Utility of circulating serum miRNAs as biomarkers of early cartilage degeneration in animal models of post-traumatic osteoarthritis and inflammatory arthritis.

OBJECTIVE: The purpose of this study was to determine if serum microRNA (miRNA) signatures were biomarkers of early cartilage degeneration in preclinical mouse models of post-traumatic osteoarthritis (OA) and inflammatory arthritis. METHODS: Cartilage degeneration was induced in 10-12 week old male C57BL6 mice by destabilization of the medial meniscus (DMM) or intra-articular injection of methylated-bovine-serum-albumin (AIA), with sham-operated or saline-injected control animals (n = 6/treatment/time). Total serum RNA and knee joints were isolated at 1, 4 and 16 weeks post-induction. Cartilage degeneration was scored histologically. Serum miRNA expression profiling was performed using Agilent microarrays and validated by qPCR. RESULTS: DMM-operated and AIA mice had characteristic cartilage degeneration (proteoglycan loss, chondrocyte hypertrophy, structural damage), that increased significantly with time compared with controls, and with distinct temporal differences between arthritis models. However, expression profiling revealed no statistically significant dysregulation of serum miRNAs between AIA vs saline-injected or DMM vs sham-operated control mice at the critical early disease stages. The inability to detect DMM or AIA serum miRNA signatures compared with controls was not due to the insensitivity of the expression profiling approach since significant changes were observed in miRNA expression between the arthritis models and between time points. CONCLUSION: While distinct patterns of progressive cartilage degradation were induced in the arthritis models, we were unable to identify any serum miRNAs that were significantly dysregulated in early stages of disease compared with controls. This suggests circulating serum miRNAs may not be useful as cartilage biomarkers in distinguishing the early or progressive stages of arthritis cartilage degeneration.

In vitro evaluation of translating and rotating plates using a robot testing system under follower load.

BACKGROUND CONTEXT: Various modifications to standard "rigid" anterior cervical plate designs (constrained plate) have been developed that allow for some degree of axial translation and/or rotation of the plate (semi-constrained plate)-theoretically promoting proper load sharing with the graft and improved fusion rates. However, previous studies about rigid and dynamic plates have not examined the influence of simulated muscle loading. PURPOSE: The objective of this study was to compare rigid, translating, and rotating plates for single-level corpectomy procedures using a robot testing system with follower load. STUDY DESIGN: In-vitro biomechanical test. METHODS: N = 15 fresh-frozen human (C3-7) cervical specimens were biomechanically tested. The follower load was applied to the specimens at the neutral position from 0 to 100 N. Specimens were randomized into a rigid plate group, a translating plate group and a rotating plate group and then tested in flexion, extension, lateral bending and axial rotation to a pure moment target of 2.0 Nm under 100N of follower load. Range of motion, load sharing, and adjacent level effects were analyzed using a repeated measures analysis of variance (ANOVA). RESULTS: No significant differences were observed between the translating plate and the rigid plate on load sharing at neutral position and C4-6 ROM, but the translating plate was able to maintain load through the graft at a desired level during flexion. The rotating plate shared less load than rigid and translating plates in the neutral position, but cannot maintain the graft load during flexion. CONCLUSIONS: This study demonstrated that, in the presence of simulated muscle loading (follower load), the translating plate demonstrated superior performance for load sharing compared to the rigid and rotating plates.

Cartilage-specific ablation of XBP1 signaling in mouse results in a chondrodysplasia characterized by reduced chondrocyte proliferation and delayed cartilage maturation and mineralization.

