A method has been developed for culturing cardiac myocytes in a collagen matrix to produce a coherently contracting 3-dimensional model heart tissue that allows direct measurement of isometric contractile force. Embryonic chick cardiomyocytes were mixed with collagen solution and allowed to gel between two Velcro-coated glass tubes. During culture, the cardiomyocytes formed spontaneously beating cardiac myocyte-populated matrices (CMPMs) anchored at opposite ends to the Velcro-covered tubes through which they could be attached to a force measuring system. Immunohistochemistry and electron microscopy revealed a highly organized tissue-like structure of alpha-actin and alpha-tropomyosin-positive cardiac myocytes exhibiting typical cross-striation, sarcomeric myofilaments, intercalated discs, desmosomes, and tight junctions. Force measurements of paced or unpaced CMPMs were performed in organ baths after 6-11 days of cultivation and were stable for up to 24 h. Force increased with frequency between 0.8 and 2.0 Hz (positive "staircase"), increasing rest length (Starling mechanism), and increasing extracellular calcium. The utility of this system as a test bed for genetic manipulation was demonstrated by infecting the CMPMs with a recombinant beta-galactosidase-carrying adenovirus. Transduction efficiency increased from about 5% (MOI 0.1) to about 50% (MOI 100). CMPMs display more physiological characteristics of intact heart tissue than monolayer cultures. This approach, simpler and faster than generation of transgenic animals, should allow functional consequences of genetic or pharmacological manipulation of cardiomyocytes in vitro to be studied under highly controlled conditions.
In patients with ischemic stroke of unknown cause cerebral vasculitis is a rare but relevant differential diagnosis, especially when signs of intracranial artery stenosis are found and laboratory findings show systemic inflammation. In such cases, high-resolution T1w vessel wall magnetic resonance imaging (MRI; 'black blood' technique) at 3 T is preferentially performed, but may not be available in every hospital. We report a case of an 84-year-old man with right hemispheric transient ischemic attack and signs of distal occlusion in the right internal carotid artery (ICA) in duplex sonography. Standard MRI with contrast agent pointed the way to the correct diagnosis since it showed an intramural contrast uptake in the right ICA and both vertebral arteries. Temporal artery biopsy confirmed the suspected diagnosis of a giant cell arteritis and dedicated vessel wall MRI performed later supported the suspected intracranial large artery inflammation. Our case also shows that early diagnosis and immunosuppressive therapy may not always prevent disease progression, as our patient suffered several infarcts in the left middle cerebral artery (MCA) territory with consecutive high-grade hemiparesis of the right side within the following four months.
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