Fluorescent in vivo tracking of hematopoietic cells. Part I. Technical considerations

We report a new technology for in vivo tracking of hematopoietic cells, using fluorescent lipophilic probes. Because the probe is irreversibly bound in the lipids of the cell membrane; substantial numbers of dye molecules can be incorporated per cell and thus substantial signal to noise can be achieved. Although this technology can be used for all hematopoietic cells, these first findings are reported on red blood cells (RBCs) owing to the importance of the membrane to RBC function and integrity. We demonstrated that labeling 10% of the RBCs of a rabbit and reinjecting them into the animal makes possible the tracking of these cells at various times after injection. Furthermore, the labeling appears not to affect in vivo cell lifetime or cellular volume changes in response to hypotonic shock. The single cell fluorescence intensity of the labeled RBCs remains relatively constant for 60 days, and an immune response appears not to be generated against labeled cells. That labeled RBCs have lifetime kinetics in vivo, as shown in other studies, indicates that the membranes are functioning normally and are unaltered by the labeling technology. The technology we present is also applicable to white blood cells, bone marrow, and platelets.     

Muscle-Activation Onset Times With Shoes and Foot Orthoses in Participants With Chronic Ankle Instability

Context

Participants with chronic ankle instability (CAI) use an altered neuromuscular strategy to shift weight from double-legged to single-legged stance. Shoes and foot orthoses may influence these muscle-activation patterns.

Objective

To evaluate the influence of shoes and foot orthoses on onset times of lower extremity muscle activity in participants with CAI during the transition from double-legged to single-legged stance.

Design

Cross-sectional study.

Setting

Musculoskeletal laboratory.

Patients or Other Participants

A total of 15 people (9 men, 6 women; age = 21.8 ± 3.0 years, height = 177.7 ± 9.6 cm, mass = 72.0 ± 14.6 kg) who had CAI and wore foot orthoses were recruited.

Intervention(s)

A transition task from double-legged to single-legged stance was performed with eyes open and with eyes closed. Both limbs were tested in 4 experimental conditions: (1) barefoot (BF), (2) shoes only, (3) shoes with standard foot orthoses, and (4) shoes with custom foot orthoses (SCFO).

Main Outcome Measure(s)

The onset of activity of 9 lower extremity muscles was recorded using surface electromyography and a single force plate.

Results

Based on a full-factorial (condition, region, limb, vision) linear model for repeated measures, we found a condition effect (F_3,91.8 = 9.39, P < .001). Differences among experimental conditions did not depend on limb or vision condition. Based on a 2-way (condition, muscle) linear model within each region (ankle, knee, hip), earlier muscle-activation onset times were observed in the SCFO than in the BF condition for the peroneus longus (P < .001), tibialis anterior (P = .003), vastus medialis obliquus (P = .04), and vastus lateralis (P = .005). Furthermore, the peroneus longus was activated earlier in the shoes-only (P = .02) and shoes-with-standard-foot-orthoses (P = .03) conditions than in the BF condition. No differences were observed for the hip muscles.

Conclusions

Earlier onset of muscle activity was most apparent in the SCFO condition for ankle and knee muscles but not for hip muscles during the transition from double-legged to single-legged stance. These findings might help clinicians understand how shoes and foot orthoses can influence neuromuscular control in participants with CAI.Key Words: footwear, insoles, ankle sprains, neuromuscular system, electromyography

Key Points

• Shoes and foot orthoses accelerated muscle-activation onset times of the ankle and knee but not the hip in participants with chronic ankle instability.
• Earlier muscle-activation onset times were most prominent in the shoes-with-custom-foot-orthoses condition.
• At the ankle, the muscle-activation onset time of the peroneus longus was earlier in the shoes-only, shoes-with-standard-foot-orthoses, and shoes-with-custom-foot-orthoses conditions than in the barefoot condition, and the muscle-activation onset time of the tibialis anterior was earlier in the shoes-with-custom-foot-orthoses condition than in the barefoot condition.
• At the knee, the muscle-activation onset times of the vastus medialis obliquus and vastus lateralis were earlier in the shoes-with-custom-foot-orthoses condition than in the barefoot condition.
• The results may help clinicians understand how shoes and foot orthoses can influence neuromuscular control of the lower extremity in participants with chronic ankle instability.

