3D morphometric analysis of calcified cartilage properties using micro-computed tomography.

OBJECTIVE: Our aim is to establish methods for quantifying morphometric properties of calcified cartilage (CC) from micro-computed tomography (µCT). Furthermore, we evaluated the feasibility of these methods in investigating relationships between osteoarthritis (OA), tidemark surface morphology and open subchondral channels (OSCCs). METHOD: Samples (n = 15) used in this study were harvested from human lateral tibial plateau (n = 8). Conventional roughness and parameters assessing local 3-dimensional (3D) surface variations were used to quantify the surface morphology of the CC. Subchondral channel properties (percentage, density, size) were also calculated. As a reference, histological sections were evaluated using Histopathological osteoarthritis grading (OARSI) and thickness of CC and subchondral bone (SCB) was quantified. RESULTS: OARSI grade correlated with a decrease in local 3D variations of the tidemark surface (amount of different surface patterns (rs = -0.600, P = 0.018), entropy of patterns (EP) (rs = -0.648, P = 0.018), homogeneity index (HI) (rs = 0.555, P = 0.032)) and tidemark roughness (TMR) (rs = -0.579, P = 0.024). Amount of different patterns (ADP) and EP associated with channel area fraction (CAF) (rp = 0.876, P < 0.0001; rp = 0.665, P = 0.007, respectively) and channel density (CD) (rp = 0.680, P = 0.011; rp = 0.582, P = 0.023, respectively). TMR was associated with CAF (rp = 0.926, P < 0.0001) and average channel size (rp = 0.574, P = 0.025). CC topography differed statistically significantly in early OA vs healthy samples. CONCLUSION: We introduced a µ-CT image method to quantify 3D CC topography and perforations through CC. CC topography was associated with OARSI grade and OSCC properties; this suggests that the established methods can detect topographical changes in tidemark and CC perforations associated with OA.

In vitro method for 3D morphometry of human articular cartilage chondrons based on micro-computed tomography.

OBJECTIVE: The aims of this study were: to 1) develop a novel sample processing protocol to visualize human articular cartilage (AC) chondrons using micro-computed tomography (µCT), 2) develop and validate an algorithm to quantify the chondron morphology in 3D, and 3) compare the differences in chondron morphology between intact and osteoarthritic AC. METHOD: The developed protocol is based on the dehydration of samples with hexamethyldisilazane (HMDS), followed by imaging with a desktop µCT. Chondron density and depth, as well as volume and sphericity, were calculated in 3D with a custom-made and validated algorithm employing semi-automatic chondron selection and segmentation. The quantitative parameters were analyzed at three AC depth zones (zone 1: 0-10%; zone 2: 10-40%; zone 3: 40-100%) and grouped by the OARSI histological grades (OARSI grades 0-1.0, n = 6; OARSI grades 3.0-3.5, n = 6). RESULTS: After semi-automatic chondron selection and segmentation, 1510 chondrons were approved for 3D morphometric analyses. The chondrons especially in the deeper tissue (zones 2 and 3) were significantly larger (P < 0.001) and less spherical (P < 0.001), respectively, in the OARSI grade 3-3.5 group compared to the OARSI grade 0-1.0 group. No statistically significant difference in chondron density between the OARSI grade groups was observed at different depths. CONCLUSION: We have developed a novel sample processing protocol for chondron imaging in 3D, as well as a high-throughput algorithm to semi-automatically quantify chondron/chondrocyte 3D morphology in AC. Our results also suggest that 3D chondron morphology is affected by the progression of osteoarthritis (OA).

Electromechanical properties of human osteoarthritic and asymptomatic articular cartilage are sensitive and early detectors of degeneration.