OBJECTIVE: To investigate the in vivo role of the IRE1/XBP1 unfolded protein response (UPR) signaling pathway in cartilage. DESIGN: Xbp1(flox/flox).Col2a1-Cre mice (Xbp1(CartΔEx2)), in which XBP1 activity is ablated specifically from cartilage, were analyzed histomorphometrically by Alizarin red/Alcian blue skeletal preparations and X-rays to examine overall bone growth, histological stains to measure growth plate zone length, chondrocyte organization, and mineralization, and immunofluorescence for collagen II, collagen X, and IHH. Bromodeoxyuridine (BrdU) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) analyses were used to measure chondrocyte proliferation and cell death, respectively. Chondrocyte cultures and microdissected growth plate zones were analyzed for expression profiling of chondrocyte proliferation or endoplasmic reticulum (ER) stress markers by Quantitative PCR (qPCR), and of Xbp1 mRNA splicing by RT-PCR to monitor IRE1 activation. RESULTS: Xbp1(CartΔEx2) displayed a chondrodysplasia involving dysregulated chondrocyte proliferation, growth plate hypertrophic zone shortening, and IRE1 hyperactivation in chondrocytes. Deposition of collagens II and X in the Xbp1(CartΔEx2) growth plate cartilage indicated that XBP1 is not required for matrix protein deposition or chondrocyte hypertrophy. Analyses of mid-gestation long bones revealed delayed ossification in Xbp1(CartΔEx2) embryos. The rate of chondrocyte cell death was not significantly altered, and only minimal alterations in the expression of key markers of chondrocyte proliferation were observed in the Xbp1(CartΔEx2) growth plate. IRE1 hyperactivation occurred in Xbp1(CartΔEx2) chondrocytes but was not sufficient to induce regulated IRE1-dependent decay (RIDD) or a classical UPR. CONCLUSION: Our work suggests roles for XBP1 in regulating chondrocyte proliferation and the timing of mineralization during endochondral ossification, findings which have implications for both skeletal development and disease.

Growth rates and energy intake of hand-reared cheetah cubs (Acinonyx jubatus) in South Africa.

Growth rate is an important factor in neonatal survival. The aim of this study was to determine growth rates in hand-reared cheetah cubs in South Africa fed a prescribed energy intake, calculated for growth in the domestic cat. Growth was then compared with previously published data from hand-reared cubs in North America and the relationship between growth and energy intake explored. Daily body weight (BW) gain, feed and energy intake data was collected from 18 hand-reared cheetah cubs up to 120 days of age. The average pre-weaning growth rate was 32 g/day, which is lower than reported in mother-reared cubs and hand-reared cubs in North American facilities. However, post-weaning growth increased to an average of 55 g/day. Growth was approximately linear prior to weaning, but over the entire age range it exhibited a sigmoidal shape with an asymptotic plateau averaging 57 kg. Energy intake associated with pre-weaning growth was 481 kJ ME/kg BW(0.75). Regression analysis described the relationship between metabolic BW, metabolisable energy (ME) intake, and hence daily weight gain. This relationship may be useful in predicting energy intake required to achieve growth rates in hand-reared cheetah cubs similar to those observed for their mother-reared counterparts.

Exposure of growing and adult captive cheetahs (Acinonyx jubatus) to dietary isoflavones: twenty years later.

Dietary isoflavones are associated with oestrogenic and anti-oestrogenic effects, and have been linked to infertility in cheetahs. This study aimed to determine the isoflavone content of commercially prepared diets consumed by captive cheetahs. Sixteen international zoological facilities provided diets, and the isoflavone content of each diet was determined by acid hydrolysis and HPLC quantification. Proximate nutritional composition was also determined. Over half the diets analysed contained detectable concentrations of isoflavones, whereby total isoflavone content ranged from 1.75-183 mg/kg dry matter. The zoo-specific diets were calculated to deliver a median isoflavone dose of 0.07 mg/kg body weight (BW) and a maximum of 1.95 mg/kg BW to captive cheetahs. On a metabolic body weight basis this equates to a maximum of 4.90-5.43 mg/kg(0.75) . Some diets prepared for hand-rearing neonatal cheetahs could expose neonates to doses of up to 4.24 mg/kg BW (or 4.24-6.33 mg/kg(0.75) for cubs under 3 months of age). Only one of six zoo-specific diets was found to deliver isoflavones in doses shown to possess biological activity in other species. Therefore, on average, dietary isoflavones were not found in commercially prepared diets consumed by captive cheetahs in concentrations predicted to cause physiological changes. However, a small proportion of these diets, including hand-rearing formulas, contained elevated isoflavones concentrations which may influence cheetah fertility, behaviour or other physiological parameters.

SOX8 expression during chick embryogenesis.