Lateral ankle sprains are estimated to account for approximately 15% of all sport injuries.¹ Even more concerning than the initial ankle sprain is the large proportion of patients with residual symptoms and recurrent ankle sprains for months to years after the initial injury.² The occurrence of repetitive ankle sprains and the feeling of the ankle “giving way” with slight or no perturbation has been defined as chronic ankle instability (CAI).³The transition task from double-legged to single-legged stance during barefoot (BF) conditions has been shown to discriminate between uninjured participants and participants with CAI. Researchers have reported that muscle-activation onset times typically were delayed^4,5 and postural stability was impaired⁶ in participants with CAI, indicating the use of another strategy to shift weight from double-legged to single-legged stance. However, it is unclear whether findings from BF tests represent typical daily situations when shoes, and for some persons foot orthoses, are often worn.The human foot is the first point of contact between the body and a supporting surface. The cutaneous mechanoreceptors on the planar surface of the foot are an important source of sensory information,⁷ which is considered essential for achieving and maintaining functional joint stability.⁸ Shoes and foot orthoses act as an interface between the body and a supporting surface and can influence the sensory feedback from these mechanoreceptors by increasing the contact area between the foot and the supporting surface.^7,9 Furthermore, the small kinematic alterations of the rear foot and tibia that have been described with the use of shoes and foot orthoses¹⁰ may put the ankle joint in a more neutral position, thereby improving the capacity of the ankle mechanoreceptors to provide more accurate proprioceptive input toward the central nervous system.¹¹ Changing the sensory input to these mechanisms consequently would change the motor output.⁷Evidence is increasing that shoes and foot orthoses can influence lower extremity muscle activation.^10,12–14 Dingenen et al¹⁴ were the first investigators to measure the influence of shoes and foot orthoses on muscle-activation onset times of the entire lower extremity in uninjured participants during the transition from double-legged to single-legged stance. Their results showed that shoes and foot orthoses can accelerate muscle-activation onset times of the peroneus longus. No differences were reported in more proximal muscles. Recently, researchers have suggested that future investigators should be focused on the influence of shoes and foot orthoses on neuromuscular control, especially in participants with injuries, such as CAI,^10,13,14 to increase our understanding of how positive clinical outcomes from the use of shoes and foot orthoses can be achieved.¹¹ Altering or improving proprioceptive information and muscle-activation patterns in participants with CAI would be clinically beneficial, given that their proprioceptive and neuromuscular deficits have been described.¹⁵To our knowledge, no investigators have focused on the influence of shoes and foot orthoses on muscle-activation onset times of the entire lower extremity in participants with CAI during the transition from double-legged to single-legged stance. Therefore, the purpose of our study was to evaluate the influence of shoes and foot orthoses on muscle-activation onset times during the transition from double-legged to single-legged stance in participants with CAI. Based on the proposed effects of shoes and foot orthoses on lower extremity neuromuscular control, we hypothesized that shoes and foot orthoses would accelerate muscle-activation onset times compared with a BF condition.     

High serum tumor necrosis factor-alpha levels are associated with lack of response to infliximab in fistulizing Crohn's disease

OBJECTIVES: Infliximab, a chimeric monoclonal antibody directed against tumor necrosis factor-alpha (anti-TNF-alpha), has been effective in the treatment of patients with active Crohn's disease and with fistulas. We investigated the effect of infliximab on circulating cytokines and acute phase proteins in patients with fistulas to determine the clinical response to anti-TNF-alpha. METHODS: A total of 36 patients with fistulizing Crohn's disease were selected for study. Serum from patients was drawn before the infusion on day 0 and at wk 2, 4, 6, 8, and 10 after completion of treatment. Circulating concentrations of TNF-alpha, interleukin-1beta (IL-1beta), and IL-6 were measured by ELISA. The functional activity of circulating TNF-alpha was assessed by the WEHI 164 TNF-alpha bioassay. Acute phase proteins were also determined. RESULTS: Elevated TNF-alpha, IL-1beta, IL-6, and acute phase proteins were observed in patients with Crohn's disease. Of the patients with fistulas, 22 (61.1%) responded to treatment. Before receiving infliximab, higher levels of serum TNF-alpha were found in patients who did not respond to infliximab compared with those who did (median interquartile range 26, 0-245 pg/ml; n = 14 vs 0, 0-22 pg/ml, n = 22). Patients showed no change in circulating levels of TNF-alpha during the course of the study. CONCLUSIONS: This treatment produces a clinical improvement in about two-thirds of CD patients with fistulas. The circulating levels of TNF-alpha are associated with the response to infliximab and could help to identify patients who would benefit from anti-TNF-alpha treatment.     

Results of a French multicentre retrospective experience with four hundred and eighteen failed unicondylar knee arthroplasties

Purpose

By means of a multicentre retrospective study based on the failure of 418 aseptic unicondylar knee arthroplasties (UKA) our aims were to present the different types of revision procedure used in failed UKAs, to establish a clear operative strategy for each type of revision and to better define the indications for each type of revision.