OBJECTIVE: To evaluate cross-correlations of ex vivo electromechanical properties with cartilage and subchondral bone plate thickness, as well as their sensitivity and specificity regarding early cartilage degeneration in human tibial plateau. METHOD: Six pairs of tibial plateaus were assessed ex vivo using an electromechanical probe (Arthro-BST) which measures a quantitative parameter (QP) reflecting articular cartilage compression-induced streaming potentials. Cartilage thickness was then measured with an automated thickness mapping technique using Mach-1 multiaxial mechanical tester. Subsequently, a visual assessment was performed by an experienced orthopedic surgeon using the International Cartilage Repair Society (ICRS) grading system. Each tibial plateau was finally evaluated with µCT scanner to determine the subchondral-bone plate thickness over the entire surface. RESULTS: Cross-correlations between assessments decreased with increasing degeneration level. Moreover, electromechanical QP and subchondral-bone plate thickness increased strongly with ICRS grade (ρ = 0.86 and ρ = 0.54 respectively), while cartilage thickness slightly increased (ρ = 0.27). Sensitivity and specificity analysis revealed that the electromechanical QP is the most performant to distinguish between different early degeneration stages, followed by subchondral-bone plate thickness and then cartilage thickness. Lastly, effect sizes of cartilage and subchondral-bone properties were established to evaluate whether cartilage or bone showed the most noticeable changes between normal (ICRS 0) and each early degenerative stage. Thus, the effect sizes of cartilage electromechanical QP were almost twice those of the subchondral-bone plate thickness, indicating greater sensitivity of electromechanical measurements to detect early osteoarthritis. CONCLUSION: The potential of electromechanical properties for the diagnosis of early human cartilage degeneration was highlighted and supported by cartilage thickness and µCT assessments.

Non-destructive electromechanical assessment (Arthro-BST) of human articular cartilage correlates with histological scores and biomechanical properties.

OBJECTIVE: The hand-held Arthro-BST™ device is used to map electromechanical properties of articular cartilage. The purpose of the study was to evaluate correlation of electromechanical properties with histological, biochemical and biomechanical properties of cartilage. METHOD: Electromechanical properties (quantitative parameter (QP)) of eight human distal femurs were mapped manually ex vivo using the Arthro-BST (1 measure/site, 5 s/measure, 3209 sites). Osteochondral cores were then harvested from different areas on the femurs and assessed with the Mankin histological score. Prior to histoprocessing, cores were tested in unconfined compression. A subset of the cores was analyzed with polarized light microscopy (PLM) to assess collagen structure. Biochemical assays were done on additional cores to obtain water content and glycosaminoglycan (GAG) content. The QP corresponding to each core was calculated by averaging all QPs collected within 6 mm of the core center. RESULTS: The electromechanical QP correlated strongly with both the Mankin score and the PLM score (r = 0.73, P < 0.0001 and r = -0.70, P < 0.0001 respectively) thus accurately reflecting tissue quality and collagen architecture. Electromechanical QP also correlated strongly with biomechanical properties including fibril modulus (r = -0.76, P < 0.0001), matrix modulus (r = -0.69, P < 0.0001), and log of permeability (r = 0.72, P < 0.0001). The QP correlated weakly with GAG per wet weight and with water content (r = -0.50, P < 0.0003 and r = 0.39, P < 0.006 respectively). CONCLUSION: Non-destructive electromechanical QP measurements correlate strongly with histological scores and biomechanical parameters providing a rapid and reliable assessment of articular cartilage quality.

Estimating ON and OFF contributions to the photopic hill: normative data and clinical applications.

BACKGROUND: With progressively brighter stimuli, the amplitude of the b-wave of the human photopic electroretinogram (ERG) first increases to a maximal value (Vmax) and then decreases to finally reach a plateau, a phenomenon known as the photopic hill (PH). A mathematical model combining a Gaussian (G) and a logistic (L) growth function was previously proposed to fit this unusual luminance-response curve, where the G and L functions were suggested to represent, respectively, the OFF and ON retinal pathway contributions to the building of the PH. METHOD: The PHs of patients presenting stationary diseases affecting specifically the ON (3 CSNB-1) or OFF (4 CPCPA) retinal pathways as well as patients affected with retinitis pigmentosa (14 RP) of different stages or etiology were analyzed using this mathematical model and compared to the PHs of a group of 28 normal subjects. RESULTS: The PH of the CSNB-1 patients had a much larger contribution from the G function compared to normal subjects, whereas the opposite was observed for the CPCPA patients. On the other hand, analysis of data from RP patients revealed variable G-L contributions to the building of their PH. CONCLUSION: In this study, we confirm the previous claim that the luminance-response function of the photopic ERG b-wave can be decomposed into a Gaussian function and a logistic growth function representing, respectively, the OFF and ON retinal pathways. Furthermore, our findings suggest that this mathematical decomposition could be useful to further segregate and potentially follow the progression of retinopathies such as RP.

