 150, 200 kGy) or different concentrations of urea (0, 2, 3 and 5 g urea/100 g DM) on in-vitro organic matter digestibility (IVOMD), digestible energy (IVDE), gross energy (GE) and cell-wall constituents: neutral-detergent fibre, acid-detergent fibre and acid-detergent lignin, have been evaluated in wheat straw, cotton seed shell, peanut shell, soybean shell, extracted olive cake and extracted unpeeled sunflower seeds. The results indicated that gamma irradiation or urea treatments increased the digestible energy values significantly ( P<0.05) and these were attributed to the increases IVOMD and decreases cell-wall constituents of treated samples. The experimental agricultural by-products do not respond to the treatments in the same amount in increasing the IVOMD. There was no significant effect of irradiation and urea treatments on GE. Combined treatments had slightly less effect in increasing IVDE as the addition of both effects. The treatment of 200 kGy and 5% urea resulted in a larger increase in the digestible energy and a better effect by reducing the concentration of the cell-wall constituents even more than what occurred using a single treatment. However, the combination of irradiation with urea treatments could reduce the applied irradiation doses for increasing the IVDE in some studied agricultural by-products.

[Biology and mechanobiology of the intervertebral disc].

PubMed

González Martínez, Emilio; García-Cosamalón, José; Cosamalón-Gan, Iván; Esteban Blanco, Marta; García-Suarez, Olivia; Vega, José A

The intervertebral disc (IVD) is noted for its low cell content, and being the largest avascular structure of human body. The low amount of cells in the disc have to adapt to an anaerobic metabolism with low oxygen pressure and acidic pH. Apart from surviving in an adverse microenvironment, they are exposed to a high level of mechanical stress. The biological adaptation of cells to acidosis and hyperosmolarity conditions are regulated by mechanoproteins, which are responsible for converting a mechanical signal into a cellular response, thus modifying its gene expression. Mechanobiology helps us to better understand the pathophysiology of IVD and its potential biological repair. Copyright © 2016 Sociedad Española de Neurocirugía. Publicado por Elsevier España, S.L.U. All rights reserved.

Quantitative T2 Magnetic Resonance Imaging Compared to Morphological Grading of the Early Cervical Intervertebral Disc Degeneration: An Evaluation Approach in Asymptomatic Young Adults

PubMed Central

Han, Zhihua; Shao, Lixin; Xie, Yan; Wu, Jianhong; Zhang, Yan; Xin, Hongkui; Ren, Aijun; Guo, Yong; Wang, Deli; He, Qing; Ruan, Dike

2014-01-01

Objective The objective of this study was to evaluate the efficacy of quantitative T2 magnetic resonance imaging (MRI) for quantifying early cervical intervertebral disc (IVD) degeneration in asymptomatic young adults by correlating the T2 value with Pfirrmann grade, sex, and anatomic level. Methods Seventy asymptomatic young subjects (34 men and 36 women; mean age, 22.80±2.11 yr; range, 18–25 years) underwent 3.0-T MRI to obtain morphological data (one T1-fast spin echo (FSE) and three-plane T2-FSE, used to assign a Pfirrmann grade (I–V)) and for T2 mapping (multi-echo spin echo). T2 values in the nucleus pulposus (NP, n = 350) and anulus fibrosus (AF, n = 700) were obtained. Differences in T2 values between sexes and anatomic level were evaluated, and linear correlation analysis of T2 values versus degenerative grade was conducted. Findings Cervical IVDs of healthy young adults were commonly determined to be at Pfirrmann grades I and II. T2 values of NPs were significantly higher than those of AF at all anatomic levels (P<0.000). The NP, anterior AF and posterior AF values did not differ significantly between genders at the same anatomic level (P>0.05). T2 values decreased linearly with degenerative grade. Linear correlation analysis revealed a strong negative association between the Pfirrmann grade and the T2 values of the NP (P = 0.000) but not the T2 values of the AF (P = 0.854). However, non-degenerated discs (Pfirrmann grades I and II) showed a wide range of T2 relaxation time. T2 values according to disc degeneration level classification were as follows: grade I (>62.03 ms), grade II (54.60–62.03 ms), grade III (<54.60 ms). Conclusions T2 quantitation provides a more sensitive and robust approach for detecting and characterizing the early stage of cervical IVD degeneration and to create a reliable quantitative in healthy young adults. PMID:24498384

Dynamic pressurization induces transition of notochordal cells to a mature phenotype while retaining production of important patterning ligands from development

PubMed Central

2013-01-01

Introduction Notochordal cells (NCs) pattern aneural and avascular intervertebral discs (IVDs), and their disappearance, is associated with onset of IVD degeneration. This study induced and characterized the maturation of nucleus pulposus (NP) tissue from a gelatinous NC-rich structure to a matrix-rich structure populated by small NP cells using dynamic pressurization in an ex vivo culture model, and also identified soluble factors from NCs with therapeutic potential. Methods Porcine NC-rich NP tissue was cultured and loaded with hydrostatic pressure (0.5 to 2 MPa at 0.1 Hz for 2 hours) either Daily, for 1 Dose, or Control (no pressurization) groups for up to eight days. Cell phenotype and tissue maturation was characterized with measurements of cell viability, cytomorphology, nitric oxide, metabolic activity, matrix composition, gene expression, and proteomics. Results Daily pressurization induced transition of NCs to small NP cells with 73.8%, 44%, and 28% NCs for Control, 1 Dose and Daily groups, respectively (P < 0.0002) and no relevant cell death. Dynamic loading matured NP tissue by significantly increasing metabolic activity and accumulating Safranin-O-stained matrix. Load-induced maturation was also apparent from the significantly decreased glycolytic, cytoskeletal (Vimentin) and stress-inducible (HSP70) proteins assessed with proteomics. Loading increased the production of bioactive proteins Sonic Hedgehog (SHH) and Noggin, and maintained Semaphorin3A (Sema3A). Discussion NP tissue maturation was induced from dynamic hydrostatic pressurization in a controlled ex vivo environment without influence from systemic effects or surrounding structures. NCs transitioned into small nonvacuolated NP cells probably via differentiation as evidenced by high cell viability, lack of nitric oxide and downregulation of stress-inducible and cytoskeletal proteins. SHH, Sema3A, and Noggin, which have patterning and neurovascular-inhibiting properties, were produced in both notochordal and matured porcine NP. Results therefore provide an important piece of evidence suggesting the transition of NCs to small NP cells is a natural part of aging and not the initiation of degeneration. Bioactive candidates identified from young porcine IVDs may be isolated and harnessed for therapies to target discogenic back pain. PMID:24427812

Intracranial pressure monitoring in severe blunt head trauma: does the type of monitoring device matter?

PubMed

Aiolfi, Alberto; Khor, Desmond; Cho, Jayun; Benjamin, Elizabeth; Inaba, Kenji; Demetriades, Demetrios

2018-03-01

OBJECTIVE Intracranial pressure (ICP) monitoring has become the standard of care in the management of severe head trauma. Intraventricular devices (IVDs) and intraparenchymal devices (IPDs) are the 2 most commonly used techniques for ICP monitoring. Despite the widespread use of these devices, very few studies have investigated the effect of device type on outcomes. The purpose of the present study was to compare outcomes between 2 types of ICP monitoring devices in patients with isolated severe blunt head trauma. METHODS This retrospective observational study was based on the American College of Surgeons Trauma Quality Improvement Program database, which was searched for all patients with isolated severe blunt head injury who had an ICP monitor placed in the 2-year period from 2013 to 2014. Extracted variables included demographics, comorbidities, mechanisms of injury, head injury specifics (epidural, subdural, subarachnoid, intracranial hemorrhage, and diffuse axonal injury), Abbreviated Injury Scale (AIS) score for each body area, Injury Severity Score (ISS), vital signs in the emergency department, and craniectomy. Outcomes included 30-day mortality, complications, number of ventilation days, intensive care unit and hospital lengths of stay, and functional independence. RESULTS During the study period, 105,721 patients had isolated severe traumatic brain injury (head AIS score ≥ 3). Overall, an ICP monitoring device was placed in 2562 patients (2.4%): 1358 (53%) had an IVD and 1204 (47%) had an IPD. The severity of the head AIS score did not affect the type of ICP monitoring selected. There was no difference in the median ISS; ISS > 15; head AIS Score 3, 4, or 5; or the need for craniectomy between the 2 device groups. Unadjusted 30-day mortality was significantly higher in the group with IVDs (29% vs 25.5%, p = 0.046); however, stepwise logistic regression analysis showed that the type of ICP monitoring was not an independent risk factor for death, complications, or functional outcome at discharge. CONCLUSIONS This study demonstrated that compliance with the Brain Trauma Foundation guidelines for ICP monitoring is poor. In isolated severe blunt head injuries, the type of ICP monitoring device does not have any effect on survival, systemic complications, or functional outcome.

45 CFR 302.13 - Plan amendments.

Code of Federal Regulations, 2010 CFR

2010-10-01

... amended whenever necessary to reflect new or revised Federal statutes or regulations, or material change in any phase of State law, organization, policy of IV-D agency operation. (b) Federal financial...

45 CFR 302.13 - Plan amendments.

Code of Federal Regulations, 2013 CFR

2013-10-01

... amended whenever necessary to reflect new or revised Federal statutes or regulations, or material change in any phase of State law, organization, policy of IV-D agency operation. (b) Federal financial...

45 CFR 302.13 - Plan amendments.

Code of Federal Regulations, 2012 CFR

2012-10-01

... amended whenever necessary to reflect new or revised Federal statutes or regulations, or material change in any phase of State law, organization, policy of IV-D agency operation. (b) Federal financial...

45 CFR 302.13 - Plan amendments.

Code of Federal Regulations, 2014 CFR

2014-10-01

... amended whenever necessary to reflect new or revised Federal statutes or regulations, or material change in any phase of State law, organization, policy of IV-D agency operation. (b) Federal financial...

45 CFR 302.39 - Standards for program operation.

Code of Federal Regulations, 2011 CFR

2011-10-01

... (CHILD SUPPORT ENFORCEMENT PROGRAM), ADMINISTRATION FOR CHILDREN AND FAMILIES, DEPARTMENT OF HEALTH AND... provide that the IV-D agency will comply with the standards for program operation and the organizational...

The effects of cyclic tensile strain on the organisation and expression of cytoskeletal elements in bovine intervertebral disc cells: an in vitro study.

PubMed

Li, S; Jia, X; Duance, V C; Blain, E J

2011-06-20

It is still relatively unclear how intervertebral disc (IVD) cells sense a mechanical stimulus and convert this signal into a biochemical response. Previous studies demonstrated that the cytoskeletal elements are mechano-responsive in many cell types and may contribute to mechano-signalling pathways. The objective of this study was to determine the response of cells from the outer annulus fibrosus (OAF) to physiological levels of cyclic tensile strain; further, cells from the nucleus pulposus (NP) were also subjected to an identical loading regime to compare biological responses across the IVD populations. We determined whether the organisation and expression of the major cytoskeletal elements and their associated accessory proteins are responsive to mechanical stimulation in these cells, and whether these changes correlated with either a catabolic or anabolic phenotype. OAF and NP cells from immature bovine IVD were seeded onto Flexcell® type I collagen coated plates. Cells were subjected to cyclic tensile strain (10 %, 1 Hz) for 60 minutes. Post-loading, cells were processed for immunofluorescence microscopy, RNA extracted for quantitative PCR and protein extracted for Western blotting analysis. F-actin reorganisation was evident in OAF and NP cells subjected to tensile strain; strain induced β-actin at the transcriptional and translational level in OAF cells. β-tubulin mRNA and protein synthesis increased in strained OAF cells, but vimentin expression was significantly inhibited. Cytoskeletal element organisation and expression were less responsive to strain in NP cells. Tensile strain increased type I collagen and differentially regulated extracellular matrix (ECM)-degrading enzymes' mRNA levels in OAF cells. Strain induced type II collagen transcription in NP cells, but had no effect on the transcription of any other genes analysed. Tensile strain induces different mechano-responses in the organisation and/or expression of cytoskeletal elements and on markers of IVD metabolism. Differential mechano-regulation of anabolic and catabolic ECM components in the OAF and NP populations reflects their respective mechanical environments in situ.

Clinical and analytical evaluation of the new Aptima Mycoplasma genitalium assay, with data on M. genitalium prevalence and antimicrobial resistance in M. genitalium in Denmark, Norway and Sweden in 2016.

PubMed

Unemo, M; Salado-Rasmussen, K; Hansen, M; Olsen, A O; Falk, M; Golparian, D; Aasterød, M; Ringlander, J; Nilsson, C Stezckó; Sundqvist, M; Schønning, K; Moi, H; Westh, H; Jensen, J S

2018-05-01

Mycoplasma genitalium (MG) causes urethritis and cervicitis, potentially causing reproductive complications. Resistance in MG to first-line (azithromycin) and second-line (moxifloxacin) treatment has increased. We examined the clinical and analytical performance of the new Conformité Européene (CE)/in vitro diagnostics (IVD) Aptima Mycoplasma genitalium assay (CE/IVD AMG; Hologic); the prevalence of MG, Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG); and MG resistance to azithromycin and moxifloxacin in Denmark, Norway and Sweden in 2016. From February 2016 to February 2017, urogenital and extragenital (only in Denmark) specimens from consecutive attendees at three sexually transmitted disease clinics were tested with the CE/IVD AMG, the research-use-only MG Alt TMA-1 assay (Hologic), Aptima Combo 2 (CT/NG) assay and a laboratory-developed TaqMan real-time mgpB quantitative real-time PCR (qPCR). Resistance-associated mutations were determined by sequencing. Strains of MG and other mycoplasma species in different concentrations were also tested. In total 5269 patients were included. The prevalence of MG was 7.2% (382/5269; 4.9-9.8% in the countries). The sensitivity of the CE/IVD AMG, MG Alt TMA-1 and mgpB qPCR ranged 99.13-100%, 99.13-100% and 73.24-81.60%, respectively, in the countries. The specificity ranged 99.57-99.96%, 100% and 99.69-100%, respectively. The prevalence of resistance-associated mutations for azithromycin and moxifloxacin was 41.4% (120/290; 17.7-56.6%) and 6.6% (18/274; 4.1-10.2%), respectively. Multidrug resistance was found in all countries (2.7%; 1.1-4.2%). Both transcription-mediated amplification (TMA)-based MG assays had a highly superior sensitivity compared to the mgpB qPCR. The prevalence of MG and azithromycin resistance was high. Validated and quality-assured molecular tests for MG, routine resistance testing of MG-positive samples and antimicrobial resistance surveillance are crucial. Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.

45 CFR 302.13 - Plan amendments.

Code of Federal Regulations, 2011 CFR

2011-10-01

... effective in the State, whichever is later. (Approved by the Office of Management and Budget under control... in any phase of State law, organization, policy of IV-D agency operation. (b) Federal financial...

45 CFR 303.0 - Scope and applicability of this part.

Code of Federal Regulations, 2011 CFR

2011-10-01

... (CHILD SUPPORT ENFORCEMENT PROGRAM), ADMINISTRATION FOR CHILDREN AND FAMILIES, DEPARTMENT OF HEALTH AND... prescribes: (a) The minimum organizational and staffing requirements the State IV-D agency must meet in...

45 CFR 303.3 - Location of noncustodial parents in IV-D cases.

Code of Federal Regulations, 2012 CFR

2012-10-01

..., unemployment insurance, income taxation, driver's licenses, vehicle registration, and criminal records and... location, whichever occurs sooner. Quarterly attempts may be limited to automated sources, but must include...

45 CFR 303.3 - Location of noncustodial parents in IV-D cases.

Code of Federal Regulations, 2011 CFR

2011-10-01

..., unemployment insurance, income taxation, driver's licenses, vehicle registration, and criminal records and... location, whichever occurs sooner. Quarterly attempts may be limited to automated sources, but must include...

45 CFR 303.3 - Location of noncustodial parents in IV-D cases.

Code of Federal Regulations, 2013 CFR

2013-10-01

..., unemployment insurance, income taxation, driver's licenses, vehicle registration, and criminal records and... location, whichever occurs sooner. Quarterly attempts may be limited to automated sources, but must include...

45 CFR 303.3 - Location of noncustodial parents in IV-D cases.

Code of Federal Regulations, 2014 CFR

2014-10-01

..., unemployment insurance, income taxation, driver's licenses, vehicle registration, and criminal records and... location, whichever occurs sooner. Quarterly attempts may be limited to automated sources, but must include...

3D segmentation of annulus fibrosus and nucleus pulposus from T2-weighted magnetic resonance images

NASA Astrophysics Data System (ADS)

Castro-Mateos, Isaac; Pozo, Jose M.; Eltes, Peter E.; Del Rio, Luis; Lazary, Aron; Frangi, Alejandro F.

2014-12-01

Computational medicine aims at employing personalised computational models in diagnosis and treatment planning. The use of such models to help physicians in finding the best treatment for low back pain (LBP) is becoming popular. One of the challenges of creating such models is to derive patient-specific anatomical and tissue models of the lumbar intervertebral discs (IVDs), as a prior step. This article presents a segmentation scheme that obtains accurate results irrespective of the degree of IVD degeneration, including pathological discs with protrusion or herniation. The segmentation algorithm, employing a novel feature selector, iteratively deforms an initial shape, which is projected into a statistical shape model space at first and then, into a B-Spline space to improve accuracy. The method was tested on a MR dataset of 59 patients suffering from LBP. The images follow a standard T2-weighted protocol in coronal and sagittal acquisitions. These two image volumes were fused in order to overcome large inter-slice spacing. The agreement between expert-delineated structures, used here as gold-standard, and our automatic segmentation was evaluated using Dice Similarity Index and surface-to-surface distances, obtaining a mean error of 0.68 mm in the annulus segmentation and 1.88 mm in the nucleus, which are the best results with respect to the image resolution in the current literature.

Total disc replacement using a tissue-engineered intervertebral disc in vivo: new animal model and initial results.

PubMed

Gebhard, Harry; Bowles, Robby; Dyke, Jonathan; Saleh, Tatianna; Doty, Stephen; Bonassar, Lawrence; Härtl, Roger

2010-08-01

 Basic science Introduction:  Chronic back pain due to degenerative disc disease (DDD) is among the most important medical conditions causing morbidity and significant health care costs. Surgical treatment options include disc replacement or fusion surgery, but are associated with significant short- and long-term risks.1 Biological tissue-engineering of human intervertebral discs (IVD) could offer an important alternative.2 Recent in vitro data from our group have shown successful engineering and growth of ovine intervertebral disc composites with circumferentially aligned collagen fibrils in the annulus fibrosus (AF) (Figure 1).3 Figure 1 Tissue-engineered composite disc a Experimental steps to generate composite tissue-engineered IVDs3b Example of different AF formulations on collagen alignment in the AF. Second harmonic generation and two-photon excited fluorescence images of seeded collagen gels (for AF) of 1 and 2.5 mg/ml over time. At seeding, cells and collagen were homogenously distributed in the gels. Over time, AF cells elongated and collagen aligned parallel to cells. Less contraction and less alignment is noted after 3 days in the 2.5 mg/mL gel. c Imaging-based creation of a virtual disc model that will serve as template for the engineered disc. Total disc dimensions (AF and NP) were retrieved from micro-computer tomography (CT) (left images), and nucleus pulposus (NP) dimensions alone were retrieved from T2-weighted MRI images (right images). Merging of MRI and micro-CT models revealed a composite disc model (middle image)-Software: Microview, GE Healthcare Inc., Princeton, NJ; and slicOmatic v4.3, TomoVision, Montreal, Canada. d Flow chart describing the process for generating multi-lamellar tissue engineered IVDs. IVDs are produced by allowing cell-seeded collagen layers to contract around a cell-seeded alginate core (NP) over time Objective:  The next step is to investigate if biological disc implants survive, integrate, and restore function to the spine in vivo. A model will be developed that allows efficient in vivo testing of tissue-engineered discs of various compositions and characteristics.  Athymic rats were anesthetized and a dorsal approach was chosen to perform a microsurgical discectomy in the rat caudal spine (Fig. 2,Fig. 3). Control group I (n = 6) underwent discectomy only, Control group II (n = 6) underwent discectomy, followed by reimplantation of the autologous disc. Two treatment groups (group III, n = 6, 1 month survival; group IV, n = 6, 6 months survival) received a tissue-engineered composite disc implant. The rodents were followed clinically for signs of infection, pain level and wound healing. X-rays and magnetic resonance imaging (MRI) were assessed postoperatively and up to 6 months after surgery (Fig. 6,Fig. 7). A 7 Tesla MRI (Bruker) was implemented for assessment of the operated level as well as the adjacent disc (hydration). T2-weighted sequences were interpreted by a semiquantitative score (0 = no signal, 1 = weak signal, 2 = strong signal and anatomical features of a normal disc). Histology was performed with staining for proteoglycans (Alcian blue) and collagen (Picrosirius red) (Fig. 4,Fig. 5). Figure 2 Disc replacement surgery a Operative situs with native disc that has been disassociated from both adjacent vertebrae b Native disc (left) and tissue-engineered implant (right) c Implant in situ before wound closureAF: Annulus fi brosus, nP: nucleus pulposus, eP: endplate, M: Muscle, T: Tendon, s: skin, art: artery, GP: Growth plate, B: BoneFigure 3 Disc replacement surgery. Anatomy of the rat caudal disc space a Pircrosirius red stained axial cut of native disc space b Saffranin-O stained sagittal cut of native disc spaceFigure 4 Histologies of three separate motion segments from three different rats. Animal one = native IVD, Animal two = status after discectomy, Animal three = tissue-engineered implant (1 month) a-c H&E (overall tissue staining for light micrsocopy) d-f Alcian blue (proteoglycans) g-i Picrosirius red (collagen I and II)Figure 5 Histology from one motion segment four months after implantation of a bio-engineered disc construct a Picrosirius red staining (collagen) b Polarized light microscopy showing collagen staining and collagen organization in AF region c Increased Safranin-O staining (proteoglycans) in NP region of the disc implant d Higher magnification of figure 5c: Integration between implanted tissue-engineered total disc replacement and vertebral body boneFigure 6 MRI a Disc space height measurements in flash/T1 sequence (top: implant (714.0 micrometer), bottom: native disc (823.5 micrometer) b T2 sequence, red circle surrounding the implant NPFigure 7 7 Tesla MRI imaging of rat tail IVDs showing axial images (preliminary pilot data) a Diffusion tensor imaging (DTI) on two explanted rat tail discs in Formalin b Higher magnification of a, showing directional alignment of collagen fibers (red and green) when compared to the color ball on top which maps fibers' directional alignment (eg, fibers directing from left to right: red, from top to bottom: blue) c Native IVD in vivo (successful imaging of top and bottom of the IVD (red) d Gradient echo sequence (GE) showing differentiation between NP (light grey) and AF (dark margin) e GE of reimplanted tail IVD at the explantation level f T1Rho sequence demonstrating the NP (grey) within the AF (dark margin), containing the yellow marked region of interest for value acquisition (preliminary data are consistent with values reported in the literature). g T2 image of native IVD in vivo for monitoring of hydration (white: NP) Results:  The model allowed reproducible and complete discectomies as well as disc implantation in the rat tail spine without any surgical or postoperative complications. Discectomy resulted in immediate collapse of the disc space. Preliminary results indicate that disc space height was maintained after disc implantation in groups II, III and IV over time. MRI revealed high resolution images of normal intervertebral discs in vivo. Eight out of twelve animals (groups III and IV) showed a positive signal in T2-weighted images after 1 month (grade 0 = 4, grade 1 = 4, grade 2 = 4). Positive staining was seen for collagen as well as proteoglycans at the site of disc implantation after 1 month in each of the six animals with engineered implants (group III). Analysis of group IV showed positive T2 signal in five out of six animals and disc-height preservation in all animals after 6 months.  This study demonstrates for the first time that tissue-engineered composite IVDs with circumferentially aligned collagen fibrils survive and integrate with surrounding vertebral bodies when placed in the rat spine for up to 6 months. Tissue-engineered composite IVDs restored function to the rat spine as indicated by maintenance of disc height and vertebral alignment. A significant finding was that maintenance of the composite structure in group III was observed, with increased proteoglycan staining in the nucleus pulposus region (Figure 4d-f). Proteoglycan and collagen matrix as well as disc height preservation and positive T2 signals in MRI are promising parameters and indicate functionality of the implants.

42 CFR 433.136 - Definitions.

Code of Federal Regulations, 2010 CFR

2010-10-01

... the organizational unit in the State that has the responsibility for administering or supervising the administration of a State plan for child support enforcement under title IV-D of the Act. [49 FR 8984, Feb. 11...

TGFβ and BMP Dependent Cell Fate Changes Due to Loss of Filamin B Produces Disc Degeneration and Progressive Vertebral Fusions

PubMed Central

Zieba, Jennifer; Forlenza, Kimberly Nicole; Khatra, Jagteshwar Singh; Sarukhanov, Anna; Duran, Ivan; Rigueur, Diana; Lyons, Karen M.; Cohn, Daniel H.; Merrill, Amy E.; Krakow, Deborah

2016-01-01

Spondylocarpotarsal synostosis (SCT) is an autosomal recessive disorder characterized by progressive vertebral fusions and caused by loss of function mutations in Filamin B (FLNB). FLNB acts as a signaling scaffold by linking the actin cytoskleteon to signal transduction systems, yet the disease mechanisms for SCT remain unclear. Employing a Flnb knockout mouse, we found morphologic and molecular evidence that the intervertebral discs (IVDs) of Flnb–/–mice undergo rapid and progressive degeneration during postnatal development as a result of abnormal cell fate changes in the IVD, particularly the annulus fibrosus (AF). In Flnb–/–mice, the AF cells lose their typical fibroblast-like characteristics and acquire the molecular and phenotypic signature of hypertrophic chondrocytes. This change is characterized by hallmarks of endochondral-like ossification including alterations in collagen matrix, expression of Collagen X, increased apoptosis, and inappropriate ossification of the disc tissue. We show that conversion of the AF cells into chondrocytes is coincident with upregulated TGFβ signaling via Smad2/3 and BMP induced p38 signaling as well as sustained activation of canonical and noncanonical target genes p21 and Ctgf. These findings indicate that FLNB is involved in attenuation of TGFβ/BMP signaling and influences AF cell fate. Furthermore, we demonstrate that the IVD disruptions in Flnb–/–mice resemble aging degenerative discs and reveal new insights into the molecular causes of vertebral fusions and disc degeneration. PMID:27019229

Reliability of macroscopic grading of intervertebral disk degeneration in dogs by use of the Thompson system and comparison with low-field magnetic resonance imaging findings.

PubMed

Bergknut, Niklas; Grinwis, Guy; Pickee, Emile; Auriemma, Edoardo; Lagerstedt, Anne-Sofie; Hagman, Ragnvi; Hazewinkel, Herman A W; Meij, Björn P

2011-07-01

To evaluate the reliability of the Thompson system for use in grading the gross pathological changes of intervertebral disk (IVD) degeneration in dogs and to investigate the agreement between gross pathological findings and low-field (0.2-T) magnetic resonance imaging (MRI) findings. Vertebral columns from cadavers of 19 dogs of various ages, breeds, and origins. 182 intervertebral segments were collected from 19 canine cadavers. Sagittal T2-weighted MRI of the T11 through S1 portion of the vertebral column was performed within 24 hours after the dogs were euthanized. The vertebral columns were subsequently divided in the midsagittal plane, and high-resolution photographs were obtained of each intervertebral segment (end plate-disk-end plate). The MRI images and photographs were graded separately in a blinded manner by 4 observers who used both Pfirrmann and Thompson grading criteria. The interobserver agreement for Thompson scores ranged from 0.76 to 0.88, and the intraobserver agreement ranged from 0.88 to 0.94 (Cohen weighted κ analysis). Agreement between scores for the Pfirrmann and Thompson grading criteria was κ = 0.70. Grading of IVD degeneration in dogs by use of the Thompson system resulted in high interobserver and intraobserver agreement, and scores for the Thompson system had substantial agreement with low-field MRI findings graded by use of the Pfirrmann system. This suggested that low-field MRI can be used to diagnose IVD degeneration in dogs.

Emergence of community-acquired methicillin-resistant Staphylococcus aureus in an Iranian referral paediatric hospital.

PubMed

Mamishi, S; Mahmoudi, S; Bahador, A; Matini, H; Movahedi, Z; Sadeghi, R H; Pourakbari, B

2015-01-01

The epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) in hospitals has been changed in recent years due to the arrival of community-associated MRSA (CA-MRSA) strains into healthcare settings. The aim of this study is to investigate the distribution of staphylococcal cassette chromosome mec (SCCmec) type V as well as SCCmec IV subtypes, which have been associated with community-acquired infection among healthcare-associated MRSA (HA-MRSA) isolates. Antimicrobial susceptibility, SCCmec type, spa type and the presence of Panton-Valentine leukocidin (PVL) genes were determined for all HA-MRSA isolates in an Iranian referral hospital. In this study of 48 HA-MRSA isolates, 13 (27%), three (6.2%), five (10.4%) and one (2%) belonged to SCCmec subtypes IVa, IVb, IVc and IVd, respectively. Only two isolates (4.2%) belonged to SCCmec types V Notably, one isolate was found to harbour concurrent SCCmec subtypes IVb and IVd. MRSA containing SCCmec subtype IVb, IVc and IVd as well as type V isolates were all susceptible to chloramphenicol, clindamycin and rifampicin, while the sensitivity to these antibiotics was lower among MRSA containing SCCmec subtype IVa. The most frequently observed spa ttype was t037, accounting for 88% (22/25). Three other spa type was t002, t1816 and t4478. Large reservoirs of MRSA containing type IV subtypes and type V now exist in patients in this Iranian hospital. Therefore, effective infection control management in order to control the spread of CA-MRSA is highly recommended.

Gellan gum-based hydrogels for intervertebral disc tissue-engineering applications.

PubMed

Silva-Correia, J; Oliveira, J M; Caridade, S G; Oliveira, J T; Sousa, R A; Mano, J F; Reis, R L

2011-06-01

Intervertebral disc (IVD) degeneration is a challenging clinical problem that urgently demands viable nucleus pulposus (NP) implant materials. The best suited biomaterial for NP regeneration has yet to be identified, but it is believed that biodegradable hydrogel-based materials are promising candidates. In this work, we have developed ionic- and photo-crosslinked methacrylated gellan gum (GG-MA) hydrogels to be used in acellular and cellular tissue-engineering strategies for the regeneration of IVDs. The physicochemical properties of the developed hydrogels were investigated by Fourier-transform infrared spectroscopy, (1) H nuclear magnetic resonance and differential scanning calorimetry. The swelling ability and degradation rate of hydrogels were also analysed in phosphate-buffered saline solution at physiological pH for a period of 30 days. Additionally, the morphology and mechanical properties of the hydrogels were assessed under a scanning electron microscope and dynamic compression, respectively. An in vitro study was carried out to screen possible cytotoxicity of the gellan gum-based hydrogels by culturing rat lung fibroblasts (L929 cells) with hydrogel leachables up to 7 days. The results demonstrated that gellan gum was successfully methacrylated. We observed that the produced GG-MA hydrogels possess improved mechanical properties and lower water uptake ability and degradation rate as compared to gellan gum. This work also revealed that GG-MA hydrogels are non-cytotoxic in vitro, thus being promising biomaterials to be used in IVD tissue-engineering strategies. Copyright © 2010 John Wiley & Sons, Ltd.

