CACNA1A mutations causing episodic and progressive ataxia alter channel trafficking and kinetics.

BACKGROUND: CACNA1A encodes CaV2.1, the pore-forming subunit of P/Q-type voltage-gated calcium channel complexes. Mutations in CACNA1A cause a wide range of neurologic disturbances variably associated with cerebellar degeneration. Functional studies to date focus on electrophysiologic defects that do not adequately explain the phenotypic findings. OBJECTIVE: To investigate whether some missense mutations might interfere with protein folding and trafficking, eventually leading to protein aggregation and neuronal injury. METHODS: The authors studied the functional consequences of two pore missense mutations, C287Y and G293R, in two families with EA2, one newly discovered and the other previously reported. Both mutations caused episodic and interictal ataxia. The biophysical properties of mutant and wild type calcium channels were examined by whole-cell patch-clamp recordings in transfected COS-7 cells. The plasma membrane targeting was visualized by confocal fluorescence imaging on CaV2.1 tagged with green fluorescent protein. RESULTS: The mutant channels exhibited a marked reduction in current expression and deficiencies in plasma membrane targeting. CONCLUSIONS: In addition to altered channel function, the deficiency in protein misfolding and trafficking associated with the C287Y and G293R mutants may contribute to the slowly progressive cerebellar ataxia.

The effects of tibial rotation on the patellofemoral joint: assessment of the changes in in situ strain in the peripatellar retinaculum and the patellofemoral contact pressures and areas.

The objective of this study was to determine the effects of tibial rotation on in situ strain in the peripatellar retinaculum and patellofemoral contact pressures and areas. Patellofemoral joint biomechanics demonstrate a strong correlation with the etiology of patellofemoral disorders, such as chondromalacia, and are significantly influenced by tibial rotation. Six human cadaveric knees were used along with a patellofemoral joint testing jig that permits physiological loading of the knee extensor muscles. Patellofemoral contact pressures and areas were measured with a Fuji pressure-sensitive film, and the changes in in situ strain in the peripatellar retinaculum were measured with four differential variable reluctance transducers. Tibial rotation had a significant effect on patellofemoral joint biomechanics. The data showed an inverse relationship between increasing knee flexion angles and the change in patellofemoral contact pressures and in situ strain with tibial rotation. At higher knee flexion angles, the patella is well-seated in the trochlear groove and the function of the peripatellar retinaculum is minimized and less affected by tibial rotations.

Deformation and strain characteristics along the length of the anterior band of the inferior glenohumeral ligament.

Efficacious surgical treatment of anterior glenohumeral instability often requires repair of the anteroinferior capsulolabral structures, including the glenoid origin of the anterior band of the inferior glenohumeral ligament. Rupture in this location, the Bankart lesion, may be accompanied by nonrecoverable stretching of the anterior band. The purpose of this study was to evaluate the amount and location of nonrecoverable stretching with tensile testing. Twelve glenoid-soft tissue-humerus complexes from fresh-frozen glenohumeral joints were studied by means of a custom jig, an Instron machine, and a video digitizing system. The joints were positioned to simulate that known to cause apprehension for anterior instability. Nonrecoverable deformation differed along the length of the anterior band but was slight in all locations. For those that failed at the glenoid insertion region, the mean nonrecoverable deformation was 0.10 +/- 0.16 mm (mean +/- SEM) at the bone-labral junction of the glenoid insertion region and 0.38 +/- 0.23 mm at the labral-ligament junction of the glenoid insertion region. It was 0.53 +/- 0.23 mm at the ligament midsubstance and 0.04 +/- 0.10 mm at the humeral insertion region. For those that failed at the glenoid insertion region, the nonrecoverable stretching was 1.4% +/- 1.9% at the bone-labral junction of the glenoid insertion region and 3.5% +/- 2.0% at the labral-ligament junction of the glenoid insertion region. It was 2.3% +/- 1.1% at the ligament midsubstance and 0.0% +/- 1.4% at the humeral insertion region. Rupture of the anterior band resulted in little nonrecoverable stretching at both the site of failure and elsewhere along the length, remote from the failure site. Surgical repairs after initial dislocation may restore the length of the anterior band of the inferior glenohumeral ligament with little shortening.

Inhibition of natural killer cell activation signals by killer cell immunoglobulin-like receptors (CD158).