Streaming potential-based arthroscopic device is sensitive to cartilage changes immediately post-impact in an equine cartilage injury model.

Models of post-traumatic osteoarthritis where early degenerative changes can be monitored are valuable for assessing potential therapeutic strategies. Current methods for evaluating cartilage mechanical properties may not capture the low-grade cartilage changes expected at these earlier time points following injury. In this study, an explant model of cartilage injury was used to determine whether streaming potential measurements by manual indentation could detect cartilage changes immediately following mechanical impact and to compare their sensitivity to biomechanical tests. Impacts were delivered ex vivo, at one of three stress levels, to specific positions on isolated adult equine trochlea. Cartilage properties were assessed by streaming potential measurements, made pre- and post-impact using a commercially available arthroscopic device, and by stress relaxation tests in unconfined compression geometry of isolated cartilage disks, providing the streaming potential integral (SPI), fibril modulus (Ef), matrix modulus (Em), and permeability (k). Histological sections were stained with Safranin-O and adjacent unstained sections examined in polarized light microscopy. Impacts were low, 17.3 ± 2.7 MPa (n = 15), medium, 27.8 ± 8.5 MPa (n = 13), or high, 48.7 ± 12.1 MPa (n = 16), and delivered using a custom-built spring-loaded device with a rise time of approximately 1 ms. SPI was significantly reduced after medium (p = 0.006) and high (p<0.001) impacts. Ef, representing collagen network stiffness, was significantly reduced in high impact samples only (p < 0.001 lateral trochlea, p = 0.042 medial trochlea), where permeability also increased (p = 0.003 lateral trochlea, p = 0.007 medial trochlea). Significant (p < 0.05, n = 68) moderate to strong correlations between SPI and Ef (r = 0.857), Em (r = 0.493), log(k) (r = -0.484), and cartilage thickness (r = -0.804) were detected. Effect sizes were higher for SPI than Ef, Em, and k, indicating greater sensitivity of electromechanical measurements to impact injury compared to purely biomechanical parameters. Histological changes due to impact were limited to the presence of superficial zone damage which increased with impact stress. Non-destructive streaming potential measurements were more sensitive to impact-related articular cartilage changes than biomechanical assessment of isolated samples using stress relaxation tests in unconfined compression geometry. Correlations between electromechanical and biomechanical methods further support the relationship between non-destructive electromechanical measurements and intrinsic cartilage properties.

A polarized light microscopy method for accurate and reliable grading of collagen organization in cartilage repair.

OBJECTIVES: Collagen organization, a feature that is critical for cartilage load bearing and durability, is not adequately assessed in cartilage repair tissue by present histological scoring systems. Our objectives were to develop a new polarized light microscopy (PLM) score for collagen organization and to test its reliability. DESIGN: This PLM score uses an ordinal scale of 0-5 to rate the extent that collagen network organization resembles that of young adult hyaline articular cartilage (score of 5) vs a totally disorganized tissue (score of 0). Inter-reader reliability was assessed using Intraclass Correlation Coefficients (ICC) for Agreement, calculated from scores of three trained readers who independently evaluated blinded sections obtained from normal (n=4), degraded (n=2) and repair (n=22) human cartilage biopsies. RESULTS: The PLM score succeeded in distinguishing normal, degraded and repair cartilages, where the latter displayed greater complexity in collagen structure. Excellent inter-reader reproducibility was found with ICCs for Agreement of 0.90 [ICC(2,1)] (lower boundary of the 95% confidence interval is 0.83) and 0.96 [ICC(2,3)] (lower boundary of the 95% confidence interval is 0.94), indicating the reliability of a single reader's scores and the mean of all three readers' scores, respectively. CONCLUSION: This PLM method offers a novel means for systematically evaluating collagen organization in repair cartilage. We propose that it be used to supplement current gold standard histological scoring systems for a more complete assessment of repair tissue quality.

[Housing and health counselling services in the management of allergic respiratory disease]. / Le Conseil Habitat-Santé dans la prise en charge des maladies allergiques respiratoires.