We have isolated the SOX8 gene from the chicken embryo. This gene shows a high degree of sequence homology to SOX9 and SOX10. Detailed analysis of SOX8 expression by whole-mount in situ shows a dynamic and restricted expression pattern during chick development. SOX8 is expressed in the somitic derivative, the dermomyotome, the developing heart, pancreas, enteric neurone system, limb and the neural tube. This is the first detailed expression analysis of SOX8 in any species

S-adenosylmethionine blood levels in major depression: changes with drug treatment.

INTRODUCTION: The relationship between plasma levels of S-adenosylmethionine (SAMe), an endogenous methyl donor, and clinical response were studied in patients with a DSM-III-R diagnosis of major depression. MATERIAL AND METHODS: A double-blind randomized protocol comparing oral SAMe with oral desipramine, involving a total of 26 patients, was employed. RESULTS: At the end of the 4-week trial, 62% of the patients treated with SAMe and 50% of the patients treated with desipramine had significantly improved. Regardless of the type of treatment, patients with a 50% decrease in their Hamilton Depression Scale (HAM-D) score showed a significant increase in plasma SAMe concentration. CONCLUSION: The significant correlation between plasma SAMe levels and the degree of clinical improvement in depressed patients regardless of the type of treatment suggests that SAMe may play an important role in regulating mood.

Decreased seasonal mesor of platelet 3H-imipramine binding in depression.

Seasonal cycles of platelet 3H-imipramine binding were compared in 49 endogenous unipolar depressed patients and 20 normal volunteers. A significant sinusoidal component was detected in the Bmax of binding in both patients and controls with similar amplitudes and seasonal peaks. However, the yearly average (mesor) of the patient group was significantly lower (20.0%) than that of the normal controls. The results support earlier claims of a diminished platelet binding in endogenous depression and indicate that this decrease was still evident in the presence of a 48.2% (controls) to 65.8% (patients) seasonal variation. Control Bmax values were normally distributed about a best-fit mean (cosinor fit). In contrast, patient values appeared to be bimodally distributed with one mode that was similar to controls and one mode that was substantially lower. In general, psychiatric symptoms failed to distinguish between patients with high and low platelet binding and no correlation was found between Bmax and severity of illness (HAM-D).

Changes in platelet 3H-imipramine binding with chronic imipramine treatment are not state-dependent.

One month of imipramine treatment increased both the Kd and Bmax of platelet 3H-imipramine binding in 11 endogenous unipolar depressed patients. Continued treatment (13 weeks) of 5 patients subsequently lowered the Bmax values of 2 patients who had initially shown the largest increases, so that binding was no longer significantly elevated after 13 weeks. The observed changes in Kd but not in Bmax, could be explained by the carryover of tightly bound drug to the binding assay, although neither of the measures were correlated with plasma imipramine levels. Posttreatment Bmax (4 weeks) values were inversely related to plasma cortisol levels, although a weak but positive correlation was found before treatment. No significant change was found in plasma cortisol with treatment. Clinical responses were not related to cortisol or Bmax changes, although optimal improvement was associated with extreme values (high and low) of pretreatment Bmax. The present results, obtained with imipramine, and similar results obtained after nortriptyline and electroconvulsive shock by others, suggest that at least some antidepressants may induce transient changes in the Bmax of platelet binding that are independent of affective state.

S-adenosylmethionine treatment of depression: a controlled clinical trial.

The antidepressant properties of S-adenosylmethionine, an endogenous methyl donor, were studied in inpatients who met the DSM-III criteria for major depression. Nine patients given intravenous S-adenosylmethionine and nine given low oral doses of imipramine were compared in a double-blind design for 14 days. The S-adenosylmethionine produced superior results by the end of the first week of treatment. By the end of the second week, 66% of the S-adenosylmethionine patients had a clinically significant improvement in depressive symptoms, compared to 22% of the imipramine patients. Side effects appeared to be fewer with S-adenosylmethionine than with imipramine during the last 5 days of the study.

Modification of tumor blood flow: current status and future directions.

