S100ß-mediated astroglial control of firing and input processing in layer 5 pyramidal neurons of the mouse visual cortex.

KEY POINTS: Inputs impinging on layer 5 pyramidal neurons perform essential operations as these cells represent one of the most important output carriers of the cerebral cortex. However, the contribution of astrocytes, a type of glial cell, to these operations is poorly documented. Here we found that optogenetic activation of astrocytes in the vicinity of layer 5 in the mouse primary visual cortex induces spiking in local pyramidal neurons through Nav1.6 ion channels and prolongs the responses elicited in these neurons by stimulation of their distal inputs in cortical layer 1. This effect partially involved glutamatergic signalling but relied mostly on the astrocytic calcium-binding protein S100ß, which regulates the concentration of calcium in the extracellular space around neurons. These findings show that astrocytes contribute to the fundamental computational operations of the cortex by acting on the ionic environment of neurons. ABSTRACT: The most complex cerebral functions are performed by the cortex, whose most important output is carried out by its layer 5 pyramidal neurons. Their firing reflects integration of the sensory and contextual information that they receive. There is evidence that astrocytes influence cortical neuron firing through the release of gliotransmitters such as ATP, glutamate or GABA. These effects have been described at the network and at the synaptic levels, but it is still unclear how astrocytes influence neuron input-output transfer function at the cellular level. Here, we used optogenetic tools coupled with electrophysiological, imaging and anatomical approaches to test whether and how astrocytic activation affected processing of distal inputs to layer 5 pyramidal neurons (L5PNs). We show that optogenetic activation of astrocytes near L5PN cell body prolonged firing induced by distal inputs to L5PNs and potentiated their ability to trigger spikes. The observed astrocytic effects on L5PN firing involved glutamatergic transmission to some extent but relied mostly on release of S100ß, an astrocytic Ca2+ -binding protein that decreases extracellular Ca2+ once released. This astrocyte-evoked decrease in extracellular Ca2+ elicited firing mediated by activation of Nav1.6 channels. Our findings suggest that astrocytes contribute to the cortical fundamental computational operations by controlling the extracellular ionic environment.

Polymer Composite with Carbon Nanofibers Aligned during Thermal Drawing as a Microelectrode for Chronic Neural Interfaces.

Microelectrodes provide a direct pathway to investigate brain activities electrically from the external world, which has advanced our fundamental understanding of brain functions and has been utilized for rehabilitative applications as brain-machine interfaces. However, minimizing the tissue response and prolonging the functional durations of these devices remain challenging. Therefore, the development of next-generation microelectrodes as neural interfaces is actively progressing from traditional inorganic materials toward biocompatible and functional organic materials with a miniature footprint, good flexibility, and reasonable robustness. In this study, we developed a miniaturized all polymer-based neural probe with carbon nanofiber (CNF) composites as recording electrodes via the scalable thermal drawing process. We demonstrated that in situ CNF unidirectional alignment can be achieved during the thermal drawing, which contributes to a drastic improvement of electrical conductivity by 2 orders of magnitude compared to a conventional polymer electrode, while still maintaining the mechanical compliance with brain tissues. The resulting neural probe has a miniature footprint, including a recording site with a reduced size comparable to a single neuron and maintained impedance that was able to capture neural activities. Its stable functionality as a chronic implant has been demonstrated with the long-term reliable electrophysiological recording with single-spike resolution and the minimal tissue response over the extended period of implantation in wild-type mice. Technology developed here can be applied to basic chronic electrophysiological studies as well as clinical implementation for neuro-rehabilitative applications.

Silicon-in-silica spheres via axial thermal gradient in-fibre capillary instabilities.

The ability to produce small scale, crystalline silicon spheres is of significant technological and scientific importance, yet scalable methods for doing so have remained elusive. Here we demonstrate a silicon nanosphere fabrication process based on an optical fibre drawing technique. A silica-cladded silicon-core fibre with diameters down to 340 nm is continuously fed into a flame defining an axial thermal gradient and the continuous formation of spheres whose size is controlled by the feed speed is demonstrated. In particular, spheres of diameter <500 nm smaller than those produced under isothermal heating conditions are shown and analysed. A fibre with dual cores, p-type and n-type silicon, is drawn and processed into spheres. Spatially coherent break-up leads to the joining of the spheres into a bispherical silicon 'p-n molecule'. The resulting device is measured to reveal a rectifying I-V curve consistent with the formation of a p-n junction.