INTRODUCTION: Input from Housing and health counselling services is advisable when a patient's health seems to be impaired by their housing conditions. METHODS: 650 home visits have been performed by our organisation since 2002. Each visit includes a questionnaire to assess respiratory as well as non-respiratory indoor risk factors, Acarex test to assess mite-allergen content in mattress dust, mould sampling and, when appropriate, air sampling for measurement of volatile organic compounds and aldehydes. RESULTS: The dwellings studied were mostly flats located in the downtown and occupied by a tenant. In most instances, several health hazards were identified. These hazards, in decreasing occurrence included: mold (74.4%), mite infestation in mattress dust (56.3%), cleaning products accessible to children's'hands (47.8%), dangerous electrical circuits (21.1%), exposure to chemical air pollutants (9.0%), exposure to an electromagnetic field (2.8%). Numerous fungal species were identified often occurring in association. CONCLUSION: This service allowed the identification of numerous and various health hazards. Its efficacy and effectiveness remains to be evaluated.

Tetrapolar measurement of electrical conductivity and thickness of articular cartilage.

A tetrapolar method to measure electrical conductivity of cartilage and bone, and to estimate the thickness of articular cartilage attached to bone, was developed. We determined the electrical conductivity of humeral head bovine articular cartilage and subchondral bone from a 1- to 2-year-old steer to be 1.14+/-0.11 S/m (mean+/-sd, n =11) and 0.306+/-0.034 S/m, (mean+/-sd, n =3), respectively. For a 4-year-old cow, articular cartilage and subchondral bone electrical conductivity were 0.88+/-0.08 S/m (mean+/-sd, n =9) and 0.179+/-0.046 S/m (mean+/-sd, n =3), respectively. Measurements on slices of cartilage taken from different distances from the articular surface of the steer did not reveal significant depth-dependence of electrical conductivity. We were able to estimate the thickness of articular cartilage with reasonable precision (<20% error) by injecting current from multiple electrode pairs with different inter-electrode distances. Requirements for the precision of this method to measure cartilage thickness include the presence of a distinct layer of calcified cartilage or bone with a much lower electrical conductivity than that of uncalcified articular cartilage, and the use of inter-electrode distances of the current injecting electrodes that are on the order of the cartilage thickness. These or similar methods present an attractive approach to the non-destructive determination of cartilage thickness, a parameter that is required in order to estimate functional properties of cartilage attached to bone, and evaluate the need for therapeutic interventions in arthritis.

Detection and analysis of cartilage degeneration by spatially resolved streaming potentials.

Cartilage molecular changes in osteoarthritis are most commonly related to the degradation and loss of proteoglycan and collagen fibrils of the extracellular matrix, which directly influence tissue stiffness and compression-generated streaming potentials. In this study, we evaluated the potential of a new technique, spatially resolved mapping of streaming potentials, to non-destructively indicate cartilage health or degeneration. Matched pairs of bovine cartilage/bone explant disks were cultured for 11 days in a serum free medium with and without interleukin-lalpha (IL-1alpha). The electromechanical properties (static stiffness, dynamic stiffness and streaming potentials) of cartilage disks were measured during unconfined compression using a mechanical tester coupled with a linear array of eight 50 microm diameter platinum-iridium microelectrodes. After 11 days of culture, the proteoglycan content of IL-1alpha treated disks was significantly reduced and the denatured and cleaved collagen content was increased compared to control disks. These biochemical alterations were concomitant with the reductions in the amplitudes of the static stiffness, the dynamic stiffness and the streaming potential profile as well as changes in the shape of the streaming potential profile. We found that spatial mapping of streaming potentials presents several advantages for the development of a clinical instrument to evaluate the degeneration of articular cartilage.

Streaming potentials maps are spatially resolved indicators of amplitude, frequency and ionic strength dependant responses of articular cartilage to load.

