High-resolution x-ray scattering from epitaxial thin films of Y/Nb on Al2O3.

During the 1990s, Roger Cowley had a strong interest in the crystal and magnetic structures of rare-earth superlattices as a means to understand the rich and exotic magnetic properties of the rare-earth metals. High-quality samples can be grown by molecular beam epitaxy on sapphire substrates by first depositing a thin epitaxial layer of niobium, then a layer of yttrium or lutetium as a seed. High-resolution x-ray scattering is an excellent probe to characterise the crystal quality and was used to study the structure of the niobium layer. However, relatively little attention was paid to the seed layer. This article summarises some of the x-ray experiments performed by the Cowley group to study the structure of epitaxial [Formula: see text] niobium on [Formula: see text] sapphire, and extends the work to report some results on the structure of thin [Formula: see text] yttrium seed layers. The structure of the yttrium films is shown to have a strong dependence on the thickness of the niobium buffer, with the buffer needing to be thicker than a critical value of ∼80 [Formula: see text] for the formation of misfit dislocations at the Nb/Al2O3 interface before highly coherent Y films can be grown. Yttrium films grown on Nb buffers thinner than ∼500 [Formula: see text] show a similar two-peak line shape in [Formula: see text] scans through their specular Bragg peaks to that seen in the specular Nb Bragg peaks, with a resolution-limited feature on a broader diffuse peak. The resolution-limited feature depends on the thickness of the yttrium film, becoming weaker and having a stronger decay with increasing [Formula: see text] as the film thickness increases, while the width of the yttrium broad peak evolves as the square root of the width of the niobium Bragg peak. The data are discussed within the context of theories describing the scattering from films with misfit dislocations.

Magnetization process in holmium: easy axis spin reorientation induced by the magnetostrictive basal plane distortion.

We report on the change of the easy axis direction in holmium, from the a to the b axis, under the application of a magnetic field in the basal plane. This spin reorientation is observed by measuring the magnetic torque in Ho(n)/Lu(15) superlattices (n and 15 are the number of atomic planes in the Ho and Lu blocks). We also observe that, at the field H0 and temperature at which the reorientation occurs, both axes are easy directions. Based on the fact that the field H0 depends on n in the same way as the field-induced magnetoelastic distortion does, we propose that this spin reorientation originates from the strong field-induced magnetoelastic deformation within the basal plane. The modulation of the alpha strains with sixfold symmetry originates a 12-fold term in the magnetic anisotropy energy.

A neuromuscular platform to extract electrophysiological signals from lesioned nerves: a technical note.

The volitional control of prosthetic devices could be greatly enhanced if the information formerly supplied by peripheral nerves to the amputated limb could be utilized. So that practical access to this information could be gained, a method was established to form a stable biological interface with fascicles of lesioned nerves. A small strip of an intact muscle was isolated in rats with the use of a silicone tube cuff electrode and innervated by the lesioned peroneal branch of the sciatic nerve. After 4 weeks survival, stimulation of the nerve fascicle produced reliable signals from the neuromuscular platform in the range of 0.5 to 2.0 mV. Histologically, myotubes remained intact and axons could be identified growing in and over the surfaces of the isolated muscle strips. These or similar interface techniques may supply electrophysiological signals of sufficient amplitude and reliability to provide peripheral nerve-based guidance information for prosthetic devices.

Resistance training and gait function in patients with Parkinson's disease.

OBJECTIVE: To determine whether patients with Parkinson's disease who are enrolled in a resistance training program can gain strength similar to that of normal control subjects and whether these gains in strength would improve their gait function. DESIGN: Subjects included 14 patients with mild-to-moderate Parkinson's disease of either gender and six normal control subjects of similar age. The training consisted of an 8-wk course of resistance training twice per week, focused primarily on the lower limbs. The primary outcome measures consisted of exercise performance monitoring and quantitative gait analysis before and after the training course. RESULTS: Both the patients with Parkinson's disease and normal control subjects significantly increased their performance with resistance training. Subjects with Parkinson's disease had gains in strength similar to those of normal elderly adults. Patients with Parkinson's disease also had significant gains in stride length, walking velocity, and postural angles compared with pretreatment values. CONCLUSIONS: Patients with mild-to-moderate Parkinson's disease can obtain increases in performance or strength similar to that of normal adults of the same age in a resistance training program. Resistance training can produce functional improvements in gait and may, therefore, be useful as part of a physical rehabilitation and/or health maintenance program for patients with Parkinson's disease.

Evidence for distinct intracellular signaling pathways in CD34+ progenitor to dendritic cell differentiation from a human cell line model.