Feasibility study for production of I-131 radioisotope using MNSR research reactor.

A feasibility study for (131)I production using a Low Power Research Reactor was conducted to predict the yield of (131)I by cyclic activation technique. A maximum activity of 5.1GBq was achieved through simulation using FORTRAN 90, for an irradiation of 6h. But experimentally only 4h irradiation could be done, which resulted in an activity of 4.0×10(5)Bq. The discrepancy in the activities was due to the fact that beta decays released during the process could not be considered.

Dietary supply of selenium for adolescents in three residential care orphanages in Southern Ghana.

Adolescents require optimum dietary supply of the essential trace mineral selenium (Se); however the absence of reliable and accurate data on the dietary supply of selenium for the adolescent population in Ghanaian residential care orphanages have made it difficult for public health nutritionists to assess the adequacy of the dietary supply. The dietary supply of selenium for adolescents (12-15 years) in three residential care orphanages, (Osu, Tutu-Akwapim and Teshie), in Southern Ghana have been evaluated by sampling their 24-h duplicate diets (including water) for 7-consecutive days using the duplicate diet sampling technique. The mass fraction of selenium in the blended lyophilized homogenates of duplicate diets was determined by radiochemical neutron activation analysis (RNAA). The validity of the RNAA method for selenium determination was checked by analyses of NIST SRM 1548a (Typical diet). The chemical yield of the radiochemical separation was determined by spectrophotometry. The average mass fractions of selenium in the blended lyophilized 24-hour duplicate diets for Osu, Tutu-Akwapim and Teshie were; 165±61 [117.2-285.2], 203±68 [110.5-304.9] and 250±92 [128.8-408.0]ng Seg(-1) lyophilized matter respectively. The average dietary supply of Se were, 57.6±17.3 [42.2-88.4], 82.0±30.7 [44.3-136.2] and 91.7±44.2 [46.0-153.4]µg Se day(-1) for Osu, Tutu-Akwapim and Teshie orphanages respectively. The data generated will help public health nutritionists in the provision of dietary advice and nutritional support for the studied orphanages, as well as other orphanages. The data will also help in the planning of institutional diets.

Predicting the yield of (177)Lu radionuclide produced by the cyclic irradiation technique.

The feasibility study on the production of (177)Lu radioisotope using a low power research reactor has been conducted. A reliable method for predicting the yield of (177)Lu produced using the cyclic activation technique based on the Westcott formalism has been established. A specific activity of 243.24 mCi/g was obtained when a (176)Lu(2)O(3) of natural abundance was irradiated for 4 h and decayed for 20 h for four cycles at GHARR-1 with a neutron flux of 5.0×10(11) ncm(-2)s(-1).

MUC1 expression and its association with other aetiological factors and localization to mitochondria in preneoplastic and neoplastic gastric tissues.

BACKGROUND: The molecular events that underlie the conversion of normal human gastric epithelium into adenocarcinoma are poorly understood. MUC1 overexpression and localization in mitochondria might confer cancer cells with attenuation of stress induced apoptosis. We studied MUC1 expression pattern, interaction with HSP70 and localization in mitochondria in preneoplastic and neoplastic human gastric tissues. METHODS: Immunohistochemistry and Western blot were used to study MUC1 expression pattern and localization in mitochondria. Coimmunoprecipitation was used to study MUC1 interaction with HSP70. MUC1 expression was correlated with other causative features including erbB2 expression. RESULTS: MUC1 was expressed in 75.8% (147/194). MUC1 overexpression was detected in 50.0% (19/38 cases) dysplasia and 58.2% (32/55 cases) adenocarcinoma tissues. MUC1-CT-HSP70 interaction was seen in 71.66% (43/60 cases) and MUC1 localized to mitochondria in 33.33% (5/15) dysplasia samples and in 47.05% (8/17) adenocarcinoma samples. MUC1 expression showed significant association with smoking (χ(2)=5.945; p<0.015), alcohol consumption (χ(2)=4.055; p<0.044) and erbB2 positivity (χ(2)=10.75; p<0.001). MUC1 expression did not show appreciable association with age (χ(2)=0.15; p<0.698), sex (χ(2)=0.22; p<0.640) or Helicobacter pylori infection (χ(2)=3.06; p<0.080). CONCLUSIONS: Significant correlation was found between MUC1 expression and smoking, alcohol and erbB2 expression. MUC1 showed aberrant expression in dysplasia and adenocarcinoma stages. MUC1 cytosolic tail was bound by HSP70 in all the stages but MUC1-CT was found to localize in mitochondria only in dysplasia and adenocarcinoma. MUC1-CT localization to mitochondria in dysplasia and adenocarcinoma might aid in the attenuation of epithelial stress response induced loss of polarity.