Suboptimal drug distribution and hypoxia, which can contribute to treatment failure, are a direct consequence of the spatial and temporal heterogeneity in perfusion that occurs in solid tumors. Therefore, improvements in tumor blood flow have wide-ranging therapeutic importance. Paradoxically, controlled decreases in tumor blood flow can also be exploited and, if permanent, induce extensive tumor cell death on their own. We review the current knowledge of the factors controlling tumor blood flow with emphasis on the roles of the endogeneous vasodilator nitric oxide and the endogenous vasoconstrictor endothelin-1. The potential importance and application of approaches that irreversibly damage vascular function, so-called vascular targeting, are also discussed. Emphasis is given to the drug-based approaches to vascular targeting that are now entering clinical evaluation. There is no doubt that increased understanding of the processes that determine blood flow in tumors, coupled with the availability of techniques to monitor blood flow noninvasively in the clinic, will enable strategies for selectively modifying tumor blood flow to be transferred from the laboratory to the clinical setting.

A performance-based incentive program for influenza immunization in the elderly.

Our objective was to implement and evaluate performance-based reimbursement for influenza immunization of the elderly in physician offices. We performed a community-based quasi-experiment with historic and concurrent comparisons, using primary care physician offices in Monroe County, New York. Participants in the intervention group included 53 primary care physicians admitting to one hospital, and the comparison group included 82 primary care physicians admitting to other hospitals. All physicians participated in a Medicare-sponsored demonstration to increase influenza immunization rates, and, during the 1990-1991 immunization season, used a target-based poster to track immunization rates. Physicians in the intervention group were enrolled in a performance-based financial incentive program that rewarded immunization rates above 70%. A survey concerning influenza immunization practices and opinions was sent to all physicians. The average physician-specific immunization rate in the incentive group was 73.1% versus 55.7% in the comparison practices (P < .001). Eligibility for incentives, practice size, sex of physician, medical specialty, reminder postcards, and practice populations including medically indigent patients were associated with immunization level. Controlling for the above variables, we completed a regression analysis showing that eligibility for the incentive was still significant (P = .003). The survey responses were not predictive of performance or significantly different between the two groups, except for the negative influence of sending postcards. This study in a community setting suggests that linking reimbursement to performance may be a successful strategy to increase influenza immunization levels for the elderly.

Effects of chronic imipramine treatment on subclasses of platelet 3H-imipramine binding sites and plasma cortisol.

A decreased density of platelet 3H-imipramine (3H-IMI) binding sites has been proposed as a putative trait marker of major depressive illness. However, subsequent studies have demonstrated that the number of such sites is increased so as to be more like normal controls upon chronic treatment with antidepressant drugs. In addition, there is some evidence to suggest that altered 3H-IMI binding may be secondary to elevated plasma cortisol levels which are common in depressed patients and which normalize with remission. The present study compares platelet 3H-imipramine binding, plasma cortisol levels, and clinical improvement of 10 endogenous depressed patients before and after 6 weeks of treatment with imipramine-HCl. Total high affinity 3H-IMI binding sites were further differentiated into two subclasses on the basis of their relative sensitivities to cyanoimipramine (CNIMI) inhibition. Treatment was associated with a significant increase (134%) in CNIMI resistant binding but a decrease (45.2%) in CNIMI sensitive binding. While the former was significantly correlated with posttreatment cortisol levels, no significant correlation was found between cortisol and CNIMI specific binding. Neither site appeared to be directly related to mood state. The significance of these findings to the evaluation of platelet binding as a trait dependent marker is discussed.

Seasonal changes in cyanoimipramine specific platelet 3H-imipramine binding in depression.

Seasonal variations in cyanoimipramine (CNIMI) sensitive and CNIMI resistant subclasses of platelet 3H-imipramine (3H-IMI) binding sites were studied in depressed patients and normal volunteers. Sinusoidal rhythms of the binding of both subclasses were found in patients and controls with peak levels in mid-February. Patient values of CNIMI sensitive binding fluctuated about a yearly average that was 32% lower than the average of controls. Patient deviations from the normal pattern were apparently bimodally distributed, whereas those of controls were normally distributed. CNIMI resistant binding was also normally distributed in controls, but not in depressed patients, although patient mesor values were not lower than those of controls. Platelet binding was not correlated with the severity of illness as measured by the Hamilton Rating Scale for Depression, and individual symptoms failed to discriminate between patients with high and low Bmax values.