Methods

Aseptic loosening was the principal cause of failure (n = 184, 44 %) of which 99 cases were isolated tibial loosening (23.5 % of the whole series and 54 % of all loosening), 25 were isolated femoral loosening (six and 13.6 %) and 60 were both femoral and tibial loosening (14.3 and 32.6 %). The next most common causes of failure were progression of arthritis (n = 56, 13.4 %), polyethylene wear (n = 53, 12.7 %), implant positioning errors (n = 26), technical difficulties (n = six) and implant failure (n = 16, 3.8 % of cases). Data collection was performed online using OrthoWave™ software (Aria, Bruay Labuissiere, France), which allows collection of all details of the primary and revision surgery to be recorded.

Results

A total of 426 revisions were performed; 371 patients underwent revision to a total knee arthroplasty (TKA) (87 %), 33 patients (7.7 %) were revised to an ipsilateral UKA, 11 (2.6 %) patients underwent contralateral UKA (ten) or patellofemoral arthroplasty (one) and 11 patients (2.6 %) underwent revision without any change in implants.

Conclusions

Before considering a revision procedure it is important to establish a definite cause of failure in order to select the most appropriate revision strategy. Revision to a TKA is by far the most common strategy for revision of failed UKA but by no means the only available option. Partial revisions either to an alternative ipsilateral UKA or contralateral UKA are viable less invasive techniques, which in carefully selected patients and in experienced hands warrant consideration.     

Evaluation of stair motion contributes to new insights into hip osteoarthritis‐related motion pathomechanics

Stair motion in the presence of hip osteoarthritis (OA) has received less attention than level walking. Its more strenuous aspect may shed the light on different locomotor strategies when compared to walking. We, therefore, aimed to define stair motion features associated to hip OA and to evaluate whether these specific features would differ from level walking and better characterize the hip pathological condition. Principal component and linear discriminant analyses were, respectively, used as data reduction and classification techniques. Our study highlighted that most of stair motion features associated to hip OA were similar to the ones of walking. Stair descent presented with the lowest misclassification error rate, ranging from 12% to 19% (estimated by cross‐validation). But, features that may be considered as a mechanism to reduce demand on the hip abductors were found to be more important in the stair ascent condition. This was reflected by both, greater importance in the classification rule and variance compared with walking, that is, decreased hip internal rotation moment at mid‐stance (72.50% vs. 57.63%) and increased trunk lateroflexion toward affected side (56.43% vs. 29.37%). This study emphasized the importance of investigating stair motion in hip osteoarthritic population by highlighting specific locomotor strategies. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:187–196, 2016.     

Video-assisted thoracic surgical procedure: management of a solitary pulmonary nodule.

Solitary pulmonary nodules continue to challenge all diagnostic skills. Herein we describe video-assisted thoracic surgical intervention, a new, minimally invasive technique that aids physicians in obtaining a definitive histologic diagnosis in a select group of patients with an indeterminate solitary pulmonary nodule.     

Matrix metalloproteinase abundance in human myocardial fibroblasts: effects of sustained pharmacologic matrix metalloproteinase inhibition

BACKGROUND: A cause-effect relationship has been established between matrix metalloproteinases (MMPs) and left ventricular (LV) myocardial remodeling through the use of pharmacologic MMP inhibitors. However, the direct effects of MMP inhibition on MMPs and endogenous tissue inhibitors of metalloproteinases (TIMPs) in LV human myocardial fibroblasts (LVHMFs) remain unknown. This study measured MMP-2, MMP-9, MMP-13, MT1-MMP, and TIMP-1 release in LVHMFs. METHODS AND RESULTS: LVHMF cultures were established from six individual patients (passages 2-5) and incubated with and without the broad-spectrum MMP inhibitor PD166793 (100 microM) for 12-36 h. While PD166793 did not influence MMP-2 release, MMP-9 levels based on substrate zymography increased at 36 h by over 30% (P < 0.05). TIMP-1 levels increased in a time-dependent manner with no effect from PD166793 incubation. However, the MMP-9/TIMP-1 ratio was increased by over 20% from time-matched values following 12-36 h of exposure to PD166793 (P < 0.05). Similar results obtained after incubation of LVHMF cultures with the broad-spectrum MMP inhibitor Batimastat (BB-94) suggest that these observations are due to a general class effect of broad-spectrum MMP inhibitors. CONCLUSIONS: This study is the first to demonstrate that a selective induction and release of an MMP species occurs with sustained exposure to pharmacologic MMP inhibition in LVHMFs. These observations may have particular importance with respect to controlling this proteolytic system in the context of LV myocardial remodeling.