Expression of PTHrP and its cognate receptor in the rheumatoid synovial microcirculation.

Parathyroid hormone-related protein (PTHrP), a multifunctional peptide that acts as a vasodilator as well as possible regulator of vascular development, is produced in increased amounts in the rheumatoid synovium. To understand whether PTHrP can contribute to the development and function of the rheumatoid microcirculation, studies were undertaken to identify and compare vascular sites of expression of PTHrP and its cognate receptor in the rheumatoid synovium and/or in cultured rheumatoid synovial endothelial cells. Endothelial cells, including apoptotic cells, as determined by TUNEL staining, were the primary site of vascular PTHrP expression in the rheumatoid synovium, a result confirmed in vitro in rheumatoid synovial microvascular endothelial cells. In contrast, the PTH/PTHrP receptor was primarily located in pericytes and smooth muscle cells within the vasculature. These results are consistent with a possible paracrine pathway for PTHrP action in the synovial microcirculation, wherein PTHrP peptides secreted by the synovial endothelium could act on surrounding PTH1R-positive pericytes and smooth muscle cells.

An experimental method for the application of lateral muscle loading and its effect on femoral strain distributions.

Experimental models that have been used to evaluate hip loading and the effect of hip implants on bone often use only a head load and abductor load. Anatomic considerations and in vivo measurements have lead several investigators to suggest that these models are inaccurate because they do not incorporate the loads imposed by additional muscles. The aim of this study was to evaluate the strains in the proximal and mid diaphysis of the femur for five hip loading models, one with a head load and abductor load only and four which incorporated lateral muscle loads as well. Head load to body weight load ratios were used to evaluate the physiologic accuracy of these models and strains were compared to determine the extent of strain changes as a function of model complexity. All models which incorporated additional lateral muscle loads more accurately simulated head load to body-weight load ratios than the simple abductor-only model. The model which incorporated a coupled vastus lateralis and iliotibial band load in addition to the abductor load provided the simplest configuration with a reasonable body-weight to head-load ratio.

Polyethylene particle morphology in synovial fluid of failed knee arthroplasty.

Synovial fluid from the knees of 16 patients undergoing revision knee arthroplasty for aseptic failure was subjected to base digestion and ultrafiltration. Filtered particles were scanned using scanning electron microscopy and analyzed with an image program. Polyethylene particles were identified visually and confirmed with the use of electron diffraction spectroscopy. Averaging more than 1500 particles per patient sample, 25,148 particles were analyzed. This corresponded to a concentration of 3000 polyethylene particles per milliliter of synovial fluid. Three populations of wear debris were identified in the fluid. Small globular particles with a mean area of 75 mu 2 represented 94% of all particles observed. The particles averaged 10 mu in diameter and often were seen in clumps. Long fibrous particles with a mean area of 1164 mu 2 made up 4% of the particle population. Large rhomboidal particles with an area of 557 mu 2 were observed least commonly and comprised the remainder of the particles visualized. All three particle types were observed in each fluid sample regardless of the wear pattern of the retrieved polyethylene liner. There were no differences in absolute particle counts, particle morphologic characteristics, or particle size between patients with and without gross polyethylene wear.

Early, incapacitating instability of posterior cruciate ligament-retaining total knee arthroplasty.

Numerous studies indicate that total knee arthroplasty (TKA) achieves excellent long-term success whether the posterior cruciate ligament (PCL) is saved or excised. In 13 patients, 16 PCL-retaining TKAs were identified with incapacitating instability secondary to early PCL deficiency. Patients with clinical PCL insufficiency present with a triad of subjective complaints: persistent swelling/effusions, anterior knee pain, and giving-way or instability episodes with activities of daily living. Of 13 patients, 12 had at least three postoperative visits with identical subjective complaints before PCL deficiency was diagnosed. On examination all patients exhibited effusion, posterior sag, positive quadriceps active test, and a visible anterior translation of the tibia on the femur while extending the leg from a seated, 90 degree flexed position. This sign has not been previously described to our knowledge but was present in all of our study patients. No patients had radiographic evidence of loosening or osteolysis. Joint aspiration was negative for infection in all patients. No patient had lateral patellofemoral maltracking. By radiographic measurement, the PCL-deficient knees had an average joint line elevation of 10.3 mm, compared with well-functioning TKAs which had an average joint line elevation of 5.0 mm. There was no correlation of PCL deficiency with excessive proximal tibial resection. Nonsurgical intervention provided no improvement in pain or instability. Six patients had improvement of pain, effusion, and stability after revision to a cruciate-substituting implant. We believe that this complication occurs more frequently than is currently being identified and should be considered in problem TKAs with normal radiographs.