Transarticular facet screw stabilization and dorsal laminectomy in 26 dogs with degenerative lumbosacral stenosis with instability.

PubMed

Hankin, Elyshia J; Jerram, Richard M; Walker, Alexander M; King, Michael D; Warman, Christopher G A

2012-07-01

To describe outcome after transarticular facet screw stabilization and dorsal laminectomy for treatment of dynamic degenerative lumbosacral stenosis (DLS) in 26 dogs. Retrospective case series. Dogs (n = 26) with dynamic DLS. Medical records (2004-2009) of dogs treated with transarticular facet screw stabilization and dorsal laminectomy were reviewed. Dogs (n = 26) were available for immediate postoperative follow-up, 21 dogs at 6 weeks, and 15 at greater than 6 months. Dogs were evaluated by radiographic assessment and owner questionnaire. Lumbosacral (LS) intervertebral disc (IVD) spaces were measured on pre and postoperative 6-week and 6-month radiographs. In 23 dogs, improvement in clinical signs occurred within 7 days of surgery. Overall postsurgical complication rate directly related to the surgical procedure was 15.4%. LS IVD space measurements taken immediately postoperatively, at 6 weeks, and ≥ 6 months were all significantly increased compared with preoperative measurements. All working dogs (4) returned to full work within 14 months. Most owners (85%) reported their dog was ambulating normally at 6 months with no perceptible lameness during normal activity. All owners perceived their dog's ability to walk, run, and jump after surgery to be improved. Transarticular facet screw stabilization and dorsal laminectomy maintains distraction of the LS IVD space for medium-to-large breed dogs with dynamic DLS with a high degree of owner satisfaction, and is comparable to other reported surgical techniques for DLS. © Copyright 2012 by The American College of Veterinary Surgeons.

45 CFR 310.10 - What are the functional requirements for the Model Tribal IV-D System?

Code of Federal Regulations, 2010 CFR

2010-10-01

... Number; and (E) Participant Identification Number; (ii) Delinquency and enforcement activities; (iii... operations and to assess program performance through the audit of financial and statistical data maintained...

26 CFR 48.4191-2 - Taxable medical device.

Code of Federal Regulations, 2013 CFR

2013-04-01

..., available at http://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfIVD/Search.cfm. (B) Devices that are... and listing database, available at http://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfrl/rl.cfm. (D...

45 CFR 301.0 - Scope and applicability of this part.

Code of Federal Regulations, 2011 CFR

2011-10-01

.... This part deals with the administration of title IV-D of the Social Security Act by the Federal... under the approved plan; review and audit of State and local expenditures; and reconsideration of...

[Diagnostic kits in parasitology: which controls?].

PubMed

Rossi, P

2004-06-01

The development of new diagnostic tools particularly for some parasitic "neglected diseases", is slowed or even hindered by limited resources assigned for basic and applied research in public institution and private sector. Even if the time-line and costs needed for developing a new In Vitro Diagnostic (IVD) test are generally lower compared to vaccines or new drugs, industry is poorly engaged in investing resources due to the perception of limited markets. To accelerate the development of diagnostics for the world's most deadly diseases, the World Health Organization's (WHO) Special Programme for Research and Training in Tropical Diseases (TDR), the United Nations Development Programme, the World Bank and the Gates Foundation, last year launched a new initiative, FIND (Foundation for Innovative New Diagnostics, www.finddiagnostics.org). The aim is to "apply the latest biotechnology innovations to develop and validate affordable diagnostic tests for diseases of the developing world". Ideally, a new diagnostic test should be accurately evaluated prior to use in medical practice. The first step would be a pre-clinical evaluation, an analytic study to determine its laboratory performance. A crucial point in this phase is the calibration of reagents (antigens, antibodies, DNA probes, etc.) against a standard reference preparation. WHO, through the WHO International Laboratories for Biological Standards, "provides International Biological Reference Preparations which serve as reference sources of defined biological activity expressed in an internationally agreed unit" (www.who.int/biologicals/IBRP/index.htm). Standardization allows "comparison of biological measurements worldwide" and ensures the reliability of diagnostic procedures. These preparations are generally intended for use in the characterization of the activity of secondary reference preparations (regional, national or in-house working standards). Unfortunately, international reference standards for parasitic diseases are not available at present, except for Toxoplasma antibodies. The first international standard reagent for Anti-Toxoplasma Serum was established in 1968 and at present, an international standard reference serum, Anti-toxoplasma serum, human TOXM is available at the National Institute for Biological Standards and Control (NIBSC) in UK. Several collaborative, multicenter studies were carried out to assess the performance of different methods and commercial tests for the diagnosis of toxoplasmosis, by providing to participating laboratories a panel of well-defined sera to be tested. A four-phase process following well-accepted methodological standards for the development of diagnostics, analogous to those internationally accepted for drugs and vaccines was recently proposed. The pre-clinical evaluation, the analytic study to assess sensitivity, specificity, predictive values in laboratory (phase I), should be followed by a proof of principle study to distinguish diseased from healthy persons in easily accessible populations (phase II). The evaluation of test performance in populations of intended use (phase III), and finally the delineation of cost-effectiveness and societal impact of new tests in comparison with existing tools (phase IV) should complete the validation procedure. In this context, national regulatory agencies play a major role in pre-market approval and post-market surveillance of IVDs. The European Community in 1998 approved a directive (Directive 98/79/EC) which rules the marketing of IVD medical devices, in order to harmonise the performance levels and standards in European countries. But, among IVDs for parasitic diseases, only those to detect congenital toxoplasmosis are submitted to defined procedures to provide the verification of products before their placing on the market and the surveillance after their marketing by a notified body, which perform appropriate examinations, tests and inspections to production facilities to verify if the device meets the requirements of the directive. In U.S.A., the Food and Drug Administration (FDA), through the Office of In Vitro Diagnostic Device Evaluation and Safety (OIVD), provides a comprehensive and regulatory activity for IVDs through pre-market evaluation and post-market surveillance. In developing countries, the scarcity of resources limits the procedures through which the national control authority can assure safety, quality and efficacy of products marketed, both imported and locally manufactured.

45 CFR 309.20 - Who submits a Tribal IV-D program application and where?

Code of Federal Regulations, 2010 CFR

2010-10-01

...) Applications must be submitted to the Office of Child Support Enforcement, Attention: Tribal Child Support Enforcement Program, 370 L'Enfant Promenade, SW., Washington, DC 20447, with a copy to the appropriate...

45 CFR 309.20 - Who submits a Tribal IV-D program application and where?

Code of Federal Regulations, 2011 CFR

2011-10-01

...) Applications must be submitted to the Office of Child Support Enforcement, Attention: Tribal Child Support Enforcement Program, 370 L'Enfant Promenade, SW., Washington, DC 20447, with a copy to the appropriate...

45 CFR 310.10 - What are the functional requirements for the Model Tribal IV-D System?

Code of Federal Regulations, 2013 CFR

2013-10-01

... Tribal financial management and expenditure information; (d) Distribute current support and arrearage..., process and monitor accounts receivable on all amounts owed, collected, and distributed with regard to: (1...

45 CFR 310.10 - What are the functional requirements for the Model Tribal IV-D System?

Code of Federal Regulations, 2012 CFR

2012-10-01

... Tribal financial management and expenditure information; (d) Distribute current support and arrearage..., process and monitor accounts receivable on all amounts owed, collected, and distributed with regard to: (1...

45 CFR 310.10 - What are the functional requirements for the Model Tribal IV-D System?

Code of Federal Regulations, 2014 CFR

2014-10-01

... Tribal financial management and expenditure information; (d) Distribute current support and arrearage..., process and monitor accounts receivable on all amounts owed, collected, and distributed with regard to: (1...

76 FR 72951 - Guidance for Industry and Food and Drug Administration Staff; Establishing the Performance...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-11-28

... recommendations for studies to establish the performance characteristics of in vitro diagnostic devices (IVDs... issuing this guidance to provide industry and Agency staff with recommendations for studies to establish...

45 CFR 310.10 - What are the functional requirements for the Model Tribal IV-D System?

Code of Federal Regulations, 2011 CFR

2011-10-01

... plan, including: (1) Identifying information such as Social Security numbers, names, dates of birth... operations and to assess program performance through the audit of financial and statistical data maintained...

[Companion diagnostics "POTELIGEO TEST IHC/FCM" used with "POTELIGEO" (mogamulizumab) for adult T-cell leukemia-lymphoma (ATL) treatment].

PubMed

Goami, Takeshi

2014-05-01

Personalized medicine offers the best treatment for individual patients, it is important for an IVD manufacturer to develop companion diagnostics in parallel with the development of new drugs through close cooperation with drug companies, and supply diagnostics companies with new drugs for physicians and patients. We received approval for a premarket approval application (PMA) for two in vitro diagnostic ("IVD") reagents, POTELIGEO TEST IHC and POTELIGEO TEST FCM ("POTELIGEO TEST"), in March 2012, and subsequently launched POTELIGEO TEST in May 2012. POTELIGEO TEST is a companion diagnostic used with POTELIGEO for which Kyowa Hakko Kirin has a new drug application (NDA) that was approved in March 2012, and is designed to help physicians identify appropriate subpopulations of adult T-cell leukemia-lymphoma(ATL) patients who are most likely to respond to POTELIGEO 20 mg (mogamulizumab) Injection ("POTELIGEO").

Spine Patterning Is Guided by Segmentation of the Notochord Sheath.

PubMed

Wopat, Susan; Bagwell, Jennifer; Sumigray, Kaelyn D; Dickson, Amy L; Huitema, Leonie F A; Poss, Kenneth D; Schulte-Merker, Stefan; Bagnat, Michel

2018-02-20

The spine is a segmented axial structure made of alternating vertebral bodies (centra) and intervertebral discs (IVDs) assembled around the notochord. Here, we show that, prior to centra formation, the outer epithelial cell layer of the zebrafish notochord, the sheath, segments into alternating domains corresponding to the prospective centra and IVD areas. This process occurs sequentially in an anteroposterior direction via the activation of Notch signaling in alternating segments of the sheath, which transition from cartilaginous to mineralizing domains. Subsequently, osteoblasts are recruited to the mineralized domains of the notochord sheath to form mature centra. Tissue-specific manipulation of Notch signaling in sheath cells produces notochord segmentation defects that are mirrored in the spine. Together, our findings demonstrate that notochord sheath segmentation provides a template for vertebral patterning in the zebrafish spine. Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.

The role of transient receptor potential channels in joint diseases.

PubMed

Krupkova, O; Zvick, J; Wuertz-Kozak, K

2017-10-10

Transient receptor potential channels (TRP channels) are cation selective transmembrane receptors with diverse structures, activation mechanisms and physiological functions. TRP channels act as cellular sensors for a plethora of stimuli, including temperature, membrane voltage, oxidative stress, mechanical stimuli, pH and endogenous, as well as, exogenous ligands, thereby illustrating their versatility. As such, TRP channels regulate various functions in both excitable and non-excitable cells, mainly by mediating Ca2+ homeostasis. Dysregulation of TRP channels is implicated in many pathologies, including cardiovascular diseases, muscular dystrophies and hyperalgesia. However, the importance of TRP channel expression, physiological function and regulation in chondrocytes and intervertebral disc (IVD) cells is largely unexplored. Osteoarthritis (OA) and degenerative disc disease (DDD) are chronic age-related disorders that significantly affect the quality of life by causing pain, activity limitation and disability. Furthermore, currently available therapies cannot effectively slow-down or stop progression of these diseases. Both OA and DDD are characterised by reduced tissue cellularity, enhanced inflammatory responses and molecular, structural and mechanical alterations of the extracellular matrix, hence affecting load distribution and reducing joint flexibility. However, knowledge on how chondrocytes and IVD cells sense their microenvironment and respond to its changes is still limited. In this review, we introduced six families of mammalian TRP channels, their mechanisms of activation, as well as, activation-driven cellular consequences. We summarised the current knowledge on TRP channel expression and activity in chondrocytes and IVD cells, as well as, the significance of TRP channels as therapeutic targets for the treatment of OA and DDD.

Three-dimensional finite element modeling of pericellular matrix and cell mechanics in the nucleus pulposus of the intervertebral disk based on in situ morphology.

PubMed

Cao, Li; Guilak, Farshid; Setton, Lori A

2011-02-01

Nucleus pulposus (NP) cells of the intervertebral disk (IVD) have unique morphological characteristics and biologic responses to mechanical stimuli that may regulate maintenance and health of the IVD. NP cells reside as single cell, paired or multiple cells in a contiguous pericellular matrix (PCM), whose structure and properties may significantly influence cell and extracellular matrix mechanics. In this study, a computational model was developed to predict the stress-strain, fluid pressure and flow fields for cells and their surrounding PCM in the NP using three-dimensional (3D) finite element models based on the in situ morphology of cell-PCM regions of the mature rat NP, measured using confocal microscopy. Three-dimensional geometries of the extracellular matrix and representative cell-matrix units were used to construct 3D finite element models of the structures as isotropic and biphasic materials. In response to compressive strain of the extracellular matrix, NP cells and PCM regions were predicted to experience volumetric strains that were 1.9-3.7 and 1.4-2.1 times greater than the extracellular matrix, respectively. Volumetric and deviatoric strain concentrations were generally found at the cell/PCM interface, while von Mises stress concentrations were associated with the PCM/extracellular matrix interface. Cell-matrix units containing greater cell numbers were associated with higher peak cell strains and lower rates of fluid pressurization upon loading. These studies provide new model predictions for micromechanics of NP cells that can contribute to an understanding of mechanotransduction in the IVD and its changes with aging and degeneration.

A Structurally and Functionally Biomimetic Biphasic Scaffold for Intervertebral Disc Tissue Engineering

PubMed Central

Choy, Andrew Tsz Hang; Chan, Barbara Pui

2015-01-01

Tissue engineering offers high hopes for the treatment of intervertebral disc (IVD) degeneration. Whereas scaffolds of the disc nucleus and annulus have been extensively studied, a truly biomimetic and mechanically functional biphasic scaffold using naturally occurring extracellular matrix is yet to be developed. Here, a biphasic scaffold was fabricated with collagen and glycosaminoglycans (GAGs), two of the most abundant extracellular matrix components in the IVD. Following fabrication, the scaffold was characterized and benchmarked against native disc. The biphasic scaffold was composed of a collagen-GAG co-precipitate making up the nucleus pulposus-like core, and this was encapsulated in multiple lamellae of photochemically crosslinked collagen membranes comprising the annulus fibrosus-like lamellae. On mechanical testing, the height of our engineered disc recovered by ~82-89% in an annulus-independent manner, when compared with the 99% recovery exhibited by native disc. The annulus-independent nature of disc height recovery suggests that the fluid replacement function of the engineered nucleus pulposus core might mimic this hitherto unique feature of native disc. Biphasic scaffolds comprised of 10 annulus fibrosus-like lamellae had the best overall mechanical performance among the various designs owing to their similarity to native disc in most aspects, including elastic compliance during creep and recovery, and viscous compliance during recovery. However, the dynamic mechanical performance (including dynamic stiffness and damping factor) of all the biphasic scaffolds was similar to that of the native discs. This study contributes to the rationalized design and development of a biomimetic and mechanically viable biphasic scaffold for IVD tissue engineering. PMID:26115332

Spinal Health during Unloading and Reloading Associated with Spaceflight

PubMed Central

Green, David A.; Scott, Jonathan P. R.

2018-01-01

Spinal elongation and back pain are recognized effects of exposure to microgravity, however, spinal health has received relatively little attention. This changed with the report of an increased risk of post-flight intervertebral disc (IVD) herniation and subsequent identification of spinal pathophysiology in some astronauts post-flight. Ground-based analogs, particularly bed rest, suggest that a loss of spinal curvature and IVD swelling may be factors contributing to unloading-induced spinal elongation. In flight, trunk muscle atrophy, in particular multifidus, may precipitate lumbar curvature loss and reduced spinal stability, but in-flight (ultrasound) and pre- and post-flight (MRI) imaging have yet to detect significant IVD changes. Current International Space Station missions involve short periods of moderate-to-high spinal (axial) loading during running and resistance exercise, superimposed upon a background of prolonged unloading (microgravity). Axial loading acting on a dysfunctional spine, weakened by anatomical changes and local muscle atrophy, might increase the risk of damage/injury. Alternatively, regular loading may be beneficial. Spinal pathology has been identified in-flight, but there are few contemporary reports of in-flight back injury and no recent studies of post-flight back injury incidence. Accurate routine in-flight stature measurements, in- and post-flight imaging, and tracking of pain and injury (herniation) for at least 2 years post-flight is thus warranted. These should be complemented by ground-based studies, in particular hyper buoyancy floatation (HBF) a novel analog of spinal unloading, in order to elucidate the mechanisms and risk of spinal injury, and to evaluate countermeasures for exploration where injury could be mission critical. PMID:29403389

Glomeruloid Microvascular Proliferation, Desmoplasia, and High Proliferative Index as Potential Indicators of High Grade Canine Choroid Plexus Tumors.

PubMed

Muscatello, Luisa Vera; Avallone, Giancarlo; Serra, Fabienne; Seuberlich, Torsten; Mandara, Maria Teresa; Sisó, Silvia; Brunetti, Barbara; Oevermann, Anna

2018-05-01

Choroid plexus tumors (CPT) are intraventricular neoplasms accounting for 10% of all primary central nervous system tumors in dogs. They are frequently classified according to the human WHO classification into choroid plexus papilloma (CPP, grade I), atypical CPP (aCPP, grade II), and choroid plexus carcinoma (CPC, grade III). Histological features observed in canine CPT such as increased vascular density (IVD) and glomeruloid microvascular proliferation (GMVP) are not part of the WHO classification. This multi-centric study aimed to investigate tumor-associated vascular hyperplasia in dogs by determining the prevalence of GMVP and IVD in 52 canine CPT and their association with tumor grade. In addition, the expression of angiogenic factors was assessed by immunohistochemistry in 25 tumors to investigate the pathogenesis of tumor-associated vascular hyperplasia. Based on the classical histological hallmarks, this study of 52 CPT identified 22 (42%) CPP (grade I) and 30 of (58%) CPC (grade III). GMVP was more prevalent in CPC (13/30; 43%) than CPP (1/22; 4%), whereas IVD occurred to a similar extent in CPP and CPC. Desmoplasia was more common in CPC (19/30; 63%) than CPP (2/22; 9%), and similarly, the proliferative index (PI) of neoplastic epithelium was significantly higher in CPC (5.14%) than CPP (0.94%). The majority of CPT expressed platelet-derived growth factor (PDGF), PDGFRα, PDGFRβ, and vascular endothelial growth factor (VEGF) irrespective of tumor grade or tumor-associated vascular hyperplasia. These results suggest that tumor-associated GMVP, desmoplasia, and PI may serve as histological indicators of malignancy in CPT.

Spinal Health during Unloading and Reloading Associated with Spaceflight.

PubMed

Green, David A; Scott, Jonathan P R

2017-01-01

Spinal elongation and back pain are recognized effects of exposure to microgravity, however, spinal health has received relatively little attention. This changed with the report of an increased risk of post-flight intervertebral disc (IVD) herniation and subsequent identification of spinal pathophysiology in some astronauts post-flight. Ground-based analogs, particularly bed rest, suggest that a loss of spinal curvature and IVD swelling may be factors contributing to unloading-induced spinal elongation. In flight, trunk muscle atrophy, in particular multifidus , may precipitate lumbar curvature loss and reduced spinal stability, but in-flight (ultrasound) and pre- and post-flight (MRI) imaging have yet to detect significant IVD changes. Current International Space Station missions involve short periods of moderate-to-high spinal (axial) loading during running and resistance exercise, superimposed upon a background of prolonged unloading (microgravity). Axial loading acting on a dysfunctional spine, weakened by anatomical changes and local muscle atrophy, might increase the risk of damage/injury. Alternatively, regular loading may be beneficial. Spinal pathology has been identified in-flight, but there are few contemporary reports of in-flight back injury and no recent studies of post-flight back injury incidence. Accurate routine in-flight stature measurements, in- and post-flight imaging, and tracking of pain and injury (herniation) for at least 2 years post-flight is thus warranted. These should be complemented by ground-based studies, in particular hyper buoyancy floatation (HBF) a novel analog of spinal unloading, in order to elucidate the mechanisms and risk of spinal injury, and to evaluate countermeasures for exploration where injury could be mission critical.

Estrus resynchronization in ewes with unknown pregnancy status.

PubMed

Miranda, Vladinis O; Oliveira, Fernando C; Dias, Jenniffer H; Vargas Júnior, Sergio F; Goularte, Karina L; Sá Filho, Manoel F; Sá Filho, Ocilon G de; Baldassarre, Hernan; Vieira, Arnaldo D; Lucia, Thomaz; Gasperin, Bernardo G

2018-01-15

Although fixed-time artificial insemination (FTAI) protocols are available for sheep, estrus resynchronization has not been previously reported. The objectives of this study were to evaluate the effect of estrus resynchronization with exogenous progestogen on endogenous progesterone levels and to compare pregnancy rates after two consecutive estrus synchronizations in ewes. In Experiment 1, ewes (n = 20) received an intravaginal device (IVD) containing 60 mg medroxyprogesterone acetate (MPA) for 10 days. At the IVD withdrawal (D0), ewes received 250 IU eCG and were allocated into two treatments: either no further treatment (Control; n = 10) or estrus resynchronization (Resynch; n = 10) from D12 to D19. Serum progesterone (P4) levels did not differ at D12 and D19 (P > 0.05), but were greater at D15 for the Control compared with the Resynch group (P < 0.05). In experiment 2, ewes (n = 250) were submitted to a first synchronization protocol followed by estrus detection and either artificial insemination (AI) or natural mating (NM). Subsequently, ewes were divided into two groups: Control (n = 104): which received no further treatment and were bred by NM; and Resynch (n = 146): which were submitted to a second synchronization starting on D14 (first IVD withdrawal = D0) and to NM after second IVD withdrawal (D20). Cumulative pregnancy rates did not differ between the Control (67.3%, 70/104) and Resynch (62.3%, 91/146) groups. In a third experiment, ewes (n = 83) were bred by two consecutive FTAI within a 20-day interval. Pregnancy rates after the first (30.1%, 25/83) and the second FTAI (36.2%, 21/58) did not differ (P > 0.05). In conclusion, although exogenous progestogen supplementation reduced circulating levels of P4, pregnancy maintenance was unaffected. Estrus resynchronization in ewes is feasible, resulting in similar fertility after the first and the second services. The use of resynchronization coupled with artificial insemination using semen from genetically superior rams may potentially accelerate genetic improvement in sheep herds by allowing a higher differential selection compared with natural breeding. Copyright © 2017 Elsevier Inc. All rights reserved.

Novel immortal human cell lines reveal subpopulations in the nucleus pulposus.

PubMed

van den Akker, Guus G H; Surtel, Don A M; Cremers, Andy; Rodrigues-Pinto, Ricardo; Richardson, Stephen M; Hoyland, Judith A; van Rhijn, Lodewijk W; Welting, Tim J M; Voncken, Jan Willem

2014-06-27

Relatively little is known about cellular subpopulations in the mature nucleus pulposus (NP). Detailed understanding of the ontogenetic, cellular and molecular characteristics of functional intervertebral disc (IVD) cell populations is pivotal to the successful development of cell replacement therapies and IVD regeneration. In this study, we aimed to investigate whether phenotypically distinct clonal cell lines representing different subpopulations in the human NP could be generated using immortalization strategies. Nondegenerate healthy disc material (age range, 8 to 15 years) was obtained as surplus surgical material. Early passage NP monolayer cell cultures were initially characterized using a recently established NP marker set. NP cells were immortalized by simian virus 40 large T antigen (SV40LTag) and human telomerase reverse transcriptase expression. Immortalized cells were clonally expanded and characterized based on collagen type I, collagen type II, α1 (COL2A1), and SRY-box 9 (SOX9) protein expression profiles, as well as on expression of a subset of established in vivo NP cell lineage markers. A total of 54 immortal clones were generated. Profiling of a set of novel NP markers (CD24, CA12, PAX1, PTN, FOXF1 and KRT19 mRNA) in a representative set of subclones substantiated successful immortalization of multiple cellular subpopulations from primary isolates and confirmed their NP origin and/or phenotype. We were able to identify two predominant clonal NP subtypes based on their morphological characteristics and their ability to induce SOX9 and COL2A1 under conventional differentiation conditions. In addition, cluster of differentiation 24 (CD24)-negative NP responder clones formed spheroid structures in various culture systems, suggesting the preservation of a more immature phenotype compared to CD24-positive nonresponder clones. Here we report the generation of clonal NP cell lines from nondegenerate human IVD tissue and present a detailed characterization of NP cellular subpopulations. Differential cell surface marker expression and divergent responses to differentiation conditions suggest that the NP subtypes may correspond to distinct maturation stages and represent distinct NP cell subpopulations. Hence, we provide evidence that the immortalization strategy that we applied is capable of detecting cell heterogeneity in the NP. Our cell lines yield novel insights into NP biology and provide promising new tools for studies of IVD development, cell function and disease.

75 FR 68610 - Proposed Information Collection Activity; Comment Request

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78 FR 32258 - Proposed Information Collection Activity; Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2013-05-29

... DEPARTMENT OF HEALTH AND HUMAN SERVICES Administration for Children and Families Proposed... Levy (FAST Levy). OMB No.: New Collection. Description: State IV-D child support enforcement agencies... institutions. To assist states in fulfilling this statutory requirement the federal Office of Child Support...

Interactive Videodisc at California State University, Fullerton.

ERIC Educational Resources Information Center

Reisman, S.

In January 1987, California State University, Fullerton (CSUF), began to explore the potential of interactive videodisk (IVD) technology on its campus. The challenge of introducing an individualized instructional technology into a cost conscious and conventional teaching environment was formidable, considering the traditional orientation of…

Patterns of patient specific dosimetry in total body irradiation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Akino, Yuichi; Department of Radiation Oncology, Osaka University Graduate School of Medicine, Suita, Osaka 565-0871; McMullen, Kevin P.

2013-04-15

Purpose: Total body irradiation (TBI) has been used for bone marrow transplant for hematologic and immune deficiency conditions. The goal of TBI is to deliver a homogeneous dose to the entire body, with a generally accepted range of dose uniformity being within {+-}10% of the prescribed dose. The moving table technique for TBI could make dose uniform in whole body by adjusting couch speed. However, it is difficult to accurately estimate the actual dose by calculation and hence in vivo dosimetry (IVD) is routinely performed. Here, the authors present patterns of patient-specific IVD in 161 TBI patients treated at ourmore » institution. Methods: Cobalt-60 teletherapy unit (Model C9 Cobalt-60 teletherapy unit, Picker X-ray Corporation) with customized moving bed (SITI Industrial Products, Inc., Fishers, IN) were used for TBI treatment. During treatment, OneDose{sup TM} (Sicel Technology, NC) Metal Oxide-silicon Semiconductor Field Effect Transistor detectors were placed at patient body surface; both entrance and exit side of the beam at patient head, neck, mediastinum, umbilicus, and knee to estimate midplane dose. When large differences (>10%) between the prescribed and measured dose were observed, dose delivery was corrected for subsequent fractions by the adjustment of couch speed and/or bolus placement. Under IRB exempt status, the authors retrospectively analyzed the treatment records of 161 patients who received TBI treatment between 2006 and 2011. Results: Across the entire cohort, the median {+-} SD (range) percent variance between calculated and measured dose for head, neck, mediastinum, umbilicus, and knee was -2.3 {+-} 10.2% (-66.2 to +35.3), 1.1 {+-} 11.5% (-62.2 to +40.3), -1.9 {+-} 9.5% (-66.4 to +46.6), -1.1 {+-} 7.2% (-35.2 to +42.9), and 3.4 {+-} 12.2% (-47.9 to +108.5), respectively. More than half of treatments were within {+-}10% of the prescribed dose for all anatomical regions. For 80% of treatments (10%-90%), dose at the umbilicus was within {+-}10%. However, some large differences greater than 35% were also found at several points. For one case, the knee received double the prescribed dose. When the dose differences for multiple fractions were averaged, compliance ({+-}10%) between the prescription and measured dose was improved compared to the dose difference of the first single fraction, for example, as at umbilicus, which improved from 83.9% to 98.5%. Conclusions: Actual dose measurement analysis of TBI patients revealed a potentially wide variance from the calculated dose. Based from their IVD method for TBI using Cobalt-60 irradiator and moving table, {+-}10% over entire body is hard to achieve. However, it can be significantly improved with immediate feedback after the first fraction prior to subsequent treatments.« less

Adding value to daily chest X-rays in the ICU through education, restricted daily orders and indication-based prompting.

PubMed

Keveson, Benjamin; Clouser, Ryan D; Hamlin, Mark P; Stevens, Pamela; Stinnett-Donnelly, Justin M; Allen, Gilman B

2017-01-01

Chest X-rays (CXRs) are traditionally obtained daily in all patients on invasive mechanical ventilation (IMV) in the intensive care unit (ICU). We sought to reduce overutilisation of CXRs obtained in the ICU, using a multifaceted intervention to eliminate automated daily studies. We first educated ICU staff about the low diagnostic yield of automated daily CXRs, then removed the 'daily' option from the electronic health records-based ordering system, and added a query (CXR indicated or not indicated) to the ICU daily rounding checklist to prompt a CXR order when clinically warranted. We built a report from billing codes, focusing on all CXRs obtained on IMV census days in the medical (MICU) and surgical (SICU) ICUs, excluding the day of admission and days that a procedure warranting CXR was performed. This generated the number of CXRs obtained every 1000 'included' ventilator days (IVDs), the latter defined as not having an 'absolute' clinical indication for CXR. The average monthly number of CXRs on an IVD decreased from 919±90 (95% CI 877 to 963) to 330±87 (95% CI 295 to 354) per 1000 IVDs in the MICU, and from 995±69 (95% CI 947 to 1055) to 649±133 (95% CI 593 to 697) in the SICU. This yielded an estimated 1830 to 2066 CXRs avoided over 2 years and an estimated annual savings of $191 600 to $224 200. There was no increase in reported adverse events. ICUs can safely transition to a higher value strategy of indication-based chest imaging by educating staff, eliminating the 'daily' order option and adding a simplified prompt to avoid missing clinically indicated CXRs.

