CCL2 and CCR2 regulate pain-related behaviour and early gene expression in post-traumatic murine osteoarthritis but contribute little to chondropathy.

OBJECTIVE: The role of inflammation in structural and symptomatic osteoarthritis (OA) remains unclear. One key mediator of inflammation is the chemokine CCL2, primarily responsible for attracting monocytes to sites of injury. We investigated the role of CCL2 and its receptor CCR2 in experimental OA. DESIGN: OA was induced in 10 weeks old male wild type (WT), Ccl2-/- and Ccr2-/- mice, by destabilisation of the medial meniscus (DMM). RNA was extracted from whole joints at 6 h and 7 days post-surgery and examined by reverse transcription polymerase chain reaction (RT-PCR). Gene expression changes between naïve and DMM-operated mice were compared. Chondropathy scores, from mice at 8, 12, 16 and 20 weeks post DMM were calculated using modified Osteoarthritis Research Society International (OARSI) grading systems. Changes in hind paw weight distribution, as a measure of pain, were assessed by Linton incapacitance. RESULTS: Absence of CCL2 strongly suppressed (>90%) selective inflammatory response genes in the joint 6 h post DMM, including arginase 1, prostaglandin synthase 2, nitric oxide synthase 2 and inhibin A. IL6, MMP3 and tissue inhibitor of metalloproteinase 1 were also significantly suppressed. Similar trends were also observed in the absence of CCR2. A lower average chondropathy score was observed in both Ccl2-/- and Ccr2-/- mice at 12, 16 and 20 weeks post DMM compared with WT mice, but this was only statistically significant at 20 weeks in Ccr2-/- mice. Pain-related behaviour in Ccl2-/- and Ccr2-/- mice post DMM was delayed in onset. CONCLUSION: The CCL2/CCR2 axis plays an important role in the development of pain in murine OA, but contributes little to cartilage damage.

Transcriptional analysis of micro-dissected articular cartilage in post-traumatic murine osteoarthritis.

OBJECTIVE: Identify gene changes in articular cartilage of the medial tibial plateau (MTP) at 2, 4 and 8 weeks after destabilisation of the medial meniscus (DMM) in mice. Compare our data with previously published datasets to ascertain dysregulated pathways and genes in osteoarthritis (OA). DESIGN: RNA was extracted from the ipsilateral and contralateral MTP cartilage, amplified, labelled and hybridized on Illumina WGv2 microarrays. Results were confirmed by real-time polymerase chain reaction (PCR) for selected genes. RESULTS: Transcriptional analysis and network reconstruction revealed changes in extracellular matrix and cytoskeletal genes induced by DMM. TGFß signalling pathway and complement and coagulation cascade genes were regulated at 2 weeks. Fibronectin (Fn1) is a hub in a reconstructed network at 2 weeks. Regulated genes decrease over time. By 8 weeks fibromodulin (Fmod) and tenascin N (Tnn) are the only dysregulated genes present in the DMM operated knees. Comparison with human and rodent published gene sets identified genes overlapping between our array and eight other studies. CONCLUSIONS: Cartilage contributes a minute percentage to the RNA extracted from the whole joint (<0.2%), yet is sensitive to changes in gene expression post-DMM. The post-DMM transcriptional reprogramming wanes over time dissipating by 8 weeks. Common pathways between published gene sets include focal adhesion, regulation of actin cytoskeleton and TGFß. Common genes include Jagged 1 (Jag1), Tetraspanin 2 (Tspan2), neuroblastoma, suppression of tumourigenicity 1 (Nbl1) and N-myc downstream regulated gene 2 (Ndrg2). The concomitant genes and pathways we identify may warrant further investigation as biomarkers or modulators of OA.

Roaming behaviour of dogs in four remote Aboriginal communities in the Northern Territory, Australia: preliminary investigations.