Contact areas and pressures between native patellas and prosthetic femoral components.

Contact areas and pressures between native patellas and a prosthetic condylar design femoral component were measured at flexion angles of 30 degrees, 60 degrees, and 90 degrees. These were compared to measurements obtained with a domed all-polyethylene patellar component. Mean native patellar contact areas were found to be fourfold greater than seen with the prosthetic patellar component. Contact stresses in the native patellas were below the yield strength of articular cartilage in 80% of the contact area. By contrast, stresses measured in the prosthetic patella exceeded the yield strength of ultrahigh molecular weight polyethylene in 64% of the measured contact area. Contact areas and stresses were not significantly effected by flexion angle. Although contact areas and stresses reflect only a part of the dynamics of the patellofemoral articulation this information would support the selective retention of the native patella in total knee arthroplasty.

Synovium as a source of increased amino-terminal parathyroid hormone-related protein expression in rheumatoid arthritis. A possible role for locally produced parathyroid hormone-related protein in the pathogenesis of rheumatoid arthritis.

Proinflammatory cytokines, including tumor necrosis factor (TNF) and interleukin 1 (IL-1), mediate the joint destruction that characterizes rheumatoid arthritis (RA). Previous studies have shown that parathyroid hormone-related protein (PTHrP) is a member of the cascade of proinflammatory cytokines induced in parenchymal organs during lethal endotoxemia. To test the hypothesis that NH2-terminal PTHrP, a potent bone resorbing agent, could also be a member of the synovial cascade of tissue-destructive cytokines whose expression is induced in RA, PTHrP expression was examined in synovium and synoviocytes obtained from patients with RA and osteoarthritis (OA). PTHrP production, as determined by measurement of immunoreactive PTHrP(1-86) in tissue explant supernatants, was increased 10-fold in RA versus OA synovial tissue. Synovial lining cells and fibroblast-like cells within the pannus expressed both PTHrP and the PTH/PTHrP receptor, findings that were confirmed by in vitro studies of cultured synoviocytes. TNF-alpha and IL-1beta stimulated PTHrP expression in synoviocytes, while dexamethasone and interferon-gamma, agents with some therapeutic efficacy in the treatment of RA, inhibited PTHrP release. Treatment of synoviocytes with PTHrP(1-34) stimulated IL-6 secretion. These results suggest that proinflammatory cytokine-stimulated production of NH2-terminal PTHrP by synovial tissue directly invading cartilage and bone in RA may mediate joint destruction through direct effects on cartilage or bone, or, indirectly, via the induction of mediators of bone resorption in the tumor-like synovium.

Safety and efficacy of bilateral total knee arthroplasty.

Recent studies have reported increased morbidity associated with bilateral simultaneous total knee arthroplasty (TKA). The purpose of this study was to evaluate the morbidity and clinical outcome associated with simultaneous bilateral TKA in contrast to unilateral TKA. All primary TKAs, either unilateral or simultaneous bilateral, performed between May 1988 and July 1993 were retrospectively reviewed. Patients were evaluated using Knee Society scores both before surgery and a minimum of 6 months after surgery. In addition to routine demographics, patients were evaluated for the incidence of both local wound and systemic complications. It is concluded that performing simultaneous bilateral TKA does not result in any significant increase in patient morbidity or compromise in postoperative function when compared with unilateral TKA.

Pullout strengths of cannulated and noncannulated cancellous bone screws.

The pullout strengths of large diameter cannulated and noncannulated cancellous screws were tested in a synthetic polyurethane foam. The foam was fabricated to have mechanical properties equivalent to human cancellous bone and was characterized by compression testing before screw pullout. Long and short thread commercially available screws from four manufacturers were tested. In screws with short threads (16-22 mm), there was no difference in holding power among the four cannulated screw designs. However, the short thread noncannulated screw performed significantly better than the short thread cannulated screw with the lowest pullout strength. There were statistically significant differences in holding power among the different long thread (32-40 mm) cannulated screw designs. Additionally, the long thread noncannulated screw had better holding power than several of the long thread cannulated screws. No differences in pullout strengths between comparably sized cannulated and noncannulated screws produced by the same manufacturer were found, and all long thread screws had significantly greater holding power than all short thread screws. There was no demonstrable effect on holding power when screws were inserted with or without tapping. Thread surface area was found to be a reasonable predictor of holding power.

