Atomic force microscopy of nonhydroxy galactocerebroside nanotubes and their self-assembly at the air-water interface, with applications to myelin.

Myelin is one of the few biological membranes to contain the lipid galactocerebrosides, although their role in myelin is unclear. To explore its structural role, we used fluorescence and atomic force microscopy (AFM) to study nonhydroxy galactocerebrosides (NCer) at the air-water interface of a Langmuir-Blodgett trough. Fluorescence microscopy at the air-water interface indicated that NCer forms micrometer scale domains of varying radii with six fractal-like extensions. Atomic force microscopy using TappingMode in water on samples transferred to mica confirmed the fractal-like domain structure in the absence of dye and showed that the domains consisted of many aggregated nanotubes with a diameter of 30 nm. The Hausdorf fractal dimension was estimated to be 1.26 and 1.11 for two domains imaged with AFM. This evidence indicates that NCer forms a bulk phase of nanotubes at the air-water interface, unlike the liquid-condensed phase of a phospholipid monolayer. That NCer forms bilayer nanotubes that aggregate strongly suggests NCer helps maintain the stability of myelin by contributing to the curvature and adhesion of the membrane. We found that NCer appears to be decreased in myelin from multiple sclerosis normal appearing white matter, which could be an important event in the loss of myelin stability.

Improving quality of life for people with chronic conditions: the example of t'ai chi and multiple sclerosis.

CONTEXT: The greatest needs of people with chronic conditions are long-term care, maximized independence, and improved quality of life. With conventional medicine becoming increasingly expensive, depersonalized, and unable to adequately meet such needs, many with chronic conditions are seeking health promotion strategies to effectively manage their symptoms. OBJECTIVE: An 8-week t'ai chi program was conducted to explore psychosocial and physical benefits for those with multiple sclerosis. DESIGN: Nonrandomized, noncontrolled pilot study. SETTING: American College of Traditional Chinese Medicine, San Francisco, Calif. PATIENTS: 19 patients with multiple sclerosis. INTERVENTION: T'ai chi. MAIN OUTCOME MEASURES: Walking speed (distance = 25 ft), hamstring flexibility, and psychosocial well-being as measured by the Medical Outcomes Study 36-item Short-form Health Survey. RESULTS: Walking speed increased by 21% and hamstring flexibility increased by 28%. Patients experienced improvements in vitality, social functioning, mental health, and ability to carry out physical and emotional roles. CONCLUSIONS: This pilot program was conducted entirely on a volunteer basis and led to the implementation of several additional t'ai chi classes for people with multiple sclerosis across the United States. T'ai chi and other health promotion programs offer help toward achieving the goals of increasing access to services, maximizing independence, and improving quality of life for people with chronic disabling conditions.

Biochemical alterations in multiple sclerosis lesions and normal-appearing white matter detected by in vivo 31P and 1H spectroscopic imaging.

The goals of the current study were threefold: first, to confirm previous single volume proton (1H) magnetic resonance spectroscopy results of reduced N-acetyl aspartate (NAA, a putative marker of neurons) in multiple sclerosis (MS) white matter lesions using multiple volume 1H magnetic resonance spectroscopic imaging (MRSI); second, to measure the phospholipid metabolites phosphomonoesters and phosphodiesters in such lesions using phosphorus (31P) MRSI; and third, to test the hypothesis that biochemical changes occur in the normal-appearing (on spin echo T2-weighted magnetic resonance images) white matter in patients with MS. Thirteen subjects with clinically definite MS were studied with both 1H and 31P MRSI, and 19 controls were studied with either 1H MRSI, 31P MRSI, or both. MS lesion, MS normal-appearing white matter, and region-matched control spectra from the centrum semiovale were analyzed. The major findings of this study were that in both white matter lesions and normal-appearing white matter in patients with MS, the metabolite ratio NAA/creatine and the total 31P peak integrals were significantly reduced compared with controls. In addition, in MS lesions NAA/choline and phosphodiesters/total 31P were significantly reduced compared with controls, and in MS normal-appearing white matter there was a trend for NAA/choline to be reduced compared with controls. In normal-appearing white matter in patients with MS, total creatine and phosphocreatine were significantly increased compared to controls, as detected with both 1H (total creatine peak integrals) and 31P (phosphocreatine/total 31P) MRSI techniques. These results suggest reduced neuronal density and altered phospholipid metabolites in white matter lesions in patients with MS.(ABSTRACT TRUNCATED AT 250 WORDS)

In vivo detection of myelin phospholipids in multiple sclerosis with phosphorus magnetic resonance spectroscopic imaging.