Intracellular signals that mediate differentiation of pluripotent hemopoietic progenitors to dendritic cells (DC) are largely undefined. We have previously shown that protein kinase C (PKC) activation (with phorbol ester (PMA) alone) specifically induces differentiation of primary human CD34+ hemopoietic progenitor cells (HPC) to mature DC. We now find that cytokine-driven (granulocyte-macrophage CSF and TNF-alpha) CD34+ HPC-->DC differentiation is preferentially blocked by inhibitors of PKC activation. To further identify intracellular signals and downstream events important in CD34+ HPC-->DC differentiation we have characterized a human leukemic cell line model of this process. The CD34+ myelomonocytic cell line KG1 differentiates into dendritic-like cells in response to granulocyte-macrophage CSF plus TNF-alpha, or PMA (with or without the calcium ionophore ionomycin, or TNF-alpha), with different stimuli mediating different aspects of the process. Phenotypic DC characteristics of KG1 dendritic-like cells include morphology (loosely adherent cells with long neurite processes), MHC I+/MHC IIbright/CD83+/CD86+/CD14- surface Ag expression, and RelB and DC-CK1 gene expression. Functional DC characteristics include fluid phase macromolecule uptake (FITC-dextran) and activation of resting T cells. Comparison of KG1 to the PMA-unresponsive subline KG1a reveals differences in expression of TNF receptors 1 and 2; PKC isoforms alpha, beta I, beta II, and mu; and RelB, suggesting that these components/pathways are important for DC differentiation. Together, these findings demonstrate that cytokine or phorbol ester stimulation of KG1 is a model of human CD34+ HPC to DC differentiation and suggest that specific intracellular signaling pathways mediate specific events in DC lineage commitment.

Standard osteopathic manipulative treatment acutely improves gait performance in patients with Parkinson's disease.

Patients with Parkinson's disease exhibit a variety of motor deficits which can ultimately result in complete disability. The primary objective of this study was to quantitatively evaluate the effect of osteopathic manipulative treatment (OMT) on the gait of patients with Parkinson's disease. Ten patients with idiopathic Parkinson's disease and a group of eight age-matched normal control subjects were subjected to an analysis of gait before and after a single session of an OMT protocol. A separate group of 10 patients with Parkinson's disease was given a sham-control procedure and tested in the same manner. In the treated group of patients with Parkinson's disease, statistically significant increases were observed in stride length, cadence, and the maximum velocities of upper and lower extremities after treatment. There were no significant differences observed in the control groups. The data demonstrate that a single session of an OMT protocol has an immediate impact on Parkinsonian gait. Osteopathic manipulation may be an effective physical treatment method in the management of movement deficits in patients with Parkinson's disease.

A comparative study of the cell cycle status and primitive cell adhesion molecule profile of human CD34+ cells cultured in stroma-free versus porcine microvascular endothelial cell cultures.

Porcine microvascular endothelial cells (PMVECs) plus cytokines support a rapid proliferation and expansion of human CD34+CD38- cells that are capable of multilineage engraftment within the bone marrow of a secondary host. CD34+CD38- cells contain the self-renewing, long-term culture-initiating cells (LTC-IC) that are ideal targets for retroviral gene transfer experiments. Previous experiments attempting retroviral infection of CD34+CD38- cells have failed partly because these cells do not enter cell cycle in response to cytokine combinations. In this study, we determined the cell cycle status and the cell adhesion molecule profile on purified CD34+ cells and the CD34+CD38- subset before and after ex vivo expansion on PMVECs. Purified human CD34+ cells were cocultured with PMVECs for 7 days in the presence of optimal concentrations of granulocyte/macrophage-colony-stimulating factor (GM-CSF) + interleukin (IL)-3 + IL-6 + stem cell factor (SCF) + Flt-3 ligand. The total CD34+ population and the CD34+CD38- subset increased 8.4- and 67-fold, respectively, with absolute increases in the number of colony-forming unit-granulocyte macrophage (CFU-GM) (28.2-fold), CFU-Mix (8.7 fold), and burst-forming unit-erythroid (BFU-E) (4.0-fold) progenitor cells. After 7 days of coculture with PMVECs, 44% of the CD34+CD38+ subset were found to be in G1, and 51% were in G2/S/M phase of the cell cycle. More remarkably, 53% of the CD34+CD38- subset were in G1, and 17% were in G2/S/M phase after 7 days of PMVEC coculture. In contrast, only 22% of the CD34+CD38- subset remaining after 7 days of stroma-free culture were in G1, and 6% were in G2/S/M phase. Despite the high level of cellular activation and proliferation induced by PMVEC coculture, the surface expression of adhesion molecules CD11a (LFA-1), CD11b, CD15s (sialyl-Lewis x), CD43, and CD44 (HCAM) on the total CD34+ population was maintained, and the surface expression of CD49d (VLA-4), CD54 (ICAM), CD58, and CD62L (L selectin) increased after ex vivo expansion. In contrast, CD34+ cells expanded on stroma-free cultures showed lower and more variable expression of CD62L and CD15s. These findings demonstrate that the primitive CD34+CD38- subset of marrow progenitor cells can be induced to enter cell cycle and can be significantly expanded ex vivo on a hematopoietic supportive microenvironment (PMVECs) while preserving the expression of cell adhesion molecules that may be important in stem cell homing and engraftment.