Expression and characterization of constitutive heat shock protein 70.1 (HSPA-1A) gene in in vitro produced and in vivo-derived buffalo (Bubalus bubalis) embryos.

PubMed

Sharma, G T; Nath, A; Prasad, S; Singhal, S; Singh, N; Gade, N E; Dubey, P K; Saikumar, G

2012-12-01

Cells are blessed with a group of stress protector molecules known as heat shock proteins (HSPs), amongst them HSP70, encoded by HSPA-1A gene, is most abundant and highly conserved protein. Variety of stresses hampers the developmental competence of embryos under in vivo and in vitro conditions. Present work was designed to study the quantitative expression of HSPA-1A mRNA in immature oocytes (IMO), matured oocytes (MO), in vitro produced (IVP) and in vivo-derived (IVD) buffalo embryos to assess the level of stress to which embryos are exposed under in vivo and in vitro culture conditions. Further, HSPA-1A gene sequence was analysed to determine its homology with other mammalian sequences. The mRNA expression analysis was carried out on 72 oocytes (40 IMO; 32 MO), 76 IVP and 55 IVD buffalo embryos. Expression of HSPA-1A was found in oocytes and throughout the developmental stages of embryos examined irrespective of the embryo source; however, higher (p < 0.05) expression was observed in 8-16 cell, morula and blastocyst stages of IVP embryos as compared to IVD embryos. Phylogenetic analysis of bubaline HSPA-1A revealed that it shares 91-98% identity with other mammalian sequences. It can be concluded that higher level of HSPA-1A mRNA in IVP embryos in comparison with in vivo-derived embryos is an indicator of cellular stress in IVP system. This study suggests need for further optimization of in vitro culture system in which HSPA-1A gene could be used as a stress biomarker during pre-implantation development. © 2012 Blackwell Verlag GmbH.

ISSLS PRIZE IN BASIC SCIENCE 2018: Growth differentiation factor-6 attenuated pro-inflammatory molecular changes in the rabbit anular-puncture model and degenerated disc-induced pain generation in the rat xenograft radiculopathy model.

PubMed

Miyazaki, Shingo; Diwan, Ashish D; Kato, Kenji; Cheng, Kevin; Bae, Won C; Sun, Yang; Yamada, Junichi; Muehleman, Carol; Lenz, Mary E; Inoue, Nozomu; Sah, Robert L; Kawakami, Mamoru; Masuda, Koichi

2018-04-01

To elucidate the effects of growth differentiation factor-6 (GDF6) on: (i) gene expression of inflammatory/pain-related molecules and structural integrity in the rabbit intervertebral disc (IVD) degeneration model, and (ii) sensory dysfunction and changes in pain-marker expression in dorsal nerve ganglia (DRGs) in the rat xenograft radiculopathy model. Forty-six adolescent rabbits received anular-puncture in two non-consecutive lumbar IVDs. Four weeks later, phosphate-buffered saline (PBS) or GDF6 (1, 10 or 100 µg) was injected into the nucleus pulposus (NP) of punctured discs and followed for 4 weeks for gene expression analysis and 12 weeks for structural analyses. For pain assessment, eight rabbits were sacrificed at 4 weeks post-injection and NP tissues of injected discs were transplanted onto L5 DRGs of 16 nude rats to examine mechanical allodynia. The rat DRGs were analyzed immunohistochemically. In GDF6-treated rabbit NPs, gene expressions of interleukin-6, tumor necrosis factor-α, vascular endothelial growth factor, prostaglandin-endoperoxide synthase 2, and nerve growth factor were significantly lower than those in the PBS group. GDF6 injections resulted in partial restoration of disc height and improvement of MRI disc degeneration grades with statistical significance in rabbit structural analyses. Allodynia induced by xenograft transplantation of rabbit degenerated NPs onto rat DRGs was significantly reduced by GDF6 injection. Staining intensities for ionized calcium-binding adaptor molecule-1 and calcitonin gene-related peptide in rat DRGs of the GDF6 group were significantly lower than those of the PBS group. GDF6 injection may change the pathological status of degenerative discs and attenuate degenerated IVD-induced pain.

A preliminary in vitro study into the use of IL-1Ra gene therapy for the inhibition of intervertebral disc degeneration

PubMed Central

Le Maitre, Christine L; Freemont, Anthony J; Hoyland, Judith A

2006-01-01

Conventional therapies for low back pain (LBP) are purely symptomatic and do not target the cause of LBP, which in approximately 40% of cases is caused by degeneration of the intervertebral disc (DIVD). Targeting therapies to inhibit the process of degeneration would be a potentially valuable treatment for LBP. There is increasing evidence for a role for IL-1 in DIVD. A natural inhibitor of IL-1 exists, IL-1Ra, which would be an ideal molecular target for inhibiting IL-1-mediated effects involved in DIVD and LBP. In this study, the feasibility of ex vivo gene transfer of IL-1Ra to the IVD was investigated. Monolayer and alginate cultures of normal and degenerate human intervertebral disc (IVD) cells were infected with an adenoviral vector carrying the IL-1Ra gene (Ad-IL-1Ra) and protein production measured using an enzyme-linked immunosorbent assay. The ability of these infected cells to inhibit the effects of IL-1 was also investigated. In addition, normal and degenerate IVD cells infected with Ad-IL-1Ra were injected into degenerate disc tissue explants and IL-1Ra production in these discs was assessed. This demonstrated that both nucleus pulposus and annulus fibrosus cells infected with Ad-IL-1Ra produced elevated levels of IL-1Ra for prolonged time periods, and these infected cells were resistant to IL-1. When the infected cells were injected into disc explants, IL-1Ra protein expression was increased which was maintained for 2 weeks of investigation. This in vitro study has shown that the use of ex vivo gene transfer to degenerate disc tissue is a feasible therapy for the inhibition of IL-1-mediated events during disc degeneration. PMID:16436110

45 CFR 303.100 - Procedures for income withholding.

Code of Federal Regulations, 2010 CFR

2010-10-01

... (CHILD SUPPORT ENFORCEMENT PROGRAM), ADMINISTRATION FOR CHILDREN AND FAMILIES, DEPARTMENT OF HEALTH AND... employer, whether the noncustodial parent has access to health insurance coverage at reasonable cost and, if so, the health insurance policy information. (b) Immediate withholding on IV-D cases. (1) In the...

45 CFR 302.39 - Standards for program operation.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 302.39 Public Welfare Regulations Relating to Public Welfare OFFICE OF CHILD SUPPORT ENFORCEMENT (CHILD SUPPORT ENFORCEMENT PROGRAM), ADMINISTRATION FOR CHILDREN AND FAMILIES, DEPARTMENT OF HEALTH AND... provide that the IV-D agency will comply with the standards for program operation and the organizational...

Mechanical Vibrations Reduce the Intervertebral Disc Swelling and Muscle Atrophy from Bed Rest

NASA Technical Reports Server (NTRS)

Holguin, Nilsson; Muir, Jesse; Evans, Harlan J.; Qin, Yi-Xian; Rubin, Clinton; Wagshul, Mark; Judex, Stefan

2007-01-01

Loss of functional weight bearing, such as experienced during space flight or bed rest (BR), distorts intervertebral disc (IVD) and muscle morphology. IVDs are avascular structures consisting of cells that may derive their nutrition and waste removal from the load induced fluid flow into and out of the disc. A diurnal cycle is produced by forces related to weight bearing and muscular activity, and comprised of a supine and erect posture over a 24 hr period. A diurnal cycle will include a disc volume change of approx. 10-13%. However, in space there are little or no diurnal changes because of the microgravity, which removes the gravitational load and compressive forces to the back muscles. The BR model and the etiology of the disc swelling and muscle atrophy could provide insight into those subjects confined to bed for chronic disease/injury and aging. We hypothesize that extremely low-magnitude, high frequency mechanical vibrations will abate the disc degeneration and muscle loss associated with long-term BR.

The rise in caesarean birth rate in Sagamu, Nigeria: reflection of changes in obstetric practice.

PubMed

Oladapo, O T; Sotunsa, J O; Sule-Odu, A O

2004-06-01

A retrospective and comparative study of women delivered by caesarean section over two different 3-year periods was conducted at Olabisi Onabanjo University Teaching Hospital, Sagamu, Nigeria. The caesarean section rate (CSR) increased from 10.3% in 1989-1991 to 23.1% in 2000-2003. The most frequent indication in both periods was different: prolonged/obstructed labour (20.0%) in 1989-1991 and antepartum haemorrhage (14.9%) in 2000-2003. Malpresentation, antepartum haemorrhage and pre-eclampsia/eclampsia were responsible for 51.7% of the difference in the CSR recorded between both periods. The CSR rose from 13.3% to 25.0% while the instrumental vaginal delivery (IVD) rate decreased significantly by 11.4% among the nulliparous women between the periods. Increase in CSR can be attributed mainly to reduction in IVD rate and alteration in the management of labour complications and induction policy. Strategies to reduce the CSR should cut across all indications and focus on encouraging instrumental vaginal deliveries, especially among nulliparous women.

Are animal models useful for studying human disc disorders/degeneration?

PubMed Central

Eisenstein, Stephen M.; Ito, Keita; Little, Christopher; Kettler, A. Annette; Masuda, Koichi; Melrose, James; Ralphs, Jim; Stokes, Ian; Wilke, Hans Joachim

2007-01-01

Intervertebral disc (IVD) degeneration is an often investigated pathophysiological condition because of its implication in causing low back pain. As human material for such studies is difficult to obtain because of ethical and government regulatory restriction, animal tissue, organs and in vivo models have often been used for this purpose. However, there are many differences in cell population, tissue composition, disc and spine anatomy, development, physiology and mechanical properties, between animal species and human. Both naturally occurring and induced degenerative changes may differ significantly from those seen in humans. This paper reviews the many animal models developed for the study of IVD degeneration aetiopathogenesis and treatments thereof. In particular, the limitations and relevance of these models to the human condition are examined, and some general consensus guidelines are presented. Although animal models are invaluable to increase our understanding of disc biology, because of the differences between species, care must be taken when used to study human disc degeneration and much more effort is needed to facilitate research on human disc material. PMID:17632738

45 CFR 301.1 - General definitions.

Code of Federal Regulations, 2012 CFR

2012-10-01

... that Act. Agent of a Child means a caretaker relative having custody of or responsibility for the child... or the child's resident parent to provide legal representation to the child or resident parent... IV-D. Federal PLS means the Parent Locator Service operated by the Office of Child Support...

45 CFR 301.1 - General definitions.

Code of Federal Regulations, 2013 CFR

2013-10-01

... that Act. Agent of a Child means a caretaker relative having custody of or responsibility for the child... or the child's resident parent to provide legal representation to the child or resident parent... IV-D. Federal PLS means the Parent Locator Service operated by the Office of Child Support...

45 CFR 301.1 - General definitions.

Code of Federal Regulations, 2014 CFR

2014-10-01

... that Act. Agent of a Child means a caretaker relative having custody of or responsibility for the child... or the child's resident parent to provide legal representation to the child or resident parent... IV-D. Federal PLS means the Parent Locator Service operated by the Office of Child Support...

45 CFR 305.65 - State cooperation in audit.

Code of Federal Regulations, 2010 CFR

2010-10-01

... PROGRAM PERFORMANCE MEASURES, STANDARDS, FINANCIAL INCENTIVES, AND PENALTIES § 305.65 State cooperation in... submitted on the Federal statistical and financial reports that will be used to calculate the State's performance. The State shall also make available personnel associated with the State's IV-D program to provide...

45 CFR 303.7 - Provision of services in interstate IV-D cases.

Code of Federal Regulations, 2010 CFR

2010-10-01

... ENFORCEMENT (CHILD SUPPORT ENFORCEMENT PROGRAM), ADMINISTRATION FOR CHILDREN AND FAMILIES, DEPARTMENT OF HEALTH AND HUMAN SERVICES STANDARDS FOR PROGRAM OPERATIONS § 303.7 Provision of services in interstate IV... central registry responsible for receiving, distributing and responding to inquiries on all incoming...

Biomarkers of Spontaneous Recovery from Traumatic Spinal Cord Injury

DTIC Science & Technology

2017-10-01

this was continued challenges with recruitment and retention, which we hope will be mitigated by our inclusion of additional sites of excellence from...Disc. Disability and pain from degenerated intervertebral discs (IVD) affects >40% of U.S adults, costs >$100 billion annually and the etiology is

45 CFR 304.30 - Public sources of State's share.

Code of Federal Regulations, 2010 CFR

2010-10-01

... funds, other than those derived from private resources, used by the IV-D agency for its child support... 304.30 Public Welfare Regulations Relating to Public Welfare OFFICE OF CHILD SUPPORT ENFORCEMENT (CHILD SUPPORT ENFORCEMENT PROGRAM), ADMINISTRATION FOR CHILDREN AND FAMILIES, DEPARTMENT OF HEALTH AND...

45 CFR 304.15 - Cost allocation.

Code of Federal Regulations, 2010 CFR

2010-10-01

... Welfare Regulations Relating to Public Welfare OFFICE OF CHILD SUPPORT ENFORCEMENT (CHILD SUPPORT... FEDERAL FINANCIAL PARTICIPATION § 304.15 Cost allocation. A State agency in support of its claims under title IV-D of the Social Security Act must have an approved cost allocation plan on file with the...

45 CFR 303.0 - Scope and applicability of this part.

Code of Federal Regulations, 2010 CFR

2010-10-01

... Section 303.0 Public Welfare Regulations Relating to Public Welfare OFFICE OF CHILD SUPPORT ENFORCEMENT (CHILD SUPPORT ENFORCEMENT PROGRAM), ADMINISTRATION FOR CHILDREN AND FAMILIES, DEPARTMENT OF HEALTH AND... prescribes: (a) The minimum organizational and staffing requirements the State IV-D agency must meet in...

Substance P Receptor Antagonist Suppresses Inflammatory Cytokine Expression in Human Disc Cells.

PubMed

Kepler, Christopher K; Markova, Dessislava Z; Koerner, John D; Mendelis, Joseph; Chen, Chiu-Ming; Vaccaro, Alexander R; Risbud, Makarand V; Albert, Todd J; Anderson, D Greg

2015-08-15

Laboratory study. To evaluate whether blockade of the Substance P (SP) NK1R attenuates its proinflammatory effect on human intervertebral disc cells (IVD), and to evaluate the signaling pathways associated with SP. SP and its receptors are expressed in human IVD cells, and cause upregulation of inflammatory mediators; however, the effects of blocking these receptors have not been studied in human IVD cells. Human annulus fibrosus (AF) and nucleus pulposus (NP) cells were expanded in monolayer, and then suspended in alginate beads. The alginate beads were treated with culture medium first containing a high affinity NK1R antagonist (L-760735) at different concentrations, and then with medium containing both NK1R antagonist and SP at 2 concentrations. Ribonucleic acid was isolated and transcribed into cDNA. Quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) was performed to evaluate expression of interleukin (IL)-1β, IL-6, and IL-8. Western blot analysis was performed to examine levels of the phosphorylated p38 mitogen-activated protein kinase (MAPK), extracellular signal regulated kinase 1/2 (ERK1/2) and nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB p65). The cells were pretreated with specific inhibitors of p38 (SB203580), ERK1/2 (PD98059), and p65 (SM7368) and then stimulated with SP. We detected expression of NK1R, neurokinin receptor 2 (NK2R), and neurokinin receptor 3 (NK3R) in AF and NP cells. Treatment of disc cells with the NK1R antagonist was able to suppress expression of IL-1β, IL-6, and IL-8 in a dose-dependent manner. SP stimulation increased phosphorylation of p38-MAPK and ERK1/2, but not of NFκB p65. This indicates that p38-MAPK and ERK1/2 control SP-induced cytokine expression independently from NF-kB p65. Inhibition of p38 and ERK1/2 activation reduced SP-induced IL-6 production in human disc cells. NK1R is responsible for the proinflammatory effect of SP on IVD cells and this effect can be blocked by preventing binding of SP to NK1R. This study shows for the first time that SP mediates signaling in disc cells through NK1R and that SP activates the proinflammatory p38-MAPK and ERK1/2 pathways. 4.

Interactive Video and Informal Learning Environments.

ERIC Educational Resources Information Center

Morrissey, Kristine A.

The Michigan State University Museum used an interactive videodisc (IVD) as an introduction to a special exhibit, "Birds in Trouble in Michigan." The hardware components included a videodisc player, a microcomputer, a video monitor, and a mouse. Software included a HyperCard program and the videodisc "Audubon Society's VideoGuide to…

75 FR 5331 - Submission for OMB Review; Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2010-02-02

... decision concerning the collection of information between 30 and 60 days after publication of this document... who are delinquent in making child support payments. Respondents: State IV-D Agencies. Annual Burden... 0.14 393.12 Certification Letter 54 1 0.40 21.60 Federal Offset Processing Menu Screens--State 176...

75 FR 82406 - Submission for OMB Review; Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2010-12-30

... interstate forms. 45 CFR 303.7 also requires CSE IV-D agencies to transmit child support case information on standard interstate forms when referring cases to other States and Territories for processing. During the... instructions have been clarified by highlighting policy information that was included with the instructions so...

45 CFR 302.14 - Fiscal policies and accountability.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 45 Public Welfare 2 2010-10-01 2010-10-01 false Fiscal policies and accountability. 302.14 Section... HUMAN SERVICES STATE PLAN REQUIREMENTS § 302.14 Fiscal policies and accountability. The State plan shall provide that the IV-D agency, in discharging its fiscal accountability, will maintain an accounting system...

45 CFR 305.62 - Disregard of a failure which is of a technical nature.

Code of Federal Regulations, 2010 CFR

2010-10-01

..., DEPARTMENT OF HEALTH AND HUMAN SERVICES PROGRAM PERFORMANCE MEASURES, STANDARDS, FINANCIAL INCENTIVES, AND... adversely affect the performance of the State's IV-D program or does not adversely affect the determination of the level of the State's paternity establishment or other performance measures percentages. ...

75 FR 69682 - Submission for OMB Review; Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2010-11-15

... OMB Review; Comment Request Title: Income Withholding for Support (IWO). OMB No.: 0970-0154. Description Use of the OMB-approved Income Withholding for Support form falls under the authority of section... the employer for income withholding in IV-D cases shall be in a standard format prescribed by the...

75 FR 57802 - Submission for OMB Review; Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2010-09-22

... OMB Review; Comment Request Title: Income Withholding for Support (IWO). OMB No.: 0970-0154. Description Use of the OMB-approved Income Withholding for Support form falls under the authority of section... the employer for income withholding in IV-D cases shall be in a standard format prescribed by the...

76 FR 27331 - Draft Guidance for Industry and Food and Drug Administration Staff; Establishing the Performance...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-05-11

... Characteristics of In Vitro Diagnostic Devices for Chlamydia Trachomatis and/or Neisseria Gonorrhoeae: Screening... entitled ``Establishing the Performance Characteristics of In Vitro Diagnostic Devices for Chlamydia... clinical performance of in vitro diagnostic devices (IVDs) intended for C. trachomatis and/or N...

45 CFR 95.601 - Scope and applicability.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 45 Public Welfare 1 2012-10-01 2012-10-01 false Scope and applicability. 95.601 Section 95.601 Public Welfare DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL ADMINISTRATION GENERAL ADMINISTRATION... IV-B, IV-D, IV-E, XIX or XXI of the Social Security Act. The conditions of approval of this subpart...

45 CFR 304.15 - Cost allocation.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 45 Public Welfare 2 2014-10-01 2012-10-01 true Cost allocation. 304.15 Section 304.15 Public Welfare Regulations Relating to Public Welfare OFFICE OF CHILD SUPPORT ENFORCEMENT (CHILD SUPPORT... title IV-D of the Social Security Act must have an approved cost allocation plan on file with the...

45 CFR 304.15 - Cost allocation.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 45 Public Welfare 2 2011-10-01 2011-10-01 false Cost allocation. 304.15 Section 304.15 Public Welfare Regulations Relating to Public Welfare OFFICE OF CHILD SUPPORT ENFORCEMENT (CHILD SUPPORT... title IV-D of the Social Security Act must have an approved cost allocation plan on file with the...

45 CFR 95.601 - Scope and applicability.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 45 Public Welfare 1 2013-10-01 2013-10-01 false Scope and applicability. 95.601 Section 95.601 Public Welfare DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL ADMINISTRATION GENERAL ADMINISTRATION... IV-B, IV-D, IV-E, XIX or XXI of the Social Security Act. The conditions of approval of this subpart...

45 CFR 1355.53 - Conditions for approval of funding.

Code of Federal Regulations, 2010 CFR

2010-10-01

... title IV-A, (ii) National Child Abuse and Neglect Data Systems (NCANDS), (iii) Systems operated under title XIX, and (iv) Systems operated under title IV-D; (3) Support the provisions of section 422(a) by... system, which is effective and efficient, to improve the program management and administration of the...

45 CFR 304.50 - Treatment of program income.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 45 Public Welfare 2 2010-10-01 2010-10-01 false Treatment of program income. 304.50 Section 304.50 Public Welfare Regulations Relating to Public Welfare OFFICE OF CHILD SUPPORT ENFORCEMENT (CHILD SUPPORT... FEDERAL FINANCIAL PARTICIPATION § 304.50 Treatment of program income. The IV-D agency must exclude from...

45 CFR 301.0 - Scope and applicability of this part.

Code of Federal Regulations, 2010 CFR

2010-10-01

... Section 301.0 Public Welfare Regulations Relating to Public Welfare OFFICE OF CHILD SUPPORT ENFORCEMENT (CHILD SUPPORT ENFORCEMENT PROGRAM), ADMINISTRATION FOR CHILDREN AND FAMILIES, DEPARTMENT OF HEALTH AND.... This part deals with the administration of title IV-D of the Social Security Act by the Federal...

45 CFR 304.50 - Treatment of program income.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 45 Public Welfare 2 2011-10-01 2011-10-01 false Treatment of program income. 304.50 Section 304.50 Public Welfare Regulations Relating to Public Welfare OFFICE OF CHILD SUPPORT ENFORCEMENT (CHILD SUPPORT... FEDERAL FINANCIAL PARTICIPATION § 304.50 Treatment of program income. The IV-D agency must exclude from...

Regulation of gene expression in intervertebral disc cells by low and high hydrostatic pressure.

PubMed

Neidlinger-Wilke, Cornelia; Würtz, Karin; Urban, Jill P G; Börm, Wolfgang; Arand, Markus; Ignatius, Anita; Wilke, Hans-Joachim; Claes, Lutz E

2006-08-01

Intervertebral disc structures are exposed to wide ranges of intradiscal hydrostatic pressure during different loading exercises and are at their minimum during lying or relaxed sitting and at maximum during lifting weights with a round back. We hypothesize that these different loading magnitudes influence the intervertebral disc (IVD) by alteration of disc matrix turnover depending on their magnitudes. Therefore the aim of this study was to assess changes in gene expression of human nucleus cells after the application of low hydrostatic pressure (0.25 MPa) and high hydrostatic pressure (2.5 MPa). IVD cells isolated from the nucleus of human (n = 18) and bovine (n = 24 from four animals) disc biopsies were seeded into three-dimensional collagen type-I matrices and exposed to the different loading magnitudes by specially developed pressure chambers. The lower pressure range (0.25 MPa, 30 min, 0.1 Hz) was applied with a recently published device by using an external compression cylinder. For the application of higher loads (2.5 MPa, 30 min, 0.1 Hz) the cell-loaded collagen gels were sealed into sterile bags with culture medium and stimulated in a newly developed water-filled compression cylinder by using a loading frame. These methods allowed the comparison of loading regimes in a wide physiological range under an equal three-dimensional culture conditions. Cells were harvested 24 h after the end of stimulation and changes in the expression of genes known to influence IVD matrix turnover (collagen-I, collagen-II, aggrecan, MMP1, MMP2, MMP3, MMP13) were analyzed by real-time RT-PCR. A Wilcoxon signed-rank test(1) and a Wilcoxon 2-sample test(2) were performed to detect differences between the stimulated and control samples(1) and differences between low and high hydrostatic pressure(2). Multiple testing was considered by adjusting the p value appropriately. Both regimes of hydrostatic pressure influenced gene expression in nucleus cells with opposite tendencies for the matrix forming proteins aggrecan and collagen type-I in response to the two different pressure magnitudes: Low hydrostatic-pressure (0.25 MPa) tended to increase collagen-I and aggrecan expression of human nucleus cells (P < 0.05) but only to a small degree. High hydrostatic pressure (2.5 MPa) tended to decrease gene expression of all anabolic proteins with significant effects on aggrecan expression of nucleus cells (P = 0.004). Low hydrostatic pressure had no influence on the expression of matrix metalloproteinases (MMP1, MMP2, MMP3 and MMP13). In contrast, high hydrostatic pressure tended to increase the expression of MMP1, MMP3 and MMP13 of human nucleus cells with high individual-individual variations. The decreased expression of aggrecan (P = 0.008) and collagen type II (P = 0.023) and the increased MMP3 expression (P = 0.008) in response to high hydrostatic pressure could be confirmed in additional experiments with bovine nucleus cells. These results suggest that hydrostatic pressure as one of the physiological stimuli of the IVD may influence matrix turnover in a magnitude dependent way. Low hydrostatic pressure (0.25 MPa) has quite small influences with a tendency to anabolic effects, whereas high hydrostatic pressure (2.5 MPa) tends to decrease the matrix protein expression with a tendency to increase some matrix-turnover enzymes. Therefore, hydrostatic pressure may regulate disc matrix turnover in a dose-dependent way.

75 FR 62404 - Submission for OMB Review; Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2010-10-08

... DEPARTMENT OF HEALTH AND HUMAN SERVICES Administration for Children and Families Submission for OMB Review; Comment Request Title: Financial Institution Data Match. OMB No.: 0970-0196. Description: Section 466(a)(17) of the Social Security Act (the Act) requires States to establish procedures under which the State Child Support Enforcement IV-D...

75 FR 17926 - Proposed Information Collection Activity; Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2010-04-08

... DEPARTMENT OF HEALTH AND HUMAN SERVICES Administration for Children and Families Proposed... and Human Services to promulgate a form for imposition of liens to be used by the State child support... interstate child support cases. Tribal IV-D agencies are not required to use this form but may choose to do...

78 FR 26051 - Proposed Information Collection Activity; Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2013-05-03

... DEPARTMENT OF HEALTH AND HUMAN SERVICES Administration for Children and Families Proposed... and Human Services to promulgate a form for imposition of liens to be used by the State child support... interstate child support cases. Tribal IV-D agencies are not required to use this form but may choose to do...

45 CFR 301.0 - Scope and applicability of this part.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 45 Public Welfare 2 2014-10-01 2012-10-01 true Scope and applicability of this part. 301.0 Section 301.0 Public Welfare Regulations Relating to Public Welfare OFFICE OF CHILD SUPPORT ENFORCEMENT (CHILD... deals with the administration of title IV-D of the Social Security Act by the Federal Government...

45 CFR 307.10 - Functional requirements for computerized support enforcement systems in operation by October 1...

Code of Federal Regulations, 2012 CFR

2012-10-01

... if the State has chosen to pay such incentives; (7) Maintaining accounts receivable on all amounts... interfacing with State financial management and expenditure information; (9) Accepting electronic case...) Providing management information on all IV-D cases under the State plan from initial referral or application...

45 CFR 307.10 - Functional requirements for computerized support enforcement systems in operation by October 1...

Code of Federal Regulations, 2014 CFR

2014-10-01

... if the State has chosen to pay such incentives; (7) Maintaining accounts receivable on all amounts... interfacing with State financial management and expenditure information; (9) Accepting electronic case...) Providing management information on all IV-D cases under the State plan from initial referral or application...

45 CFR 307.10 - Functional requirements for computerized support enforcement systems in operation by October 1...

Code of Federal Regulations, 2013 CFR

2013-10-01

... if the State has chosen to pay such incentives; (7) Maintaining accounts receivable on all amounts... interfacing with State financial management and expenditure information; (9) Accepting electronic case...) Providing management information on all IV-D cases under the State plan from initial referral or application...

42 CFR 435.115 - Individuals deemed to be receiving AFDC.

Code of Federal Regulations, 2011 CFR

2011-10-01

... collection of child or spousal support under title IV-D of the Social Security Act results in the termination of AFDC eligibility in accordance with section 406(h) of the Social Security Act. States must... eligible spouse of such relative as described in section 406(b) of the Social Security Act) who: (1...

Detection and Identification of Viruses using the Integrated Virus Detection System (IVDS)

DTIC Science & Technology

2005-11-01

Hepatitis Virus (MHV) or Coronaviridae ................................. 5 3.2.5 M VM Parvovirus ...Acetate ........... 29 24. Mouse Hepatitus Virus MHV-A59 Diluted with Potassium Phosphate .......... 30 25. Scan of MVM Parvovirus - Neat...31 26. Scan of MVM Parvovirus -Diluted in Ammonium Acetate, Expanded Scale ...... 32 27. Scan of

Essentials for Attorneys in Child Support Enforcement.

ERIC Educational Resources Information Center

Henry, Michael R.; And Others

This handbook presents a course developed to provide a national perspective for attorneys who represent state and local child support enforcement agencies operating under Title IV-D of the Social Security Act. The introduction provides an overview of the child support problem in the United States, citing causes and effects of the problem and…

75 FR 18016 - Notice of Allocation Availability (NOAA) Inviting Applications for the CY 2010 Allocation Round...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-04-08

... Availability (NOAA) Inviting Applications for the CY 2010 Allocation Round of the New Markets Tax Credit Program Funding Opportunity Title: Notice of Allocation Availability (NOAA) Inviting Applications for the... form (see Section IV.D. of this NOAA for more details). Applications must meet all eligibility and...

45 CFR 303.11 - Case closure criteria.

Code of Federal Regulations, 2011 CFR

2011-10-01

... the best interests of the child to establish paternity in a case involving incest or forcible rape, or... 45 Public Welfare 2 2011-10-01 2011-10-01 false Case closure criteria. 303.11 Section 303.11... STANDARDS FOR PROGRAM OPERATIONS § 303.11 Case closure criteria. (a) The IV-D agency shall establish a...

Biomarkers in the Diagnosis and Prognosis of Alzheimer's Disease.

