Variation of the neurofilament medium KSP repeat sub-domain across mammalian species: implications for altering axonal structure.

The evolution of larger mammals resulted in a corresponding increase in peripheral nerve length. To ensure optimal nervous system functionality and survival, nerve conduction velocities were likely to have increased to maintain the rate of signal propagation. Increases of conduction velocities may have required alterations in one of the two predominant properties that affect the speed of neuronal transmission: myelination or axonal diameter. A plausible mechanism to explain faster conduction velocities was a concomitant increase in axonal diameter with evolving axonal length. The carboxy terminal tail domain of the neurofilament medium subunit is a determinant of axonal diameter in large caliber myelinated axons. Sequence analysis of mammalian orthologs indicates that the neurofilament medium carboxy terminal tail contains a variable lysine-serine-proline (KSP) repeat sub-domain flanked by two highly conserved sub-domains. The number of KSP repeats within this region of neurofilament medium varies among species. Interestingly, the number of repeats does not change within a species, suggesting that selective pressure conserved the number of repeats within a species. Mapping KSP repeat numbers onto consensus phylogenetic trees reveals independent KSP expansion events across several mammalian clades. Linear regression analyses identified three subsets of mammals, one of which shows a positive correlation in the number of repeats with head-body length. For this subset of mammals, we hypothesize that variations in the number of KSP repeats within neurofilament medium carboxy terminal tail may have contributed to an increase in axonal caliber, increasing nerve conduction velocity as larger mammals evolved.

Collective inquiry: understanding the essence of best practice construction in mental health.

Delivering quality services has become a committed aspiration of mental health services over the past decade. Service planners look to validated care models to give guidance on what constitutes best practice. While there are many different views of what is 'best', there is a growing acknowledgement that services need to listen to the experiences of a network of frontline stakeholders in order to create quality mental health services. This paper describes an exploratory study within a regional mental health service that aimed to understand the meaning and enactment of best practice from the perspectives of a representative sample of service users and providers. A number of themes emerged as important and include the inherent value placed on consistent familial style relationships between service user and provider. This was deemed pivotal to the provision of expertise, good clinical decision-making, choice and collaboration. The study also highlighted stakeholder preference for autonomy and openness to inquiry into developing practice. In deconstructing the meaning of best practice, the study prompts a closer consideration of how best practice is created and suggests a view of best practice as a fluid dialogic process that is co-constructed by its participants in ongoing dialogic communication and reflection.

Healing after arterial dilatation with radiofrequency thermal and nonthermal balloon angioplasty systems.

Thermal balloon angioplasty has been proposed as a means of reducing acute and delayed reclosure of arteries after percutaneous transluminal balloon angioplasty. A radiofrequency (rf) balloon catheter was used to perform thermal balloon angioplasty on canine arteries in vivo. The histologic appearance of rf-treated sites was compared with that of control sites treated by conventional percutaneous transluminal angioplasty. Acutely, rf-treated sites showed a reduced medial cellularity with preservation of internal elastic lamina except at the transitional zone between thermal injury and normal artery, where localized internal elastic lamina disruption was found. Nonthermal sites showed generalized disruption of internal elastic lamina and normal medial cellularity. Both thermal and nonthermal sites displayed a return of intimal cover commencing at 1 to 2 weeks and completed by 4 weeks. Diffuse myointimal hyperplasia appeared by 2 weeks after injury at breaks in the internal elastic lamina along the nonthermal vessels but was localized to the transitional zone in thermal injury sites. In rf-treated vessels, repopulation of the acellular thermally modified media had commenced by 4 weeks, and by 8 weeks the media was diffusely repopulated by spindle-shaped cells resembling smooth muscle cells lying between and aligned with preserved connective tissue laminae. Overall, the distribution and extent of the proliferative response after rf thermal balloon angioplasty were less than those seen after nonthermal balloon angioplasty. Thermal sites, which underwent reintimalization before medial cells returned, were considerably less prone to the development of myointimal hyperplasia. These results suggest that this modality may have beneficial effects on arterial healing after angioplasty.

Potential of radio-frequency balloon angioplasty: weld strengths, dose-response relationship, and correlative histology.