Effect of combined administration of insulin-like growth factor and platelet-derived growth factor on the regeneration of transected and anastomosed sciatic nerve in rats.

OBJECTIVE: To evaluate the efficacy of combined administration of platelet-derived growth factor (PDGF) and insulin-like growth factor (IGF-I) on the regeneration of transected and anastomosed sciatic nerves. ANIMALS: 25 male Lewis rats. PROCEDURE: Transection and anastomosis was performed of the left sciatic nerve of all rats. At the time of surgery, treatment group rats received a combination of PDGF (0.75 microgram) and IGF-I (1.5 micrograms) along with vehicle (2% methyl cellulose gel) at the anastomosis site. Control rats received vehicle alone. Evaluation consisted of motor evoked potentials and nerve conduction velocity, quantitative histologic examination, sciatic functional index, and visual assessment of repair organization (lesion scores). RESULTS: Significant differences in motor evoked potential latency, nerve conduction velocity, axon number, sciatic functional index, or lesion scores were not found between groups. CONCLUSIONS: Combined administration of PDGF and IGF-I did not enhance peripheral nerve regeneration in a transection and anastomosis model as determined 6 weeks after surgery. CLINICAL RELEVANCE: These findings support the assumption that mechanical, versus trophic, effects predominate when nerve ends are closely apposed.

Gel matrix vehicles for growth factor application in nerve gap injuries repaired with tubes: a comparison of biomatrix, collagen, and methylcellulose.

The repair of nerve gap injuries with tubular nerve guides has been used extensively as an in vivo test model in identifying substances which may enhance nerve regeneration. The model has also been used clinical nerve repair. The objective of this study was to compare three different gel matrix-forming materials as potential vehicles for growth factors in this system. The vehicles included a laminin containing extracellular matrix preparation (Biomatrix), collagen, and a 2% methylcellulose gel. The growth factor test substance consisted of a combination of platelet-derived growth factor BB (PDGF-BB) and insulin-like growth factor I (IGF-I). An 8-mm gap in rat sciatic nerve was repaired with a silicone tube containing each of the vehicles alone or with a combination of each vehicle plus PDGF-BB and IGF-I. At 4 weeks after injury, the application of the growth factor combination significantly stimulated axonal regeneration when applied in methylcellulose or collagen, but not in Biomatrix. A similar trend was present between the vehicle control groups. By 8 weeks after injury, nerves repaired with methylcellulose as a vehicle had significantly greater conduction velocity than either collagen or Biomatrix. It was concluded that a 2% methylcellulose gel was the best of the three matrices tested, both in its effects on nerve regeneration and flexibility of formulation.

Partial regeneration of the sciatic nerve in rats enhances motor excitability to magnetic stimulation.

Motor-evoked potentials elicited from transcranial magnetic stimulation were recorded from both cranial tibial muscles of 10 Lewis rats. One sciatic nerve was then transected and surgically reanastomosed and allowed to regenerate for 6 weeks. Walking function was tested weekly using a footprint test giving a functional sciatic nerve index. At the end of the study motor-evoked potentials were again performed, as were nerve conduction velocities and supramaximal stimulation of operated and unoperated sciatic nerves. The nerve was evaluated for myelinated axon numbers. Functional sciatic index, numbers of myelinated axons, nerve conduction velocities, and supramaximal stimulation demonstrated the regenerated nerve to be functional, but to a lesser extent than the unoperated nerve. In all cases the amplitude of the compound muscle action potential evoked from transcranial magnetic stimulation of the motor cortex was greater in the operated motor pathway. This finding demonstrates hyperexcitability of the motor pathway to magnetic stimulation during peripheral nerve regeneration.

Wire mesh as a post-operative physiotherapy assistive device following peripheral nerve graft repair in the rat.