OBJECTIVE: To estimate the home range (HR) and investigate the potential predictors for roaming of 58 dogs in four Aboriginal communities in the Northern Territory. DESIGN: Prospective study. PROCEDURE: Global positioning system (GPS) collars were attached to the dogs for 1-4 days, recording location fixes every 1-3 min. Utilisation distributions (UDs) and extended (95% isopleth) and core (50% isopleth) HRs of dogs were determined. Potential predictors of roaming were assessed. RESULTS: Estimated core (median, 0.27 ha) and extended (median, 3.1 ha) HRs differed significantly (P = 0.0225 and 0.0345, respectively) between the four communities; dogs in the coastal community travelled significantly (P < 0.0001) more per day than dogs in the three inland communities studied. Significant associations were found between extended HR size and sex (P = 0.0050) and sex + neuter (P = 0.0218), and between core HR size and sex (P = 0.0010), neuter status (P = 0.0255) and sex + neuter (P = 0.0025). Entire males roamed more than neutered females. The core HR of dogs with poor/fair body condition scores (BCSs) was larger than dogs with ideal/obese BCSs (P = 0.0394). Neutered male dogs also travelled more per day than entire female dogs (P = 0.0475). CONCLUSIONS: Roaming information can be used to inform the management of dogs in remote communities and to design disease control programs. Widespread data collection across the Northern Territory should be undertaken to further investigate the associations found in this study, considering that data were collected during relatively short periods of time in one season.

Endochondral Growth Defect and Deployment of Transient Chondrocyte Behaviors Underlie Osteoarthritis Onset in a Natural Murine Model.

OBJECTIVE: To explore whether aberrant transient chondrocyte behaviors occur in the joints of STR/Ort mice (which spontaneously develop osteoarthritis [OA]) and whether they are attributable to an endochondral growth defect. METHODS: Knee joints from STR/Ort mice with advanced OA and age-matched CBA (control) mice were examined by Affymetrix microarray profiling, multiplex polymerase chain reaction (PCR) analysis, and immunohistochemical labeling of endochondral markers, including sclerostin and MEPE. The endochondral phenotype of STR/Ort mice was analyzed by histologic examination, micro-computed tomography, and ex vivo organ culture. A novel protocol for quantifying bony bridges across the murine epiphysis (growth plate fusion) using synchrotron x-ray computed microtomography was developed and applied. RESULTS: Meta-analysis of transcription profiles showed significant elevation in functions linked with endochondral ossification in STR/Ort mice (compared to CBA mice; P < 0.05). Consistent with this, immunolabeling revealed increased matrix metalloproteinase 13 (MMP-13) and type X collagen expression in STR/Ort mouse joints, and multiplex quantitative reverse transcriptase-PCR showed differential expression of known mineralization regulators, suggesting an inherent chondrocyte defect. Support for the notion of an endochondral defect included accelerated growth, increased zone of growth plate proliferative chondrocytes (P < 0.05), and widespread type X collagen/MMP-13 labeling beyond the expected hypertrophic zone distribution. OA development involved concomitant focal suppression of sclerostin/MEPE in STR/Ort mice. Our novel synchrotron radiation microtomography method showed increased numbers (P < 0.001) and mean areal growth plate bridge densities (P < 0.01) in young and aged STR/Ort mice compared to age-matched CBA mice. CONCLUSION: Taken together, our data support the notion of an inherent endochondral defect that is linked to growth dynamics and subject to regulation by the MEPE/sclerostin axis and may represent an underlying mechanism of pathologic ossification in OA.

Owned dog and cat populations in remote Indigenous communities in the Northern Territory: a retrospective study.