The killer cell immunoglobulin-like receptor (KIR) family includes receptors that bind to HLA class I molecules on target cells and inhibit natural killer (NK)-cell cytotoxicity, and receptors such as KIR3DL7 with no known ligand and function. Inhibitory KIR recruit the tyrosine phosphatase SHP-1 to block signals transduced by any one of a number of activation receptors. Inhibition of overall protein tyrosine phosphorylation by SHP-1 during binding of KIR to MHC class I on target cells is selective, suggesting that a limited number of substrates are dephosphorylated by SHP-1. We have chosen to study KIR inhibition as it occurs during binding of KIR to MHC class I on target cells, despite the technical limitations inherent to studies of processes regulated by cell contact. KIR binding to MHC class I on target cells inhibits tyrosine phosphorylation of the activation receptor 2B4 (CD244) and disrupts adhesion of NK cells to target cells. Inhibition of proximal events in NK activation may increase the availability of NK cells by liberating them from non-productive interactions with resistant target cells. As the receptors and the signaling pathways that induce NK cytotoxicity are not fully characterized, elucidation of the inhibitory mechanism employed by KIR may provide insight into NK activation.

The healing effects on the biomechanical properties of joint capsular tissue treated with Ho:YAG laser: An in vivo rabbit study.

PURPOSE: The objective of this study was to evaluate the healing response, after thermal treatment with a Ho:YAG laser, on the biomechanical properties of capsular soft tissue. TYPE OF STUDY: Before and after trial. METHODS: Forty-five New Zealand white rabbits were used in this study. A medial peripatellar retinacular thermal capsuloplasty using a Ho:YAG laser and a lateral peripatellar retinacular release was performed on 1 knee of each rabbit. The contralateral knee served as a control and had a lateral release of the retinaculum only. The temperature of the medial retinaculum was maintained at 55 degrees C +/- 5 degrees C during treatment. The medial peripatellar retinaculum was evaluated at 0, 6, and 12 weeks postoperatively. Tensile testing of the medial retinaculum and a biomechanical assessment evaluating the structural and material properties were performed. RESULTS: The ultimate load (force) of the medial retinaculum was 70%, 56%, and 84% of control at 0, 6, and 12 weeks, respectively, after the procedure. The stiffness (force/deformation) of the medial retinaculum was 83% of control at 0 weeks, 54% at 6 weeks, and 85% at 12 weeks. The ultimate stress (force/area) of the medial retinaculum also showed a significant reduction at 0 and 6 weeks postoperatively, 63% and 62% of control, respectively. By 12 weeks, the ultimate stress was 83% of control. CONCLUSIONS: Thermal treatment of the medial retinaculum with a Ho:YAG laser results in soft tissue with significantly diminished biomechanical properties after treatment. The results of this study suggest that a 12-week period of minimal stress on the capsular tissues should follow a thermal capsuloplasty procedure.

Glenohumeral translation after arthroscopic thermal capsuloplasty with a radiofrequency probe.

The purpose of this study was to determine whether there are changes in anterior and posterior glenohumeral translation after arthroscopic thermal capsuloplasty with a radiofrequency probe. Anteriorly directed loads of 15 N and 20 N were sequentially applied to the humerus of each of 5 cadaveric glenohumeral joints, and anterior translation on the glenoid was measured through use of a customized translation apparatus and an electromagnetic tracking device. The tests were then repeated with posteriorly directed forces, and posterior translation was measured. During testing, the glenoid was rigidly fixed and the glenohumeral joint was positioned to simulate 90 degrees of shoulder abduction and 90 degrees of external rotation. By means of the radiofrequency probe, thermal energy was then applied to the anteroinferior capsuloligamentous structures; anterior and posterior translation measurements were repeated. The results showed a significant reduction in anterior and posterior translations after thermal capsuloplasty (P < .05). Anterior translation decreased from 6.8 to 4.0 mm (a 41% decrease) with the 15-N load and from 8.6 to 4.9 mm (a 42% decrease) with the 20-N load. Posterior translation decreased from 9.3 to 5.8 mm (a 36% decrease) with the 15-N load and from 10.4 to 6.5 mm (a 35% decrease) with the 20-N load. The results of this study indicate that the radiofrequency probe can be used to decrease both anterior and posterior glenohumeral translation in vitro. The biological effect on heat-treated tissues over time needs to be studied to prove that this is a satisfactory treatment for glenohumeral instability.

Age related biomechanical properties of the glenoid-anterior band of the inferior glenohumeral ligament-humerus complex.