The goal of this study was to investigate myelin phospholipids in vivo in multiple sclerosis lesions and normal-appearing white matter by evaluating the spectral broad component from phosphorus 31 magnetic resonance spectroscopic imaging data. The phospholipid broad component was reduced nearly 35% (p < 0.001) in both lesions and in normal-appearing white matter in multiple sclerosis subjects compared to control subjects, suggesting reduced myelin phospholipid concentration or altered relaxation times.

Contributions of neuroimaging to diagnosis and monitoring of multiple sclerosis.

Recent advances in magnetic resonance neuroimaging have resulted in an increased ability to distinguish acute, potentially reversible lesions from chronic, irreversible lesions in multiple sclerosis. Refinements of magnetic resonance imaging techniques, such as fluid-attenuated inversion recovery, diffusion imaging, and magnetization transfer imaging, as well as magnetic resonance spectroscopic imaging, are providing increased sensitivity and allowing detection of changes in multiple sclerosis white matter that appears normal on standard spin-echo magnetic resonance images. Increased neuroimaging specificity and sensitivity enhance the ability to diagnose, monitor, and understand the progression of multiple sclerosis. Magnetic resonance spectroscopy and magnetic resonance spectroscopic imaging detect metabolites in vivo and have even greater potential for elucidating the biochemical pathology of demyelination in multiple sclerosis.

Molar quantitation of in vivo proton metabolites in human brain with 3D magnetic resonance spectroscopic imaging.

A method for molar quantitation of in vivo proton metabolites in human brain with three-dimensional (3D) proton magnetic resonance spectroscopic imaging (MRSI) is described. The method relies on comparison of brain and calibration phantom measurements, with corrections for coil loading, and spin-lattice and spin-spin relaxation times. A 3D proton MRSI pulse sequence was developed which acquires two echoes and enables acquisition of both the TMS coil loading reference phantom and proton metabolite signals from a single experiment. With the aqueous fraction (tissue water) taken into account, the calculated molar concentrations from 24 centrum semiovale white matter voxels from 4 control subjects were (mmol/l +/- SD): N-acetyl aspartate = 14.6 +/- 2.8, total creatine+phosphocreatine = 6.0 +/- 1.2, total choline = 1.9 +/- 0.4. These values are equivalent to previously reported results obtained from single volume localized proton magnetic resonance spectroscopy.

Electroacupuncture in anaesthesia for hysterectomy.

We have studied the effects of electroacupuncture at classical acupuncture points, applied before and during surgery in patients undergoing hysterectomy, on postoperative pain and metabolic stress responses in a prospective, randomized and patient-blinded manner. Fifty otherwise healthy women were allocated randomly to receive or not receive electroacupuncture. Electroacupuncture was begun 20 min before skin incision and continued to the end of surgery. All patients received similar general anaesthesia and all received patient-controlled analgesia (PCA) after operation. Postoperative pain in the two groups was evaluated by recording analgesic requirements by PCA and by pain-rating performed by patients and nursing staff. There were no significant differences between the two groups in postoperative analgesic requirements, pain-rating or metabolic stress responses.

Editing 13C-NMR spectra of membranes.