PubMed

Schaffer, Cole; Sarad, Nakia; DeCrumpe, Ashton; Goswami, Disha; Herrmann, Sara; Morales, Jose; Patel, Parth; Osborne, Jim

2015-10-01

Alzheimer's disease (AD) is a neurodegenerative disease that inhibits cognitive functions and has no cure. This report reviews the current diagnostic standards for AD with an emphasis on early diagnosis using the cerebrospinal fluid (CSF) biomarkers amyloid-beta, t-tau, and p-tau and fluorodeoxyglucose positron emission tomography imaging. Abnormal levels of these CSF biomarkers and decreased cerebral uptake of glucose have recently been used in the early diagnosis of AD in experimental studies. These promising biomarkers can be measured using immunoassays performed in singleplex or multiplex formats. Although presently, there are no Food and Drug Administration-approved in vitro diagnostics (IVDs) for early detection of AD, a multiplex immunoassay measuring a panel of promising AD biomarkers in CSF may be a likely IVD candidate for the clinical AD diagnostic market. Specifically, the INNO-BIA AlzBio3 immunoassay kit, performed using bead arrays on the xMAP Luminex analyzer, allows simultaneous quantification of amyloid-beta, t-tau, and p-tau biomarkers. AD biomarkers can also be screened using enzyme-linked immunosorbent assays that are offered as laboratory-developed tests. © 2014 Society for Laboratory Automation and Screening.

Ion Dynamics in a Single and Dual Radio Frequency Sheath Measured by Laser-Induced Fluorescence

NASA Astrophysics Data System (ADS)

Moore, Nathaniel Breckenridge

Ion dynamics are investigated in a single and dual radio frequency sheath as a function of radius above a 30 cm diameter biased silicon wafer for the first time in an industrial inductively coupled (440 kHz, 500 W) plasma etch tool. Ion velocity distribution (IVD) function measurements in the argon plasma are taken using laser induced fluorescence (LIF). Planar sheets of laser light enter the chamber both parallel and perpendicular to the surface of the wafer in order to measure both parallel and perpendicular IVDs at thousands of spatial positions. A fast (30 ns exposure) CCD camera measures the resulting fluorescence with a spatial resolution of 0.4 mm. The dual-frequency bias on the wafer is comprised of a 2 MHz low frequency (LF) bias and a 19 MHz high frequency (HF) bias. The laser is phase locked to the LF bias and IVD measurements are taken at several different LF phases. Ion energy distribution (IED) function measurements and calculated moments are compared for several cases. For the LF case (no HF), the IEDs were found to be highly phase dependent and were varied radially up to 10%. Calculated mean velocity vectors showed large impact angles near the surface of the wafer with the largest angles observed near the wafer edge. The LF experimental results are compared with simulations designed specifically for this particular plasma tool and showed good qualitative agreement. For the dual frequency case, IEDs were measured at two disparate phases of the phase-locked LF bias. IEDs were found to be multi-peaked and were well-approximated by a sum of Maxwellian distributions. The calculated fluxes in the dual frequency case were found to be substantially more radially uniform than the single frequency bias case. For industrial applications, this radially uniform ion flux is evidently a trade off with the undesirable multi-peaked structure in the IEDs.

Age-related differences in the response of the L5-S1 intervertebral disc to spinal traction.

PubMed

Mitchell, Ulrike H; Beattie, Paul F; Bowden, Jennifer; Larson, Robert; Wang, Haonan

2017-10-01

Lumbar traction is a common treatment for low back pain; however its mechanisms of action are poorly understood. It has been hypothesized that a key effect of lumbar traction is its capacity to influence fluid movement within the intervertebral disc (IVD). To determine differences in the apparent diffusion coefficient (ADC) obtained with lumbar diffusion-weighted imaging (DWI) of the L5-S1 IVD before, and during, the application of lumbar traction. Case series, repeated measures. A static traction load of ∼50% of body-weight was applied to the low back using a novel "MRI-safe" apparatus. DWI of the lumbar spine was performed prior to, and during the application of the traction load. Participants were currently asymptomatic and included a young adult group (n = 18) and a middle-aged group (n = 15). The young adult group had a non-significant 2.2% increase in ADC (mean change = 0.03 × 10 -3  mm 2 /s, SD = 0.24, 95% CI = -0.09, 0.15). The ADC for the middle-aged group significantly increased by 20% (mean change of 0.18 × 10 -3  mm 2 /s, SD = 0.19; 95% CI = 0.07, 0.28; p = 0.003; effect size = 0.95). There was an inverse relationship between the ADC obtained before traction and the percent increase in ADC that was measured during traction. Static traction was associated with an increase in diffusion of water within the L5-S1 IVDs of middle-age individuals, but not in young adults, suggesting age-related differences in the diffusion response. Further study is needed to assess the relationship between these findings and the symptoms of back pain. 4. Copyright © 2017 Elsevier Ltd. All rights reserved.

Novel Immortal Cell Lines Support Cellular Heterogeneity in the Human Annulus Fibrosus.

PubMed

van den Akker, Guus G H; Surtel, Don A M; Cremers, Andy; Richardson, Stephen M; Hoyland, Judith A; van Rhijn, Lodewijk W; Voncken, Jan Willem; Welting, Tim J M

2016-01-01

Loss of annulus fibrosus (AF) integrity predisposes to disc herniation and is associated with IVD degeneration. Successful implementation of biomedical intervention therapy requires in-depth knowledge of IVD cell biology. We recently generated unique clonal human nucleus pulposus (NP) cell lines. Recurring functional cellular phenotypes from independent donors provided pivotal evidence for cell heterogeneity in the mature human NP. In this study we aimed to generate and characterize immortal cell lines for the human AF from matched donors. Non-degenerate healthy disc material was obtained as surplus surgical material. AF cells were immortalized by simian virus Large T antigen (SV40LTAg) and human telomerase (hTERT) expression. Early passage cells and immortalized cell clones were characterized based on marker gene expression under standardized culturing and in the presence of Transforming Growth factor β (TGFβ). The AF-specific expression signature included COL1A1, COL5A1, COL12A1, SFRP2 and was largely maintained in immortal AF cell lines. Remarkably, TGFβ induced rapid 3D sheet formation in a subgroup of AF clones. This phenotype was associated with inherent differences in Procollagen type I processing and maturation, and correlated with differential mRNA expression of Prolyl 4-hydroxylase alpha polypeptide 1 and 3 (P4HA1,3) and Lysyl oxidase (LOX) between clones and differential P4HA3 protein expression between AF cells in histological sections. We report for the first time the generation of representative human AF cell lines. Gene expression profile analysis and functional comparison of AF clones revealed variation between immortalized cells and suggests phenotypic heterogeneity in the human AF. Future characterization of AF cellular (sub-)populations aims to combine identification of additional specific AF marker genes and their biological relevance. Ultimately this knowledge will contribute to clinical application of cell-based technology in IVD repair.

Lactoferricin Enhances BMP7-Stimulated Anabolic Pathways in Intervertebral Disc Cells

PubMed Central

Ellman, Michael B; Kim, Jaesung; An, Howard S; Chen, Di; Kc, Ranjan; Li, Xin; Xiao, Guozhi; Yan, Dongyao; Suh, Joon; van Wijnen, Andre J.; Wang, James H-C; Kim, Su-Gwan; Im, Hee-Jeong

2013-01-01

Bone-morphogenetic protein-7 (BMP7) is a well-known anabolic and anti-catabolic growth factor on intervertebral (IVD) matrix and cell homeostasis. Similarly, lactoferricin B (LfcinB) has recently been shown to have pro-anabolic, anti-catabolic, anti-oxidative and/or anti-inflammatory effects in bovine disc cells in vitro. In this study, we investigated the potential benefits of using combined peptide therapy with LfcinB and BMP7 for intervertebral disc (IVD) matrix repair and to understand cellular and signaling mechanisms controlled by these factors. We studied the effects of BMP7 and LfcinB as individual treatments and combined therapy on bovine nucleus pulposus (NP) cells by assessing proteoglycan (PG) accumulation and synthesis, and the expression of matrix protein aggrecan and transcription factor SOX-9. We also analyzed the role of noggin, a BMP antagonist, in IVD tissue and examined its effect after stimulation with LfcinB. To understand the molecular mechanisms by which LfcinB synergizes with BMP7, we investigated the ERK-SP1 axis as a downstream intracellular signaling regulator involved in BMP7 and LfcinB-mediated activities. Treatment of bovine NP cells cultured in alginate with LfcinB plus BMP7 synergistically stimulates PG synthesis and accumulation in part by upregulation of aggrecan gene expression. The synergism results from LfcinB-mediated activation of Sp1 and SMAD signaling pathways by (i) phosphorylation of SMAD 1/5/8; (ii) downregulation of SMAD inhibitory factors [i.e., noggin (BMP receptor antagonist) and SMAD6 (inhibitory SMAD)]; and (iii) upregulation of SMAD4 (universal co-SMAD). These data indicate that LfcinB-suppression of noggin may eliminate the negative feedback of BMP7, thereby maximizing biological activity of BMP7 and ultimately shifting homeostasis to a pro-anabolic state in disc cells. We propose that combination growth factor therapy using BMP7 and LfcinB may be beneficial for treatment of disc degeneration. PMID:23644135

A novel synthetic single crystal diamond device for in vivo dosimetry

DOE Office of Scientific and Technical Information (OSTI.GOV)

Marinelli, Marco; Prestopino, G., E-mail: giuseppe.prestopino@uniroma2.it; Tonnetti, A.

Purpose: Aim of the present work is to evaluate the synthetic single crystal diamond Schottky photodiode developed at the laboratories of “Tor Vergata” University in Rome in a new dosimeter configuration specifically designed for offline wireless in vivo dosimetry (IVD) applications. Methods: The new diamond based dosimeter, single crystal diamond detector (SCDD-iv), consists of a small unwired detector and a small external reading unit that can be connected to commercial electrometers for getting the detector readout after irradiation. Two nominally identical SCDD-iv dosimeter prototypes were fabricated and tested. A basic dosimetric characterization of detector performances relevant for IVD application wasmore » performed under irradiation with {sup 60}Co and 6 MV photon beams. Preirradiation procedure, response stability, short and long term reproducibility, leakage charge, fading effect, linearity with dose, dose rate dependence, temperature dependence, and angular response were investigated. Results: The SCDD-iv is simple, with no cables linked to the patient and the readout is immediate. The range of response with dose has been tested from 1 up to 12 Gy; the reading is independent of the accumulated dose and dose rate independent in the range between about 0.5 and 5 Gy/min; its temperature dependence is within 0.5% between 25 and 38 °C, and its directional dependence is within 2% from 0° to 90°. The combined relative standard uncertainty of absorbed dose to water measurements is estimated lower than the tolerance and action level of 5%. Conclusions: The reported results indicate the proposed novel offline dosimeter based on a synthetic single crystal diamond Schottky photodiode as a promising candidate for in vivo dosimetry applications with photon beams.« less

Comparison of B0 versus B0 and B1 field inhomogeneity correction for glycosaminoglycan chemical exchange saturation transfer imaging.

PubMed

Müller-Lutz, Anja; Ljimani, Alexandra; Stabinska, Julia; Zaiss, Moritz; Boos, Johannes; Wittsack, Hans-Jörg; Schleich, Christoph

2018-05-14

The study compares glycosaminoglycan chemical exchange saturation transfer (gagCEST) imaging of intervertebral discs corrected for solely B 0 inhomogeneities or both B 0 and B 1 inhomogeneities. Lumbar intervertebral discs of 20 volunteers were examined with T 2 -weighted and gagCEST imaging. Field inhomogeneity correction was performed with B 0 correction only and with correction of both B 0 and B 1 . GagCEST effects measured by the asymmetric magnetization transfer ratio (MTR asym ) and signal-to-noise ratio (SNR) were compared between both methods. Significant higher MTR asym and SNR values were obtained in the nucleus pulposus using B 0 and B 1 correction compared with B 0 -corrected gagCEST. The GagCEST effect was significantly different in the nucleus pulposus compared with the annulus fibrosus for both methods. The B 0 and B 1 field inhomogeneity correction method leads to an improved quality of gagCEST imaging in IVDs compared with only B 0 correction.

45 CFR 303.72 - Requests for collection of past-due support by Federal tax refund offset.

Code of Federal Regulations, 2010 CFR

2010-10-01

... the accuracy of the past-due support amount. If the State IV-D agency has verified this information...) The amount of past-due support owed; (iv) The State codes as contained in the Federal Information Processing Standards (FIPS) publication of the National Bureau of Standards and also promulgated by the...

45 CFR 309.100 - What procedures for the establishment of paternity must a Tribe or Tribal organization include in...

Code of Federal Regulations, 2011 CFR

2011-10-01

... ENFORCEMENT PROGRAM), ADMINISTRATION FOR CHILDREN AND FAMILIES, DEPARTMENT OF HEALTH AND HUMAN SERVICES TRIBAL... paternity, and setting forth facts establishing a reasonable possibility of the requisite sexual contact... of the nonexistence of sexual contact between the parties. (b) The Tribal IV-D agency need not...

45 CFR 309.100 - What procedures for the establishment of paternity must a Tribe or Tribal organization include in...

Code of Federal Regulations, 2013 CFR

2013-10-01

... ENFORCEMENT PROGRAM), ADMINISTRATION FOR CHILDREN AND FAMILIES, DEPARTMENT OF HEALTH AND HUMAN SERVICES TRIBAL... paternity, and setting forth facts establishing a reasonable possibility of the requisite sexual contact... of the nonexistence of sexual contact between the parties. (b) The Tribal IV-D agency need not...

45 CFR 309.100 - What procedures for the establishment of paternity must a Tribe or Tribal organization include in...

Code of Federal Regulations, 2010 CFR

2010-10-01

... ENFORCEMENT PROGRAM), ADMINISTRATION FOR CHILDREN AND FAMILIES, DEPARTMENT OF HEALTH AND HUMAN SERVICES TRIBAL... paternity, and setting forth facts establishing a reasonable possibility of the requisite sexual contact... of the nonexistence of sexual contact between the parties. (b) The Tribal IV-D agency need not...

45 CFR 309.100 - What procedures for the establishment of paternity must a Tribe or Tribal organization include in...

Code of Federal Regulations, 2012 CFR

2012-10-01

... ENFORCEMENT PROGRAM), ADMINISTRATION FOR CHILDREN AND FAMILIES, DEPARTMENT OF HEALTH AND HUMAN SERVICES TRIBAL... paternity, and setting forth facts establishing a reasonable possibility of the requisite sexual contact... of the nonexistence of sexual contact between the parties. (b) The Tribal IV-D agency need not...

45 CFR 309.100 - What procedures for the establishment of paternity must a Tribe or Tribal organization include in...

Code of Federal Regulations, 2014 CFR

2014-10-01

... ENFORCEMENT PROGRAM), ADMINISTRATION FOR CHILDREN AND FAMILIES, DEPARTMENT OF HEALTH AND HUMAN SERVICES TRIBAL... paternity, and setting forth facts establishing a reasonable possibility of the requisite sexual contact... of the nonexistence of sexual contact between the parties. (b) The Tribal IV-D agency need not...

45 CFR 302.33 - Services to individuals not receiving title IV-A assistance.

Code of Federal Regulations, 2013 CFR

2013-10-01

..., DEPARTMENT OF HEALTH AND HUMAN SERVICES STATE PLAN REQUIREMENTS § 302.33 Services to individuals not... services and is no longer eligible for assistance under the title IV-A, IV-E foster care, and Medicaid... IV-A, IV-E foster care, and Medicaid programs, the IV-D agency must notify the family, within five...

45 CFR 302.33 - Services to individuals not receiving title IV-A assistance.

Code of Federal Regulations, 2012 CFR

2012-10-01

..., DEPARTMENT OF HEALTH AND HUMAN SERVICES STATE PLAN REQUIREMENTS § 302.33 Services to individuals not... services and is no longer eligible for assistance under the title IV-A, IV-E foster care, and Medicaid... IV-A, IV-E foster care, and Medicaid programs, the IV-D agency must notify the family, within five...

45 CFR 302.33 - Services to individuals not receiving title IV-A assistance.

Code of Federal Regulations, 2014 CFR

2014-10-01

..., DEPARTMENT OF HEALTH AND HUMAN SERVICES STATE PLAN REQUIREMENTS § 302.33 Services to individuals not... services and is no longer eligible for assistance under the title IV-A, IV-E foster care, and Medicaid... IV-A, IV-E foster care, and Medicaid programs, the IV-D agency must notify the family, within five...

45 CFR 302.33 - Services to individuals not receiving title IV-A assistance.

Code of Federal Regulations, 2011 CFR

2011-10-01

..., DEPARTMENT OF HEALTH AND HUMAN SERVICES STATE PLAN REQUIREMENTS § 302.33 Services to individuals not... services and is no longer eligible for assistance under the title IV-A, IV-E foster care, and Medicaid... IV-A, IV-E foster care, and Medicaid programs, the IV-D agency must notify the family, within five...

45 CFR 302.33 - Services to individuals not receiving title IV-A assistance.

Code of Federal Regulations, 2010 CFR

2010-10-01

..., DEPARTMENT OF HEALTH AND HUMAN SERVICES STATE PLAN REQUIREMENTS § 302.33 Services to individuals not... services and is no longer eligible for assistance under the title IV-A, IV-E foster care, and Medicaid... IV-A, IV-E foster care, and Medicaid programs, the IV-D agency must notify the family, within five...

Cadmium Alternatives High Strength Steel JTP

DTIC Science & Technology

2005-03-01

Cadmium Alternatives HSS JTP Alternative Coatings Selected by JCAT: Primary Repair LHE Cadmium (control) IVD Aluminum (control) Zinc-Nickel, acid...Tin-Zinc Cadmium Alternatives HSS JTP Demonstration Tests Selected by JCAT: Phase I Phase II Hydrogen Embrittlement Hydrogen Re-Embrittlement (NRB...immersion, exposed C-ring (Army)) Bend Adhesion (Q/A) Appearance Throwing Power Composition Uniformity Strippability Galvanic Potential Bend Adhesion

76 FR 1169 - Agency Information Collection Activities; Submission for Office of Management and Budget Review...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-01-07

... Labels and in Labeling of In Vitro Diagnostic Devices Intended for Professional Use AGENCY: Food and Drug... Support Use of Symbols on Labels and in Labeling of In Vitro Diagnostic Devices Intended for Professional... required for: (1) In vitro diagnostic devices (IVDs), intended for professional use under 21 CFR 809.10...

45 CFR 305.63 - Standards for determining substantial compliance with IV-D requirements.

Code of Federal Regulations, 2013 CFR

2013-10-01

...) Separation of cash handling and accounting functions, § 302.20 of this chapter; and (4) Notice of collection... must use or attempt to use at least one enforcement technique available under State law in addition to... is not appropriate in a particular case, and the State uses at least one enforcement technique...

45 CFR 305.63 - Standards for determining substantial compliance with IV-D requirements.

Code of Federal Regulations, 2012 CFR

2012-10-01

...) Separation of cash handling and accounting functions, § 302.20 of this chapter; and (4) Notice of collection... must use or attempt to use at least one enforcement technique available under State law in addition to... is not appropriate in a particular case, and the State uses at least one enforcement technique...

45 CFR 305.63 - Standards for determining substantial compliance with IV-D requirements.

Code of Federal Regulations, 2014 CFR

2014-10-01

...) Separation of cash handling and accounting functions, § 302.20 of this chapter; and (4) Notice of collection... must use or attempt to use at least one enforcement technique available under State law in addition to... is not appropriate in a particular case, and the State uses at least one enforcement technique...

45 CFR 305.63 - Standards for determining substantial compliance with IV-D requirements.

Code of Federal Regulations, 2011 CFR

2011-10-01

...) Separation of cash handling and accounting functions, § 302.20 of this chapter; and (4) Notice of collection... must use or attempt to use at least one enforcement technique available under State law in addition to... is not appropriate in a particular case, and the State uses at least one enforcement technique...

Continuous "in vitro" Evolution of a Ribozyme Ligase: A Model Experiment for the Evolution of a Biomolecule

ERIC Educational Resources Information Center

Ledbetter, Michael P.; Hwang, Tony W.; Stovall, Gwendolyn M.; Ellington, Andrew D.

2013-01-01

Evolution is a defining criterion of life and is central to understanding biological systems. However, the timescale of evolutionary shifts in phenotype limits most classroom evolution experiments to simple probability simulations. "In vitro" directed evolution (IVDE) frequently serves as a model system for the study of Darwinian…

42 CFR 436.114 - Individuals deemed to be receiving AFDC.

Code of Federal Regulations, 2011 CFR

2011-10-01

... increased collection of child or spousal support under title IV-D of the Social Security Act results in the termination of AFDC eligibility in accordance with section 406(h) of the Social Security Act. States must... eligible spouse of such relative as described in section 406(b) of the Social Security Act) who: (1...

76 FR 32392 - Notice of Allocation Availability (NOAA) Inviting Applications for the CY 2011 Allocation Round...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-06-06

... Availability (NOAA) Inviting Applications for the CY 2011 Allocation Round of the New Markets Tax Credit... charts) in electronic form (see Section IV.D. of this NOAA for more details). Applications must meet all eligibility and other requirements and deadlines, as applicable, set forth in this NOAA. Allocation applicants...

78 FR 70306 - Distribution of In Vitro Diagnostic Products Labeled for Research Use Only or Investigational Use...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-11-25

... for Devices and Radiological Health (CDRH) Food and Drug Administration, 10903 New Hampshire Ave...''). This guidance is intended to provide the current thinking of CDRH and CBER on when IVD products are... capability for all CDRH guidance documents is available at http://www.fda.gov/MedicalDevices/Device...

Commercial Molecular Tests for Fungal Diagnosis from a Practical Point of View.

PubMed

Lackner, Michaela; Lass-Flörl, Cornelia

2017-01-01

The increasing interest in molecular diagnostics is a result of tremendously improved knowledge on fungal infections in the past 20 years and the rapid development of new methods, in particular polymerase chain reaction. High expectations have been placed on molecular diagnostics, and the number of laboratories now using the relevant technology is rapidly increasing-resulting in an obvious need for standardization and definition of laboratory organization. In the past 10 years, multiple new molecular tools were marketed for the detection of DNA, antibodies, cell wall components, or other antigens. In contrast to classical culture methods, molecular methods do not detect a viable organisms, but only molecules which indicate its presence; this can be nucleic acids, cell components (antigens), or antibodies (Fig. 1). In this chapter, an overview is provided on commercially available detection tools, their strength and how to use them. A main focus is laid on providing tips and tricks that make daily life easier. We try to focus and mention methodical details which are not highlighted in the manufacturer's instructions of these test kits, but are based on our personal experience in the laboratory. Important to keep in mind is that molecular tools cannot replace culture, microscopy, or a critical view on patients' clinical history, signs, and symptoms, but provide a valuable add on tool. Diagnosis should not be based solely on a molecular test, but molecular tools might deliver an important piece of information that helps matching the diagnostic puzzle to a diagnosis, in particular as few tests are in vitro diagnostic tests (IVD) or only part of the whole test carries the IVD certificate (e.g., DNA extraction is often not included). Please be aware that the authors do not claim to provide a complete overview on all commercially available diagnostic assays being currently marketed for fungal detection, as those are subject to constant change. A main focus is put on commonly used panfungal assays and pathogen-specific assays, including Aspergillus-specific, Candida-specific, Cryptococcus specific, Histoplasma-specific, and Pneumocystis-specific assays. Assays are categorized according to their underlying principle in either antigen-detecting or antibody-detecting or DNA-detecting (Fig. 1). Other non-DNA-detecting nucleic acid methods such as FISH and PNA FISH are not summarized in this chapter and an overview on test performance, common false positives, and the clinical evaluation of commercial tests in studies is provided already in a previous book series by Javier Yugueros Marcos and David H. Pincus (Marcos and Pincus, Methods Mol Biol 968:25-54, 2013).

45 CFR 305.62 - Disregard of a failure which is of a technical nature.

Code of Federal Regulations, 2014 CFR

2014-10-01

... nature. 305.62 Section 305.62 Public Welfare Regulations Relating to Public Welfare OFFICE OF CHILD... PENALTIES § 305.62 Disregard of a failure which is of a technical nature. A State subject to a penalty under... the noncompliance with one or more of the IV-D requirements, is of a technical nature which does not...

Testing the Suitability of Mediation of Child Support Orders in Title IV-D Cases

ERIC Educational Resources Information Center

Schraufnagel, Scot; Li, Quan

2010-01-01

Objectives: The purpose of this study is to test mediation versus a traditional court process for the establishment or modification of child support orders. The intention is to determine which dispute resolution process is associated with greater client satisfaction and compliance. An auxiliary objective is to test the type of cases which are most…

26 CFR 1.414(r)-5 - Qualified separate line of business-administrative scrutiny requirement-safe harbors.

Code of Federal Regulations, 2014 CFR

2014-04-01

...-administrative scrutiny requirement-safe harbors. 1.414(r)-5 Section 1.414(r)-5 Internal Revenue INTERNAL REVENUE..., Profit-Sharing, Stock Bonus Plans, Etc. § 1.414(r)-5 Qualified separate line of business—administrative...(r)-3 satisfies the administrative scrutiny requirement of § 1.414(r)-1(b)(2)(iv)(D) for a testing...

26 CFR 1.414(r)-5 - Qualified separate line of business-administrative scrutiny requirement-safe harbors.

Code of Federal Regulations, 2013 CFR

2013-04-01

...-administrative scrutiny requirement-safe harbors. 1.414(r)-5 Section 1.414(r)-5 Internal Revenue INTERNAL REVENUE..., Profit-Sharing, Stock Bonus Plans, Etc. § 1.414(r)-5 Qualified separate line of business—administrative...(r)-3 satisfies the administrative scrutiny requirement of § 1.414(r)-1(b)(2)(iv)(D) for a testing...

26 CFR 1.414(r)-5 - Qualified separate line of business-administrative scrutiny requirement-safe harbors.

Code of Federal Regulations, 2012 CFR

2012-04-01

...-administrative scrutiny requirement-safe harbors. 1.414(r)-5 Section 1.414(r)-5 Internal Revenue INTERNAL REVENUE..., Profit-Sharing, Stock Bonus Plans, Etc. § 1.414(r)-5 Qualified separate line of business—administrative...(r)-3 satisfies the administrative scrutiny requirement of § 1.414(r)-1(b)(2)(iv)(D) for a testing...

26 CFR 1.414(r)-5 - Qualified separate line of business-administrative scrutiny requirement-safe harbors.

Code of Federal Regulations, 2011 CFR

2011-04-01

...-administrative scrutiny requirement-safe harbors. 1.414(r)-5 Section 1.414(r)-5 Internal Revenue INTERNAL REVENUE..., Profit-Sharing, Stock Bonus Plans, Etc. § 1.414(r)-5 Qualified separate line of business—administrative...(r)-3 satisfies the administrative scrutiny requirement of § 1.414(r)-1(b)(2)(iv)(D) for a testing...

45 CFR 303.70 - Requests by the State Parent Locator Service (SPLS) for information from the Federal Parent...

Code of Federal Regulations, 2010 CFR

2010-10-01

...), ADMINISTRATION FOR CHILDREN AND FAMILIES, DEPARTMENT OF HEALTH AND HUMAN SERVICES STANDARDS FOR PROGRAM... information: (1) The parent's name; (2) The parent's social security number (SSN). If the SSN is unknown, the... noncustodial parent who owes a support obligation to a family on whose behalf the IV-D agency is providing...

45 CFR 305.63 - Standards for determining substantial compliance with IV-D requirements.

Code of Federal Regulations, 2010 CFR

2010-10-01

...) Separation of cash handling and accounting functions, § 302.20 of this chapter; and (4) Notice of collection...-custodial parents under § 303.3 of this chapter; establishment of paternity under § 303.5(a) and (f) of this... (c)(8) through (10) of this chapter; location of non-custodial parents under § 303.3 of this chapter...

45 CFR 305.62 - Disregard of a failure which is of a technical nature.

Code of Federal Regulations, 2011 CFR

2011-10-01

... nature. 305.62 Section 305.62 Public Welfare Regulations Relating to Public Welfare OFFICE OF CHILD... PENALTIES § 305.62 Disregard of a failure which is of a technical nature. A State subject to a penalty under... the noncompliance with one or more of the IV-D requirements, is of a technical nature which does not...

45 CFR 305.62 - Disregard of a failure which is of a technical nature.

Code of Federal Regulations, 2013 CFR

2013-10-01

... nature. 305.62 Section 305.62 Public Welfare Regulations Relating to Public Welfare OFFICE OF CHILD... PENALTIES § 305.62 Disregard of a failure which is of a technical nature. A State subject to a penalty under... the noncompliance with one or more of the IV-D requirements, is of a technical nature which does not...

45 CFR 305.62 - Disregard of a failure which is of a technical nature.

Code of Federal Regulations, 2012 CFR

2012-10-01

... nature. 305.62 Section 305.62 Public Welfare Regulations Relating to Public Welfare OFFICE OF CHILD... PENALTIES § 305.62 Disregard of a failure which is of a technical nature. A State subject to a penalty under... the noncompliance with one or more of the IV-D requirements, is of a technical nature which does not...

45 CFR 309.45 - When and how may a Tribe or Tribal organization request reconsideration of a disapproval action?

Code of Federal Regulations, 2012 CFR

2012-10-01

... supportive of its application, plan or plan amendment; and (2) A written response to each ground for... the application, plan, or plan amendment as well as the Tribe or Tribal organization's response. The... disapproval of a Tribal IV-D application, plan or plan amendment by filing a written Request for...

45 CFR 309.45 - When and how may a Tribe or Tribal organization request reconsideration of a disapproval action?

Code of Federal Regulations, 2011 CFR

2011-10-01

... supportive of its application, plan or plan amendment; and (2) A written response to each ground for... the application, plan, or plan amendment as well as the Tribe or Tribal organization's response. The... disapproval of a Tribal IV-D application, plan or plan amendment by filing a written Request for...

45 CFR 309.45 - When and how may a Tribe or Tribal organization request reconsideration of a disapproval action?

Code of Federal Regulations, 2010 CFR

2010-10-01

... supportive of its application, plan or plan amendment; and (2) A written response to each ground for... the application, plan, or plan amendment as well as the Tribe or Tribal organization's response. The... disapproval of a Tribal IV-D application, plan or plan amendment by filing a written Request for...

75 FR 20437 - Amended Authorization of Emergency Use of the Antiviral Product Peramivir Accompanied by...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-04-19

... manufactured for BioCryst Pharmaceuticals, Inc. (BioCryst) for intravenous (IV) administration in certain adult... peramivir is as follows: Peramivir injection: 200mg/20mL (10 mg/mL) single use vial manufactured for BioCryst Pharmaceuticals, Inc. (BioCryst). (See Section IV.D.3. of this letter). 1. The above peramivir...

Point-of-care diagnostics: market trends and growth drivers.

PubMed

Rajan, Aruna; Glorikian, Harry

2009-01-01

There is a significant demand for in vitro diagnostic (IVD) testing to move closer to the patient point-of-care diagnostics [POC]), whether in the hospital, physician's office, rapid clinic or the home, effectively cutting time to results and helping patients make better informed decisions about their health. To analyze the point-of-care market and its trends and growth drivers. In 2007, POC made up 30% of the IVD market and is expected to grow at 9% a year. Although the overall POC market is expected to grow steadily, infectious POC is now the most attractive segment. Availability of rapid random access molecular diagnostic system for critical care infectious diseases such as MRSA and sepsis in the near future is likely to be a significant driver of infectious POC post 2012. Owing to the extraordinary increase in the cost of care, healthcare delivery is moving to increasingly decentralized settings such as rapid clinics and the home, driven by point-of-care diagnostics that provide accurate and directional results. We are evolving from the analog testing world to the digital testing world, where diagnosis is exact and therapy can be administered and be predictably effective.