OBJECTIVE: To quantify the biomechanical properties of the glenoid-anterior band of the inferior glenohumeral ligament-humerus complex for the two age groups. DESIGN: In vitro human cadaver study evaluating the biomechanical properties of the glenoid-anterior band of the inferior glenohumeral ligament-humerus complex for a younger group (n=5, mean age 38.5, SD 0.5 years) and an older group (n=7, mean age 74.8, SD 5.3 years). BACKGROUND: Glenohumeral instability is more of a problem in younger than in older individuals, primarily because recurrence is much more common at a young age. METHODS: Tensile testing was performed on the glenoid-anterior band of the inferior glenohumeral ligament-humerus complex in the shoulder apprehension position using a custom jig, Instron machine and a video digitizing system. RESULTS: In the younger individuals disruption of the complex most often occurred at the glenoid-labrum region of the glenoid insertion site. In the older individual, disruption most often occurred at the midsubstance region. The load and the stress at failure of the glenoid-anterior band of the inferior glenohumeral ligament-humerus complex showed that the older group was only 61% and 46% of the younger group, respectively. CONCLUSIONS: The structural properties of the glenoid-anterior band of the inferior glenohumeral ligament-humerus complex and the material characteristics of the anterior band of the inferior glenohumeral ligament for the younger group were significantly superior than the older group. RELEVANCE: A stronger and more extensive repair, such as the traditional open technique, may be necessary for younger individuals with glenohumeral instability whereas in older individuals, a different repair technique, such as an arthroscopic technique, may be sufficient.

The anterior band of the inferior glenohumeral ligament. Assessment of its permanent deformation and the anatomy of its glenoid attachment.

Surgical treatment for traumatic, anterior glenohumeral instability requires repair of the anterior band of the inferior glenohumeral ligament, usually at the site of glenoid insertion, often combined with capsuloligamentous plication. In this study, we determined the mechanical properties of this ligament and the precise anatomy of its insertion into the glenoid in fresh-frozen glenohumeral joints of cadavers. Strength was measured by tensile testing of the glenoid-soft-tissue-humerus (G-ST-H) complex. Two other specimens of the complex were frozen in the position of apprehension, serially sectioned perpendicular to the plane containing the anterior and posterior rims of the glenoid, and stained with Toluidine Blue. On tensile testing, eight G-ST-H complexes failed at the site of the glenoid insertion, representing a Bankart lesion, two at the insertion into the humerus, and two at the midsubstance. For those which failed at the glenoid attachment the mean yield load was 491.0 N and the mean ultimate load, 585.0 N. At the glenoid region, stress at yield was 7.8 +/- 1.3 MPa and stress at failure, 9.2 +/- 1.5 MPa. The permanent deformation, defined as the difference between yield and ultimate deformation, was only 2.3 +/- 0.8 mm. The strain at yield was 13.0 +/- 0.7% and at failure, 15.4 +/- 1.2%; therefore permanent strain was only 2.4 +/- 1.1%. Histological examination showed that there were two attachments of the anterior band of the inferior glenohumeral ligament at the site of the glenoid insertion. In one, poorly organised collagen fibres inserted into the labrum. In the other, dense collagen fibres were attached to the front of the neck of the glenoid.

Glenohumeral joint translation after arthroscopic, nonablative, thermal capsuloplasty with a laser.

The purpose of this study was to determine whether there are changes in anterior and posterior glenohumeral translation after arthroscopic, nonablative, thermal capsuloplasty with a laser. Two anteriorly and two posteriorly directed loads were sequentially applied to the humerus of nine cadaveric glenohumeral joints, and anterior and posterior translation of the humerus on the glenoid was measured. The glenoid was rigidly fixed, and the glenohumeral joint was positioned simulating 90 degrees of shoulder abduction and 90 degrees of external rotation. Using the holmium:yttrium-aluminum-garnet laser, thermal energy was then applied to the anterior capsuloligamentous structures and anterior and posterior translation measurements were then repeated. The results showed a significant reduction in anterior and posterior translation after laser anterior capsuloplasty. Anterior translation decreased from 10.9 +/- 2.0 mm (mean +/- SEM) to 6.4 +/- 1.5 mm with the 15-N load; and from 13.4 +/- 2.1 mm to 8.9 +/- 1.8 mm with the 20-N load. Posterior translation decreased from 7.2 +/- 1.2 mm to 4.4 +/- 0.6 mm with the 15-N load and from 10.4 +/- 1.4 mm to 6.5 +/- 0.9 mm with the 20-N load. These results indicate that the holmium:yttrium-aluminum-garnet laser can be used to decrease glenohumeral joint translation and may be an effective treatment for glenohumeral joint instability.

