Clinical assessment of specific enzyme immunoassay for the human cardiac myosin light chain II (MLC II) with use of monoclonal antibodies

A highly specific enzyme-linked "sandwich" immunoassay was developed for determining cardiac myosin light chain II (MLC II) in serum by using an anticardiac MLC II monoclonal antibody and a solid phase consisting of glass rods coated with another monoclonal antibody. We can detect as little as 0.2 ng of cardiac MLC II per assay. The measurable range of cardiac MLC II concentration in serum is 1 to 30 micrograms/L. The assay demonstrated no cross-reactivity with a skeletal muscle MLC within the measurable range. The mean coefficients of variation were 6.1% within assay and 5.1% between assay. The concentration of cardiac MLC II in sera from healthy subjects ranged from 0 to 4.0 micrograms/L (mean 0.75 micrograms/L and median 0 micrograms/L). The concentrations of cardiac MLC II in serum of patients with skeletal muscle disease due to various causes (n = 15) and patients with effort angina (n = 25), in general, were not significantly elevated above normal. In all patients with myocardial infarction, the concentrations of cardiac MLC II were over 4.0 micrograms/L at 12 h after onset. The mean (+/- 1 SD) peak concentration of cardiac MLC II was 16.2 (+/- 4.4) micrograms/L at 90 h (mean) after onset. On the 5th day, the cardiac MLC II concentrations in all patients with myocardial infarction were significantly elevated above normal; none showed abnormal MB-creatine kinase (CK-MB) activity at this time. Thus, the measurement of cardiac MLC II concentration in serum may be useful to provide a specific and sensitive diagnosis of myocardial necrosis at any time period following myocardial infarction.     

Localization of palatal area in human somatosensory cortex

To determine the 'hard palate representing' area in the primary somatosensory cortex, we recorded somatosensory-evoked magnetic fields from the cortex in ten healthy volunteers, using magnetoencephalography. Following electrical stimulation of 3 sites on the hard palate (the first and third transverse palatine ridges, and the greater palatine foramen), magnetic responses showed peak latencies of 15, 65, and 125 ms. Equivalent current dipoles for early magnetic responses were found along the posterior wall of the inferior part of the central sulcus. These dipoles were localized anterior-inferiorly, compared with locations for the hand area in the cortex. However, there were no significant differences in three-dimensional locations among the 3 selected regions for hard palate stimulation. These results demonstrated the precise location of palatal representation in the primary somatosensory cortex, the actual area being small.     

Middle ear mucosal regeneration with three‐dimensionally tissue‐engineered autologous middle ear cell sheets in rabbit model

The likelihood of recurrent retraction and adhesion of newly formed tympanic membrane is high when middle ear mucosa is extensively lost during cholesteatoma and adhesive otitis media surgery. If rapid postoperative regeneration of the mucosa on the exposed bone surface can be achieved, prevention of recurrent eardrum adhesion and cholesteatoma formation, for which there has been no definitive treatment, can be expected. Suture‐less transplantation of tissue‐engineered mucosal cell sheets was examined immediately after the operation of otitis media surgery in order to quickly regenerate middle ear mucosa lost during surgery in a rabbit model. Transplantable middle ear mucosal cell sheets with a three‐dimensional tissue architecture very similar to native middle ear mucosa were fabricated from middle ear mucosal tissue fragments obtained in an autologous manner from middle ear bulla on temperature‐responsive culture surfaces. Immediately after the mucosa was resected from middle ear bone bulla inner cavity, mucosal cell sheets were grafted at the resected site. Both bone hyperplasia and granulation tissue formation were inhibited and early mucosal regeneration was observed in the cell sheet‐grafted group, compared with the control group in which only mucosal removal was carried out and the bone surface exposed. This result indicates that tissue engineered mucosal cell sheets would be useful to minimize complications after the surgical operation on otitis media and future clinical application is expected. Copyright © 2013 John Wiley & Sons, Ltd.     

Early activation of gammadelta T lymphocytes in patients with severe systemic inflammatory response syndrome

Innate immunity plays an important role in host defense after severe insult. gammadelta T lymphocytes are recognized as the first line of defense against microbial invasion. In this study, we evaluated gammadelta T lymphocytes in the peripheral blood of patients with severe systemic inflammatory response syndrome (SIRS), and examined on role of these cells. Thirty-seven patients with severe SIRS (SIRS criteria and serum C-reactive protein > or = 10 mg/dL) and 27 healthy volunteers were studied. Severe SIRS was caused by trauma in 14 patients (Injury Severity Score of 30.1 +/- 10.8) and by sepsis in 23 patients. The counts of gammadelta and alphabeta T lymphocytes were determined by flow cytometry of cells stained with monoclonal antibodies to gammadelta and alphabeta T lymphocyte receptors. The activation of these cells was evaluated by flow cytometry of cells stained with monoclonal antibodies to CD69 and HLA-DR. Serial counts and activation of gammadelta and alphabeta T lymphocytes were also determined in eight trauma patients (Injury Severity Score of 31.0 +/- 13.5) during a 2-week observation period. The count of gammadelta T lymphocytes in the peripheral blood of SIRS patients (30.1 +/- 6.0/microL) was significantly lower (P < 0.05) than that of the healthy volunteers (104.3 +/- 10.9/microL). The expression of CD69, an index of early activation of T lymphocytes, was significantly greater on gammadelta T lymphocytes from SIRS patients (patients 23.9% +/- 3.4%, healthy controls 4.8% +/- 0.6%, P < 0.05). In trauma patients, the expression of CD69 on gammadelta T lymphocytes increased rapidly within 48 h after injuries. In conclusion, gammadelta T lymphocytes are activated and decreased in the peripheral blood of severe SIRS patients. In trauma patients, the activation of gammadelta T lymphocytes occurs in the fairly acute phase after injuries. These results suggest a significant role for gammadelta T lymphocytes as early responders after severe insult.     

