Purification of the cardiac sarcoplasmic reticulum membrane protein phospholamban from recombinant Escherichia coli

Phospholamban (PLN) was expressed in Escherichia coli as a protein fusion with glutathione S-transferase (GST). GST-PLN was mostly present in the insoluble protein fraction and accounted for approximately 50% of total insoluble protein. Attempts to suppress inclusion body formation or to use GST as an affinity-purification tag failed. A successful purification method is based on preparative SDS/PAGE and electrodialysis. From 1 g cells we typically purified 13.5 mg fusion protein with a PLN content of 2.8 mg. We genetically inserted an enterokinase (EK) protease site just in front of the PLN sequence and demonstrated the proteolytical liberation of PLN from the carrier protein. The approach described represents a substantial advancement in PLN expression and purification.     

Stability of motor-unit force thresholds in the decerebrate cat

To further test the hypothesis that some fixed property of motoneurons determines their recruitment order, we quantified the variation in force threshold (FT) for motoneurons recruited in muscle stretch reflexes in the decerebrate cat. Motor axons supplying the medial gastrocnemius (MG) muscle were penetrated with micropipettes and physiological properties of the motoneuron and its muscle fibers, i.e., the motor unit, were measured. FT, defined as the amount of MG force produced when the isolated motor unit was recruited, was measured from 20 to 93 consecutive stretch trials for 29 motor units. Trials were selected for limited variation in base force and rate of rise of force, which have been shown to covary with FT, and in peak stretch force, which gives some index of motor-pool excitability. Under these restricted conditions, large variation in FT would have been inconsistent with the hypothesis. Analysis of the variation in FT employed the coefficient of variation (CV), because of the tendency for FT variance and mean to increase together. We found that CV was distributed with a median value of 10% and with only 2 of 29 units exceeding 36%. Some of this variation was associated with measurement error and with intertrial fluctuations in base, peak, and the rate of change of muscle force. CV was not significantly correlated with motor-unit axonal conduction velocity, contraction time, or force. In three cases FT was measured simultaneously from two motor units in the same stretch trials. Changes in recruitment order were rarely observed (5 of 121 stretch trials), even when FT ranges for units in a pair overlapped. We suggest that the large variation in recruitment threshold observed in some earlier studies resulted not from wide variation in the recruitment ranking of motoneurons within one muscle, but rather from variation in the relative activity of different pools of motoneurons. Our findings are consistent with the hypothesis that recruitment order is determined by some fixed property of alpha-motoneurons and/or by some unvarying combination of presynaptic inputs that fluctuate in parallel.     

Bronchiolo-alveolar cell carcinoma with apparent response to chemotherapy to anti-tuberculosis

Pigmented vulvar lesions were observed in a child during a sexual abuse evaluation. Gross examination of the lesions appeared most consistent with bowenoid papulosis; however, biopsy confirmed the lesions to be pigmented apocrine hamartomas. To our knowledge, these rare and benign tumors have never been described as pigmented, but should be added to the differential diagnosis of pigmented vulvar lesions.     

Dissolved organic carbon dynamics in the upper rhone: The influence of side-arms?

Dissolved organic carbon (DOC) dynamics were investigated over a two year period in the Upper Rhǒne River to examine the role of side-arms in providing DOC to the main channel in relation to discharge fluctuations, especially floods. Concentrations of DOC are shown to remain low in space and in time (average 1.5 mg ^?1) and to be more related to global hydrological events (precipitation) than to any local flushing for backwaters during floods. The results do not support the general assumption that side-arms are the providers of DOC to the main channel.     

Traumatic vertebral arterial dissection and vertebrobasilar arterial thrombosis successfully treated with endovascular thrombolysis and stenting

A case of traumatic extracranial vertebral arterial dissection leading to vertebrobasilar thrombosis and respiratory compromise requiring mechanical ventilation was managed with intraarterial thrombolysis and stenting of the vertebral intimal dissection. In contrast to similar, previously reported cases, this critically ill patient made a full recovery, returning to his job as a secondary school teacher.     

Principles of lymphogenic and hematogenous metastasis and metastasis classification

A subpopulation of rabbit polyclonal anti-idiotypic antibody (anti-Id) was previously produced to a murine monoclonal antibody (mAb) (M1875) specific for the bluetongue virus core protein VP7. In this report, mimicry of VP7 by this anti-Id (designated RAb2-A) was functionally analyzed through immunization of Balb/c mice with RAb2-A or purified VP7. Animals immunized with RAb2-A were able to produce an M1875-like Ab3 antibody response with idiotype and epitope specificity resembling that of M1875 without subsequent exposure to the nominal antigen. This conclusion was supported by experiments showing that the RAb2-A-induced Ab3 antibodies (i) reacted specifically with the immunizing anti-Id; (ii) were capable of binding VP7; (iii) inhibited M1875 from binding to VP7; and (iv) inhibited M1875 from binding to RAb2-A. Similarly, mice immunized with purified VP7 also produced antibodies that exhibited characteristics such as idiotype and epitope specificity in common with M1875. No antibody response to VP7 was detected in control groups of mice immunized with either normal rabbit IgG or BHK-21 cell components. Therefore, it can be concluded that rabbit anti-Id RAb-2-A mimics an M1875-defined VP7 epitope sufficiently to function as a surrogate antigen for inducing an anti-bluetongue virus response.     

Control of stroma-dependent hematopoiesis by basic fibroblast growth factor: stromal phenotypic plasticity and modified myelopoietic functions

The immunological agglutination reactions of physically absorbed F(ab')2 molecules onto anionic and cationic latex particles have been investigated by means of optical absorbance measurements. These measurements have been conducted under different conditions to determine the most influential factors. Surface F(ab')2 and BSA densities, particle concentration in the reaction medium and polyethylene glycol concentration are some of these factors. Sensitized cationic and anionic latexes differ considerably with respect to their colloidal stability and reactivity. As a general rule, the sensitized cationic latex has a relatively higher colloidal stability and hence, it provides reagents with a better optical response. Less than 0.025 microgram/ml of C-reactive protein has been detected using this particle enhanced optical immunoassay.     

Position-sensitive superheated-liquid-droplet neutron dosimeter

The one-dimensional (1D) position-sensitive superheated-liquid-droplet dosimeter (SLDD) has been fabricated and tested in the laboratory. The 1D SLDD is fabricated from a 9.525-mm OD, 6.35-mm ID, 20-cm long, Plexiglas-walled tube filled with a mixture of superheated-liquid Freon droplets and host medium glycerol. Washer-shaped piezoelectric acoustic transducers are positioned at both ends of the tube; they determine the number and positions of the acoustic events when the superheated-liquid droplets evaporate upon neutron irradiation. The SLDD is irradiated with the ¹³⁷Cs and ⁶⁰Co γ-sources, as well as ²⁵²Cf neutron source to test for its radiation response and spatial resolution. The SLDD based on the Freon-134a superheated-liquid droplets operating at 20°C and 1 atm is found to be ideal for measuring absorbed neutron dose. This study also proves that the positions of the radiation-induced nucleation acoustic events can be linearly determined from the differences in the transmission times received by the acoustic transducers on the 1D SLDD. The spatial resolution of the neutron depth-dose is 1 mm due the finite response time (1 μs) of the piezoelectric acoustic transducers.     

Thermal expansion of semiconductor materials

The thermal expansion of tellurides of germanium, bismuth, and intermetallic compounds is investigated over the temperature range 293-973 K.Belarusian Agricultural Technical University, Minsk. Translated from Inzhenerno-Fizicheskii Zhurnal, Vol. 66, No. 5, pp. 612–616, May 1994.