Michigan Medicaid recipients' perceptions of medication counseling as required by OBRA '90.

OBJECTIVE: To assess pharmacist counseling under OBRA '90 from the Medicaid recipient's perspective. Specifically, the study was designed to (1) assess pharmacists' compliance with counseling requirements, (2) assess recipients' level of satisfaction with the information provided during counseling and whether the information provided increased their comfort level in taking medication correctly, and (3) determine relationships between variables associated with pharmacist counseling and recipient satisfaction and comfort level. DESIGN: Cross-sectional telephone survey of Medicaid recipients. Medicaid recipient or caregiver was the unit of measure. SETTING: Michigan. PATIENTS: 408 recipients who received new prescriptions during a one-week period in November 1995. INTERVENTIONS: Telephone survey. MAIN OUTCOME MEASURE: Recipients' perception of whether an offer to counsel was made. RESULTS: Only 104 (25.5%) recipients indicated that someone offered counseling for their new prescription, and only 62 (15.2%) recipients indicated they knew of the requirement; 163 (40.0%) indicated someone offered counseled them. Counseled recipients were satisfied with the amount, quality, and way the information was presented, and were more likely to assign a higher level of importance to pharmacist counseling. The majority of respondents indicated high levels of comfort in using their medications safely, with those who were counseled expressing a higher level of comfort. CONCLUSION: From the perspective of the Medicaid recipient, pharmacies are failing to offer counseling for most new prescriptions. The results indicate that counseling improves measures of recipient comfort in using medications safely and enhances the level of importance patients assign to pharmacist counseling.

Complete genome sequence of Treponema pallidum, the syphilis spirochete.

The complete genome sequence of Treponema pallidum was determined and shown to be 1,138,006 base pairs containing 1041 predicted coding sequences (open reading frames). Systems for DNA replication, transcription, translation, and repair are intact, but catabolic and biosynthetic activities are minimized. The number of identifiable transporters is small, and no phosphoenolpyruvate:phosphotransferase carbohydrate transporters were found. Potential virulence factors include a family of 12 potential membrane proteins and several putative hemolysins. Comparison of the T. pallidum genome sequence with that of another pathogenic spirochete, Borrelia burgdorferi, the agent of Lyme disease, identified unique and common genes and substantiates the considerable diversity observed among pathogenic spirochetes.

The minimal gene complement of Mycoplasma genitalium.

The complete nucleotide sequence (580,070 base pairs) of the Mycoplasma genitalium genome, the smallest known genome of any free-living organism, has been determined by whole-genome random sequencing and assembly. A total of only 470 predicted coding regions were identified that include genes required for DNA replication, transcription and translation, DNA repair, cellular transport, and energy metabolism. Comparison of this genome to that of Haemophilus influenzae suggests that differences in genome content are reflected as profound differences in physiology and metabolic capacity between these two organisms.

Michigan drug utilization review and OBRA 90.

The DUR Board's role in the Medicaid DUR process is significant. With the requirements for membership on the DUR Board and the responsibilities directed by OBRA 90, the standards will ensure that they reflect the accepted medical practice and are clinically relevant to the geographical area represented. The DUR Board has the potential to improve drug utilization review and ultimately improve the quality of care. The approval of criteria for both patient retrospective and prospective DUR and the addition of education intervention will help us to improve the quality of care. DSS encourages your input and participation on the DUR regional committees. There are currently three committees that meet monthly and are located in: Detroit, Ann Arbor, Lansing.

Repair of sublethal and potentially lethal radiation damage by rat embryos exposed to gamma rays or helium lons.

Embryonic survival was examined in rats exposed to a 24-hour split-dose regimen of gamma rays or extended-Bragg-peak (EBP) helium ions on the fifth and sixth days of gestation. The data indicate that EBP helium ions, which are known to have a single-dose relative biological effectiveness (RBE) of 1.0, exhibit a split-dose RBE of 1.5 with respect to embryo killing. Using an experimental rat embryo system, delayed implantation, it was also noted that the embryocidal damage induced by EBP helium ions contains a smaller potentially lethal component than that induced by gamma rays.