Streaming potential distributions were measured on the surface of articular cartilage in uniaxial unconfined compression using a linear array of microelectrodes. Potential profiles were obtained for sinusoidal and ramp/stress-relaxation displacements and exhibited dependencies on radial position, sinusoidal amplitude and frequency, time during stress relaxation, and on ionic strength. The measurements agreed with trends predicted by biphasic and related models. In particular, the absolute potential amplitude was maximal at the disk center, as was the predicted fluid pressure and the potential gradient (the electric field) was seen to be maximal at the disk periphery, as was the predicted fluid velocity. We also observed a similarity between non-linear behavior of streaming potential amplitude and load amplitude with respect to sinusoidal displacement amplitude. Taken together, these results support many of the phenomena concerning relative fluid-solid movement and fluid pressurization predicted by biphasic and related models, and they indicate the general utility of spatially resolved measurements of streaming potentials for the investigation of electromechanical phenomena in tissues. For example, these streaming potential maps could be used to non-destructively diagnose cartilage extracellular matrix composition and function, as well as to quantify spatially and temporally varying physical signals in cartilage that can induce cellular and extracellular biological responses to load.

Glucanolytic Actinomycetes Antagonistic to Phytophthora fragariae var. rubi, the Causal Agent of Raspberry Root Rot.

A collection of about 200 actinomycete strains was screened for the ability to grow on fragmented Phytophthora mycelium and to produce metabolites that inhibit Phytophthora growth. Thirteen strains were selected, and all produced (beta)-1,3-, (beta)-1,4-, and (beta)-1,6-glucanases. These enzymes could hydrolyze glucans from Phytophthora cell walls and cause lysis of Phytophthora cells. These enzymes also degraded other glucan substrates, such as cellulose, laminarin, pustulan, and yeast cell walls. Eleven strains significantly reduced the root rot index when inoculated on raspberry plantlets.

Contributions of Martha Rogers to the development of nursing knowledge.

Martha Rogers' greatest contribution may be her introduction into nursing of a questioning stance about the prevailing models of science, leading to increased acceptance of alternative paradigms, a variety of research methods, and explorations of topics such as existentialism and Eastern philosophy. Nursing is a richer, more diverse profession because of Martha Rogers.

Mouth protectors and oral trauma: a study of adolescent football players.

A total of 754 junior high school and high school football players from the Birmingham, AL, area were studied to determine the extent of sports-related oral trauma among players. More than half the oral injuries and more than a third of the concussions were reported in sports other than football. Basketball and baseball players had a high prevalence of hard tissue oral trauma with virtually none of the players wearing mouth protectors. The results of this study indicate that current mouth protector designs are helpful in preventing hard tissue trauma, yet additional soft tissue protection is indicated in football mouth protector design.

A self-help program for childhood asthma in a residential treatment center.

A structured program designed to enhance self-treatment was successfully implemented in a residential center for asthmatic children. The ultimate objective of the program was to improve compliance with therapeutic regimens, which was felt to be a factor that had necessitated placement of many of the patients. The program was designed to educate the patient and the patient's family regarding the nature of asthma, its treatment and the importance of self-help. Efforts were also made to enhance the emotional maturity of the child. Patients remembered to take their medication over 90% of the time within 1 month of implementation of the program. a similar program was instituted for outpatient use.

Changes in the beta-endorphin content of discrete hypothalamic nuclei during the estrous cycle of the rat.

Concentrations of beta-endorphin were measured in discrete brain nuclei of 4-day cycling rats on each day of the estrous cycle. On the afternoon of proestrus beta-endorphin levels were significantly higher in median eminence and suprachiasmatic nucleus, and lower in arcuate nucleus, when compared to levels found on other days of the cycle. These changes coincided with the peak of plasma prolactin, which was blocked by prior administration of naloxone.

Changes of pituitary thyrotropin releasing hormone (TRH) receptor level and prolactin response to TRH during the rat estrous cycle.

The plasma PRL and TSH responses to TRH injected iv at different stages of the estrous cycle in normal rats under Surital anesthesia were maximal during the afternoon of proestrus and morning of estrus and lowest on diestrus I. As calculated from the areas under the plasma response curves, a 10-fold difference was found between the maximal and minimal PRL responses while a 2-fold difference was measured for TSH. The plasma PRL and TSH responses to TRH showed a correlation with the binding of [3H]TRH to anterior pituitary gland, a 3-fold difference being observed between the minimal binding measured on the morning of diestrus II and the maximal value found on the evening of proestrus. Contrary to findings with LHRH and LH, repeated injections of a small dose (10 ng) of TRH in the afternoon of proestrus abolished PRL and TSH responses to subsequent injection of the neurohormone.