In vivo gene expression revealed by cDNA arrays: the pattern in relapsing-remitting multiple sclerosis patients compared with normal subjects.

OBJECTIVES: To use DNA arrays to identify differences in gene expression associated with relapsing-remitting (RR) MS. METHODS: Total RNA was isolated from monocyte depleted peripheral blood mononuclear cells of 15 RR MS patients and 15 age- and sex-matched controls. The RNA was reverse transcribed to radiolabeled cDNA and the resultant cDNA was used to probe a DNA array containing over 4000 named human genes. The binding of radiolabeled cDNA to the probes on the array was measured by phosphorimager. RESULTS: Of more than 4000 genes tested, only 34 were significantly different in RR-MS patients from controls. Of these, 25 were significantly increased and 9 significantly decreased in the RR MS patients. Twelve of these genes have inflammatory and/or immunological functions that could be relevant to the MS disease process. The potentially relevant genes that were elevated (15% to 28%) were P protein, LCK, cAMP responsive element modulator, IL-7 receptor, matrix metalloproteinase-19, M130 antigen, and peptidyl-prolyl isomerase. Those that were significantly decreased (15% to 35%) were SAS transmembrane 4 superfamily protein, STRL22 (C-C chemokine receptor 6), AFX protein, DNA fragmentation factor-45 and immunoglobulin gamma 3 (Gm marker). CONCLUSIONS: The RR-MS disease effect was relatively restricted and most of the mRNAs tested were not different from the normal controls. However, there were significant differences identified in the expression of a subset of mRNAs, including 13 with inflammatory/immune functions that could be relevant to MS. The systematic use of DNA arrays can provide insight into the dynamic cellular pathways involved in MS pathogenesis and its phenotypic heterogeneity.

Duplex venous imaging: role for a comprehensive lower extremity examination.

Real-time compression ultrasound (CU) along with venous duplex imaging is the most commonly performed noninvasive vascular examination. It has become the definitive diagnostic test for most patients with deep venous thrombosis (DVT). Some practioners have recommended that CU alone of the common femoral vein (CFV) and of the popliteal vein (PV) are all that is required since a complete examination is time consuming and calf veins are difficult to visualize. However, if only the CFV and PV are examined, all patients with isolated superficial femoral vein (SFV) and calf DVT remain undiagnosed. The purpose of this study is to establish the value of a comprehensive venous duplex examination compared to CFV and PV compression alone for detecting both proximal and infrapopliteal DVT. From January 1996 through December 1997, the initial venous duplex examinations of 5767 extremities in 3067 patients were reviewed and results tabulated according to presence and location of clot. The ATL 3000 with a 7-14 mHz probe was utilized. Studies were interpreted as normal, proximal DVT (popliteal and above, with or without calf DVT), isolated calf, or isolated SFV deep venous thrombosis. If only the CFV and PV had been examined, 30.3% (isolated SFV + isolated calf vein DVT) of all DVT and 4.5% of proximal DVT would have been missed. A complete venous duplex examination altered the care in 288 (30.3%) of all patients examined who had DVT, and is therefore recommended as the standard noninvasive examination when evaluating patients for acute DVT.

Cholesterol movement between bovine rod outer segment disk membranes and phospholipid vesicles.

The ability of cholesterol to move between bovine rod outer segment disk membranes and phospholipid membranes was examined. Disk membranes were incubated with small unilamellar phospholipid vesicles containing varying amounts of cholesterol. Aliquots were removed at specific times, and then the disks and the vesicles were separated by centrifugation and assayed for phospholipid and cholesterol content. When incubated with vesicles containing no cholesterol, the cholesterol to phospholipid ratio in the disk membrane was reduced due to migration of cholesterol from the disks into the vesicles. The cholesterol content of these cholesterol depleted disks could be readily returned to the normal disk cholesterol content by incubation of the cholesterol-depleted disks with small unilamellar vesicles containing high cholesterol. An apparent partition coefficient K was calculated as the quotient of the cholesterol/phospholipid mole ratio in the donor membranes and the cholesterol/phospholipid mole ratio in the acceptor membranes. The value of K was approximately 1 at cholesterol levels below normal disk cholesterol content, for disk membranes and phosphatidylcholine small unilamellar vesicles. Inclusion of phosphatidylethanolamine in the small unilamellar vesicle acceptor raised K, indicating that phosphatidylethanolamine creates an unfavourable environment for cholesterol. The cholesterol to phospholipid ratio of native disks could be increased by incubation with phosphatidylcholine small unilamellar vesicles (donor) which contained higher amounts of cholesterol than the disk membrane acceptor. In these experiments the distribution of cholesterol between disks and small unilamellar vesicles always favored the vesicles. The apparent partition coefficient was 1.7 at several cholesterol levels above the native disk cholesterol content. Liposomes made from lipid extracted from the disk membrane behaved in the same manner as intact disks with respect to cholesterol distribution at equilibrium. The phospholipid content of the disk membrane may be an important factor in determining the cholesterol content of the disk membrane.