Assessment of pain‐related behavior and pro‐inflammatory cytokine levels in the rat rotator cuff tear model

The cause of pain following rotator cuff tear has not been fully elucidated. The purpose of this study was to evaluate behavior and inflammatory cytokines in a rat unstabilized rotator cuff defect (UCD) model. Forty‐five Sprague–Dawley rats were divided into three groups: sham; UCD; and stabilized rotator cuff defect (SCD). Gait analysis was examined using CatWalk. Tumor necrosis factor (TNF)‐α, interleukin(IL)‐1β, and IL‐6 were measured within the subacromial bursa and the glenohumeral joint synovium at 21 and 56 days after surgery using an enzyme‐linked immunosorbent assay (ELISA). Stride length, print area and contact intensity in the UCD group was significantly lower than in the sham group after surgery. Stride length, print area and contact intensity in the SCD group was significantly higher than in the UCD group. In contrast, TNF‐α, IL‐1β, and IL‐6 in the UCD group was significantly higher than in the sham group at days 21 and 56. However, TNF‐α, IL‐1β, and IL‐6 in the SCD group was significantly lower than in the UCD group at days 21 and 56. The present results suggest that SCD is effective not only in improving shoulder function but also in reducing inflammatory cytokines, which may serve as one source of pain due to rotator cuff tear. © 2013 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 32:286–290, 2014.     

Awareness and use of diagnostic support tools for lumbar spinal stenosis in Japan

BackgroundLumbar spinal stenosis (LSS) is a major clinical problem associated with back pain, intermittent claudication, leg pain, and leg numbness. Diagnostic support tools for LSS such as the self-administered, selfreported history questionnaire (SSHQ) and developmental clinical diagnosis support tool (ST) have been validated in Japan. However, the degree of awareness and use of these two diagnostic support tools for LSS in Japan has not been clarified. The aims of the current study were to determine the degree of awareness and use of these two diagnostic support tools by Japanese physicians. Furthermore, we compared these results among nonorthopedic general practitioner (GP), orthopedic GP, and hospital-based orthopedic physicians.MethodsThe LSS Diagnosis Support Tool (DISTO) Project was conducted to evaluate the degree of awareness and use of these two diagnostic support tools in Japan from 2011 to 2012. A total of 1,811 answers were obtained from physicians including nonorthopedic general practitioners (GP), orthopedic GPs, and hospital-based orthopedic physicians. Questions were (1) Do you know about these two diagnostic tools? and (2) If you know about these two diagnostic tools, have you used them?ResultsThe degree of awareness of ST and SSHQ was about 30 and 26 % by nonorthopedic GPs, 70 and 46 % by orthopedic GPs, and 68 and 41 % by hospital-based orthopedic physicians. The degree of awareness of ST and SSHQ by nonorthopedic GPs was significantly lower than by orthopedic GPs or hospital-based orthopedic physicians (p < 0.001). For physicians who were aware of ST and SSHQ, the degree of use of ST or SSHQ was < 50 % by nonorthopedic GPs, orthopedic GPs, and hospital-based orthopedic physicians.ConclusionsWe expect that use of the ST or SSHQ tools in primary care will improve the accuracy of diagnosis and lead to improved quality of patient care. The low proportion of awareness by nonorthopedic GPs (< 30 %) and use by all physicians (< 50 %) indicate a need to encourage physicians to use ST and SSHQ more frequently.     

Mini-Open Anterior Retroperitoneal Lumbar Interbody Fusion: Oblique Lateral Interbody Fusion for Lumbar Spinal Degeneration Disease

Purpose

Surgery for lumbar spinal degeneration disease is widely performed. While posterior decompression and fusion are popular, anterior lumbar interbody fusion (ALIF) is also used for treatment. Extreme lateral interbody fusion (XLIF) is commonly used for noninvasive ALIF; however, several complications, such as spinal nerve and psoas muscle injury, have been reported. In the current study, we examined the clinical efficacy and complications of oblique lateral interbody fusion (OLIF) for lumbar spinal degeneration disease.

Materials and Methods

Thirty-five patients with degenerated spondylolisthesis, discogenic pain, and kyphoscoliosis were examined. All patients underwent OLIF surgery (using a cage and bone graft from the iliac crest) with or without posterior decompression, without real-time electromyography monitoring. Posterior screws were used in all patients. Visual analog scale (VAS) score and Oswestry Disability Index (ODI) were evaluated before and 6 months after surgery. Surgical complications were also evaluated.

Results

Pain scores significantly improved after surgery, compared to those before surgery (p<0.05). There was no patient who underwent revision surgery. There was no spinal nerve, major vessel, peritoneal, or urinary injury. Few patients showed symptoms from psoas invasion.

Conclusion

OLIF surgery produced good surgical results without any major complication.