Average and peak contact stress distribution evaluation of total knee arthroplasties.

Seven total knee arthroplasty systems were tested to determine contact stress patterns and contact areas using a calibrated Fuji film stress analysis technique. Knees were loaded to 2,000 N (204 kg) at 15 degrees, 60 degrees, 90 degrees, and 135 degrees flexion at 24 and 37 degrees C. Evaluation of stresses at 37 degrees C at 15 degrees and 60 degrees using an average contact stress assessment technique indicated that the LCS meniscal bearing knee system, (DePuy, Warsaw, IN), the AMK knee with a constrained insert (DePuy), and the PFC knee with a posterior-lipped insert (Johnson and Johnson, Raynham, MA) had the lowest average contact stresses (near or below 10 MPa). The PFC with a regular insert (Johnson and Johnson) the Ortholoc II (Dow Corning Wright, Arlington, TN), and the AMK with a regular insert (DePuy) had intermediate contact stresses. The AMK with a Hylamer-M insert (DePuy) and the MG II (Zimmer, Warsaw, IN) had the highest average contact stresses (near or above 20 MPa). A stress-calibrated Fuji film measurement technique has shown that an assessment of ranges of contact stress provides much more information about regions of expected wear than an assessment of average contact stresses. Testing of the tibiofemoral articulation of artificial knees revealed that all knees had some areas of contact with maximum stresses in excess of 15 MPa. As the yield strength of ultrahigh-molecular-weight polyethylene is approximately 15 MPa, all tibial inserts could wear to some extent. Peak contact stresses at four test angles of the AMK, Series 7000 (Osteonics, Allendale, NJ:) Genesis (Smith & Nephew Orthopaedics, Memphis, TN), and MG II patellofemoral articulations were high (above 30 MPa). Contact areas varied from line-shaped to bilateral circular or elliptical shapes. The LCS knee system experienced substantially lower patellofemoral contact stresses and larger contact areas. Changes in conformity of knee designs are warranted to overcome wear problems. Peak contact stresses measured from the LCS meniscal bearing tibiofemoral and patellofemoral joint were in excess of 30 MPa in some areas at low flexion angles. This design does create large areas of contact at very low contact pressures, however, and for this reason is expected to wear less than other designs.

Characterization of three formulations of a synthetic foam as models for a range of human cancellous bone types.

Porous polyurethane foams were prepared from Daro foam components with a range of mechanical properties to simulate human trabecular bone. Ratios of 10.0:5.0, 10.0:7.9, and 10.0:10.0 isocyanate to resin were mixed, cured, and cut into cubes. Properties were determined from uniaxial compression to 50% of the original cube height at a strain rate of 1.2 mm/s. Electron microscopy was used to characterize the foam structure. Average compressive yield stress values, ultimate compressive stresses, and elastic moduli ranged from 4.44 to 2.79, 5.61 to 3.28, and 134.0 to 110.1 MPa, respectively, for the three formulations. The foam materials showed a similar morphology of spherical bubbles, and the average bubble size tended to decrease as the ratio of isocyanate to resin increased even though the bubble size differences were not statistically significant. The results indicate that large blocks of foam can be prepared with consistent mechanical properties simulating a range of trabecular bone properties so that implants can be tested for various patient populations.

A biomechanical evaluation of three forms of internal fixation used in ankle arthrodesis.

A biomechanical study was undertaken to evaluate the relative stability of three types of internal fixation used for ankle arthrodesis. Crossed screw fixation, RAF fibular strut fixation, and T-plate fixation were tested in 30 cadaver ankles using an MTS machine. T-plate fixation consistently provided the stiffest construct when compared with the other types of fixation. Failure occurred by distraction of bony surfaces, posterior to the plane of fixation, in the crossed screw and RAF groups. In contrast, failure in the T-plate group occurred through compression of bone anterior to the midcoronal plane of the tibia. Although the stability of fixation is only one factor in determining the success or failure of ankle arthrodesis, the results of this study would support T-plate fixation over the other forms tested.