Traditional methods of assessing nerve recovery following injury, including histomorphometry and electrophysiology, do not necessarily correlate with return of motor and sensory function. Accordingly, many investigators have used walking track analysis as an assessment of global functional recovery following sciatic, peroneal or tibial nerve injury. However, walking track reliability may be compromised by the development of flexion contractures secondary to neurologic loss. To prevent this, regular manual physiotherapy is recommended which is a time-consuming and often frustrating process for both the animal and investigator. We report the use of a wire mesh as a simple post-operative assistive device to provide constant physiotherapy. Twelve Lewis rats were randomized to one of two experimental groups. Each rat received a 2 cm posterior tibial nerve autograft. Postoperatively, animals in group 1 received manual physiotherapy, consisting of repeated flexion and extension exercises of the ankle, knee and hip every two weeks. Group 2 rats were permitted to climb freely on a 30 x 18 cm piece of wire mesh placed at a 45 degree angle within their cage. Group 2 rats received no manual physiotherapy throughout the course of the study. Serial walking tracks were performed every four weeks until sacrifice at sixteen weeks. There was no development of flexion contractures in the injured hind limbs of either group. There was no morbidity such as blisters associated with the use of the wire mesh. There was no statistical difference in walking track recovery between groups at any time period. However, a trend towards better functional recovery was seen in the group receiving constant physiotherapy via the wire mesh. The use of a wire mesh as a post-operative assistive device is an inexpensive, simple and reliable method to provide continuous physiotherapy to animals following denervation.

Oligodendrocyte survival and function in the long-lived strain of the myelin deficient rat.

This study has examined cellular and molecular aspects of glial cell function in a newly described long-lived myelin deficient rat mutant. In contrast to the shorter-lived mutants which died at 25-30 days, the longer-lived mutant rats lived to 75-80 days of age. Despite living longer, these mutants had a similar frequency of seizures to their younger counterparts. In the spinal cord and optic nerves of the older mutants, myelinated fibres in similar numbers to those seen in the younger myelin deficient rats were present. However, the total glial cell numbers were markedly reduced with few remaining normal appearing oligodendrocytes, and very few microglia compared to the younger mutants. In addition, little or no cell death or division was seen in the longer-lived rats. However, there was some evidence of ongoing myelination and the persistence of immature oligodendrocytes or their progenitors in the older mutant. There was some continued myelin gene expression, although this was at much reduced levels compared to normal, with proteolipid protein and myelin basic protein being most affected. In situ hybridization analysis for proteolipid protein mRNA showed that few proteolipid protein expressing oligodendrocytes remained in the 70-80-day-old mutant. Polymerase chain reaction analysis of exon 3 of the long-lived mutant revealed the same point mutation as described in the younger myelin deficient rat.

Glycogen storage disease type Ia in two littermate Maltese puppies.

Glycogen storage disease type Ia (GSD-Ia) (von Gierke's disease) was identified in two 47-day-old littermate Maltese puppies. The puppies were presented for necropsy with a history of failure to thrive, mental depression, and poor body condition. Gross findings included small body size and emaciation (212 and 246 g versus 595 g for normal littermate), severely enlarged pale livers (48 and 61 g), and pale kidneys. Histologically, there was marked diffuse vacuolation of hepatocytes with large amounts of glycogen and small amounts of lipid. Renal tubular epithelium was mildly to moderately vacuolated. Soft tissue mineralization was present in renal tubules and pulmonary alveolar septa. Biochemical analysis showed that levels of glucose-6-phosphatase were markedly reduced in liver (0.3 and 0.4 microM/minute/g tissue versus 4.7 +/- 1.5 microM/minute/g tissue for controls) and kidney (0.45 and 0.4 microM/minute/g tissue versus 4.1 microM/minute/g tissue for controls) and that glycogen content was increased in liver (9.4% and 9.4% versus 1.3% +/- 1.4% for controls). This is the first confirmed report of animals with glycogen storage disease type Ia.

RNA transcription in axotomized dorsal root ganglion neurons.

Levels of RNA transcription were examined in L5 sensory ganglion neurons after unilateral crush injuries of the sciatic nerve using an in situ autoradiographic technique. Neuronal transcription increased in a biphasic pattern on the lesioned side within the first two weeks after injury. The timing and pattern of increases in transcription corresponded to previously reported increases RNA synthesis. The data demonstrate a dynamic regulation of RNA transcription in regenerating neuronal populations. This technique may prove to be useful in the study of factors controlling transcriptional activity in injured neurons.

Bilateral phasic increases in dorsal root ganglia nerve growth factor synthesis after unilateral sciatic nerve crush.