45 CFR 309.10 - Who is eligible to apply for and receive Federal funding to operate a Tribal IV-D program?

Code of Federal Regulations, 2010 CFR

2010-10-01

... Tribal law or code, in the population subject to the jurisdiction of the Tribal court or administrative... defined by Tribal laws or codes, in the population of the Tribes subject to the jurisdiction of the Tribal... and provide justification for operating a program with less than the minimum number of children may be...

45 CFR 309.10 - Who is eligible to apply for and receive Federal funding to operate a Tribal IV-D program?

Code of Federal Regulations, 2011 CFR

2011-10-01

... Tribal law or code, in the population subject to the jurisdiction of the Tribal court or administrative... defined by Tribal laws or codes, in the population of the Tribes subject to the jurisdiction of the Tribal... and provide justification for operating a program with less than the minimum number of children may be...

45 CFR 309.10 - Who is eligible to apply for and receive Federal funding to operate a Tribal IV-D program?

Code of Federal Regulations, 2012 CFR

2012-10-01

... Tribal law or code, in the population subject to the jurisdiction of the Tribal court or administrative... defined by Tribal laws or codes, in the population of the Tribes subject to the jurisdiction of the Tribal... and provide justification for operating a program with less than the minimum number of children may be...

45 CFR 309.10 - Who is eligible to apply for and receive Federal funding to operate a Tribal IV-D program?

Code of Federal Regulations, 2013 CFR

2013-10-01

... Tribal law or code, in the population subject to the jurisdiction of the Tribal court or administrative... defined by Tribal laws or codes, in the population of the Tribes subject to the jurisdiction of the Tribal... and provide justification for operating a program with less than the minimum number of children may be...

45 CFR 309.10 - Who is eligible to apply for and receive Federal funding to operate a Tribal IV-D program?

Code of Federal Regulations, 2014 CFR

2014-10-01

... Tribal law or code, in the population subject to the jurisdiction of the Tribal court or administrative... defined by Tribal laws or codes, in the population of the Tribes subject to the jurisdiction of the Tribal... and provide justification for operating a program with less than the minimum number of children may be...

Materials Research Center, University of Pittsburgh

DTIC Science & Technology

1994-04-29

porosities could be synthesized with active protein contained within the material. The incorporation of proteins into polyacrylates seres as a model system...enzyme with PEG and incorporate the modified enzyme into polyacrylates (Figure IV.D.3.1). The activity and stability of the functionalized enzyme have been... polyacrylate polymer. By varying the ratio of solvent (chloroform) and non-solvent (carbon tetrachloride) during the free-radical initiated

Lab-on-a-Chip Proteomic Assays for Psychiatric Disorders.

PubMed

Peter, Harald; Wienke, Julia; Guest, Paul C; Bistolas, Nikitas; Bier, Frank F

2017-01-01

Lab-on-a-chip assays allow rapid identification of multiple parameters on an automated user-friendly platform. Here we describe a fully automated multiplex immunoassay and readout in less than 15 min using the Fraunhofer in vitro diagnostics (ivD) platform to enable inexpensive point-of-care profiling of sera or a single drop of blood from patients with various diseases such as psychiatric disorders.

Quantitative T2 magnetic resonance imaging compared to morphological grading of the early cervical intervertebral disc degeneration: an evaluation approach in asymptomatic young adults.

PubMed

Chen, Chun; Huang, Minghua; Han, Zhihua; Shao, Lixin; Xie, Yan; Wu, Jianhong; Zhang, Yan; Xin, Hongkui; Ren, Aijun; Guo, Yong; Wang, Deli; He, Qing; Ruan, Dike

2014-01-01

The objective of this study was to evaluate the efficacy of quantitative T2 magnetic resonance imaging (MRI) for quantifying early cervical intervertebral disc (IVD) degeneration in asymptomatic young adults by correlating the T2 value with Pfirrmann grade, sex, and anatomic level. Seventy asymptomatic young subjects (34 men and 36 women; mean age, 22.80±2.11 yr; range, 18-25 years) underwent 3.0-T MRI to obtain morphological data (one T1-fast spin echo (FSE) and three-plane T2-FSE, used to assign a Pfirrmann grade (I-V)) and for T2 mapping (multi-echo spin echo). T2 values in the nucleus pulposus (NP, n = 350) and anulus fibrosus (AF, n = 700) were obtained. Differences in T2 values between sexes and anatomic level were evaluated, and linear correlation analysis of T2 values versus degenerative grade was conducted. Cervical IVDs of healthy young adults were commonly determined to be at Pfirrmann grades I and II. T2 values of NPs were significantly higher than those of AF at all anatomic levels (P<0.000). The NP, anterior AF and posterior AF values did not differ significantly between genders at the same anatomic level (P>0.05). T2 values decreased linearly with degenerative grade. Linear correlation analysis revealed a strong negative association between the Pfirrmann grade and the T2 values of the NP (P = 0.000) but not the T2 values of the AF (P = 0.854). However, non-degenerated discs (Pfirrmann grades I and II) showed a wide range of T2 relaxation time. T2 values according to disc degeneration level classification were as follows: grade I (>62.03 ms), grade II (54.60-62.03 ms), grade III (<54.60 ms). T2 quantitation provides a more sensitive and robust approach for detecting and characterizing the early stage of cervical IVD degeneration and to create a reliable quantitative in healthy young adults.

SU-E-T-287: Patterns of Patient Specific Dosimetry in Total Body Irradiation.

PubMed

Akino, Y; McMullen, K; Das, I

2012-06-01

Total body irradiation (TBI) is commonly used for conditioning prior to transplant in hematologic and immunologic diseases. Due to variability in body thickness, achieving dose uniformity across body within ±10% of the prescribed dose is challenging. The dose uniformity is further complicated by, techniques and beam energy used, lung shielding and selection of detector. The translational table technique for TBI could compensate for estimated delivered dose to whole body by adjusting couch speed during treatment. However, it is difficult to accurately estimate the dose by calculation and hence in vivo dosimetry (IVD) is routinely performed for TBI. The patterns of patient specific dosimetry, IVD are presented in this study. Under IRB exempt status, 161 patients who received TBI treatment between 2006 and 2011 were retrospectively analyzed using the treatment records from Cobalt-60 teletherapy unit and translational treatment couch. During treatment, IVD detectors (TLD, diode, or MOSFET) were placed on patient surface; both entrance and exit dose were recorded at the patient's head, neck, mediastinum, umbilicus, and knee. When large differences between prescribed and measured dose were observed, the dose delivery was corrected for subsequent fractions by adjustment in couch speed and/or bolus placement. Across the entire cohort, the mean (range) percent variance between calculated and measured dose were -2.3% (-66.2 - 35.3), 1.1% (-62.2 - 40.3), -1.9% (-66.4 - 46.6), -1.1% (-35.2 - 42.9), and 3.4% (-47.9 - 108.5) for head, neck, mediastinum, umbilicus, and knee, respectively. When the dose differences for multiple fractions were averaged, the compliance (±10%) between prescription and measured dose was improved as at umbilicus from 83.9% to 98.5%. Actual dose measurement analysis of TBI patients reveals a potentially wide variance from calculated dose. Dose uniformity can be significantly improved with immediate feedback after the first fraction prior to subsequent treatments. This work was supported by the JSPS Core-to-Core Program No. 23003. © 2012 American Association of Physicists in Medicine.

A research protocol for developing a Point-Of-Care Key Evidence Tool 'POCKET': a checklist for multidimensional evidence reporting on point-of-care in vitro diagnostics.

PubMed

Huddy, Jeremy R; Ni, Melody; Mavroveli, Stella; Barlow, James; Williams, Doris-Ann; Hanna, George B

2015-07-10

Point-of-care in vitro diagnostics (POC-IVD) are increasingly becoming widespread as an acceptable means of providing rapid diagnostic results to facilitate decision-making in many clinical pathways. Evidence in utility, usability and cost-effectiveness is currently provided in a fragmented and detached manner that is fraught with methodological challenges given the disruptive nature these tests have on the clinical pathway. The Point-of-care Key Evidence Tool (POCKET) checklist aims to provide an integrated evidence-based framework that incorporates all required evidence to guide the evaluation of POC-IVD to meet the needs of policy and decisionmakers in the National Health Service (NHS). A multimethod approach will be applied in order to develop the POCKET. A thorough literature review has formed the basis of a robust Delphi process and validation study. Semistructured interviews are being undertaken with POC-IVD stakeholders, including industry, regulators, commissioners, clinicians and patients to understand what evidence is required to facilitate decision-making. Emergent themes will be translated into a series of statements to form a survey questionnaire that aims to reach a consensus in each stakeholder group to what needs to be included in the tool. Results will be presented to a workshop to discuss the statements brought forward and the optimal format for the tool. Once assembled, the tool will be field-tested through case studies to ensure validity and usability and inform refinement, if required. The final version will be published online with a call for comments. Limitations include unpredictable sample representation, development of compromise position rather than consensus, and absence of blinding in validation exercise. The Imperial College Joint Research Compliance Office and the Imperial College Hospitals NHS Trust R&D department have approved the protocol. The checklist tool will be disseminated through a PhD thesis, a website, peer-reviewed publication, academic conferences and formal presentations. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

Dynamic pressurization induces transition of notochordal cells to a mature phenotype while retaining production of important patterning ligands from development.

PubMed

Purmessur, Devina; Guterl, Clare C; Cho, Samuel K; Cornejo, Marisa C; Lam, Ying W; Ballif, Bryan A; Laudier, James C Iatridis; Iatridis, James C

2013-01-01

Notochordal cells (NCs) pattern aneural and avascular intervertebral discs (IVDs), and their disappearance, is associated with onset of IVD degeneration. This study induced and characterized the maturation of nucleus pulposus (NP) tissue from a gelatinous NC-rich structure to a matrix-rich structure populated by small NP cells using dynamic pressurization in an ex vivo culture model, and also identified soluble factors from NCs with therapeutic potential. Porcine NC-rich NP tissue was cultured and loaded with hydrostatic pressure (0.5 to 2 MPa at 0.1 Hz for 2 hours) either Daily, for 1 Dose, or Control (no pressurization) groups for up to eight days. Cell phenotype and tissue maturation was characterized with measurements of cell viability, cytomorphology, nitric oxide, metabolic activity, matrix composition, gene expression, and proteomics. Daily pressurization induced transition of NCs to small NP cells with 73.8%, 44%, and 28% NCs for Control, 1 Dose and Daily groups, respectively (P < 0.0002) and no relevant cell death. Dynamic loading matured NP tissue by significantly increasing metabolic activity and accumulating Safranin-O-stained matrix. Load-induced maturation was also apparent from the significantly decreased glycolytic, cytoskeletal (Vimentin) and stress-inducible (HSP70) proteins assessed with proteomics. Loading increased the production of bioactive proteins Sonic Hedgehog (SHH) and Noggin, and maintained Semaphorin3A (Sema3A). NP tissue maturation was induced from dynamic hydrostatic pressurization in a controlled ex vivo environment without influence from systemic effects or surrounding structures. NCs transitioned into small nonvacuolated NP cells probably via differentiation as evidenced by high cell viability, lack of nitric oxide and downregulation of stress-inducible and cytoskeletal proteins. SHH, Sema3A, and Noggin, which have patterning and neurovascular-inhibiting properties, were produced in both notochordal and matured porcine NP. Results therefore provide an important piece of evidence suggesting the transition of NCs to small NP cells is a natural part of aging and not the initiation of degeneration. Bioactive candidates identified from young porcine IVDs may be isolated and harnessed for therapies to target discogenic back pain.

Response to benzodiazepines and the clinical course in malignant catatonia associated with schizophrenia: A case report.

PubMed

Ohi, Kazutaka; Kuwata, Aki; Shimada, Takamitsu; Yasuyama, Toshiki; Nitta, Yusuke; Uehara, Takashi; Kawasaki, Yasuhiro

2017-04-01

Malignant catatonia (MC) is a disorder consisting of catatonic symptoms, hyperthermia, autonomic instability, and altered mental status. Neuroleptic malignant syndrome (NMS) caused by antipsychotics is considered a variant of MC. Benzodiazepine (BZD) medications are safe and effective treatments providing rapid relief from MC. This case study reports a detailed clinical course of a case of MC associated with schizophrenia initially diagnosed as NMS that responded successfully to BZDs but not to dantrolene. A 53-year-old man with schizophrenia was admitted to the psychiatric hospital because of excitement, monologue, muscle rigidity, and insomnia. In the 3 days before admission, the patient had discontinued his medications after his family member's death. He presented with hyperthermia, tachycardia, hypertension, excessive sweating, and an elevated serum creatine phosphokinase (CPK) level. On the basis of these features, he was suspected to have NMS. The patient was treated with dantrolene for 7 days without improvement despite having a normalized serum CPK level. The patient was transferred to our university hospital for an in-depth examination and treatment of his physical status. Infection and pulmonary embolism were excluded as possible causes. To treat his excitement and auditory hallucination, an intravenous drip (IVD) of haloperidol was initiated, but this treatment increased the patient's catatonic and psychotic symptoms, although his serum CPK level had remained within a normal range. As a result, the treatment was changed to diazepam. After an IVD of diazepam, the patient's symptoms rapidly improved, and the IVD was subsequently replaced with oral administration of lorazepam. Eventually, the patient was diagnosed with MC associated with schizophrenia. BZD therapy was dramatically effective. Catatonia, MNS, and MC may be due to a common brain pathophysiology and these conditions may be in a spectrum, although uncertainty in the boundaries among conditions, and the BZD treatment may be useful. Most importantly, catatonia has not been described as a subtype of schizophrenia on the basis of the Diagnostic and Statistical Manual of Mental Disorders (DSM)-5 criteria, and the medications for catatonia and schizophrenia are different. Antipsychotics are not effective in relieving catatonia, or they may induce NMS, whereas BZDs are effective for treating both MC and NMS.

Comparison of species identification of endocarditis associated viridans streptococci using rnpB genotyping and 2 MALDI-TOF systems.

PubMed

Isaksson, Jenny; Rasmussen, Magnus; Nilson, Bo; Stadler, Liselott Svensson; Kurland, Siri; Olaison, Lars; Ek, Elisabeth; Herrmann, Björn

2015-04-01

Streptococcus spp. are important causes of infective endocarditis but challenging in species identification. This study compared identification based on sequence determination of the rnpB gene with 2 systems of matrix-assisted laser desorption ionization-time of flight mass spectrometry, MALDI Biotyper (Bruker) and VITEK MS IVD (bioMérieux). Blood culture isolates of viridans streptococci from 63 patients with infective endocarditis were tested. The 3 methods showed full agreement for all 36 isolates identified in the Anginosus, Bovis, and Mutans groups or identified as Streptococcus cristatus, Streptococcus gordonii, or Streptococcus sanguinis. None of the methods could reliably identify the 23 isolates to the species level when designated as Streptococcus mitis, Streptococcus oralis, or Streptococcus tigurinus. In 7 isolates classified to the Mitis group, the rnpB sequences deviated strikingly from all reference sequences, and additional analysis of sodA and groEL genes indicated the occurrence of yet unidentified Streptococcus spp. Copyright © 2015 Elsevier Inc. All rights reserved.

Safety and Accuracy of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry for Identification of Highly Pathogenic Organisms.

PubMed

Rudrik, James T; Soehnlen, Marty K; Perry, Michael J; Sullivan, Maureen M; Reiter-Kintz, Wanda; Lee, Philip A; Pettit, Denise; Tran, Anthony; Swaney, Erin

2017-12-01

Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) sample preparation methods, including the direct, on-plate formic acid, and ethanol/formic acid tube extraction methods, were evaluated for their ability to render highly pathogenic organisms nonviable and safe for handling in a biosafety level 2 laboratory. Of these, the tube extraction procedure was the most successful, with none of the tested strains surviving this sample preparation method. Tube extracts from several agents of bioterrorism and their near neighbors were analyzed in an eight-laboratory study to examine the utility of the Bruker Biotyper and Vitek MS MALDI-TOF MS systems and their in vitro diagnostic (IVD), research-use-only, and Security-Relevant databases, as applicable, to accurately identify these agents. Forty-six distinct strains of Bacillus anthracis , Yersinia pestis , Francisella tularensis , Burkholderia mallei , Burkholderia pseudomallei , Clostridium botulinum , Brucella melitensis , Brucella abortus , Brucella suis , and Brucella canis were extracted and distributed to participating laboratories for analysis. A total of 35 near-neighbor isolates were also analyzed. Copyright © 2017 Rudrik et al.

45 CFR 302.32 - Collection and disbursement of support payments by the IV-D Agency.

Code of Federal Regulations, 2010 CFR

2010-10-01

... collections must be sent to the family within 2 business days of the date of receipt by the SDU. (ii) Except... collections to the family within 2 business days of the date of receipt by the SDU. (iii) Except as specified... SDU must send any payment to the title IV-A family or title IV-E agency within 15 calendar days of the...

45 CFR 303.73 - Applications to use the courts of the United States to enforce court orders.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 45 Public Welfare 2 2010-10-01 2010-10-01 false Applications to use the courts of the United... Applications to use the courts of the United States to enforce court orders. The IV-D agency may apply to the Secretary for permission to use a United States district court to enforce a support order of a court of...

Evaluation of three automated nucleic acid extraction systems for identification of respiratory viruses in clinical specimens by multiplex real-time PCR.

PubMed

Kim, Yoonjung; Han, Mi-Soon; Kim, Juwon; Kwon, Aerin; Lee, Kyung-A

2014-01-01

A total of 84 nasopharyngeal swab specimens were collected from 84 patients. Viral nucleic acid was extracted by three automated extraction systems: QIAcube (Qiagen, Germany), EZ1 Advanced XL (Qiagen), and MICROLAB Nimbus IVD (Hamilton, USA). Fourteen RNA viruses and two DNA viruses were detected using the Anyplex II RV16 Detection kit (Seegene, Republic of Korea). The EZ1 Advanced XL system demonstrated the best analytical sensitivity for all the three viral strains. The nucleic acids extracted by EZ1 Advanced XL showed higher positive rates for virus detection than the others. Meanwhile, the MICROLAB Nimbus IVD system was comprised of fully automated steps from nucleic extraction to PCR setup function that could reduce human errors. For the nucleic acids recovered from nasopharyngeal swab specimens, the QIAcube system showed the fewest false negative results and the best concordance rate, and it may be more suitable for detecting various viruses including RNA and DNA virus strains. Each system showed different sensitivity and specificity for detection of certain viral pathogens and demonstrated different characteristics such as turnaround time and sample capacity. Therefore, these factors should be considered when new nucleic acid extraction systems are introduced to the laboratory.

Unique glycosignature for intervertebral disc and articular cartilage cells and tissues in immaturity and maturity.

PubMed

Collin, E C; Kilcoyne, M; White, S J; Grad, S; Alini, M; Joshi, L; Pandit, A S

2016-03-11

In this study, on/off markers for intervertebral disc (IVD) and articular cartilage (AC) cells (chondrocytes) and distinct glycoprofiles of cell and tissue-types were identified from immaturity to maturity. Three and eleven month-old ovine IVD and AC tissues were histochemically profiled with a panel of lectins and antibodies. Relationships between tissue and cell types were analysed by hierarchical clustering. Chondroitin sulfate (CS) composition of annulus fibrosus (AF), nucleus pulposus (NP) and AC tissues was determined by HPLC analysis. Clear on/off cell type markers were identified, which enabled the discrimination of chondrocytes, AF and NP cells. AF and NP cells were distinguishable using MAA, SNA-I, SBA and WFA lectins, which bound to both NP cells and chondrocytes but not AF cells. Chondrocytes were distinguished from NP and AF cells with a specific binding of LTA and PNA lectins to chondrocytes. Each tissue showed a unique CS composition with a distinct switch in sulfation pattern in AF and NP tissues upon disc maturity while cartilage maintained the same sulfation pattern over time. In conclusion, distinct glycoprofiles for cell and tissue-types across age groups were identified in addition to altered CS composition and sulfation patterns for tissue types upon maturity.

The performance of the SEPT9 gene methylation assay and a comparison with other CRC screening tests: A meta-analysis.

PubMed

Song, Lele; Jia, Jia; Peng, Xiumei; Xiao, Wenhua; Li, Yuemin

2017-06-08

The SEPT9 gene methylation assay is the first FDA-approved blood assay for colorectal cancer (CRC) screening. Fecal immunochemical test (FIT), FIT-DNA test and CEA assay are also in vitro diagnostic (IVD) tests used in CRC screening. This meta-analysis aims to review the SEPT9 assay performance and compare it with other IVD CRC screening tests. By searching the Ovid MEDLINE, EMBASE, CBMdisc and CJFD database, 25 out of 180 studies were identified to report the SEPT9 assay performance. 2613 CRC cases and 6030 controls were included, and sensitivity and specificity were used to evaluate its performance at various algorithms. 1/3 algorithm exhibited the best sensitivity while 2/3 and 1/1 algorithm exhibited the best balance between sensitivity and specificity. The performance of the blood SEPT9 assay is superior to that of the serum protein markers and the FIT test in symptomatic population, while appeared to be less potent than FIT and FIT-DNA tests in asymptomatic population. In conclusion, 1/3 algorithm is recommended for CRC screening, and 2/3 or 1/1 algorithms are suitable for early detection for diagnostic purpose. The SEPT9 assay exhibited better performance in symptomatic population than in asymptomatic population.

Accuracy of five serologic tests for the follow up of Strongyloides stercoralis infection.

PubMed

Buonfrate, Dora; Sequi, Marco; Mejia, Rojelio; Cimino, Ruben O; Krolewiecki, Alejandro J; Albonico, Marco; Degani, Monica; Tais, Stefano; Angheben, Andrea; Requena-Mendez, Ana; Muñoz, José; Nutman, Thomas B; Bisoffi, Zeno

2015-02-01

Traditional faecal-based methods have poor sensitivity for the detection of S. stercoralis, therefore are inadequate for post-treatment evaluation of infected patients who should be carefully monitored to exclude the persistence of the infection. In a previous study, we demonstrated high accuracy of five serology tests for the screening and diagnosis of strongyloidiasis. Aim of this study is to evaluate the performance of the same five tests for the follow up of patients infected with S. stercoralis. Retrospective study on anonymized, cryo-preserved samples available at the Centre for Tropical Diseases (Negrar, Verona, Italy). Samples were collected before and from 3 to 12 months after treatment. The samples were tested with two commercially-available ELISA tests (IVD, Bordier), two techniques based on a recombinant antigen (NIE-ELISA and NIE-LIPS) and one in-house IFAT. The results of each test were evaluated both in relation to the results of fecal examination and to those of a composite reference standard (classifying as positive a sample with positive stools and/or at least three positive serology tests). The associations between the independent variables age and time and the dependent variable value of serological test (for all five tests), were analyzed by linear mixed-effects regression model. A high proportion of samples demonstrated for each test a seroreversion or a relevant decline (optical density/relative light units halved or decrease of at least two titers for IFAT) at follow up, results confirmed by the linear mixed effects model that showed a trend to seroreversion over time for all tests. In particular, IVD-ELISA (almost 90% samples demonstrated relevant decline) and IFAT (almost 87%) had the best performance. Considering only samples with a complete negativization, NIE-ELISA showed the best performance (72.5% seroreversion). Serology is useful for the follow up of patients infected with S. stercoralis and determining test of cure.

The Development and Evaluation of a Computer-Based System for Managing the Design and Pilot-Testing of Interactive Videodisc Programs. Training and Development Research Center, Project Number Forty-Three.

ERIC Educational Resources Information Center

Sayre, Scott Alan

The purpose of this study was to develop and validate a computer-based system that would allow interactive video developers to integrate and manage the design components prior to production. These components of an interactive video (IVD) program include visual information in a variety of formats, audio information, and instructional techniques,…

Response to benzodiazepines and the clinical course in malignant catatonia associated with schizophrenia

PubMed Central

Ohi, Kazutaka; Kuwata, Aki; Shimada, Takamitsu; Yasuyama, Toshiki; Nitta, Yusuke; Uehara, Takashi; Kawasaki, Yasuhiro

2017-01-01

Abstract Background: Malignant catatonia (MC) is a disorder consisting of catatonic symptoms, hyperthermia, autonomic instability, and altered mental status. Neuroleptic malignant syndrome (NMS) caused by antipsychotics is considered a variant of MC. Benzodiazepine (BZD) medications are safe and effective treatments providing rapid relief from MC. This case study reports a detailed clinical course of a case of MC associated with schizophrenia initially diagnosed as NMS that responded successfully to BZDs but not to dantrolene. Case presentation: A 53-year-old man with schizophrenia was admitted to the psychiatric hospital because of excitement, monologue, muscle rigidity, and insomnia. In the 3 days before admission, the patient had discontinued his medications after his family member's death. He presented with hyperthermia, tachycardia, hypertension, excessive sweating, and an elevated serum creatine phosphokinase (CPK) level. On the basis of these features, he was suspected to have NMS. The patient was treated with dantrolene for 7 days without improvement despite having a normalized serum CPK level. The patient was transferred to our university hospital for an in-depth examination and treatment of his physical status. Infection and pulmonary embolism were excluded as possible causes. To treat his excitement and auditory hallucination, an intravenous drip (IVD) of haloperidol was initiated, but this treatment increased the patient's catatonic and psychotic symptoms, although his serum CPK level had remained within a normal range. As a result, the treatment was changed to diazepam. After an IVD of diazepam, the patient's symptoms rapidly improved, and the IVD was subsequently replaced with oral administration of lorazepam. Eventually, the patient was diagnosed with MC associated with schizophrenia. BZD therapy was dramatically effective. Conclusion: Catatonia, MNS, and MC may be due to a common brain pathophysiology and these conditions may be in a spectrum, although uncertainty in the boundaries among conditions, and the BZD treatment may be useful. Most importantly, catatonia has not been described as a subtype of schizophrenia on the basis of the Diagnostic and Statistical Manual of Mental Disorders (DSM)-5 criteria, and the medications for catatonia and schizophrenia are different. Antipsychotics are not effective in relieving catatonia, or they may induce NMS, whereas BZDs are effective for treating both MC and NMS. PMID:28422845

Age-related accumulation of pentosidine in aggrecan and collagen from normal and degenerate human intervertebral discs

PubMed Central

Sivan, Sarit Sara; Tsitron, Eve; Wachtel, Ellen; Roughley, Peter; Sakkee, Nico; van der Ham, Frits; Degroot, Jeroen; Maroudas, Alice

2006-01-01

During aging and degeneration, many changes occur in the structure and composition of human cartilaginous tissues, which include the accumulation of the AGE (advanced glycation end-product), pentosidine, in long-lived proteins. In the present study, we investigated the accumulation of pentosidine in constituents of the human IVD (intervertebral disc), i.e. collagen, aggrecan-derived PG (proteoglycan) (A1) and its fractions (A1D1–A1D6) in health and pathology. We found that, after maturity, pentosidine accumulates with age. Over the age range studied, a linear 6-fold increase was observed in pentosidine accumulation for A1 and collagen with respective rates of 0.12 and 0.66 nmol·(g of protein)−1·year−1. Using previously reported protein turnover rate constants (kT) obtained from measurements of the D-isomer of aspartic residue in collagen and aggrecan of human IVD, we could calculate the pentosidine formation rate constants (kF) for these constituents [Sivan, Tsitron, Wachtel, Roughley, Sakkee, van der Ham, DeGroot, Roberts and Maroudas (2006) J. Biol. Chem. 281, 13009–13014; Tsitron (2006) MSc Thesis, Technion-Israel Institute of Technology, Haifa, Israel]. In spite of the comparable formation rate constants obtained for A1D1 and collagen [1.81±0.25 compared with 3.71±0.26 μmol of pentosidine·(mol of lysine)−1·year−1 respectively], the higher pentosidine accumulation in collagen is consistent with its slower turnover (0.005 year−1 compared with 0.134 year−1 for A1D1). Pentosidine accumulation increased with decreasing buoyant density and decreasing turnover of the proteins from the most glycosaminoglycan-rich PG components (A1D1) to the least (A1D6), with respective kF values of 1.81±0.25 and 3.18±0.37 μmol of pentosidine·(mol of lysine)−1·year−1. We concluded that protein turnover is an important determinant of pentosidine accumulation in aggrecan and collagen of human IVD, as was found for articular cartilage. Correlation of pentosidine accumulation with protein half-life in both normal and degenerate discs further supports this finding. PMID:16787390

Human umbilical cord mesenchymal stromal cells exhibit immature nucleus pulposus cell phenotype in a laminin-rich pseudo-three-dimensional culture system.

PubMed

Chon, Brian H; Lee, Esther J; Jing, Liufang; Setton, Lori A; Chen, Jun

2013-10-02

Cell supplementation to the herniated or degenerated intervertebral disc (IVD) is a potential strategy to promote tissue regeneration and slow disc pathology. Human umbilical cord mesenchymal stromal cells (HUCMSCs) - originating from the Wharton's jelly - remain an attractive candidate for such endeavors with their ability to differentiate into multiple lineages. Previously, mesenchymal stem cells (MSCs) have been studied as a potential source for disc tissue regeneration. However, no studies have demonstrated that MSCs can regenerate matrix with unique characteristics matching that of immature nucleus pulposus (NP) tissues of the IVD. In our prior work, immature NP cells were found to express specific laminin isoforms and laminin-binding receptors that may serve as phenotypic markers for evaluating MSC differentiation to NP-like cells. The goal of this study is to evaluate these markers and matrix synthesis for HUCMSCs cultured in a laminin-rich pseudo-three-dimensional culture system. HUCMSCs were seeded on top of Transwell inserts pre-coated with Matrigel™, which contained mainly laminin-111. Cells were cultured under hypoxia environment with three differentiation conditions: NP differentiation media (containing 2.5% Matrigel™ solution to provide for a pseudo-three-dimensional laminin culture system) with no serum, or the same media supplemented with either insulin-like growth factor-1 (IGF-1) or transforming growth factor-β1 (TGF-β1). Cell clustering behavior, matrix production and the expression of NP-specific laminin and laminin-receptors were evaluated at days 1, 7, 13 and 21 of culture. Data show that a pseudo-three-dimensional culture condition (laminin-1 rich) promoted HUCMSC differentiation under no serum conditions. Starting at day 1, HUCMSCs demonstrated a cell clustering morphology similar to that of immature NP cells in situ and that observed for primary immature NP cells within the similar laminin-rich culture system (prior study). Differentiated HUCMSCs under all conditions were found to contain glycosaminoglycan, expressed extracellular matrix proteins of collagen II and laminin α5, and laminin receptors (integrin α3 and β4 subunits). However, neither growth factor treatment generated distinct differences in NP-like phenotype for HUCMSC as compared with no-serum conditions. HUCMSCs have the potential to differentiate into cells sharing features with immature NP cells in a laminin-rich culture environment and may be useful for IVD cellular therapy.