OBJECTIVE: To determine the population of owned dogs and cats in Indigenous communities in the Northern Territory (NT), and compare the data with those for the average Australian household. METHODS: Results of 20 Indigenous community animal health programs were analysed for species present and dog and cat numbers. The female breeding and puppy populations were also identified. RESULTS: The average dog population density was significantly higher than the average Australian household, with an average of 24.4 dogs per 10 households, but the average cat population density was similar (3.3 cats per 10 households). Numbers of other species were not determined. The average percentage of puppies in these communities was 17.6% of the treated canine population, the average percentage of breeding canine females was 18.6% of the treated canine population, and the average percentage of breeding feline females was 19.7% of the total feline population. CONCLUSIONS: Dog populations in NT Indigenous communities were at least 6.3-fold higher per household compared with data for the rest of Australia. Cat populations per household were similar to the overall population. Factors contributing to the relatively high dog populations in remote Indigenous communities include a lack of veterinary presence, community remoteness, poor socioeconomic factors, poor house and yard designs, cultural reasons, communal beliefs, lack of community animal management and a lack of funding. We believe that animal health programs are an important way of addressing a number of these issues. Other elements that should be addressed include improving house and yard design, increasing education regarding animal health, care and welfare, and increasing the training and presence of health and animal professionals.

Innovation in immediate neonatal care: development of the Bedside Assessment, Stabilisation and Initial Cardiorespiratory Support (BASICS) trolley.

OBJECTIVE: Babies receive oxygen through their umbilical cord while in the uterus and for a few minutes after birth. Currently, if the baby is not breathing well at birth, the cord is cut so as to transfer the newborn to a resuscitation unit. We sought to develop a mobile resuscitation trolley on which newly born babies can be resuscitated while still receiving oxygenated blood and the 'placental transfusion' through the umbilical cord. This would also prevent separation of the mother and baby in the first minutes after birth. DESIGN: Multidisciplinary iterative product development. SETTING: Clinical Engineering Department of a University Teaching Hospital. METHODS: Following an initial design meeting, a series of prototypes were developed. At each stage, the prototype was reviewed by a team of experts in the laboratory and in the hospital delivery suite to determine ease of use and fitness for purpose. A commercial company was identified to collaborate on the trolley's development and secure marking with the Conformité Européenne mark, allowing the trolley to be introduced into clinical practice. RESULTS: The trolley is a small mobile resuscitation unit based on the concept of an overbed hospital table. It can be manoeuvred to within 50 cm of the mother's pelvis so that the umbilical cord can remain intact during resuscitation, irrespective of whether the baby is born naturally, by instrumental delivery or by caesarean section. Warmth for the newborn comes from a heated mattress and the trolley has the facility to provide suction, oxygen and air. CONCLUSIONS: This is the first mobile resuscitation device designed specifically to facilitate newborn resuscitation at the bedside and with an intact cord. The next step is to assess its safety, its acceptability to clinicians and parents, and to determine whether it allows resuscitation with an intact cord.

Canine tick-borne pathogens and associated risk factors in dogs presenting with and without clinical signs consistent with tick-borne diseases in northern Australia.

OBJECTIVES: To estimate the proportion of canine tick-borne disease (CTBD) pathogens in dogs from northern states of Australia presenting with and without clinical signs/laboratory abnormalities suggestive of CTBD and to evaluate associated risk factors. DESIGN: Client-owned dogs presented to a general practice clinic in the Northern Territory (NT; n = 138) and five referral hospitals in south-east Queensland (SEQ; n = 100) were grouped into CTBD-suspect and -control groups based on clinical and laboratory criteria. Blood and sera were screened for haemotropic Mycoplasma spp., Babesia spp., Anaplasma spp., Ehrlichia spp. and Hepatozoon spp. using microscopic examination, in-clinic ELISA testing and PCR assays. Dog-specific risk factors associated with the presence of CTBD pathogens were evaluated. RESULTS: Overall, 24.4% of the suspect group and 12.2% of the control group dogs were infected. The proportions of M. haemocanis, B. vogeli, A. platys, Candidatus Mycoplasma haematoparvum, and C. Mycoplasma haemobos were 7.1%, 5.0%, 3.8%, 1.7% and 0.4%, respectively. Dogs originating from the NT were 3.6-fold (95% confidence interval (CI) 1.51-8.62; P = 0.004) more likely to be infected with CTBD pathogens than those from SEQ. Male dogs were 2.3-fold (95% CI 1.17-4.80, P = 0.024) more likely to be PCR-positive to CTBD pathogens than female dogs. Dogs presenting with clinical signs consistent with CTBD and thrombocytopenia were more likely to be infected by CTBD pathogens (odds ratio 2.85; 95% CI 1.16, 7.02; P = 0.019). CONCLUSIONS: Haemotropic mycoplasmas were the most common tick-borne pathogen infecting client-owned dogs. Subclinical cases were common in dogs from the NT. Veterinary practitioners should be aware of the proportion of CTBD pathogens and the presenting features of clinical and subclinical disease in their area.