Previous studies have established the feasibility of combining tissue heat generated by radio-frequency (RF) current and mechanical pressure to manage problems of percutaneous transluminal angioplasty (PTA) that are thought to cause postangioplasty restenosis (PARS). In the current in vitro study of normal and atherosclerotic human aortic layers (intima-media and media-adventitia) separated artificially, the purposes were to identify a dose-response relationship between the total RF energy delivered and the resultant weld strength between vascular tissues and to study the histologic correlates. Twenty-eight control and 100 experimental specimens were evaluated. The mean weld strength for all specimens with no RF current (at 0 J) was 4.1 g +/- 2.0; at 100 J, 5.9 g +/- 2.8; at 200 J, 28.5 g +/- 3.3; at 300 J, 50.0 g +/- 5.5; and at 500 J, 82.0 g +/- 8.2. Inspection of treated specimens revealed vascular molding (depression in the surface corresponding in dimensions to those of the electrode gap in the treatment chamber). Histologic examination revealed necrosis in the region underlying the electrode gap, a hypocellular fusion zone, indistinct boundaries between welded specimen parts, and transition zones to normal histologic characteristics at either end of each specimen. RF energy combined with mechanical pressure produces dose-dependent thermal welds in artificially dissected vascular tissues, molding of vascular tissue, and cellular destruction in the media. These findings may have clinical application in the management or prevention of all forms of PARS in humans and in the treatment of spontaneous aortic dissection.

A rabbit model for evaluation of an extracorporeal LDL removal system.

Elevated levels of plasma low density lipoprotein cholesterol (LDL) are associated with an increased incidence of atherosclerosis and its sequelae. A decrease in LDL levels correlates with a reduction in atherosclerosis. Drug therapy and diet have been moderately successful in lowering cholesterol levels, but require significant periods of treatment. Furthermore, a patient with genetically high LDL levels, as in familial hypercholesterolemia, may need a more rapid and reliable method of decreasing serum LDL. The present study was designed to test a device which can rapidly remove LDL from the circulation. The device consists of a filter cartridge filled with semipermeable hollow fibers which have a proprietary acidic polymer (pap) immobilized on the blood/plasma contacting surfaces; this polymer has been reported to selectively bind LDL cholesterol. The device was evaluated in a hypercholesterolemic rabbit model. Reductions in LDL serum concentration, ranging from 1.4-25.3%, occurred in 6/6 studies. All animals survived the procedure. Autopsy results at 10 days showed normal histology. Binding of other proteins (HDL, albumin) were minimal and no hemolysis was observed. Results of this preliminary study demonstrate the feasibility of reducing serum LDL levels with a hollow fiber device.

Thermal compression and molding of atherosclerotic vascular tissue with use of radiofrequency energy: implications for radiofrequency balloon angioplasty.

The combined delivery of pressure and thermal energy may effectively remodel intraluminal atherosclerotic plaque and fuse intimal tears. To test these hypotheses with use of a non-laser thermal energy source, radiofrequency energy was delivered to postmortem human atherosclerotic vessels from a metal "hot-tip" catheter, block-mounted bipolar electrodes and from a prototype radiofrequency balloon catheter. Sixty-two radiofrequency doses delivered from a metal electrode tip produced dose-dependent ablation of atherosclerotic plaque, ranging from clean and shallow craters with histologic evidence of thermal compression at doses less than 40 J to tissue charring and vaporization at higher (greater than 80 J) doses. Lesion dimensions ranged between 3.14 and 3.79 mm in diameter and 0.20 and 0.47 mm in depth. Tissue perforation was not observed. To test the potential for radiofrequency fusion of intimal tears, 5 atm of pressure and 200 J radiofrequency energy were delivered from block-mounted bipolar electrodes to 48 segments of human atherosclerotic aorta, which had been manually separated into intima-media and media-adventitial layers. Significantly stronger tissue fusion resulted (28.5 +/- 3.3 g) with radiofrequency compared with that with pressure alone (4.8 +/- 0.26 g; p less than 0.0001). A prototype radiofrequency balloon catheter was used to deliver 3 atm of balloon pressure with or without 200 J radiofrequency energy to 20 postmortem human atherosclerotic arterial segments. In 10 of 10 radiofrequency-treated vessels, thermal "molding" of both normal and atherosclerotic vessel wall segments resulted with increased luminal diameter and histologic evidence of medial myocyte damage.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of radiofrequency-generated thermal energy on the mechanical and histologic characteristics of the arterial wall in vivo: implications for radiofrequency angioplasty.

Abrupt reclosure of atherosclerotic vessels after percutaneous transluminal balloon angioplasty has been blamed on traumatic dissections and elastic recoil of the vessel wall. Thermal energy with compression produces fusion of separated arterial wall layers, and heat appears to alter the elastic recoil of the vessel wall. Radiofrequency (RF) thermal energy has been used to perform vascular anastomoses and thermal angioplasty. A simple in vivo experiment was designed to describe and quantitate vascular tissue weld strength produced by a range of RF thermal energy levels. Canine carotid arteries were compressed between a pair of modified bipolar forceps that applied RF energy, causing occlusive tissue welds between the apposed intimal surfaces. The strength of the welds was evaluated by measuring the perfusion pressure required to reopen the vessel lumen. A dosimetry range of 0 to 205 joules showed a typical dose-response curve for the relationship between energy applied and bond strength, plateauing at approximately 300 mm Hg. Light microscopy showed fusion of the inner surfaces of the vessel with preservation of vessel wall architecture. Additionally inflation of a bipolar RF balloon catheter in the normal canine carotid lumen produced an alteration of vessel profile angiographically and histologically. Results of these preliminary experiments suggest that balloon angioplasty with adjunctive RF thermal energy may have benefits in reducing the factors causing acute failure of conventional percutaneous transluminal balloon angioplasty.