Cadmium (Tank) Electroplating Alternative

DTIC Science & Technology

2011-08-01

ASTM F519 HE: 75% NFS 200 hrs HRE : 45% NFS 150 hrs Threshold limit greater than /equal to LHE Cd (AMS 2417G) ASETS Defense Focused Workshop (2011...Test Specimens  Reporting Sustained/Threshold load (%NFS), Time to failure. HRE Testing Cd Zn-Ni IVD Al LHE Cd Re-Embrittlement Test Fluids:  DI...Hydrogen Embrittlement/ HRE ASTM F519 A5, Type 1.a.1 Brush Plating ASETS Defense Focused Workshop (2011) Luzmarie G. Santiago Materials Engineer Naval Air

Development of Ultrasound to Measure In-Vivo Dynamic Cervical Spine Intervertebral Disc Mechanics

DTIC Science & Technology

2016-01-01

Award Number: W81XWH-13-1-0050 TITLE: Development of Ultrasound to Measure In-vivo Dynamic Cervical Spine Intervertebral Disc Mechanics PRINCIPAL...CONTRACT NUMBER W81XWH-13-1-0050 Development of Ultrasound to Measure In-vivo Dynamic Cervical Spine Intervertebral Disc Mechanics 5b. GRANT NUMBER 5c...elasticity during compression or tension. As a portable, low cost imaging modality, the dual ultrasound system quantified cervical spine IVD displacement and

Fatigue Testing of AA7050-T7451 with Various Corrosion Prevention Surface Treatments

DTIC Science & Technology

2013-06-01

UNCLASSIFIED Authors Marcus McDonald Air Vehicles Division Marcus McDonald initially spent 3 years working in the oil & gas, steel and...treatments: caustic degreasing, de-oxidising or anodising. The pits in the anodised coupons were larger than the pits caused by the pre-IVD etching...those fatigue cracks that grew from them spent less time in growing whilst they were small, because the crack started at what was a larger effective

Magnetic resonance imaging versus computed tomography to plan hemilaminectomies in chondrodystrophic dogs with intervertebral disc extrusion.

PubMed

Noyes, Julie A; Thomovsky, Stephanie A; Chen, Annie V; Owen, Tina J; Fransson, Boel A; Carbonneau, Kira J; Matthew, Susan M

2017-10-01

To determine the influence of preoperative computed tomography (CT) versus magnetic resonance (MR) on hemilaminectomies planned to treat thoracolumbar (TL) intervertebral disc (IVD) extrusions in chondrodystrophic dogs. Prospective clinical study. Forty chondrodystrophic dogs with TL IVD extrusion and preoperative CT and MR studies. MR and CT images were randomized and reviewed by 4 observers masked to the dog's identity and corresponding imaging studies. Observers planned the location along the spine, side, and extent (number of articular facets to be removed) based on individual reviews of CT and MR studies. Intra-observer agreement was determined between overall surgical plan, location, side, and size of the hemilaminectomy planned on CT versus MR of the same dog. Similar surgical plans were developed based on MR versus CT in 43.5%-66.6% of dogs, depending on the observer. Intra-observer agreement in location, side, and size of the planned hemilaminectomy based on CT versus MR ranged between 48.7%-66.6%, 87%-92%, and 51.2%-71.7% of dogs, respectively. Observers tended to plan larger laminectomy defects based on MR versus CT of the same dog. Findings from this study indicated considerable differences in hemilaminectomies planned on preoperative MR versus CT imaging. Surgical location and size varied the most; the side of planned hemilaminectomies was most consistent between imaging modalities. © 2017 The American College of Veterinary Surgeons.

2-dimensional ion velocity distributions measured by laser-induced fluorescence above a radio-frequency biased silicon wafer

NASA Astrophysics Data System (ADS)

Moore, Nathaniel B.; Gekelman, Walter; Pribyl, Patrick; Zhang, Yiting; Kushner, Mark J.

2013-08-01

The dynamics of ions traversing sheaths in low temperature plasmas are important to the formation of the ion energy distribution incident onto surfaces during microelectronics fabrication. Ion dynamics have been measured using laser-induced fluorescence (LIF) in the sheath above a 30 cm diameter, 2.2 MHz-biased silicon wafer in a commercial inductively coupled plasma processing reactor. The velocity distribution of argon ions was measured at thousands of positions above and radially along the surface of the wafer by utilizing a planar laser sheet from a pulsed, tunable dye laser. Velocities were measured both parallel and perpendicular to the wafer over an energy range of 0.4-600 eV. The resulting fluorescence was recorded using a fast CCD camera, which provided resolution of 0.4 mm in space and 30 ns in time. Data were taken at eight different phases during the 2.2 MHz cycle. The ion velocity distributions (IVDs) in the sheath were found to be spatially non-uniform near the edge of the wafer and phase-dependent as a function of height. Several cm above the wafer the IVD is Maxwellian and independent of phase. Experimental results were compared with simulations. The experimental time-averaged ion energy distribution function as a function of height compare favorably with results from the computer model.

In vivo quality assurance of volumetric modulated arc therapy for ano-rectal cancer with thermoluminescent dosimetry and image-guidance.

PubMed

Dipasquale, Giovanna; Nouet, Philippe; Rouzaud, Michel; Dubouloz, Angèle; Miralbell, Raymond; Zilli, Thomas

2014-06-01

To assess in vivo dose distribution using cone-beam computed tomography scans (CBCTs) and thermoluminescent dosimeters (TLDs) in patients with anal or rectal cancer treated with volumetric modulated arc therapy (VMAT). Intracavitary (IC) in vivo dosimetry (IVD) was performed in 11 patients using adapted endorectal probes containing TLDs, with extra measurements at the perianal skin (PS) for anal margin tumors. Measured doses were compared to calculated ones obtained from image fusion of CBCT with CT treatments plans. A total of 55 IC and 6 PS measurements were analyzed. IC TLD median planned and measured doses were 1.81 Gy (range, 0.25-2.02 Gy) and 1.82 Gy (range, 0.19-2.12 Gy), respectively. In comparison to the planned doses all IC TLD dose measurements differed by a median dose of 0.02 Gy (range, -0.11/+0.19 Gy, p=0.102) (median difference of 1.1%, range -6.1%/+10.6%). Overall, 95% of IC measurements were within ±7.7% of the expected percentage doses and only 1 value was above +10%. For PS measurements, only one was not within ±7.7% of expected values (i.e., -8.9%). Image guidance using CBCT for IVD with TLDs is helpful to validate the delivered doses in patients treated with VMAT for ano-rectal tumors. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

Indian Hedgehog signaling pathway members are associated with magnetic resonance imaging manifestations and pathological scores in lumbar facet joint osteoarthritis.

PubMed

Shuang, Feng; Zhou, Ying; Hou, Shu-Xun; Zhu, Jia-Liang; Liu, Yan; Zhang, Chun-Li; Tang, Jia-Guang

2015-05-20

Indian Hedgehog (HH) has been shown to be involved in osteoarthritis (OA) in articular joints, where there is evidence that Indian HH blockade could ameliorate OA. It seems to play a prominent role in development of the intervertebral disc (IVD) and in postnatal maintenance. There is little work on IHH in the IVD. Hence the aim of the current study was to investigate the role of Indian Hedgehog in the pathology of facet joint (FJ) OA. 24 patients diagnosed with lumbar intervertebral disk herniation or degenerative spinal stenosis were included. Preoperative magnetic resonance imaging (MRI) and Osteoarthritis Research Society International (OARSI) histopathology grading system was correlated to the mRNA levels of GLI1, PTCH1, and HHIP in the FJs. The Weishaupt grading and OARSI scores showed high positive correlation (r = 0.894) (P < 0.01). MRI Weishaupt grades showed positive correlation with GLI1 (r = 0.491), PTCH1 (r = 0.444), and HHIP (r = 0.654) mRNA levels (P < 0.05 in each case). OARSI scores were also positively correlated with GLI1 (r = 0. 646), PTCH1 (r = 0. 518), and HHIP (r = 0.762) mRNA levels (P < 0.01 in each case). Cumulatively our findings indicate that Indian HH signaling is increased in OA and is perhaps a key component in OA pathogenesis and progression.

Guidelines on the implementation of diode in vivo dosimetry programs for photon and electron external beam therapy.

PubMed

Alecu, R; Loomis, T; Alecu, J; Ochran, T

1999-01-01

Semiconductor diodes offer many advantages for clinical dosimetry: high sensitivity, real-time readout, simple instrumentation, robustness and air pressure independence. The feasibility and usefulness of in vivo dosimetry with diodes has been shown by numerous publications, but very few, if any, refer to the utilization of diodes in electron beam dosimetry. The purpose of this paper is to present our methods for implementing an effective IVD program for external beam therapy with photons and electrons and to evaluate a new type of diodes. Methods of deciding on reasonable action levels along with calibration procedures, established according to the type of measurements intended to be performed and the action limits, are discussed. Correction factors to account for nonreference clinical conditions for new types of diodes (designed for photon and electron beams) are presented and compared with those required by older models commercially available. The possibilities and limitations of each type of diode are presented, emphasizing the importance of using the appropriate diode for each task and energy range.

Parallel Unsteady Overset Mesh Methodology for a Multi-Solver Paradigm with Adaptive Cartesian Grids

DTIC Science & Technology

2008-08-21

Engineer, U.S. Army Research Laboratory ., Matthew.W.Floros@nasa.gov, AIAA Member ‡Senior Research Scientist, Scaled Numerical Physics LLC., awissink...IV.E and IV.D). Good linear scalability was observed for all three cases up to 12 processors. Beyond that the scalability drops off depending on grid...Research Laboratory for the usage of SUGGAR module and Yikloon Lee at NAVAIR for the usage of the NAVAIR-IHC code. 13 of 22 American Institute of

Effect of Repeated Normobaric Hypoxia Exposures during Sleep on Acute Mountain Sickness, Exercise Performance, and Sleep during Exposure to Terrestrial Altitude

DTIC Science & Technology

2011-01-01

physiological adjustments that compensate for hypoxemia, with augmented ventilation being one of the most important and consistently reported (17, 18, 22, 28... physiological outcomes were affected favorably relative to no treatment utilized HH treatment prior to HH residence (2–4, 18) or NH treatment prior to NH...erythropoietin (EPO; Quantikine IVD ELISA, R & D Systems, Minneapolis, MN), epinephrine and norepinephine (HPLC; Bio-Rad), and cortisol and aldosterone

Development of Ultrasound to Measure In-vivo Dynamic Cervical Spine Intervertebral Disc Mechanics

DTIC Science & Technology

2014-01-01

The deformation between C4 and C6 measured by the US probe was affected by bulging of the IVD and soft tissues during compressive loading as...endplates of the vertebrae and cartilaginous endplate of the discs were added to all segments. Figure 28 Coronal views of the updated C4-T1 FEM (a...the ligaments and soft tissue connections that provide stability to the cervical spine FSUs were added (Figures 30 and 31). For the anterior

Distinction between the extracellular matrix of the nucleus pulposus and hyaline cartilage: a requisite for tissue engineering of intervertebral disc.

PubMed

Mwale, F; Roughley, P; Antoniou, J

2004-12-15

Tissue engineering of intervertebral discs (IVD) using mesenchymal stem cells (MSCs) induced to differentiate into a disc-cell phenotype has been considered as an alternative treatment for disc degeneration. However, since there is no unique marker characteristic of discs and since hyaline cartilage and immature nucleus pulposus (NP) possess similar macromolecules in their extracellular matrix, it is currently difficult to recognize MSC conversion to a disc cell. This study was performed to compare the proteoglycan to collagen ratio (measured as GAG to hydroxyproline ratio) in the NP of normal disc to that of the hyaline cartilage of the endplate within the same group of individuals and test the hypothesis that this ratio can be used for in vivo studies to distinguish between a normal NP and hyaline cartilage phenotype. Whole human lumbar spine specimens from fresh cadavers, ranging in age from 12 weeks to 79 years, were used to harvest the IVDs and adjacent endplates. The GAG to hydroxyproline ratio within the NP of young adults is approximately 27:1, whereas the ratio within the hyaline cartilage endplate of the same aged individuals is about 2:1. The production of an extracellular matrix with a high proteoglycan to collagen ratio can be used in vivo to distinguish NP cells from chondrocytes, and could help in identifying a NP-like phenotype in vivo as opposed to a chondrocyte when MSCs are induced to differentiate for tissue engineering of a disc.

Indian Hedgehog signaling pathway members are associated with magnetic resonance imaging manifestations and pathological scores in lumbar facet joint osteoarthritis

NASA Astrophysics Data System (ADS)

Shuang, Feng; Zhou, Ying; Hou, Shu-Xun; Zhu, Jia-Liang; Liu, Yan; Zhang, Chun-Li; Tang, Jia-Guang

2015-05-01

Indian Hedgehog (HH) has been shown to be involved in osteoarthritis (OA) in articular joints, where there is evidence that Indian HH blockade could ameliorate OA. It seems to play a prominent role in development of the intervertebral disc (IVD) and in postnatal maintenance. There is little work on IHH in the IVD. Hence the aim of the current study was to investigate the role of Indian Hedgehog in the pathology of facet joint (FJ) OA. 24 patients diagnosed with lumbar intervertebral disk herniation or degenerative spinal stenosis were included. Preoperative magnetic resonance imaging (MRI) and Osteoarthritis Research Society International (OARSI) histopathology grading system was correlated to the mRNA levels of GLI1, PTCH1, and HHIP in the FJs. The Weishaupt grading and OARSI scores showed high positive correlation (r = 0.894) (P < 0.01). MRI Weishaupt grades showed positive correlation with GLI1 (r = 0.491), PTCH1 (r = 0.444), and HHIP (r = 0.654) mRNA levels (P < 0.05 in each case). OARSI scores were also positively correlated with GLI1 (r = 0. 646), PTCH1 (r = 0. 518), and HHIP (r = 0.762) mRNA levels (P < 0.01 in each case). Cumulatively our findings indicate that Indian HH signaling is increased in OA and is perhaps a key component in OA pathogenesis and progression.

Lactoferricin mediates anabolic and anti-catabolic effects in the intervertebral disc.

PubMed

Kim, Jae-Sung; Ellman, Michael B; An, Howard S; Yan, Dongyao; van Wijnen, Andre J; Murphy, Gillian; Hoskin, David W; Im, Hee-Jeong

2012-04-01

Lactoferricin (LfcinB) antagonizes biological effects mediated by angiogenic and catabolic growth factors, in addition to pro-inflammatory cytokines and chemokines in human endothelial cells and tumor cells. However, the effect of LfcinB on intervertebral disc (IVD) cell metabolism has not yet been investigated. Using bovine nucleus pulposus (NP) cells, we analyzed the effect of LfcinB on proteoglycan (PG) accumulation, PG synthesis, and anabolic gene expression. We assessed expression of genes for matrix-degrading enzymes such as matrix metalloproteases (MMPs) and a disintegrin-like and metalloprotease with thrombospondin motifs (ADAMTS family), as well as their endogenous inhibitors, tissue inhibitor of metalloproteases (TIMPs). In order to understand the specific molecular mechanisms by which LfcinB exerts its biological effects, we investigated intracellular signaling pathways in NP cells. LfcinB increased PG accumulation mainly via PG synthesis in a dose-dependent manner. Simultaneously, LfcinB dose-dependently downregulated catabolic enzymes. LfcinB's anti-catabolic effects were further demonstrated by a dose-dependent increase in multiple TIMP family members. Our results demonstrate that ERK and/or p38 mitogen-activated protein kinase pathways are the key signaling cascades that exert the biological effects of LfcinB in NP cells, regulating transcription of aggrecan, SOX-9, TIMP-1, TIMP-2, TIMP-3, and iNOS. Our results suggest that LfcinB has anabolic and potent anti-catabolic biological effects on bovine IVD cells that may have considerable promise in the treatment of disc degeneration in the future. Copyright © 2011 Wiley Periodicals, Inc.

Correlation of matrix metalloproteinase (MMP)-1, -2, -3, and -9 expressions with demographic and radiological features in primary lumbar intervertebral disc disease.

PubMed

Basaran, Recep; Senol, Mehmet; Ozkanli, Seyma; Efendioglu, Mustafa; Kaner, Tuncay

2017-07-01

Degeneration of IVD is a progressive and irreversible process and can be evaluated with immunohistochemical examination or radiological grading. MMPs are a family of proteolytic enzymes and involved in the degradation of the matrix components of the IVD. We aimed to compare MMP-1, -2, -3, and -9 expressions with demographic features, visual analogue scale (VAS), Oswestry Disability Index (ODI) and radiological (MRI) grades. The study involved 60 participants. We recorded data about age, complaint, radiological imaging, expression levels of MMP-1, -2, -3, and -9, ODI and VAS for back pain retrospectively. Intervertebral disc degeneration was graded on a 0-5 scale according to the Pfirrmann classification. As a result of the study, the median age was 52.09±12.74years. There were statistical significances between age and MMP-1, and MMP-2. There was a close correlation between grade and MMP-9. We found correlation between the VAS and the MMP-9 expression. In addition, there was relationship between expression of MMP-2 and MMP-1, MMP-3, MMP-9. In conclusion, the expressions of MMP-1 and -2 are increased with aging. There was no relationship between radiological evaluation of IVDD and aging. Increased expression of MMPs affected IVDD positively. The relationship with MMPs is not explained. This study adds to our understanding of the interaction between MMPs and IVDD. Copyright © 2017 Elsevier Ltd. All rights reserved.

International Symposium on Gas Kinetics (8th) Held in Nottingham, England on 15-20 July 1984. Abstracts

DTIC Science & Technology

1984-07-20

Hall 16.00 -17.30 Tea, Hugh Stewart Hall 19.00 Buffet/Reception, Hugh Stewart Hall 19.00- 24.00 Cash bar in Hugh Stewart Hall Accesion For STATEMENT ...transfer involving highly vibrationally excited molecules J. R. Barker (SRI), T. C. Brown and K. D. King (Adelaide) 10.45-11.15 Coffee SESSION IV...D. King (Adelaide) and R. G. Gilbert (Sydney) 15.15-15.35 J4 Quantitative intermolecular energy transfer efficiencies from thermal studies C. D. Eley

SensiScreen® KRAS exon 2-sensitive simplex and multiplex real-time PCR-based assays for detection of KRAS exon 2 mutations

PubMed Central

Guldmann-Christensen, Mariann; Hauge Kyneb, Majbritt; Voogd, Kirsten; Andersen, Christina; Epistolio, Samantha; Merlo, Elisabetta; Yding Wolff, Tine; Hamilton-Dutoit, Stephen; Lorenzen, Jan; Christensen, Ulf Bech

2017-01-01

Activating mutations in codon 12 and codon 13 of the KRAS (Kirsten rat sarcoma viral oncogene homolog) gene are implicated in the development of several human cancer types and influence their clinical evaluation, treatment and prognosis. Numerous different methods for KRAS genotyping are currently available displaying a wide range of sensitivities, time to answer and requirements for laboratory equipment and user skills. Here we present SensiScreen® KRAS exon 2 simplex and multiplex CE IVD assays, that use a novel real-time PCR-based method for KRAS mutation detection based on PentaBase’s proprietary DNA analogue technology and designed to work on standard real-time PCR instruments. By means of the included BaseBlocker™ technology, we show that SensiScreen® specifically amplifies the mutated alleles of interest with no or highly subdued amplification of the wild type allele. Furthermore, serial dilutions of mutant DNA in a wild type background demonstrate that all SensiScreen® assays display a limit of detection that falls within the range of 0.25–1%. Finally, in three different colorectal cancer patient populations, SensiScreen® assays confirmed the KRAS genotype previously determined by commonly used methods for KRAS mutation testing, and notably, in two of the populations, SensiScreen® identified additional mutant positive cases not detected by common methods. PMID:28636636

Photo-crosslinked alginate hydrogels support enhanced matrix accumulation by nucleus pulposus cells in vivo.

PubMed

Chou, A I; Akintoye, S O; Nicoll, S B

2009-10-01

Intervertebral disc (IVD) degeneration is a major health concern in the United States. Replacement of the nucleus pulposus (NP) with injectable biomaterials represents a potential treatment strategy for IVD degeneration. The objective of this study was to characterize the extracellular matrix (ECM) assembly and functional properties of NP cell-encapsulated, photo-crosslinked alginate hydrogels in comparison to ionically crosslinked alginate constructs. Methacrylated alginate was synthesized by esterification of hydroxyl groups with methacrylic anhydride. Bovine NP cells were encapsulated in alginate hydrogels by ionic crosslinking using CaCl(2) or through photo-crosslinking upon exposure to long-wave UV light in the presence of a photoinitiator. The hydrogels were evaluated in vitro by gross and histological analysis and in vivo using a murine subcutaneous pouch model. In vivo samples were analyzed for gene expression, ECM localization and accumulation, and equilibrium mechanical properties. Ionically crosslinked hydrogels exhibited inferior proteoglycan accumulation in vitro and were unable to maintain structural integrity in vivo. In further studies, photo-crosslinked alginate hydrogels were implanted for up to 8 weeks to examine NP tissue formation. Photo-crosslinked hydrogels displayed temporal increases in gene expression and assembly of type II collagen and proteoglycans. Additionally, hydrogels remained intact over the duration of the study and the equilibrium Young's modulus increased from 1.24+/-0.09 kPa to 4.31+/-1.39 kPa, indicating the formation of functional matrix with properties comparable to those of the native NP. These findings support the use of photo-crosslinked alginate hydrogels as biomaterial scaffolds for NP replacement.

[Evidence level and grade of recommendation of the Ponseti method for arthrogryposis-related syndromic talipes equinovarus and Moebius syndrome: a systematic review].

PubMed

Guerra-Jasso, J J; Valcarce-León, J A; Quíntela-Núñez-Del Prado, H M

2017-01-01

Adduct congenital talipes equinovarus (CTE) or clubfoot is a common musculoskeletal malformation affecting between 1 and 4.5 out of every 1000 live births. It is usually associated with arthrogryposis and Moebius syndrome with the latter two being more difficult to treat as they require aggressive surgery to achieve a plantigrade foot. We therefore ask what is the evidence level and grade of recommendation of the Ponseti method to treat syndromic talipes equinovarus resulting from arthrogryposis and Moebius syndrome. Systematic review according to the recommendations of the Cochrane group identifying medical subheadings (MeSH) and of the Boolean method to identify the articles that met the selection criteria through the search of primary sources such as OVID, Cochrane, EBSCOhost, Elsevier, Medigraphic. A total of six papers met the selection criteria through the search in the primary sources. Five of them address the treatment of adduct CTE with the Ponseti method in patients with arthrogryposis, and one in patients with Moebius. Evidence levels and grades of recommendation of these papers were: IV-D (3), III-D (1) y II (B). The Ponseti method is an first-line treatment for arthrogryposis and Moebius syndrome. It is unexpensive, non-invasive and does not require a physician to perform it (however, it does require trained staff). Its use is recommended as it avoids extensive surgeries and medical complications that go beyond the hospitals economic possibilities.

RNA sample preparation applied to gene expression profiling for the horse biological passport.

PubMed

Bailly-Chouriberry, Ludovic; Baudoin, Florent; Cormant, Florence; Glavieux, Yohan; Loup, Benoit; Garcia, Patrice; Popot, Marie-Agnès; Bonnaire, Yves

2017-09-01

The improvement of doping control is an ongoing race. Techniques to fight doping are usually based on the direct detection of drugs or their metabolites by analytical methods such as chromatography hyphenated to mass spectrometry after ad hoc sample preparation. Nowadays, omic methods constitute an attractive development and advances have been achieved particularly by application of molecular biology tools for detection of anabolic androgenic steroids (AAS), erythropoiesis-stimulating agent (ESA), or to control human growth hormone misuses. These interesting results across different animal species have suggested that modification of gene expression offers promising new methods of improving the window of detection of banned substances by targeting their effects on blood cell gene expression. In this context, the present study describes the possibility of using a modified version of the dedicated Human IVD (in vitro Diagnostics) PAXgene® Blood RNA Kit for horse gene expression analysis in blood collected on PAXgene® tubes applied to the horse biological passport. The commercial kit was only approved for human blood samples and has required an optimization of specific technical requirements for equine blood samples. Improvements and recommendations were achieved for sample collection, storage and RNA extraction procedure. Following these developments, RNA yield and quality were demonstrated to be suitable for downstream gene expression analysis by qPCR techniques. Copyright © 2017 John Wiley & Sons, Ltd. Copyright © 2017 John Wiley & Sons, Ltd.

Rapid Differentiation of Haemophilus influenzae and Haemophilus haemolyticus by Use of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry with ClinProTools Mass Spectrum Analysis.

PubMed

Chen, Jonathan H K; Cheng, Vincent C C; Wong, Chun-Pong; Wong, Sally C Y; Yam, Wing-Cheong; Yuen, Kwok-Yung

2017-09-01

Haemophilus influenzae is associated with severe invasive disease, while Haemophilus haemolyticus is considered part of the commensal flora in the human respiratory tract. Although the addition of a custom mass spectrum library into the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) system could improve identification of these two species, the establishment of such a custom database is technically complicated and requires a large amount of resources, which most clinical laboratories cannot afford. In this study, we developed a mass spectrum analysis model with 7 mass peak biomarkers for the identification of H. influenzae and H. haemolyticus using the ClinProTools software. We evaluated the diagnostic performance of this model using 408 H. influenzae and H. haemolyticus isolates from clinical respiratory specimens from 363 hospitalized patients and compared the identification results with those obtained with the Bruker IVD MALDI Biotyper. The IVD MALDI Biotyper identified only 86.9% of H. influenzae (311/358) and 98.0% of H. haemolyticus (49/50) clinical isolates to the species level. In comparison, the ClinProTools mass spectrum model could identify 100% of H. influenzae (358/358) and H. haemolyticus (50/50) clinical strains to the species level and significantly improved the species identification rate (McNemar's test, P < 0.0001). In conclusion, the use of ClinProTools demonstrated an alternative way for users lacking special expertise in mass spectrometry to handle closely related bacterial species when the proprietary spectrum library failed. This approach should be useful for the differentiation of other closely related bacterial species. Copyright © 2017 American Society for Microbiology.

Rapid Differentiation of Haemophilus influenzae and Haemophilus haemolyticus by Use of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry with ClinProTools Mass Spectrum Analysis

PubMed Central

Cheng, Vincent C. C.; Wong, Chun-Pong; Wong, Sally C. Y.; Yam, Wing-Cheong

2017-01-01

ABSTRACT Haemophilus influenzae is associated with severe invasive disease, while Haemophilus haemolyticus is considered part of the commensal flora in the human respiratory tract. Although the addition of a custom mass spectrum library into the matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) system could improve identification of these two species, the establishment of such a custom database is technically complicated and requires a large amount of resources, which most clinical laboratories cannot afford. In this study, we developed a mass spectrum analysis model with 7 mass peak biomarkers for the identification of H. influenzae and H. haemolyticus using the ClinProTools software. We evaluated the diagnostic performance of this model using 408 H. influenzae and H. haemolyticus isolates from clinical respiratory specimens from 363 hospitalized patients and compared the identification results with those obtained with the Bruker IVD MALDI Biotyper. The IVD MALDI Biotyper identified only 86.9% of H. influenzae (311/358) and 98.0% of H. haemolyticus (49/50) clinical isolates to the species level. In comparison, the ClinProTools mass spectrum model could identify 100% of H. influenzae (358/358) and H. haemolyticus (50/50) clinical strains to the species level and significantly improved the species identification rate (McNemar's test, P < 0.0001). In conclusion, the use of ClinProTools demonstrated an alternative way for users lacking special expertise in mass spectrometry to handle closely related bacterial species when the proprietary spectrum library failed. This approach should be useful for the differentiation of other closely related bacterial species. PMID:28637909

In vitro and biomechanical screening of polyethylene glycol and poly(trimethylene carbonate) block copolymers for annulus fibrosus repair.

PubMed

Long, Rose G; Rotman, Stijn G; Hom, Warren W; Assael, Dylan J; Illien-Jünger, Svenja; Grijpma, Dirk W; Iatridis, James C

2018-02-01

Herniated intervertebral discs (IVDs) are a common cause of back and neck pain. There is an unmet clinical need to seal annulus fibrosus (AF) defects, as discectomy surgeries address acute pain but are complicated by reherniation and recurrent pain. Copolymers of polyethylene glycol with trimethylene carbonate (TMC) and hexamethylene diisocyanate (HDI) end-groups were formulated as AF sealants as the HDI form covalent bonds with native AF tissue. TMC adhesives were evaluated and optimized using the design criteria: stable size, strong adherence to AF tissue, high cytocompatibility, restoration of IVD biomechanics to intact levels following in situ repair, and low extrusion risk. TMC adhesives had high adhesion strength as assessed with a pushout test (150 kPa), and low degradation rates over 3 weeks in vitro. Both TMC adhesives had shear moduli (220 and 490 kPa) similar to, but somewhat higher than, AF tissue. The adhesive with three TMC moieties per branch (TMC3) was selected for additional in situ testing because it best matched AF shear properties. TMC3 restored torsional stiffness, torsional hysteresis area and axial range of motion to intact states. However, in a failure test of compressive deformation under fixed 5 ° flexion, some herniation risk was observed with failure strength of 5.9 MPa compared with 13.5 MPa for intact samples; TMC3 herniated under cyclic organ culture testing. These TMC adhesives performed well during in vitro and in situ testing, but additional optimization to enhance failure strength is required to further this material to advanced screening tests, such as long-term degradation. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

Fibrotic-like changes in degenerate human intervertebral discs revealed by quantitative proteomic analysis.