We report the carbon-13 'magic-angle' sample-spinning nuclear magnetic resonance (NMR) spectra of several lipid-water systems, under a variety of radiofrequency excitation conditions. Our results show that complex lipid or membrane spectra can be greatly simplified by using 'spectral editing' techniques. For example, in a 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC)-water mesophase, the glycerol (C-1, C-2 and C-3) carbons are readily distinguished from the headgroup C alpha, C beta and C gamma carbons, on the basis of their mix-time behavior in a cross-polarization (CP) experiment, while in the more complex DMPC/cholesterol-water system, many of the more rigid cholesterol carbon resonances can be edited from the phospholipid peaks. In very complex systems, such as human myelin membranes, editing permits the unambiguous observation of the mobile lipid headgroup carbon resonances, as well as the much more rigid sterol ring carbons. We also report the observation of a large differential CP due to C-H vector 'magic-angle' orientational effects in the DMPC/desipramine system. Thus, both motional or orientational reduction of the C-H dipolar interaction can lead to considerable simplifications of complex membrane spectra, and are of interest from both spectral assignment and membrane dynamics aspects.

Carbon-13 "magic-angle" sample-spinning nuclear magnetic resonance studies of human myelin, and model membrane systems.

We have obtained high-field (11.7 Tesla), high-resolution carbon-13 solid-state "magic-angle" sample-spinning nuclear magnetic resonance (NMR) spectra of a variety of phospholipids, sphingolipids, myelin and white matter samples, resolving and assigning over 40 resonances in the spectra of human and bovine myelin. The NMR results indicated no large spectral changes due to sample preparation, sample freezing, or brain location, and also no changes in myelin structure detectable via light microscopy, electron microscopy, thin layer chromatography, or sodium dodecyl sulfate-polyacrylamide gel electrophoresis, attributable to the sometimes lengthy NMR data acquisition process. Human myelin and white matter chemical shift assignments were made based on 13C "magic angle" sample spinning (MAS) NMR spectra of individual model lipids, as well as on spectra of lipid mixtures. In all myelin samples there were essentially no features attributable to membrane proteins, with the exception of one small feature due to C zeta of Arg residues, primarily in the myelin basic proteins. The general similarity between the model lipid and intact myelin spectra suggested no major effects of protein on lipid mobility. We have also investigated human myelin samples as a function of developmental age (4, 15, 48 months and adult), and our results showed only small changes in overall lipid composition, although there were significant decreases in lipid hydrocarbon chain unsaturation with age, as determined by computer line-shape simulations of myelin and model compounds. The spectrum of an infant leukoencephalopathy myelin showed marked decreases in galactocerebrosides. Overall, the ability to resolve and assign over 40 resonances in the 13C MAS NMR spectra of myelin, and to detect changes as a function of development and disease, should provide a useful starting point for further more detailed studies of myelin membrane molecular motions, and function.

Lateralization of human focal epilepsy by 31P magnetic resonance spectroscopic imaging.

We attempted to lateralize the epileptogenic focus (seven temporal lobe hippocampal foci, one frontal lobe focus) in medically refractory unilateral complex partial seizures, using noninvasive 31P magnetic resonance spectroscopic imaging (MRSI) blindly and interictally to compare hippocampal or frontal regions. The seizure foci were more alkaline (intracellular pH = 7.17 +/- 0.03) compared with the contralateral region (7.06 +/- 0.02, p < 0.01) in all eight cases; the inorganic phosphate was relatively increased (240 +/- 50% of contralateral, seven of eight cases, p < 0.01); and phosphomonoesters were relatively reduced (68 +/- 9% of contralateral, seven of eight cases, p < 0.01). Other phosphorus metabolites were symmetric (+/- 10%). 31P MRSI correctly lateralized the seizure focus in all eight cases. By comparison, imaging correctly lateralized four cases and SPECT, two cases. In conclusion, 31P MRSI is a useful tool for the noninvasive clinical assessment of focal epilepsy and can accurately lateralize the epileptogenic focus.

Antitumor effects of alpha-interferon and gamma-interferon on a murine renal cancer (Renca) in vitro and in vivo.