Distribution of 5-HT1 binding sites in cat spinal cord.

Quantitative analysis of high affinity [3H]5-HT binding to 5-HT1 receptors in the cervical, thoracic, lumbar, and sacral spinal cord of the cat revealed specific binding throughout the grey matter, with the highest levels of binding in laminae II and III, and the lowest levels in laminae I and VII. Relatively high levels were also observed in the thoracic intermediolateral cell column. There were no significant differences in the degree of binding between various segmental levels. Comparison of these data with published maps of 5-HT immunoreactivity reveals--with the exception of lamina I--a close correspondence between the degree of immunoreactivity and the degree of 5-HT binding. These results suggest that 5-HT plays an important role in a variety of spinal cord sensory, motor and autonomic functions.

Levodopa reduces muscle tone and lower extremity tremor in Parkinson's disease.

OBJECTIVE: We have quantified the effects of levodopa treatment in Parkinsonian subjects during maintained stance. METHODS: Electromyographic muscle activity during quiet stance was assessed in subjects with Parkinson's disease, who exhibited a fluctuating response to levodopa, and in age-matched control subjects. Stance stability was also assessed from mean displacement and velocity of the center of pressure excursions during stance. RESULTS: Lower extremity and trunk muscles showed high amplitude activity in all Parkinson's subjects when "off", and a 4-5 Hz tremor in three of these subjects. When "on", the amplitude of muscle activity was reduced in the distal muscles more than the proximal, while tremor was suppressed in all muscles. Corresponding to the excessive muscle activity, the Parkinson's subjects had increased velocity and variability of velocity in the anterior-posterior center of foot pressure excursions, but the mean displacement of the center of pressure excursion was not different from the controls. The velocity of center of pressure excursions in the Parkinson's subjects "on", approached those of the control subjects suggesting that the excessive distal muscle amplitude and tremor contributed to the high velocity of the center of pressure. CONCLUSIONS: These findings suggest that dopaminergic systems are involved in the regulation of muscle tone during stance. Depletion of dopaminergic transmission results in increased muscle tone and tremor in the lower extremities which may contribute to changes in posture and stability.

Influence of instruction, prediction, and afferent sensory information on the postural organization of step initiation.