Synthesis of phosphatidylethanol--a potential marker for adult males at risk for alcoholism.

Phosphatidylethanol, whose synthesis is catalyzed by a phospholipase D in a transphosphatidylation reaction, is a unique metabolite of ethanol. Phorbol 12-tetradecanoate 13-acetate, a tumor-promoting phorbol ester and stimulator of protein kinase C, activates this enzyme in peripheral blood lymphocytes. This system has been developed into an assay for measuring the potential of this pathway in human subjects. A pilot study of phosphatidylethanol synthesis in lymphocytes of adult males who have both an alcohol dependency and a family history of alcoholism has revealed that the average potential for phosphatidylethanol synthesis in this population is significantly elevated over that of control subjects.

Radiofrequency balloon angioplasty. Rationale and proof of principle.

Post-angioplasty restenosis (PARS) in atherosclerotic lesions of medium and small arteries occurs in about one-third of cases in the first year following percutaneous transluminal angioplasty (PTA) (early PARS). PARS includes acute spasm, dissection with reclosure, elastic recoil, fibrocellular proliferative response, and progressive atheromatous disease. Fibrocellular proliferation (possibly initiated by platelet derived growth factor) is felt to be culpable in many cases of early PARS (months). Pharmacologic regimens, stents, and thermal welding of the intimal-medial cracks of PTA are among the interventions being developed to deal with PARS. Radiofrequency (RF) current as a source of thermal energy may be useful in combination with balloon angioplasty to reduce PARS. Ideally, this combination would (1) weld intimal-medial cracks of PTA; (2) mold plaque and normal vessel to increase lumen diameters without creating intimal-medial cracks; and (3) destroy medial smooth muscle cells and multipotential cells (cellular substrate of PARS). Canine in vivo studies have established the feasibility of RF-mediated vascular tissue welding. Human aortic specimens (N = 28) were manually dissected into intima-media and media-adventitia layers. Bipolar RF energy (650 KHz, total 300 J) and mechanical pressure (1 atm) (experimental group, N = 24) or mechanical pressure alone (control group, N = 4) were applied to the reapposed specimen layers in a special chamber. The chamber was modified with a bipolar electrode designed to reproduce that planned for an RF balloon angioplasty catheter. Welding was demonstrated in normal and atherosclerotic treated specimens (23/24 or 96%) but not controls (0/4).(ABSTRACT TRUNCATED AT 250 WORDS)

The isolation and characterization of Ca++-accumulating subcellular membrane fractions from cerebral arteries.

A study was undertaken using differential centrifugation methods to isolate from rabbit cerebral arteries the subcellular microsomal protein fractions capable of actively sequestering Ca++. One isolated protein fraction displayed a relatively large adenosine triphosphate (ATP)-dependent Ca++-accumulating capacity that was completely inhibited by NaN3, and was therefore designated the "mitochondrial fraction." Electron microscopy confirmed that this fraction consisted of numerous mitochondrial elements. Another isolated membrane fraction possessed a Ca++-accumulating capacity dependent on ATP and oxalate and only partially sensitive to NaN3. In the presence of mersalyl acid or the Ca++ ionophore, A23187, Ca++ uptake by this fraction was inhibited 98.0% and 87.4%, respectively. Electron microscopy revealed that this fraction consisted of numerous membrane vesicles, and measurements of Na+-K+-ATPase (adenosine triphosphatase) activity indicated minimal plasma membrane contamination. It was concluded that this microsomal fraction consisted primarily of sarcoplasmic reticulum. At physiological free [Ca++] levels, Ca++ uptake by this fraction was inhibited by norepinephrine through a process sensitive to tolazoline but not propranolol. The effects on Ca++ uptake of added cyclic adenosine monophosphate (cAMP) alone or with rabbit or bovine protein kinase were inconclusive. The organic Ca++ channel blockers, nifedipine and verapamil, significantly inhibited Ca++ uptake by sarcoplasmic reticulum.

The effects of epsilon aminocaproic acid on contractile properties and catecholamine content of rat carotid arteries.