Locked nailing of humeral shaft fractures. Experience in Edinburgh over a two-year period.

We report the results of locked Seidel nailing for 30 fractures of the humerus. There were frequent technical difficulties at operation especially with the locking mechanisms. Protrusion of the nail above the greater tuberosity occurred in 12 cases, usually due to inadequate locking, and resulted in shoulder pain and poor function. Poor shoulder function was also seen in five patients with no nail protrusion, presumably because of local rotator cuff damage during insertion. Our results suggest that considerable modifications are required to the nail, and possibly to its site of insertion, before its use can be advocated.

Primary knee arthroplasty for distal femoral fractures in elderly patients.

We discuss the role of primary knee arthroplasty in supracondylar and intercondylar fractures of the femur in elderly patients with reference to 13 cases. This method of treatment is shown to be effective and to have good results. It is recommended for all type C and some type A supracondylar fractures in old people.

Physician profiling: 12 critical points.

Physician profilers encompass an array of technological products that purport to evaluate individual clinician performance on utilization and other measures. Prior to installation, an organization should be familiar with a number of major concepts that encompass understanding of clinical practice processes, claims data limitations and idiosyncrasies, ethical issues, and mathematical principles. Twelve specific elements are described to assure that these concepts are addressed.

Analysis of power law models for the creep of nucleus pulposus tissue.

Tissue engineering approaches are now being investigated for altering the course of intervertebral disc degeneration (IDD). Because the disease changes the mechanical properties of the load bearing tissues of the disc, viscoelastic tissue behavior is a key measure for comparing the efficacy of treatments. To investigate the basic viscoelastic behavior of nucleus pulposus tissue, tissue from the rabbit disc was tested in torsional creep. Both the Andrade and Nutting creep models had a good fit to the data, however, the Andrade creep model gave a much better prediction of the longer term creep. This is the first application of Andrade creep to biological tissue and results indicate that this model may be particularly well suited for characterizing the viscoelastic behavior of very soft biological tissues.

The isoflavone content of commercially-available feline diets in New Zealand.

AIM: To identify and quantify concentrations of the isoflavones genistein, daidzein, biochanin A and formononetin in commercially- prepared feline diets sold in New Zealand. METHODS: Feline diets (n=138) were collected from supermarkets, pet stores and veterinary clinics in New Zealand. Diets were classified into five categories based on the following criteria: the presence/absence of soy, the presence/absence of non-soy plant material, and dry matter (DM) content. A high performance liquid chromatography (HPLC)-based assay was developed and validated to identify and quantify concentrations of the isoflavones genistein, daidzein, biochanin A and formononetin. RESULTS: Isoflavones were detected in all categories of diet, and at quantifiable concentrations in 104/138 (75%) of the diets tested. More dry diets (127/138; 92%) contained isoflavones at quantifiable concentrations than moist diets (83/138; 60%, p<0.001). Of the isoflavone-positive diets, moist diets that contained soy had the highest median isoflavone content (71.1 mg/kg DM and 0.018 mg/kcal metabolisable energy; ME) whilst moist meat-only diets had the lowest (3.24 mg/kg DM and 0.0004 mg/kcal ME). Isoflavone contents varied similarly between categories of diet whether evaluated on a DM or ME basis. High isoflavone contents were associated with diets containing soy and those of low cost. Biochanin A and formononetin were found at concentrations above the detection limit of the assay in 25/138 (18%) and 7/138 (5%) of the diets analysed, respectively. The estimated median daily isoflavone exposure for cats consuming feline diets sold in New Zealand ranged from 0.03 mg/kg bodyweight (BW) for moist meat-only diets to 1.47 mg/kg BW for moist diets containing soy, whilst the diet with the highest isoflavone content would provide daily exposure of 8.13 mg/kg BW. CONCLUSIONS: Isoflavones are present in commercially-prepared feline diets sold in New Zealand at concentrations that have elicited physiological responses in the reproductive, endocrine, and immune systems of other mammalian species. Since isoflavones appear to be a common constituent of feline diets, further investigation of the biological activities of these dietary compounds in cats is warranted.