ABSTRACT
AIM: At the interface of tissue and capillaries, pericytes (PC) may generate electrical signals to be conducted along the skeletal muscle vascular network, but they are functionally not well characterized. We aimed to isolate and cultivate muscle PC allowing to analyse functional properties considered important for signal generation and conduction. METHODS: Pericytes were enzymatically isolated from hamster thigh muscles and further selected during a 16-30 days' cultivation period. PC markers were studied by fluorescence activated cell scanning (FACS) and immunocytochemistry. Electrical properties of the cultured PC were investigated by patch clamp technique as well as the membrane potential sensitive dye DiBAC(4) (3). RESULTS: The cultured cells showed typical PC morphology and were positive for NG2, alpha smooth muscle actin, PDGFR-ß and the gap junction protein Cx43. Expressions of at least one single or combinations of several markers were found in 80-90% of subpopulations. A subset of the patched cells expressed channel activities consistent with a Kv1.5 channel. In vivo presence of the channels was confirmed in sections of hamster thigh muscles. Interleukin-8, a myokine known to be released from exercising muscle, increased the expression but not the activity of this channel. Pharmacologic stimulation of the channel activity by flufenamic acid induced hyperpolarization of PC alone but not of endothelial cells [human umbilical vein endothelial cells (HUVEC)] alone. However, hyperpolarization was observed in HUVEC adjacent to PC when kept in co-culture. CONCLUSION: We established a culture method for PC from skeletal muscle. A first functional characterization revealed properties which potentially enable these cells to generate hyperpolarizing signals and to communicate them to endothelial cells.
Subject(s)
Cell Separation/methods , Immunomagnetic Separation/methods , Muscle, Skeletal/cytology , Pericytes/cytology , Pericytes/physiology , Animals , Biomarkers/metabolism , Cells, Cultured , Connexin 43/metabolism , Cricetinae , Gap Junctions/metabolism , HeLa Cells , Humans , Interleukin-8/pharmacology , Kv1.5 Potassium Channel/metabolism , Membrane Potentials/physiology , Mesocricetus , Muscle, Skeletal/blood supply , Patch-Clamp Techniques , Pericytes/drug effectsABSTRACT
Oxygen equilibrium curves and the relationships between the partial pressure of CO2 and pH were determined for the haemolymph of the arachnids Eurypelma californicum, Pandinus imperator and Cupiennius salei. A new type of experimental apparatus was constructed, tested and used to make these measurements on small undiluted cell-free haemolymph samples. Most of its components were made in our workshop and were inexpensive. The apparatus proved to be very reliable as demonstrated by control (oxygen concentration) experiments. In previous reports on O2 equilibrium curves of E. californicum haemocyanin, TrisHCl buffer was used to set the pH, whereas in our experiments, pH was adjusted by altering the partial pressure of CO2. The O2 concentration measurements demonstrate an increase in oxygen affinity of E. californicum haemocyanin when using TrisHCl buffer. The position and shape of the O2 equilibrium curves of E. californicum and P. imperator haemocyanin show a distinct dependency on pH. Oxygen affinity is lower in E. californicum and cooperativity changes with pH in E. californicum, but not in P. imperator. Oxygen transport in the haemolymph of E. californicum during rest, activity and recovery was calculated on the basis of the O2 equilibrium curves of undiluted haemolymph. Apart from oxygen transport, there are indications that haemocyanin is also involved in other physiological processes. For example, it may function as a storage protein.
ABSTRACT
A multichannel biotelemetry system using pulse-width modulation-frequency modulation (PWM-FM) is described in detail for laboratory construction. Its application in a kinematic gait-analysis system is demonstrated, employing minimally encumbering electrogoniometry and foot-contact switches. The triaxial electrogoniometers sense rotational joint motion, and four foot-switches under the sole of each foot provide information on placement and temporal contact. Signals from the multiple sensors are amplified, encoded by pulse-width modulation, and transmitted at an FM radio frequency of 107 MHz. Received data are decoded and then sampled by a minicomputer for analysis. Results from a comparative study of kinematic gait in five normal subjects and five children with cerebral palsy demonstrate system effectiveness in providing quantitative data and various intrasubject and intersubject gait differences. Factors reviewed in the analysis include swing and stance times; cadence; hip-joint motion in sagittal, coronal, and transverse planes; and sequence of foot placement.
Subject(s)
Electrodiagnosis/instrumentation , Gait , Telemetry/instrumentation , Analog-Digital Conversion , Biomechanical Phenomena , Cerebral Palsy/diagnosis , Child , Equipment Design , Female , Humans , Male , Movement , Radio/instrumentation , TransducersABSTRACT
The activity of 82Rb produced from a 82Sr/82Rb generator is dependent on elution conditions (volume and eluent flow rate) and sampling conditions (time and position of collection). Assays for 82Rb in generator eluates are described using a commercial dose calibrator in a static procedure and a plastic scintillator in a dynamic procedure. Dynamic assays more accurately reflect the 82Rb administered when the eluate is injected directly. Radionuclidic contaminants which may be present with 82Sr are identified and procedures for their measurement are described.