The prevalent use of epsilon aminocaproic acid (EACA) to prevent rebleeding in the preoperative management of ruptured intracranial aneurysms raises the question of a relationship between this drug and cerebral vasospasm. The effects of EACA on the contractile properties of the rat common carotid artery were measured in vitro, and the effects of EACA on the catecholamine content of these arteries were determined qualitatively. When carotid artery segments from rats that had been infused with lactated Ringer's solution alone were incubated in the presence of 1 mM EACA, they exhibited a decrease in contractile activity brought about by the presence of either serotonin or norepinephrine, and an increase in contractile activity when potassium chloride was used. These changes were not witnessed if the arterial segments came from rats that had been infused with lactated Ringer's solution containing EACA. No effect of epsilon aminocaproic acid was found on the median effective dose values for each vasoconstrictor used. Fluorescence histochemistry showed no qualitative changes in the catecholamine content of carotid arteries following EACA treatment.

Measurement of fibrinolytic activity after epsilon-aminocaproic acid administration in rats (Rattus norvegicus).

The antifibrinolytic agent, epsilon-aminocaproic acid was administered to rats via either intraperitoneal injection or intravenous infusion. Time-response and dose-response studies of the resultant plasma levels of fibrinolytic activity were measured by a modification of a standard fibrin plate technique. The modified technique involved the addition of human plasma to rat plasma in a ratio of 10:1 (rat:human). The plasma levels of fibrinolytic activity reflected plasma concentrations of epsilon-aminocaproic acid.

Heparin and small caliber polytetrafluoroethylene grafts in the carotid arteries of rats.

Polytetrafluoroethylene (PTFE) grafts having an internal diameter of 1 mm were implanted in the common carotid arteries of rats with or without the simultaneous application of various heparin regiments. Graft patency was determined from postoperative angiograms, and grafts were patent 40 days after surgery were harvested for light microscopic examination. The level of anticoagulation produced by the various heparin regimens was measured in separate groups of rats and found to be clinically therapeutic. All of the grafts from rats that did not receive heparin were occluded at 24 hours. In the two groups of rats that received high doses of heparin, 5 of 10 and 4 of 11 grafts were patent 40 days postoperatively. There were no patent grafts three days postoperatively in the group receiving low doses of heparin. The grafts that were patent 40 days postoperatively had lumens that were completely lined with presumptive endothelial cells. Further laboratory experimentation is needed before PTFE small caliber grafts can be recommended for use in humans.

Effect of intravenous ethanol on cerebral vasospasm produced by subarachnoid blood.

The cat basilar artery was exposed using the transclival approach. After administration of 5% ethanol via intravenous infusion, vasospasm was produced by applying the animal's fresh arterial blood to the exposed artery. The resultant vasospasm was of markedly reduced intensity and duration as compared to vasospasm in control animals. In ethanol-treated animals with spasm induced from non-autogenous fresh arterial bloof free of ethanol, a reduction in the duration of vasospasm was noted although the initial intensity of spasm was similar to control animals. There was no anti-spasm effect if the ethanol infusion followed the production of vasospasm.

Small animal model for investigation of subarachnoid hemorrhage and cerebral vasospasm.

A method for induction of subarachnoid hemorrhage (SAH) in a rat model is described. Resolution of the hemorrhage was documented photographically and microscopically at intervals from 1 hr to 8 days. Photographs indicated that most of the hemorrhage was resorbed within 3 days, an observation confirmed microscopically by the amount of red blood cells in the subarachnoid space. Significant cerebral vasospasm was documented within the first 2 days after the induction of hemorrhage with the basilar artery returning to baseline values at an average of 3 days followed by moderate dilatation at 5 to 8 days. The suitability of the rat as an animal model for further investigation of subarachnoid hemorrhage is discussed.

The interaction of lanthanides with isolated sarcoplasmic reticulum vesicles from rabbit skeletal muscle.

The interaction of lanthanides with isolated sarcoplasmic reticulum (SR) vesicles from rabbit skeletal muscle and the effects of lanthanides on 45Ca2+ uptake by the vesicles were studied. 153Gd3+ was taken up by the vesicles in the absence of ATP and oxalate in a time-dependent manner, reaching a maximum total accumulation of 380 nmol 153Gd3+/mg protein after 20 min with 200 microM 153Gd3+. This 153Gd3+ accumulation was not washed out by 1 mM EGTA. The addition of ATP induced the release of 87% of the bound 153Gd3+, leaving behind irreversibly-accumulated 153Gd3+. Pre-incubation of the vesicles with lanthanides in the absence of ATP and oxalate inhibited 45Ca2+ uptake without affecting Ca2+-ATPase activity. The percent inhibition of 45Ca2+ uptake increased with length of pre-incubation of the vesicles with lanthanides, reaching 33% after 20 min of pre-incubation. Increasing the 45Ca2+ concentration or adding ATP or oxalate to the preincubation medium abolished these inhibitory effects on 45Ca2+ uptake.