The amount of nerve growth factor (NGF) in the L5, L6, and cervical dorsal root ganglia of rats was examined from 1 to 30 days after a unilateral crush lesion of the sciatic nerve and adjacent branches of the lumbar plexus at the level of the sciatic notch. Unilateral nerve crush produced increases in NGF content of lumbar ganglia at 1, 4, and 7-8 days after injury, with increased NGF mRNA at 4 and 7-8 days. Increases in NGF at 1 and 4 days were most pronounced on the unlesioned side while increases at days 7 and 8 were most pronounced on the lesioned side. NGF content increased in cervical ganglia of nerve-lesioned animals at 3 and 7 days after injury and in lumbar and cervical ganglia of sham-operated animals 3-5 days after surgery, with no comparable changes in NGF mRNA. Elevations of ganglionic NGF coincide temporally with some of the alterations in metabolism and morphology which occur in dorsal root ganglion neurons after sciatic nerve crush. However, the bilateral nature of increases in NGF demonstrates that the factor(s) producing the response is not restricted to ganglia axotomized by the injury. The data suggest that ganglionic NGF may be regulated by systemic factors, produced during stress or trauma, as well as by factors from the denervated target tissue and/or regenerating axons.

Hypereosinophilia associated with transitional cell carcinoma in a cat.

A 14-year-old spayed female domestic shorthair cat was evaluated because of an abdominal mass and eosinophilia. Widely disseminated, transitional cell carcinoma of the urinary bladder was diagnosed histologically. To further characterize the eosinophilia, eosinophils were separated from other leukocytes and cultured in vitro. Harvested cells were evaluated for density and for in vitro survivability. Results of these tests, hyperplasia of bone marrow eosinophil precursors, and lack of tumor tissue eosinophilic infiltrates suggested that an eosinophilopoietic stimulus of undetermined origin was likely the cause of this cat's hypereosinophilia.

Changes in plasma cytokines associated with peripheral nerve injury.

Plasma levels of interleukins 1, 2, 4 and 6 and tumor necrosis factor (TNF) were measured from 0 to 30 days in rats after a unilateral crush of the sciatic nerve at the level of the sciatic notch and after sham operations without nerve crush. Interleukin-6 was observed to peak and return to baseline levels within 24 h and remained at baseline for the duration of the experiment. An initial sharp rise in interleukin-1 and TNF was observed in all animals 1-2 days after the operation. A transient increase in interleukin-1 and TNF was also observed only in nerve-injured animals between 10 and 14 days after injury. A large increase in interleukin-2 appeared only in nerve-injured animals beginning at 11 days after injury and remained elevated for the remaining study period. No alterations in plasma interleukin-4 were observed at any time point. The experiments provide preliminary evidence for significant trauma-induced alterations in plasma cytokines which could provide a basis for some of the diffuse responses of peripheral neurons to trauma. The biphasic nature changes in plasma cytokines suggest that the immune system may participate in tissue reactions involved in the recovery from nerve injury.

Circadian rhythms of chemotaxis to ammonium and of methylammonium uptake in chlamydomonas.

Chlamydomonas reinhardtii expresses a well-documented circadian rhythm of phototaxis, which peaks in the subjective daytime. We find that vegetative cells also express circadian rhythms of chemotaxis to ammonium and ammonium uptake (as gauged by uptake of [(14)C]methylammonium). The chemotaxis rhythm peaks in the subjective night. Methylammonium uptake is light dependent, and its rhythm peaks at subjective dawn. Unlike vegetative cells, gametes are not attracted to ammonium. We believe this to be the first report of a circadian rhythm of chemotaxis.

Morphological alterations in dorsal root ganglion neurons after peripheral axon injury: association with changes in metabolism.

Axotomized rat sensory ganglion neurons have been shown to undergo rapid metabolic changes in the first 2 weeks after injury. The present study examined selected morphological features of these neurons over the same time period. Parameters studied included the position of the cell nucleus (eccentricity) and soma, nuclear, and nucleolar size over time periods primarily in the first 2 weeks after a unilateral crush injury of the sciatic nerve. Comparisons were made with normal ganglia and ganglia contralateral to the injury. The eccentricity of the nucleus in injured neurons was significantly altered within 1 day after injury and remained so over the entire time period studied. Alterations in neuron soma included an initial decrease in size at 1 day followed by a significant bilateral increase at 3 days after injury. Nuclear and nucleolar size changes were phasic with significant increases size peaking at 3-4 and 8-11 days after injury. These alterations coincided temporally with known changes in RNA synthesis occurring in these neurons after injury. Significant alterations in all parameters were observed on the uninjured side. Preliminary studies of the bilateral changes suggested that the trauma of the operation may be the major factor in this response. The data suggest that significant morphological alterations parallel the rapidly fluctuating change in neuronal metabolism after axon injury.