PubMed

Yee, A; Lam, M P Y; Tam, V; Chan, W C W; Chu, I K; Cheah, K S E; Cheung, K M C; Chan, D

2016-03-01

Intervertebral disc degeneration (IDD) can lead to symptomatic conditions including sciatica and back pain. The purpose of this study is to understand the extracellular matrix (ECM) changes in disc biology through comparative proteomic analysis of degenerated and non-degenerated human intervertebral disc (IVD) tissues of different ages. Seven non-degenerated (11-46 years of age) and seven degenerated (16-53 years of age) annulus fibrosus (AF) and nucleus pulposus (NP) samples were used. Proteins were extracted using guanidine hydrochloride, separated from large proteoglycans (PGs) by caesium chloride (CsCl) density gradient ultracentrifugation, and identified using liquid chromatography (LC) coupled with tandem mass spectrometry (MS/MS). For quantitative comparison, proteins were labeled with iTRAQ reagents. Collagen fibrils in the NP were assessed using scanning electron microscopy (SEM). In the AF, quantitative analysis revealed increased levels of HTRA1, COMP and CILP in degeneration when compared with samples from older individuals. Fibronectin showed increment with age and degeneration. In the NP, more CILP and CILP2 were present in degenerated samples of younger individuals. Reduced protein solubility was observed in degenerated and older non-degenerated samples correlated with an accumulation of type I collagen in the insoluble fibers. Characterization of collagen fibrils in the NP revealed smaller mean fibril diameters and decreased porosity in the degenerated samples. Our study identified distinct matrix changes associated with aging and degeneration in the intervertebral discs (IVDs). The nature of the ECM changes, together with observed decreased in solubility and changes in fibril diameter is consistent with a fibrotic-like environment. Copyright © 2015 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.

Prediction of glycosaminoglycan synthesis in intervertebral disc under mechanical loading.

PubMed

Gao, Xin; Zhu, Qiaoqiao; Gu, Weiyong

2016-09-06

The loss of glycosaminoglycan (GAG) content is a major biochemical change during intervertebral disc (IVD) degeneration. Abnormal mechanical loading is one of the major factors causing disc degeneration. In this study, a multiscale mathematical model was developed to quantify the effect of mechanical loading on GAG synthesis. This model was based on a recently developed cell volume dependent GAG synthesis theory that predicts the variation of GAG synthesis rate of a cell under the influence of mechanical stimuli, and the biphasic theory that describes the deformation of IVD under mechanical loading. The GAG synthesis (at the cell level) was coupled with the mechanical loading (at the tissue level) via a cell-matrix unit approach which established a relationship between the variation of cell dilatation and the local tissue dilatation. This multiscale mathematical model was used to predict the effect of static load (creep load) on GAG synthesis in bovine tail discs. The predicted results are in the range of experimental results. This model was also used to investigate the effect of static (0.2MPa) and diurnal loads (0.1/0.3MPa and 0.15/0.25MPa in 12/12 hours shift with an average of 0.2MPa over a cycle) on GAG synthesis. It was found that static load and diurnal loads have different effects on GAG synthesis in a diurnal cycle, and the diurnal load effects depend on the amplitude of the load. The model is important to understand the effect of mechanical loading at the tissue level on GAG synthesis at the cellular level, as well as to optimize the mechanical loading in growing engineered tissue. Copyright © 2016 Elsevier Ltd. All rights reserved.

A novel synthetic single crystal diamond device for in vivo dosimetry.

PubMed

Marinelli, Marco; Prestopino, G; Tonnetti, A; Verona, C; Verona-Rinati, G; Falco, M D; Bagalà, P; Pimpinella, M; Guerra, A S; De Coste, V

2015-08-01

Aim of the present work is to evaluate the synthetic single crystal diamond Schottky photodiode developed at the laboratories of "Tor Vergata" University in Rome in a new dosimeter configuration specifically designed for offline wireless in vivo dosimetry (IVD) applications. The new diamond based dosimeter, single crystal diamond detector (SCDD-iv), consists of a small unwired detector and a small external reading unit that can be connected to commercial electrometers for getting the detector readout after irradiation. Two nominally identical SCDD-iv dosimeter prototypes were fabricated and tested. A basic dosimetric characterization of detector performances relevant for IVD application was performed under irradiation with (60)Co and 6 MV photon beams. Preirradiation procedure, response stability, short and long term reproducibility, leakage charge, fading effect, linearity with dose, dose rate dependence, temperature dependence, and angular response were investigated. The SCDD-iv is simple, with no cables linked to the patient and the readout is immediate. The range of response with dose has been tested from 1 up to 12 Gy; the reading is independent of the accumulated dose and dose rate independent in the range between about 0.5 and 5 Gy/min; its temperature dependence is within 0.5% between 25 and 38 °C, and its directional dependence is within 2% from 0° to 90°. The combined relative standard uncertainty of absorbed dose to water measurements is estimated lower than the tolerance and action level of 5%. The reported results indicate the proposed novel offline dosimeter based on a synthetic single crystal diamond Schottky photodiode as a promising candidate for in vivo dosimetry applications with photon beams.

Comparison of an In Vitro Diagnostic Next-Generation Sequencing Assay with Sanger Sequencing for HIV-1 Genotypic Resistance Testing.

PubMed

Tzou, Philip L; Ariyaratne, Pramila; Varghese, Vici; Lee, Charlie; Rakhmanaliev, Elian; Villy, Carolin; Yee, Meiqi; Tan, Kevin; Michel, Gerd; Pinsky, Benjamin A; Shafer, Robert W

2018-06-01

The ability of next-generation sequencing (NGS) technologies to detect low frequency HIV-1 drug resistance mutations (DRMs) not detected by dideoxynucleotide Sanger sequencing has potential advantages for improved patient outcomes. We compared the performance of an in vitro diagnostic (IVD) NGS assay, the Sentosa SQ HIV genotyping assay for HIV-1 genotypic resistance testing, with Sanger sequencing on 138 protease/reverse transcriptase (RT) and 39 integrase sequences. The NGS assay used a 5% threshold for reporting low-frequency variants. The level of complete plus partial nucleotide sequence concordance between Sanger sequencing and NGS was 99.9%. Among the 138 protease/RT sequences, a mean of 6.4 DRMs was identified by both Sanger and NGS, a mean of 0.5 DRM was detected by NGS alone, and a mean of 0.1 DRM was detected by Sanger sequencing alone. Among the 39 integrase sequences, a mean of 1.6 DRMs was detected by both Sanger sequencing and NGS and a mean of 0.15 DRM was detected by NGS alone. Compared with Sanger sequencing, NGS estimated higher levels of resistance to one or more antiretroviral drugs for 18.2% of protease/RT sequences and 5.1% of integrase sequences. There was little evidence for technical artifacts in the NGS sequences, but the G-to-A hypermutation was detected in three samples. In conclusion, the IVD NGS assay evaluated in this study was highly concordant with Sanger sequencing. At the 5% threshold for reporting minority variants, NGS appeared to attain a modestly increased sensitivity for detecting low-frequency DRMs without compromising sequence accuracy. Copyright © 2018 American Society for Microbiology.

Lactoferricin mediates Anti-Inflammatory and Anti-Catabolic Effects via Inhibition of IL-1 and LPS Activity in the Intervertebral Disc†

PubMed Central

Kim, Jae-Sung; Ellman, Michael B.; Yan, Dongyao; An, Howard S.; Kc, Ranjan; Li, Xin; Chen, Di; Xiao, Guozhi; Cs-Zabo, Gabriella; Hoskin, David W.; Buechter, D.D.; Van Wijnen, Andre J.; Im, Hee-Jeong

2013-01-01

The catabolic cytokine interleukin-1 (IL-1) and endotoxin lipopolysaccharide (LPS) are well-known inflammatory mediators involved in degenerative disc disease, and inhibitors of IL-1 and LPS may potentially be used to slow or prevent disc degeneration in vivo. Here, we elucidate the striking anti-catabolic and anti-inflammatory effects of bovine lactoferricin (LfcinB) in the intervertebral disc (IVD) via antagonism of both IL-1 and LPS-mediated catabolic activity using in vitro and ex vivo analyses. Specifically, we demonstrate the biological counteraction of LfcinB against IL-1 and LPS-mediated proteoglycan (PG) depletion, matrix-degrading enzyme production and enzyme activity in long-term (alginate beads) and short-term (monolayer) culture models using bovine and human nucleus pulposus (NP) cells. LfcinB significantly attenuates the IL-1 and LPS-mediated suppression of PG production and synthesis, and thus restores PG accumulation and pericellular matrix formation. Simultaneously, LfcinB antagonizes catabolic factor mediated induction of multiple cartilage-degrading enzymes, including MMP-1, MMP-3, MMP-13, ADAMTS-4, and ADAMTS-5, in bovine NP cells at both mRNA and protein levels. LfcinB also suppresses the catabolic factor-induced stimulation of oxidative and inflammatory factors such as iNOS, IL-6, and toll-like receptor-2 (TLR-2) and TLR-4. Finally, the ability of LfcinB to antagonize IL-1 and LPS-mediated suppression of PG is upheld in an en bloc intradiscal microinjection model followed by ex vivo organ culture using both mouse and rabbit IVD tissue, suggesting a potential therapeutic benefit of LfcinB on degenerative disc disease in the future. PMID:23460134

Lactoferricin mediates anti-inflammatory and anti-catabolic effects via inhibition of IL-1 and LPS activity in the intervertebral disc.

PubMed

Kim, Jae-Sung; Ellman, Michael B; Yan, Dongyao; An, Howard S; Kc, Ranjan; Li, Xin; Chen, Di; Xiao, Guozhi; Cs-Szabo, Gabriella; Hoskin, David W; Buechter, Doug D; Van Wijnen, Andre J; Im, Hee-Jeong

2013-09-01

The catabolic cytokine interleukin-1 (IL-1) and endotoxin lipopolysaccharide (LPS) are well-known inflammatory mediators involved in degenerative disc disease, and inhibitors of IL-1 and LPS may potentially be used to slow or prevent disc degeneration in vivo. Here, we elucidate the striking anti-catabolic and anti-inflammatory effects of bovine lactoferricin (LfcinB) in the intervertebral disc (IVD) via antagonism of both IL-1 and LPS-mediated catabolic activity using in vitro and ex vivo analyses. Specifically, we demonstrate the biological counteraction of LfcinB against IL-1 and LPS-mediated proteoglycan (PG) depletion, matrix-degrading enzyme production, and enzyme activity in long-term (alginate beads) and short-term (monolayer) culture models using bovine and human nucleus pulposus (NP) cells. LfcinB significantly attenuates the IL-1 and LPS-mediated suppression of PG production and synthesis, and thus restores PG accumulation and pericellular matrix formation. Simultaneously, LfcinB antagonizes catabolic factor mediated induction of multiple cartilage-degrading enzymes, including MMP-1, MMP-3, MMP-13, ADAMTS-4, and ADAMTS-5, in bovine NP cells at both mRNA and protein levels. LfcinB also suppresses the catabolic factor-induced stimulation of oxidative and inflammatory factors such as iNOS, IL-6, and toll-like receptor-2 (TLR-2) and TLR-4. Finally, the ability of LfcinB to antagonize IL-1 and LPS-mediated suppression of PG is upheld in an en bloc intradiscal microinjection model followed by ex vivo organ culture using both mouse and rabbit IVD tissue, suggesting a potential therapeutic benefit of LfcinB on degenerative disc disease in the future. Copyright © 2013 Wiley Periodicals, Inc.

Conformal coating of highly structured surfaces

DOEpatents

Ginley, David S.; Perkins, John; Berry, Joseph; Gennett, Thomas

2012-12-11

Method of applying a conformal coating to a highly structured substrate and devices made by the disclosed methods are disclosed. An example method includes the deposition of a substantially contiguous layer of a material upon a highly structured surface within a deposition process chamber. The highly structured surface may be associated with a substrate or another layer deposited on a substrate. The method includes depositing a material having an amorphous structure on the highly structured surface at a deposition pressure of equal to or less than about 3 mTorr. The method may also include removing a portion of the amorphous material deposited on selected surfaces and depositing additional amorphous material on the highly structured surface.

Development of an Aerosol Surface Inoculation Method for Bacillus Spores ▿

PubMed Central

Lee, Sang Don; Ryan, Shawn P.; Snyder, Emily Gibb

2011-01-01

A method was developed to deposit Bacillus subtilis spores via aerosolization onto various surface materials for biological agent decontamination and detection studies. This new method uses an apparatus coupled with a metered dose inhaler to reproducibly deposit spores onto various surfaces. A metered dose inhaler was loaded with Bacillus subtilis spores, a surrogate for Bacillus anthracis. Five different material surfaces (aluminum, galvanized steel, wood, carpet, and painted wallboard paper) were tested using this spore deposition method. This aerosolization method deposited spores at a concentration of more than 107 CFU per coupon (18-mm diameter) with less than a 50% coefficient of variation, showing that the aerosolization method developed in this study can deposit reproducible numbers of spores onto various surface coupons. Scanning electron microscopy was used to probe the spore deposition patterns on test coupons. The deposition patterns observed following aerosol impaction were compared to those of liquid inoculation. A physical difference in the spore deposition patterns was observed to result from the two different methods. The spore deposition method developed in this study will help prepare spore coupons via aerosolization fast and reproducibly for bench top decontamination and detection studies. PMID:21193670

Development of an aerosol surface inoculation method for bacillus spores.

PubMed

Lee, Sang Don; Ryan, Shawn P; Snyder, Emily Gibb

2011-03-01

A method was developed to deposit Bacillus subtilis spores via aerosolization onto various surface materials for biological agent decontamination and detection studies. This new method uses an apparatus coupled with a metered dose inhaler to reproducibly deposit spores onto various surfaces. A metered dose inhaler was loaded with Bacillus subtilis spores, a surrogate for Bacillus anthracis. Five different material surfaces (aluminum, galvanized steel, wood, carpet, and painted wallboard paper) were tested using this spore deposition method. This aerosolization method deposited spores at a concentration of more than 10(7) CFU per coupon (18-mm diameter) with less than a 50% coefficient of variation, showing that the aerosolization method developed in this study can deposit reproducible numbers of spores onto various surface coupons. Scanning electron microscopy was used to probe the spore deposition patterns on test coupons. The deposition patterns observed following aerosol impaction were compared to those of liquid inoculation. A physical difference in the spore deposition patterns was observed to result from the two different methods. The spore deposition method developed in this study will help prepare spore coupons via aerosolization fast and reproducibly for bench top decontamination and detection studies.

Optical control of multi-stage thin film solar cell production

DOE Office of Scientific and Technical Information (OSTI.GOV)

Li, Jian; Levi, Dean H.; Contreras, Miguel A.

2016-05-17

Embodiments include methods of depositing and controlling the deposition of a film in multiple stages. The disclosed deposition and deposition control methods include the optical monitoring of a deposition matrix to determine a time when at least one transition point occurs. In certain embodiments, the transition point or transition points are a stoichiometry point. Methods may also include controlling the length of time in which material is deposited during a deposition stage or controlling the amount of the first, second or subsequent materials deposited during any deposition stage in response to a determination of the time when a selected transitionmore » point occurs.« less

2014 Bone and Muscle Risks Standing Review Panel

NASA Technical Reports Server (NTRS)

Steinberg, Susan; Glowacki, Julie; Gregor, Robert; Cullen, Diane; Drake, Almond; Enoka, Roger; Hanley, Edward Jr.; Kraemer, William; Raven, Peter; Sumner, Rick, D.

2014-01-01

The 2014 Bone and Muscle Risks Standing Review Panel (from here on referred to as the SRP) met for a site visit in Houston, TX on December 17 - 18, 2014. The SRP reviewed the updated research plans for the Risk of Impaired Performance Due to Reduced Muscle Mass, Strength and Endurance (Muscle Risk) and the Risk of Reduced Physical Performance Capabilities Due to Reduced Aerobic Capacity (Aerobic Risk). The SRP also received a status update on the Risk of Bone Fracture (Bone Risk), the Risk of Early Onset Osteoporosis Due To Spaceflight (Osteo Risk), the Risk of Intervertebral Disc Damage (IVD Risk), and the Risk of Renal Stone Formation (Renal Risk).

Cherenkov imaging for Total Skin Electron Therapy (TSET)

NASA Astrophysics Data System (ADS)

Xie, Yunhe; Petroccia, Heather; Maity, Amit; Miao, Tianshun; Zhu, Yihua; Bruza, Petr; Pogue, Brian W.; Andreozzi, Jacqueline M.; Plastaras, John P.; Dong, Lei; Zhu, Timothy C.

2018-03-01

Total Skin Electron Therapy (TSET) utilizes high-energy electrons to treat cancers on the entire body surface. The otherwise invisible radiation beam can be observed via the optical Cherenkov photons emitted from interaction between the high-energy electron beam and tissue. Using a specialized camera-system, the Cherenkov emission can thus be used to evaluate the dose uniformity on the surface of the patient in real-time. Each patient was also monitored during TSET via in-vivo detectors (IVD) in nine locations. Patients undergoing TSET in various conditions (whole body and half body) were imaged and analyzed, and the viability of the system to provide clinical feedback was established.

A suitable deposition method of CdS for high performance CdS-sensitized ZnO electrodes: Sequential chemical bath deposition

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chen, Haining; Li, Weiping; Liu, Huicong

2010-07-15

A suitable deposition method of CdS is necessary for the high performance CdS-sensitized ZnO electrodes. In this paper, chemical bath deposition (CBD) and sequential chemical bath deposition (S-CBD) methods were used to deposit CdS on ZnO mesoporous films for ZnO/CdS electrodes. The analysis results of XRD patterns and UV-vis spectroscopy indicated that CBD deposition method leaded to the dissolving of ZnO mesoporous films in deposition solution and thickness reduction of ZnO/CdS electrodes. Absorption in visible region by the ZnO/CdS electrodes with CdS deposition by S-CBD was enhanced as deposition cycles increased due to the stability of ZnO mesoporous films inmore » the S-CBD deposition solutions. The results of photocurrent-voltage (I-V) measurement showed that the performance of ZnO/CdS electrodes with CdS deposition by CBD first increased and then decreased as deposition time increased, and the greatest short-circuit current (J{sub sc}) was obtained at the deposition time of 4 min. The performance of ZnO/CdS electrodes with CdS deposition by S-CBD increased as deposition cycles increased, and both open-circuit voltage (V{sub oc}) and J{sub sc} were greater than those electrodes with CdS deposition by CBD when the deposition cycles of S-CBD were 10 or greater. These results indicated that S-CBD is a more suitable method for high performance ZnO/CdS electrodes. (author)« less

Method and apparatus for rapidly growing films on substrates using pulsed supersonic jets

DOEpatents

Eres, Diula; Lowndes, Douglas H.

1992-01-01

A method and apparatus for the rapid and economical deposition of uniform and high quality films upon a substrate for subsequent use in producing electronic devices, for example. The resultant films are either epitaxial (crystalline) or amorphous depending upon the incidence rate and the temperature and structure of the substrate. The deposition is carried out in a chamber maintained at about 10.sup.-6 Torr. A gaseous source of the material for forming the deposit is injected into the deposition chamber in the form of a pulsed supersonic jet so as to obtain a high incidence rate. The supersonic jet is produced by a pulsed valve between a relatively high presure reservoir, containing the source gaseous molecules, and the deposition chamber; the valve has a small nozzle orifice (e.g., 0.1-1.0 mm diameter). The type of deposit (crystalline amorphous) is then dependent upon the temperature and structure of the substrate. Very high deposition rates are achieved, and the deposit is very smooth and of uniform thickness. Typically the deposition rate is about 100 times that of much more expensive conventional molecular beam methods for deposition, and comparable to certain expensive plasma-assisted CVD methods of the art. The high growth rate of this method results in a reduced contamination of the deposit from other elements in the environment. The method is illustrated by the deposition of epitaxial and amorphour germanium films upon GaAs substrates.

Aqueous, Room Temperature Deposition of Silicon, Molybdenum and Germanium onto Aluminum Substrates

NASA Astrophysics Data System (ADS)

Krishnamurthy, Aarti Krishna

Electrochemical deposition of active materials such as Si, Mo and Ge is notoriously difficult, so they are typically deposited using expensive vacuum methods such as chemical vapor deposition (CVD), plasma-enhanced chemical vapor deposition (PECVD), and magnetron sputtering. However, for most materials, electrochemical deposition has significant advantages of cost, scalability, and manufacturability. There are two main challenges in depositing these materials from aqueous electrolytes at room temperature, namely their highly cathodic standard reduction potential and the formation of native oxides. This has led researchers to use non-aqueous electrolytes such as organic solvents, room temperature ionic liquids (RTILs), and high temperature molten salts. However, these have drawbacks over aqueous electrolytes such as high cost, low conductivity, flammability, and corrosive behavior. During my PhS studies, these two challenges were overcome by using the galvanic method of deposition and by including HF in the electrolyte. Si thin films are employed in a variety of technologies, including microelectronic and photovoltaic devices, Li ion battery anodes, and corrosion-resistant coatings. A galvanic and a combined galvanic/electroless method of Si deposition were developed using aqueous electrolytes at room temperature to obtain nanoporous and compact films, respectively. These films were characterized to understand the surface morphology, thickness, crystallinity, growth rate, composition and nucleation behavior. Approximately 7-10 µm thick compact Si films were achieved with a deposition time of around 28 hours. The galvanic method of deposition was also extended to deposit compact Mo films. Mo thin films have a number of technological applications, including back contacts for CIGS/CZTS photovoltaic devices and corrosion-resistant coatings. Mo thin films were also thoroughly characterized and approximately 4.5 µm thick films were obtained after 3 hours. Similar to Si depostion, a galvanic method of deposition and the galvanic/electroless method of deposition was tested for the deposition of Ge. However no Ge deposit could be consistently obtained, probably due to oxyanion formation in aqueous hexaflurogermante solution.

Ethanol catalytic optical driven deposition for 1D and 2D materials with ultra-low power threshold of 0 dBm

NASA Astrophysics Data System (ADS)

Wang, Hao; Chen, Bohua; Xiao, Xu; Guo, Chaoshi; Zhang, Xiaoyan; Wang, Jun; Jiang, Meng; Wu, Kan; Chen, Jianping

2018-01-01

We have demonstrated a generalized optical driven deposition method, ethanol catalytic deposition (ECD) method, which is widely applicable to the deposition of a broad range of one-dimensional (1D) and two-dimensional (2D) materials with common deposition parameters. Using ECD method, deposition of 1D material carbon nanotubes and 2D materials MoS2, MoSe2, WS2 and WSe2 on tapered fiber has been demonstrated with the threshold power as low as 0 dBm. To our knowledge, this is the lowest threshold power ever reported in optical driven deposition, noting that the conventional optical driven deposition has a threshold typically near 15 dBm. It means ECD method can significantly reduce the power requirement and simplify the setup of the optical driven deposition as well as its wide applicability to different materials, which benefits the research on optical nonlinearity and ultrafast photonics of 1D and 2D materials.

Multicomponent aerosol particle deposition in a realistic cast of the human upper respiratory tract.

PubMed

Nordlund, Markus; Belka, Miloslav; Kuczaj, Arkadiusz K; Lizal, Frantisek; Jedelsky, Jan; Elcner, Jakub; Jicha, Miroslav; Sauser, Youri; Le Bouhellec, Soazig; Cosandey, Stephane; Majeed, Shoaib; Vuillaume, Grégory; Peitsch, Manuel C; Hoeng, Julia

2017-02-01

Inhalation of aerosols generated by electronic cigarettes leads to deposition of multiple chemical compounds in the human airways. In this work, an experimental method to determine regional deposition of multicomponent aerosols in an in vitro segmented, realistic human lung geometry was developed and applied to two aerosols, i.e. a monodisperse glycerol aerosol and a multicomponent aerosol. The method comprised the following steps: (1) lung cast model preparation, (2) aerosol generation and exposure, (3) extraction of deposited mass, (4) chemical quantification and (5) data processing. The method showed good agreement with literature data for the deposition efficiency when using a monodisperse glycerol aerosol, with a mass median aerodynamic diameter (MMAD) of 2.3 μm and a constant flow rate of 15 L/min. The highest deposition surface density rate was observed in the bifurcation segments, indicating inertial impaction deposition. The experimental method was also applied to the deposition of a nebulized multicomponent aerosol with a MMAD of 0.50 μm and a constant flow rate of 15 L/min. The deposited amounts of glycerol, propylene glycol and nicotine were quantified. The three analyzed compounds showed similar deposition patterns and fractions as for the monodisperse glycerol aerosol, indicating that the compounds most likely deposited as parts of the same droplets. The developed method can be used to determine regional deposition for multicomponent aerosols, provided that the compounds are of low volatility. The generated data can be used to validate aerosol deposition simulations and to gain insight in deposition of electronic cigarette aerosols in human airways.

Laser Synthesis of Supported Catalysts for Carbon Nanotubes

NASA Technical Reports Server (NTRS)

VanderWal, Randall L.; Ticich, Thomas M.; Sherry, Leif J.; Hall, Lee J.; Schubert, Kathy (Technical Monitor)

2003-01-01

Four methods of laser assisted catalyst generation for carbon nanotube (CNT) synthesis have been tested. These include pulsed laser transfer (PLT), photolytic deposition (PLD), photothermal deposition (PTD) and laser ablation deposition (LABD). Results from each method are compared based on CNT yield, morphology and structure. Under the conditions tested, the PLT was the easiest method to implement, required the least time and also yielded the best pattemation. The photolytic and photothermal methods required organometallics, extended processing time and partial vacuums. The latter two requirements also held for the ablation deposition approach. In addition to control of the substrate position, controlled deposition duration was necessary to achieve an active catalyst layer. Although all methods were tested on both metal and quartz substrates, only the quartz substrates proved to be inactive towards the deposited catalyst particles.

Studies of mist deposition for the formation of quantum dot CdSe films

NASA Astrophysics Data System (ADS)

Price, S. C.; Shanmugasundaram, K.; Ramani, S.; Zhu, T.; Zhang, F.; Xu, J.; Mohney, S. E.; Zhang, Q.; Kshirsagar, A.; Ruzyllo, J.

2009-10-01

Films of CdSe(ZnS) colloidal nanocrystalline quantum dots (NQDs) were deposited on bare silicon, glass and polymer coated silicon using mist deposition. This effort is a part of an exploratory investigation in which this deposition technique is studied for the first time as a method to form semiconductor NQD films. The process parameters, including deposition time, solution concentration and electric field, were varied to change the thickness of the deposited film. Blanket films and films deposited through a shadow mask were created to investigate the method's ability to pattern films during the deposition process. The differences between these deposition modes in terms of film morphology were observed. Overall, the results show that mist deposition of quantum dots is a viable method for creating thin, patterned quantum dot films using colloidal solution as the precursor. It is concluded that this technique shows very good promise for quantum dot (light emitting diode, LED) fabrication.

Argon thermochronology of mineral deposits; a review of analytical methods, formulations, and selected applications

USGS Publications Warehouse

Snee, Lawrence W.

2002-01-01

40Ar/39Ar geochronology is an experimentally robust and versatile method for constraining time and temperature in geologic processes. The argon method is the most broadly applied in mineral-deposit studies. Standard analytical methods and formulations exist, making the fundamentals of the method well defined. A variety of graphical representations exist for evaluating argon data. A broad range of minerals found in mineral deposits, alteration zones, and host rocks commonly is analyzed to provide age, temporal duration, and thermal conditions for mineralization events and processes. All are discussed in this report. The usefulness of and evolution of the applicability of the method are demonstrated in studies of the Panasqueira, Portugal, tin-tungsten deposit; the Cornubian batholith and associated mineral deposits, southwest England; the Red Mountain intrusive system and associated Urad-Henderson molybdenum deposits; and the Eastern Goldfields Province, Western Australia.

Electrodeposition of reduced graphene oxide with chitosan based on the coordination deposition method

PubMed Central

Liu, Mingyang; Qin, Chaoran; Zhang, Zheng; Ma, Shuai; Cai, Xiuru; Li, Xueqian

2018-01-01

The electrodeposition of graphene has drawn considerable attention due to its appealing applications for sensors, supercapacitors and lithium-ion batteries. However, there are still some limitations in the current electrodeposition methods for graphene. Here, we present a novel electrodeposition method for the direct deposition of reduced graphene oxide (rGO) with chitosan. In this method, a 2-hydroxypropyltrimethylammonium chloride-based chitosan-modified rGO material was prepared. This material disperses homogenously in the chitosan solution, forming a deposition solution with good dispersion stability. Subsequently, the modified rGO material was deposited on an electrode through codeposition with chitosan, based on the coordination deposition method. After electrodeposition, the homogeneous, deposited rGO/chitosan films can be generated on copper or silver electrodes or substrates. The electrodeposition method allows for the convenient and controlled creation of rGO/chitosan nanocomposite coatings and films of different shapes and thickness. It also introduces a new method of creating films, as they can be peeled completely from the electrodes. Moreover, this method allows for a rGO/chitosan film to be deposited directly onto an electrode, which can then be used for electrochemical detection. PMID:29765797

Method and apparatus for conducting variable thickness vapor deposition

DOEpatents

Nesslage, G.V.

1984-08-03

A method of vapor depositing metal on a substrate in variable thickness comprises conducting the deposition continuously without interruption to avoid formation of grain boundaries. To achieve reduced deposition in specific regions a thin wire or ribbon blocking body is placed between source and substrate to partially block vapors from depositing in the region immediately below.

Electroless atomic layer deposition

DOEpatents

Robinson, David Bruce; Cappillino, Patrick J.; Sheridan, Leah B.; Stickney, John L.; Benson, David M.

2017-10-31

A method of electroless atomic layer deposition is described. The method electrolessly generates a layer of sacrificial material on a surface of a first material. The method adds doses of a solution of a second material to the substrate. The method performs a galvanic exchange reaction to oxidize away the layer of the sacrificial material and deposit a layer of the second material on the surface of the first material. The method can be repeated for a plurality of iterations in order to deposit a desired thickness of the second material on the surface of the first material.

Spinal Tissue Loading Created by Different Methods of Spinal Manipulative Therapy Application

PubMed Central

Funabashi, Martha; Nougarou, François; Descarreaux, Martin; Prasad, Narasimha; Kawchuk, Gregory N.

2017-01-01

Study Design. Comparative study using robotic replication of spinal manipulative therapy (SMT) vertebral kinematics together with serial dissection. Objective. The aim of this study was to quantify loads created in cadaveric spinal tissues arising from three different forms of SMT application. Summary of Background Data. There exist many distinct methods by which to apply SMT. It is not known presently whether different forms of SMT application have different effects on spinal tissues. Should the method of SMT application modulate spinal tissue loading, quantifying this relation may help explain the varied outcomes of SMT in terms of effect and safety. Methods. SMT was applied to the third lumbar vertebra in 12 porcine cadavers using three SMT techniques: a clinical device that applies forces through a hand-held instrument (INST), a manual technique of applying SMT clinically (MAN) and a research device that applies parameters of manual SMT through a servo-controlled linear actuator motor (SERVO). The resulting kinematics from each SMT application were tracked optically via indwelling bone pins. The L3/L4 segment was then removed, mounted in a parallel robot and the resulting kinematics from SMT replayed for each SMT application technique. Serial dissection of spinal structures was conducted to quantify loading characteristics of discrete spinal tissues. Results. In terms of load magnitude, SMT application with MAN and SERVO created greater forces than INST in all conditions (P < 0.05). Additionally, MAN and SERVO created comparable posterior forces in the intact specimen, but MAN created greater posterior forces on IVD structures compared to SERVO (P < 0.05). Conclusion. Specific methods of SMT application create unique vertebral loading characteristics, which may help explain the varied outcomes of SMT in terms of effect and safety. Level of Evidence: N/A PMID:28146021

Alloy vapor deposition using ion plating and flash evaporation

NASA Technical Reports Server (NTRS)

Spalvins, T.