1. Our previous study showed that two distinct postural modifications occurred when subjects were instructed to step, rather than maintain stance, in response to a backward surface translation: 1) the automatic postural responses to the surfaces perturbation were reduced in magnitude and 2) the anticipatory postural adjustments promoting foot-off were shortened in duration. This study investigates the extent to which task instruction, prediction of perturbation velocity, and afferent sensory information related to perturbation velocity are responsible for these postural modification. 2. Eleven human subjects were instructed in advance, to either maintain stance or step forward in response to a backward surface translation. Four different velocities of translation were used to perturb equilibrium. To assess the influence of predicted versus actual velocity information, the surface translations were presented in both a blocked order of increasing perturbation velocity (predictable) and a random order (unpredictable). Lower-extremity electromyographs (EMGs), ground reaction forces, and movement kinematics were quantified for both the automatic postural responses to perturbation and the anticipatory postural adjustments for step initiation. 3. The instruction to step was not solely responsible for the suppression of the automatic postural response. Prediction of perturbation velocity was required for significant suppression of the early automatic postural response when subjects stepped in response to the perturbation. When compared with the stance condition, the magnitude of the initial 50 ms of the automatic response in bilateral soleus and the left limb gastrocnemius (initial stance limb) was significantly reduced only when the perturbation velocities were presented in a blocked order. The magnitude of the automatic response was not reduced in the gastrocnemius of the right limb, which was always the initial swing limb and recruited for heel-off in the step conditions. This asymmetrical reduction of the gastrocnemius suggests that modification of the response was specific to the instruction, rather than a general decrease in the extensor muscle excitability. 4. The suppression of the early automatic postural response involved a change in the bias of the response. Despite the reduced magnitude during the predictable velocity step condition, the slope (i.e., gain) of the response with increasing velocities was not different from that of the stance condition. Thus the excitability of the automatic response was reduced by a relatively constant amount for each velocity when the perturbation velocity was predictable. 5. In contrast to the importance of velocity prediction for modification of the automatic postural response, actual velocity information was used for modification of the anticipatory postural adjustments when step was initiated in response to the surface perturbation. Regardless of whether the perturbation velocities were presented in a blocked or random order, the anticipatory postural adjustments were rapidly initiated and the duration of the postural adjustments for step initiation was shortened as the velocity of perturbation increased. 6. We conclude that the CNS uses prediction of perturbation velocity to modify the excitability of early automatic postural responses when the postural goal changes. In contrast, actual afferent velocity information can be used to modify the duration of the anticipatory postural adjustments for a voluntary step in response to perturbation. Thus the CNS utilizes feed-forward prediction to modify peripherally triggered postural responses, and utilizes immediate afferent information to modify the centrally initiated postural adjustments associated with voluntary movement.

Modification of postural responses and step initiation: evidence for goal-directed postural interactions.

1. In this study, the interaction between anticipatory postural adjustments for step initiation and automatic postural responses to an external perturbation were investigated by having subjects initiate a voluntary forward step while perturbed by a backward surface translation, which caused forward sway of the body. The postural adjustments for step initiation act to move the body center of mass (COM) forward, whereas the automatic postural responses act to move the COM backward to restore stance equilibrium. Because the postural behaviors are in opposition, we asked whether a temporal hierarchy exists in which automatic postural responses are executed to restore equilibrium and followed by stereotypic postural adjustments for step initiation, or whether the interaction between these two postural behaviors is more dynamic. 2. Lower extremity electromyographs (EMGs), ground reaction forces, and kinematics were recorded from 10 subjects during three conditions: to quantify the anticipatory postural adjustments for step initiation, subjects stepped forward as soon as they felt a proprioceptive cue; to quantify the automatic postural responses to perturbation, subjects maintained stance equilibrium in response to a backward surface translation under both feet; and to quantify the interaction between the postural adjustments for the voluntary step and the automatic responses to the perturbation, subjects were exposed to a backward surface translation and instructed to step forward as soon as they felt the platform begin to move. 3. The anticipatory adjustments for step initiation included tibialis activation [stance limb = 163 +/- 28 (SE) ms; swing limb = 173 +/- 33 ms] and soleus inhibition resulting in center of foot pressure (COP) moving backward and lateral toward the swing limb to propel the COM forward over the stance limb. Subsequently, activation of the swing limb gastrocnemius resulted in heel-off. In contrast, the automatic postural adjustments for maintenance of stance equilibrium during a backward surface translation included activation of soleus and gastrocnemius (104 +/- 23 ms and 115 +/- 14 ms, respectively) resulting in a symmetrical forward displacement of the COP that moved the COM back to its original position with respect to the feet. 4. When a forward step was initiated in response to the translation, the automatic postural responses were reduced in amplitude bilaterally in soleus and in the stance limb gastrocnemius. When present the postural response occurred at the same latency when the goal was to initiate a step as when the goal was to maintain standing.(ABSTRACT TRUNCATED AT 400 WORDS)

Human services workers: a study of perceptions of their role.