Previous studies have shown that established murine renal cancer (Renca) can be successfully treated with the investigational drug flavone acetic acid (FAA) used in combination with recombinant interleukin 2 (IL-2). Additional experiments demonstrated that the in vivo administration of FAA rapidly induced the expression of the genes, as well as the biologically active proteins, for alpha- and beta-interferons (IFNs) as well as tumor necrosis factor alpha. Both IFN-alpha and IFN-gamma have been shown to have direct antiproliferative effects against some tumors, as well as being potent immunodulators for the induction of antitumor effector cells. Thus, the present study was designed to investigate the ability of IFN-alpha and/or IFN-gamma to mediate direct antiproliferative effects against Renca in vitro as well as to cause regression of Renca in vivo. The present study confirms that RAA and/or IL-2 are inactive against Renca in vitro, further suggesting an indirect mechanism for FAA-induced antitumor effects in vivo. However, the exposure of Renca in vitro to recombinant human IFN-alpha A/D, murine IFN-alpha or murine IFN-beta resulted in a dose dependent growth inhibition of Renca as assessed by the microculture tetrazolium dye incorporation assay. Very little growth inhibition was induced by recombinant murine IFN-gamma. Interestingly, IFN-alpha (100-1000 units/ml) when combined with very low doses of recombinant murine IFN-gamma (1-10 units/ml) yielded significantly more pronounced growth inhibition than either cytokine alone. This effect was most evident by 5 days of culture where combinations of 100-1000 units/ml recombinant human IFN-alpha A/D with 1-5 units/ml recombinant murine IFN-gamma yielded growth inhibition in the range of 45-99%. In order to determine whether the mechanisms for the antitumor activity of recombinant human IFN-alpha A/D and recombinant murine IFN-gamma was due to their direct antiproliferative effects, we also studied the efficacy of these combinations against i.p. Renca in athymic mice. In contrast to the potent antitumor effects observed in euthymic mice, the combination of IFN-alpha and IFN-gamma only slightly increased mean survival times in athymic mice and no long term survivors were obtained. Subsequent studies demonstrated that most mice (77%) cured of peritoneal Renca by recombinant human IFN-alpha A/D plus recombinant murine IFN-gamma were immune to rechallenge. Therefore the combination of IFN-alpha and IFN-gamma may directly inhibit the growth of Renca, but a major effect of IFNs in vivo must be to contribute to the induction of an anti-Renca immune response.

Technique of cast wedging in long bone fractures.

A method to determine the width of a cast wedge to correct an angular deformity of a long bone is presented. The trigonometric theory behind the method is presented, but the method does not require any trigonometric calculations and can be done easily by drawing on the x-ray, with only a pencil and ruler-protractor required. Accurate reduction of angular deformities of long bones can be done with cast wedging when the width of the opening wedge can be predetermined.

Failure of polyethylene acetabular cups. Two case reports.

Fatigue fracture involving the metallic femoral stem is well recognized after total hip arthroplasty. Two cases of Charnley-Mueller polyethylene acetabular cup failure, in a 54-year-old-woman and a 77-year-old woman with abnormal wear patterns, were diagnosed prior to operation. The fracture of the acetabular cup was recognized by the fragmented, crushed appearance of the cup. The arthrogram clearly showed the radiographic contrast agent passing through the substance of the cup. The wear of the cups was measured by micrometer calipers which have an accuracy of 0.001 mm. The polyethylene acetabular cup was analyzed in four zones formed by three circumferential grooves. Markedly increased wear of the components occurred in the superolateral dome area. This wear rate was as high as 0.8 mm/year, which is four times the average wear of 0.2 mm/year. This marked wear associated with repetitive cyclic fatigue or static load resulted in fracture of the polyethylene acetabular cup. Improper machining of the cup using low- rather than high-density polyethylene in the manufacturing process and the heavy weight of these patients may have been factors in the marked fissures and crack lines revealed by microscopic study of the fractured area after sputter coating with gold palladium. Fracture of the polyethylene acetabular component, although rare, may be encountered more often in longer follow-up studies of patients who have undergone total hip arthroplasty. Proper selection of patients, acetabular spacers, and pressure injection techniques may prevent early loosening of the acetabular polyethylene components.

Traumatic disruption of the manubriosternal joint. A case report.

A case of manubriosternal dislocation is presented. The possible mechanism of injury was hyperflexion of the spine which resulted in chin to chest contact, disrupting the manubriosternal joint. If the dislocation is Type I and the patient has compression symptoms on the trachea or major vessels, surgical treatment by wiring may be needed. In Type II dislocations, the best management is closed reduction and elastoplast strapping.