1971-01-01

Method extends scope of ion plating technique to include deposition of alloy films without changing composition of plating alloy. Coatings flow with specimen material without chipping or peeling. Technique is most effective vacuum deposition method for depositing alloys for strong and lasting adherence.

Method for deposition of a conductor in integrated circuits

DOEpatents

Creighton, J. Randall; Dominguez, Frank; Johnson, A. Wayne; Omstead, Thomas R.

1997-01-01

A method is described for fabricating integrated semiconductor circuits and, more particularly, for the selective deposition of a conductor onto a substrate employing a chemical vapor deposition process. By way of example, tungsten can be selectively deposited onto a silicon substrate. At the onset of loss of selectivity of deposition of tungsten onto the silicon substrate, the deposition process is interrupted and unwanted tungsten which has deposited on a mask layer with the silicon substrate can be removed employing a halogen etchant. Thereafter, a plurality of deposition/etch back cycles can be carried out to achieve a predetermined thickness of tungsten.

Nanostructure templating using low temperature atomic layer deposition

DOEpatents

Grubbs, Robert K [Albuquerque, NM; Bogart, Gregory R [Corrales, NM; Rogers, John A [Champaign, IL

2011-12-20

Methods are described for making nanostructures that are mechanically, chemically and thermally stable at desired elevated temperatures, from nanostructure templates having a stability temperature that is less than the desired elevated temperature. The methods comprise depositing by atomic layer deposition (ALD) structural layers that are stable at the desired elevated temperatures, onto a template employing a graded temperature deposition scheme. At least one structural layer is deposited at an initial temperature that is less than or equal to the stability temperature of the template, and subsequent depositions made at incrementally increased deposition temperatures until the desired elevated temperature stability is achieved. Nanostructure templates include three dimensional (3D) polymeric templates having features on the order of 100 nm fabricated by proximity field nanopatterning (PnP) methods.

Influence of low glucose supply on the regulation of gene expression by nucleus pulposus cells and their responsiveness to mechanical loading.

PubMed

Rinkler, Christina; Heuer, Frank; Pedro, Maria Teresa; Mauer, Uwe Max; Ignatius, Anita; Neidlinger-Wilke, Cornelia

2010-10-01

Environmental alterations resulting in a decrease in the nutrient supply have been associated with intervertebral disc (IVD) degeneration, particularly of the nucleus pulposus (NP). The goal of the present study was to examine the hypothesis that glucose deprivation alters the metabolism of NP cells and their responsiveness to mechanical loading. A possible interaction of glucose supply and hydrostatic pressure (HP) with gene expression by NP cells has not been investigated. The influence of glucose supply (physiological concentration: 5 mM, reduction: 0 or 0.5 mM) and cyclic HP loading (2.5 MPa, 0.1 Hz, 30 minutes) on bovine and human NP cell matrix turnover was analyzed by quantitative real-time reverse transcriptase–polymerase chain reaction. Glucose-dependent effects on cell viability were determined by trypan blue exclusion. A glycosaminoglycan (GAG) assay was performed to determine nutritional effects on the protein level. Glucose reduction resulted in significant downregulations (p < 0.05) of aggrecan, collagen-I, and collagen-II gene expression by bovine NP cells. Exemplary human donors also displayed a similar trend for aggrecan and collagen-II, whereas matrix metalloproteinases (MMPs) tended to be upregulated under glucose deprivation. After HP loading, human NP cells showed individual upregulations of collagen-I and collagen-II expression, while MMP expression tended to be downregulated under glucose reduction relative to a normal glucose supply. Cell viability decreased with glucose deprivation. The GAG content was similar in all groups at Day 1, whereas at Day 3 there was a significant increase under physiological conditions. Glucose deprivation strongly affected NP cell metabolism. The effects of an altered glucose supply on gene expression were more pronounced than the mechanically induced effects. Data in this study demonstrate that the glucose environment is more critical for disc cell metabolism than mechanical loads. In individual human donors, however, adequate mechanical stimuli might have a beneficial effect on matrix turnover during IVD degeneration.

Spatiotemporal analysis of putative notochordal cell markers reveals CD24 and keratins 8, 18, and 19 as notochord‐specific markers during early human intervertebral disc development

PubMed Central

Rodrigues‐Pinto, Ricardo; Berry, Andrew; Piper‐Hanley, Karen; Hanley, Neil; Richardson, Stephen M.

2016-01-01

ABSTRACT In humans, the nucleus pulposus (NP) is composed of large vacuolated notochordal cells in the fetus but, soon after birth, becomes populated by smaller, chondrocyte‐like cells. Although animal studies indicate that notochord‐derived cells persist in the adult NP, the ontogeny of the adult human NP cell population is still unclear. As such, identification of unique notochordal markers is required. This study was conducted to determine the spatiotemporal expression of putative human notochordal markers to aid in the elucidation of the ontogeny of adult human NP cells. Human embryos and fetuses (3.5–18 weeks post‐conception (WPC)) were microdissected to isolate the spine anlagens (notochord and somites/sclerotome). Morphology of the developing IVD was assessed using hematoxylin and eosin. Expression of keratin (KRT) 8, KRT18, KRT19, CD24, GAL3, CD55, BASP1, CTGF, T, CD90, Tie2, and E‐cadherin was assessed using immunohistochemistry. KRT8, KRT18, KRT19 were uniquely expressed by notochordal cells at all spine levels at all stages studied; CD24 was expressed at all stages except 3.5 WPC. While GAL3, CD55, BASP1, CTGF, and T were expressed by notochordal cells at specific stages, they were also co‐expressed by sclerotomal cells. CD90, Tie2, and E‐cadherin expression was not detectable in developing human spine cells at any stage. This study has identified, for the first time, the consistent expression of KRT8, KRT18, KRT19, and CD24 as human notochord‐specific markers during early IVD development. Thus, we propose that these markers can be used to help ascertain the ontogeny of adult human NP cells. © 2016 The Authors. Journal of Orthopaedic Research Published by Wiley Periodicals, Inc. J Orthop Res 34:1327–1340, 2016. PMID:26910849

Spatiotemporal analysis of putative notochordal cell markers reveals CD24 and keratins 8, 18, and 19 as notochord-specific markers during early human intervertebral disc development.

PubMed

Rodrigues-Pinto, Ricardo; Berry, Andrew; Piper-Hanley, Karen; Hanley, Neil; Richardson, Stephen M; Hoyland, Judith A

2016-08-01

In humans, the nucleus pulposus (NP) is composed of large vacuolated notochordal cells in the fetus but, soon after birth, becomes populated by smaller, chondrocyte-like cells. Although animal studies indicate that notochord-derived cells persist in the adult NP, the ontogeny of the adult human NP cell population is still unclear. As such, identification of unique notochordal markers is required. This study was conducted to determine the spatiotemporal expression of putative human notochordal markers to aid in the elucidation of the ontogeny of adult human NP cells. Human embryos and fetuses (3.5-18 weeks post-conception (WPC)) were microdissected to isolate the spine anlagens (notochord and somites/sclerotome). Morphology of the developing IVD was assessed using hematoxylin and eosin. Expression of keratin (KRT) 8, KRT18, KRT19, CD24, GAL3, CD55, BASP1, CTGF, T, CD90, Tie2, and E-cadherin was assessed using immunohistochemistry. KRT8, KRT18, KRT19 were uniquely expressed by notochordal cells at all spine levels at all stages studied; CD24 was expressed at all stages except 3.5 WPC. While GAL3, CD55, BASP1, CTGF, and T were expressed by notochordal cells at specific stages, they were also co-expressed by sclerotomal cells. CD90, Tie2, and E-cadherin expression was not detectable in developing human spine cells at any stage. This study has identified, for the first time, the consistent expression of KRT8, KRT18, KRT19, and CD24 as human notochord-specific markers during early IVD development. Thus, we propose that these markers can be used to help ascertain the ontogeny of adult human NP cells. © 2016 The Authors. Journal of Orthopaedic Research Published by Wiley Periodicals, Inc. J Orthop Res 34:1327-1340, 2016. © 2016 The Authors. Journal of Orthopaedic Research Published by Wiley Periodicals, Inc.

Detection of Cytomegalovirus (CMV) DNA in EDTA Whole-Blood Samples: Evaluation of the Quantitative artus CMV LightCycler PCR Kit in Conjunction with Automated Sample Preparation▿

PubMed Central

Michelin, Birgit D. A.; Hadžisejdić, Ita; Bozic, Michael; Grahovac, Maja; Hess, Markus; Grahovac, Blaženka; Marth, Egon; Kessler, Harald H.

2008-01-01

Whole blood has been found to be a reliable matrix for the detection and quantitation of cytomegalovirus (CMV) DNA. In this study, the performance of the artus CMV LightCycler (LC) PCR kit in conjunction with automated sample preparation on a BioRobot EZ1 workstation was evaluated. The accuracy, linearity, analytical sensitivity, and inter- and intra-assay variations were determined. A total of 102 clinical EDTA whole-blood samples were investigated, and results were compared with those obtained with the in vitro diagnostics (IVD)/Conformité Européene (CE)-labeled CMV HHV6,7,8 R-gene quantification kit. When the accuracy of the new kit was tested, seven of eight results were found to be within ±0.5 log10 unit of the expected panel results. Determination of linearity resulted in a quasilinear curve over more than 5 log units. The lower limit of detection of the assay was determined to be 139 copies/ml in EDTA whole blood. The interassay variation ranged from 15 to 58%, and the intra-assay variation ranged from 7 to 35%. When clinical samples were tested and the results were compared with those of the routinely used IVD/CE-labeled assay, 53 samples tested positive and 13 samples tested negative by both of the assays. One sample was found to be positive with the artus CMV LC PCR kit only, and 35 samples tested positive with the routinely used assay only. The majority of discrepant results were found with low-titer samples. In conclusion, use of the artus CMV LC PCR kit in conjunction with automated sample preparation on the BioRobot EZ1 workstation may be suitable for the detection and quantitation of CMV DNA in EDTA whole blood in the routine low-throughput laboratory; however, low-positive results may be missed by this assay. PMID:18272703

Detection of cytomegalovirus (CMV) DNA in EDTA whole-blood samples: evaluation of the quantitative artus CMV LightCycler PCR kit in conjunction with automated sample preparation.

PubMed

Michelin, Birgit D A; Hadzisejdic, Ita; Bozic, Michael; Grahovac, Maja; Hess, Markus; Grahovac, Blazenka; Marth, Egon; Kessler, Harald H

2008-04-01

Whole blood has been found to be a reliable matrix for the detection and quantitation of cytomegalovirus (CMV) DNA. In this study, the performance of the artus CMV LightCycler (LC) PCR kit in conjunction with automated sample preparation on a BioRobot EZ1 workstation was evaluated. The accuracy, linearity, analytical sensitivity, and inter- and intra-assay variations were determined. A total of 102 clinical EDTA whole-blood samples were investigated, and results were compared with those obtained with the in vitro diagnostics (IVD)/Conformité Européene (CE)-labeled CMV HHV6,7,8 R-gene quantification kit. When the accuracy of the new kit was tested, seven of eight results were found to be within +/-0.5 log(10) unit of the expected panel results. Determination of linearity resulted in a quasilinear curve over more than 5 log units. The lower limit of detection of the assay was determined to be 139 copies/ml in EDTA whole blood. The interassay variation ranged from 15 to 58%, and the intra-assay variation ranged from 7 to 35%. When clinical samples were tested and the results were compared with those of the routinely used IVD/CE-labeled assay, 53 samples tested positive and 13 samples tested negative by both of the assays. One sample was found to be positive with the artus CMV LC PCR kit only, and 35 samples tested positive with the routinely used assay only. The majority of discrepant results were found with low-titer samples. In conclusion, use of the artus CMV LC PCR kit in conjunction with automated sample preparation on the BioRobot EZ1 workstation may be suitable for the detection and quantitation of CMV DNA in EDTA whole blood in the routine low-throughput laboratory; however, low-positive results may be missed by this assay.

Lactoferricin enhances BMP7-stimulated anabolic pathways in intervertebral disc cells.

PubMed

Ellman, Michael B; Kim, Jaesung; An, Howard S; Chen, Di; Kc, Ranjan; Li, Xin; Xiao, Guozhi; Yan, Dongyao; Suh, Joon; van Wjnen, Andre J; Wang, James H-C; Kim, Su-Gwan; Im, Hee-Jeong

2013-07-25

Bone-morphogenetic protein-7 (BMP7) is a well-known anabolic and anti-catabolic growth factor on intervertebral disc (IVD) matrix and cell homeostasis. Similarly, Lactoferricin B (LfcinB) has recently been shown to have pro-anabolic, anti-catabolic, anti-oxidative and/or anti-inflammatory effects in bovine disc cells in vitro. In this study, we investigated the potential benefits of using combined peptide therapy with LfcinB and BMP7 for intervertebral disc matrix repair and to understand cellular and signaling mechanisms controlled by these factors. We studied the effects of BMP7 and LfcinB as individual treatments and combined therapy on bovine nucleus pulposus (NP) cells by assessing proteoglycan (PG) accumulation and synthesis, and the gene expression of matrix protein aggrecan and transcription factor SOX-9. We also analyzed the role of Noggin, a BMP antagonist, in IVD tissue and examined its effect after stimulation with LfcinB. To understand the molecular mechanisms by which LfcinB synergizes with BMP7, we investigated the ERK-SP1 axis as a downstream intracellular signaling regulator involved in BMP7 and LfcinB-mediated activities. Treatment of bovine NP cells cultured in alginate with LfcinB plus BMP7 synergistically stimulates PG synthesis and accumulation in part by upregulation of aggrecan gene expression. The synergism results from LfcinB-mediated activation of Sp1 and SMAD signaling pathways by (i) phosphorylation of SMAD 1/5/8; (ii) downregulation of SMAD inhibitory factors [i.e., noggin and SMAD6 (inhibitory SMAD)]; and (iii) upregulation of SMAD4 (universal co-SMAD). These data indicate that LfcinB-suppression of Noggin may eliminate the negative feedback of BMP7, thereby maximizing biological activity of BMP7 and ultimately shifting homeostasis to a pro-anabolic state in disc cells. We propose that combination growth factor therapy using BMP7 and LfcinB may be beneficial for treatment of disc degeneration. Copyright © 2013 Elsevier B.V. All rights reserved.

Liquid precursor for deposition of copper selenide and method of preparing the same

DOEpatents

Curtis, Calvin J.; Miedaner, Alexander; Franciscus Antonius Maria Van Hest, Marinus; Ginley, David S.; Hersh, Peter A.; Eldada, Louay; Stanbery, Billy J.

2015-09-08

Liquid precursors containing copper and selenium suitable for deposition on a substrate to form thin films suitable for semiconductor applications are disclosed. Methods of preparing such liquid precursors and methods of depositing a precursor on a substrate are also disclosed.

FAST TRACK COMMUNICATION Understanding adhesion at as-deposited interfaces from ab initio thermodynamics of deposition growth: thin-film alumina on titanium carbide

NASA Astrophysics Data System (ADS)

Rohrer, Jochen; Hyldgaard, Per

2010-12-01

We investigate the chemical composition and adhesion of chemical vapour deposited thin-film alumina on TiC using and extending a recently proposed nonequilibrium method of ab initio thermodynamics of deposition growth (AIT-DG) (Rohrer and Hyldgaard 2010 Phys. Rev. B 82 045415). A previous study of this system (Rohrer et al 2010 J. Phys.: Condens. Matter 22 015004) found that use of equilibrium thermodynamics leads to predictions of a non-binding TiC/alumina interface, despite its industrial use as a wear-resistant coating. This discrepancy between equilibrium theory and experiment is resolved by the AIT-DG method which predicts interfaces with strong adhesion. The AIT-DG method combines density functional theory calculations, rate-equation modelling of the pressure evolution of the deposition environment and thermochemical data. The AIT-DG method was previously used to predict prevalent terminations of growing or as-deposited surfaces of binary materials. Here we extend the method to predict surface and interface compositions of growing or as-deposited thin films on a substrate and find that inclusion of the nonequilibrium deposition environment has important implications for the nature of buried interfaces.

Thin-film preparation by back-surface irradiation pulsed laser deposition using metal powder targets

NASA Astrophysics Data System (ADS)

Kawasaki, Hiroharu; Ohshima, Tamiko; Yagyu, Yoshihito; Ihara, Takeshi; Yamauchi, Makiko; Suda, Yoshiaki

2017-01-01

Several kinds of functional thin films were deposited using a new thin-film preparation method named the back-surface irradiation pulsed laser deposition (BIPLD) method. In this BIPLD method, powder targets were used as the film source placed on a transparent target holder, and then a visible-wavelength pulsed laser was irradiated from the holder side to the substrate. Using this new method, titanium oxide and boron nitride thin films were deposited on the silicon substrate. Surface scanning electron microscopy (SEM) images suggest that all of the thin films were deposited on the substrate with some large droplets irrespective of the kind of target used. The deposition rate of the films prepared by using this method was calculated from film thickness and deposition time to be much lower than that of the films prepared by conventional PLD. X-ray diffraction (XRD) measurement results suggest that rutile and anatase TiO2 crystal peaks were formed for the films prepared using the TiO2 rutile powder target. Crystal peaks of hexagonal boron nitride were observed for the films prepared using the boron nitride powder target. The crystallinity of the prepared films was changed by annealing after deposition.

Liquid precursor for deposition of indium selenide and method of preparing the same

DOEpatents

Curtis, Calvin J.; Miedaner, Alexander; van Hest, Marinus Franciscus Antonius Maria; Ginley, David S.; Hersh, Peter A.; Eldada, Louay; Stanbery, Billy J.

2015-09-22

Liquid precursors containing indium and selenium suitable for deposition on a substrate to form thin films suitable for semiconductor applications are disclosed. Methods of preparing such liquid precursors and method of depositing a liquid precursor on a substrate are also disclosed.

Annealing dependence of residual stress and optical properties of TiO2 thin film deposited by different deposition methods.

PubMed

Chen, Hsi-Chao; Lee, Kuan-Shiang; Lee, Cheng-Chung

2008-05-01

Titanium oxide (TiO(2)) thin films were prepared by different deposition methods. The methods were E-gun evaporation with ion-assisted deposition (IAD), radio-frequency (RF) ion-beam sputtering, and direct current (DC) magnetron sputtering. Residual stress was released after annealing the films deposited by RF ion-beam or DC magnetron sputtering but not evaporation, and the extinction coefficient varied significantly. The surface roughness of the evaporated films exceeded that of both sputtered films. At the annealing temperature of 300 degrees C, anatase crystallization occurred in evaporated film but not in the RF ion-beam or DC magnetron-sputtered films. TiO(2) films deposited by sputtering were generally more stable during annealing than those deposited by evaporation.

Method for deposition of a conductor in integrated circuits

DOEpatents

Creighton, J.R.; Dominguez, F.; Johnson, A.W.; Omstead, T.R.

1997-09-02

A method is described for fabricating integrated semiconductor circuits and, more particularly, for the selective deposition of a conductor onto a substrate employing a chemical vapor deposition process. By way of example, tungsten can be selectively deposited onto a silicon substrate. At the onset of loss of selectivity of deposition of tungsten onto the silicon substrate, the deposition process is interrupted and unwanted tungsten which has deposited on a mask layer with the silicon substrate can be removed employing a halogen etchant. Thereafter, a plurality of deposition/etch back cycles can be carried out to achieve a predetermined thickness of tungsten. 2 figs.

Properties of zinc tin oxide thin film by aerosol assisted chemical vapor deposition (AACVD)

NASA Astrophysics Data System (ADS)

Riza, Muhammad Arif; Rahman, Abu Bakar Abd; Sepeai, Suhaila; Ludin, Norasikin Ahmad; Teridi, Mohd Asri Mat; Ibrahim, Mohd Adib

2018-05-01

This study focuses on the properties of ZTO which have been deposited by a low-cost method namely aerosol assisted chemical vapor deposition (AACVD). The precursors used in this method were zinc acetate dihidrate and tin chloride dihydrate for ZTO thin film deposition. Both precursors were mixed and stirred until fully dissolved before deposition. The ZTO was deposited on borosilicate glass substrate for the investigation of optical properties. The films deposited have passed the scotch tape adherence test. XRD revealed that the crystal ZTO is slightly in the form of perovskite structure but several deteriorations were also seen in the spectrum. The UV-Vis analysis showed high transmittance of ˜85% and the band gap was calculated to be 3.85 eV. The average thickness of the film is around 284 nm. The results showed that the ZTO thin films have been successfully deposited by the utilization of AACVD method.

The Development of an Environmentally Compliant, Multi-Functional Aerospace Coating Using Molecular- and Nano-Engineering Methods

DTIC Science & Technology

2006-10-02

Al -TM-RE) alloy which could by spray applied using various deposition routes or deposited as a powder that is...corrosion properties of various spray deposited alloys from their properties as defective coatings on 2024-T3. "* HVOF spray deposited and cold spray ...layer. "* A method has been developed to distinguish the intrinsic corrosion properties of various spray deposited

Method and apparatus for fabricating a thin-film solar cell utilizing a hot wire chemical vapor deposition technique

DOEpatents

Wang, Qi; Iwaniczko, Eugene

2006-10-17

A thin-film solar cell is provided. The thin-film solar cell comprises an a-SiGe:H (1.6 eV) n-i-p solar cell having a deposition rate of at least ten (10) .ANG./second for the a-SiGe:H intrinsic layer by hot wire chemical vapor deposition. A method for fabricating a thin film solar cell is also provided. The method comprises depositing a n-i-p layer at a deposition rate of at least ten (10) .ANG./second for the a-SiGe:H intrinsic layer.

Method for in situ carbon deposition measurement for solid oxide fuel cells

NASA Astrophysics Data System (ADS)

Kuhn, J.; Kesler, O.

2014-01-01

Previous methods to measure carbon deposition in solid oxide fuel cell (SOFC) anodes do not permit simultaneous electrochemical measurements. Electrochemical measurements supplemented with carbon deposition quantities create the opportunity to further understand how carbon affects SOFC performance and electrochemical impedance spectra (EIS). In this work, a method for measuring carbon in situ, named here as the quantification of gasified carbon (QGC), was developed. TGA experiments showed that carbon with a 100 h residence time in the SOFC was >99.8% gasified. Comparison of carbon mass measurements between the TGA and QGC show good agreement. In situ measurements of carbon deposition in SOFCs at varying molar steam/carbon ratios were performed to further validate the QGC method, and suppression of carbon deposition with increasing steam concentration was observed, in agreement with previous studies. The technique can be used to investigate in situ carbon deposition and gasification behavior simultaneously with electrochemical measurements for a variety of fuels and operating conditions, such as determining conditions under which incipient carbon deposition is reversible.

The increasing application of multiplex nucleic acid detection tests to the diagnosis of syndromic infections.

PubMed

Gray, J; Coupland, L J

2014-01-01

On 14 January 2013, the US Food and Drug Administration (FDA) announced permission for a multiplex nucleic acid test, the xTAG® Gastrointestinal Pathogen Panel (GPP) (Luminex Corporation, USA), which simultaneously detects 11 common viral, bacterial and parasitic causes of infectious gastroenteritis, to be marketed in the USA. This announcement reflects the current move towards the development and commercialization of detection technologies based on nucleic acid amplification techniques for diagnosis of syndromic infections. We discuss the limitations and advantages of nucleic acid amplification techniques and the recent advances in Conformité Européene - in-vitro diagnostic (CE-IVD)-approved multiplex real-time PCR kits for the simultaneous detection of multiple targets within the clinical diagnostics market.

Uniform deposition of uranium hexafluoride (UF6): Standardized mass deposits and controlled isotopic ratios using a thermal fluorination method.

PubMed

McNamara, Bruce K; O'Hara, Matthew J; Casella, Andrew M; Carter, Jennifer C; Addleman, R Shane; MacFarlan, Paul J

2016-07-01

We report a convenient method for the generation of volatile uranium hexafluoride (UF6) from solid uranium oxides and other U compounds, followed by uniform deposition of low levels of UF6 onto sampling coupons. Under laminar flow conditions, UF6 is shown to interact with surfaces within a fixed reactor geometry to a highly predictable degree. We demonstrate the preparation of U deposits that range between approximately 0.01 and 500ngcm(-2). The data suggest the method can be extended to creating depositions at the sub-picogramcm(-2) level. The isotopic composition of the deposits can be customized by selection of the U source materials and we demonstrate a layering technique whereby two U solids, each with a different isotopic composition, are employed to form successive layers of UF6 on a surface. The result is an ultra-thin deposit that bears an isotopic signature that is a composite of the two U sources. The reported deposition method has direct application to the development of unique analytical standards for nuclear safeguards and forensics. Further, the method allows access to very low atomic or molecular coverages of surfaces. Copyright © 2016 Elsevier B.V. All rights reserved.

Methods for producing films using supercritical fluid

DOEpatents

Yonker, Clement R.; Fulton, John L.

2004-06-15

A method for forming a continuous film on a substrate surface that involves depositing particles onto a substrate surface and contacting the particle-deposited substrate surface with a supercritical fluid under conditions sufficient for forming a continuous film from the deposited particles. The particles may have a mean particle size of less 1 micron. The method may be performed by providing a pressure vessel that can contain a compressible fluid. A particle-deposited substrate is provided in the pressure vessel and the compressible fluid is maintained at a supercritical or sub-critical state sufficient for forming a film from the deposited particles. The T.sub.g of particles may be reduced by subjecting the particles to the methods detailed in the present disclosure.

Assessment of nasal spray deposition pattern in a silicone human nose model using a color-based method.

PubMed

Kundoor, Vipra; Dalby, Richard N

2010-01-01

To develop a simple and inexpensive method to visualize and quantify droplet deposition patterns. Deposition pattern was determined by uniformly coating the nose model with Sar-Gel (a paste that changes from white to purple on contact with water) and subsequently discharging sprays into the nose model. The color change was captured using a digital camera and analyzed using Adobe Photoshop. Several tests were conducted to validate the method. Deposition patterns of different nasal sprays (Ayr, Afrin, and Zicam) and different nasal drug delivery devices (Afrin nasal spray and PARI Sinustar nasal nebulizer) were compared. We also used the method to evaluate the effect of inhaled flow rate on nasal spray deposition. There was a significant difference in the deposition area for Ayr, Afrin, and Zicam. The deposition areas of Afrin nasal spray and PARI Sinustar nasal nebulizer (2 min and 5 min) were significantly different. Inhaled flow rate did not have a significant effect on the deposition pattern. Lower viscosity formulations (Ayr, Afrin) provided greater coverage than the higher viscosity formulation (Zicam). The nebulizer covered a greater surface area than the spray pump we evaluated. Aerosol deposition in the nose model was not affected by air flow conditions.

A simplified method for assessing particle deposition rate in aircraft cabins

NASA Astrophysics Data System (ADS)

You, Ruoyu; Zhao, Bin

2013-03-01

Particle deposition in aircraft cabins is important for the exposure of passengers to particulate matter, as well as the airborne infectious diseases. In this study, a simplified method is proposed for initial and quick assessment of particle deposition rate in aircraft cabins. The method included: collecting the inclined angle, area, characteristic length, and freestream air velocity for each surface in a cabin; estimating the friction velocity based on the characteristic length and freestream air velocity; modeling the particle deposition velocity using the empirical equation we developed previously; and then calculating the particle deposition rate. The particle deposition rates for the fully-occupied, half-occupied, 1/4-occupied and empty first-class cabin of the MD-82 commercial airliner were estimated. The results show that the occupancy did not significantly influence the particle deposition rate of the cabin. Furthermore, the simplified human model can be used in the assessment with acceptable accuracy. Finally, the comparison results show that the particle deposition rate of aircraft cabins and indoor environments are quite similar.

Method of fabricating an optoelectronic device having a bulk heterojunction

DOEpatents

Shtein, Max [Ann Arbor, MI; Yang, Fan [Princeton, NJ; Forrest, Stephen R [Princeton, NJ

2008-10-14

A method of fabricating an optoelectronic device comprises: depositing a first layer having protrusions over a first electrode, in which the first layer comprises a first organic small molecule material; depositing a second layer on the first layer such that the second layer is in physical contact with the first layer; in which the smallest lateral dimension of the protrusions are between 1 to 5 times the exciton diffusion length of the first organic small molecule material; and depositing a second electrode over the second layer to form the optoelectronic device. A method of fabricating an organic optoelectronic device having a bulk heterojunction is also provided and comprises: depositing a first layer with protrusions over an electrode by organic vapor phase deposition; depositing a second layer on the first layer where the interface of the first and second layers forms a bulk heterojunction; and depositing another electrode over the second layer.

Uniform deposition of uranium hexafluoride (UF6): Standardized mass deposits and controlled isotopic ratios using a thermal fluorination method

DOE Office of Scientific and Technical Information (OSTI.GOV)

McNamara, Bruce K.; O’Hara, Matthew J.; Casella, Andrew M.

2016-07-01

Abstract: We report a convenient method for the generation of volatile uranium hexafluoride (UF6) from solid uranium oxides and other uranium compounds, followed by uniform deposition of low levels of UF6 onto sampling coupons. Under laminar flow conditions, UF6 is shown to interact with surfaces within the chamber to a highly predictable degree. We demonstrate the preparation of uranium deposits that range between ~0.01 and 470±34 ng∙cm-2. The data suggest the method can be extended to creating depositions at the sub-picogram∙cm-2 level. Additionally, the isotopic composition of the deposits can be customized by selection of the uranium source materials. Wemore » demonstrate a layering technique whereby two uranium solids, each with a different isotopic composition, are employed to form successive layers of UF6 on a surface. The result is an ultra-thin deposit of UF6 that bears an isotopic signature that is a composite of the two uranium sources. The reported deposition method has direct application to the development of unique analytical standards for nuclear safeguards and forensics.« less

Methods of degrading napalm B

DOEpatents

Tyndall, Richard L.; Vass, Arpad

1995-01-01

Methods of degrading napalm and/or trinitrotoluene involve contacting the waste with specific intra-amoebic isolates of ATCC 40908 and/or dispersants derived therefrom. Useful isolates include is deposited as ATCC 77529, NAP-1 deposited as ATCC 77526 and 13 deposited as ATCC 77527.

Methods of degrading napalm B

DOEpatents

Tyndall, R.L.; Vass, A.

1995-09-12

Methods of degrading napalm and/or trinitrotoluene involve contacting the waste with specific intra-amoebic isolates of ATCC 40908 and/or dispersants derived therefrom. Useful isolates are deposited as ATCC 77529, NAP-1 deposited as ATCC 77526 and 13 deposited as ATCC 77527.