Human services workers at the Veterans Administration Hospital, Brentwood, are involved with patients from the time they are admitted to the hospital until they are discharged and adjust to the community. As part of an evaluation of their role, a study was done of their tasks as perceived by themselves, by their supervisors, by other paraprofessionals, and by patients. In general, human services workers were perceived to be most involved in assisting patients in the hospital, followed by involvement in treatment planning; discharge; therapy; assisting patients in the community; support activities; community activities; patient admission and evaluation; and research, education, and program evaluation.

Effects of thalamic stimulation on tremor, balance, and step initiation: a single subject study.

This study was conducted to evaluate the clinically apparent balance improvements in a patient with Parkinson's disease who had stimulating electrodes surgically implanted to the VIM nucleus of the right thalamus for control of left-upper-extremity tremor. Experiments were conducted to determine if balance improved simply because the large-amplitude upper-extremity tremor was reduced or if the neural control of balance improved. Using EMGs and forceplate recordings, we quantified the effects of the thalamic stimulation on the contralateral upper-extremity tremor and on the lower-extremity postural muscle activations for quiet stance, step initiation, and equilibrium responses to surface displacements. The results demonstrated that, beside reducing the amplitude and destabilizing effects of the upper-extremity tremor, the thalamic stimulation was also effective in reducing tremor activity of the trunk and contralateral lower-extremity muscles. In addition, the contralateral lower-extremity muscle activation patterns, strengths, and durations for the balance tasks were enhanced during stimulation. These results suggest that thalamic stimulation improved this patient's balance by reducing tremor in the contralateral extremities and by increasing burst duration and magnitude of the tibialis anterior, which functions as the postural prime mover for the step initiation and balance tasks.

Influence of cryotherapy on spasticity at the human ankle.

The application of cryotherapy to temporarily reduce spasticity is a widespread clinical practice. A method of quantifying spasticity, based on viscoelastic stiffness measurements at the ankle, was applied to objectively determine the efficacy of cryotherapy in reducing spasticity of the calf. Baseline, cryotherapy and one-hour postcryotherapy measurements of spasticity were performed in 25 subjects with clinical signs of spasticity secondary to traumatic brain injury, spinal cord injury, and stroke. A statistically significant reduction in spasticity occurred during cryotherapy. Postcryotherapy results were equivocal, although there was a tendency for diminished spasticity relative to the baseline measurement. Two subjects showed a clear aggravation of spasticity following cryotherapy, thus leading to the conclusion that dichotomous results are possible.

Quantitative evaluation of sway as an indicator of functional balance in post-traumatic brain injury.

The test of sway, using different conditions of stance with measurements of the average radial deviation of the center of pressure and its path length of sway per unit of time, has been shown to be a useful clinical tool in determining balance problems in traumatic brain injury (TBI) patients. Normative values were established to determine if an individual patient's sway values fell within the normal range (mean +/- 2SD). The tests have shown good test-retest reliability for TBI patients. In addition, it has been shown that the sensitivity of the test is sufficient to identify changes in patients' performances as their clinical conditions change. It has been demonstrated that the different stance conditions of the battery of tests become progressively more difficult to perform (from comfortable stance, eyes open and eyes closed, through narrow stance, eyes open and eyes closed, to tandem stance with right or left foot forward, eyes open and eyes closed). By using these subtests, it is easy to distinguish between the performances of able-bodied patients and TBI patients with very mild balance problems. The validity of the measure has been documented by correlating the sway performance with clinical functional performance tests. The test performance also correlates with the patient's own assessment of his or her gait difficulties. The limited data available suggest that the test of sway relates difficulties in static balance to the frequency of falls. Finally, subtests permit identification of specific problems in maintaining balance as a basis for therapeutic intervention.(ABSTRACT TRUNCATED AT 250 WORDS)