Isolation and partial characterization of surface components of cell line MDA-MB-231 derived from a human metastatic breast carcinoma.

Neuraminidase (Vibrio cholerae) treatment of human metastatic mammary carcinoma MDA-MB-231 cells grown in culture released 0.60-0.63 mg of N-acetylneuraminic acid from 10(9) cells. Incubation of intact cells with a modified trypsin and fractionation by gel filtration gave mainly O-glycopeptides. The presence of O-glycosyl-linked chains having one or two carbohydrate residues was confirmed by treatment of the glycopeptide fractions with galactose oxidase, followed by reduction with alkaline sodium borotritide and fractionation. The major glycopeptide fraction, which consisted of 53% carbohydrate and 47% protein, and a minor glycopeptide fraction each inhibited hemagglutination by peanut lectin. These results suggest the presence of O-beta-D-galactopyranosyl-(1 leads to 3)-O-(2-acetamido-2-deoxy-alpha-D-galactopyranosyl)-(1 leads to 3)-L-serine (threonine) residues. The absorptive capacities for anti-HLA-A2 and anti-HLA-B8 antisera were slightly greater for intact than for lyophilized cells, which suggested that masking of these major histocompatibility antigens did not occur in intact cells.

[Influence of age on optical aberrations of the human eye]. / Einfluss des Alters auf die optischen Aberrationen des menschlichen Auges.

BACKGROUND: Currently the influence of age on corneal and ocular aberrations is still insufficiently known. The aim of this clinical study was to compare age-related aberrations of human eyes. METHODS: In a prospective study 98 eyes of 49 healthy patients ranging from 17 to 65 years of age (38.6+/-10.0 years) were consecutively examined. The best corrected visual acuity ranged from 0.8 to 1.6; 48 eyes were emmetropic (SE+/-0.5 D), 42 eyes myopic (SE <-0.5 to -6.75 D), and 8 eyes hyperopic (SE >+0.5 to +3.88 D). The corneal aberrations were derived from corneal topography (Keratron Scout, Optikon). The measurement of ocular aberrations was performed with a Tscherning wavefront aberrometer (ORK, Schwind). The aberrations of the Zernike coefficients and RMS values (1st to 4th order) were determined. RESULTS: The mean corneal and ocular Zernike coefficients of higher order were smaller than 0.2 microm. There was an evident decrease of wavefront aberrations with increasing order. Higher order corneal aberrations were larger than the corresponding ocular aberrations. With increasing age higher optical errors increased in complexity, and the correlation of corneal and ocular aberrations decreased with significant differences. Although the corneal ocular RMS value of the 3rd and 4th order correlated in the younger group (r=0.51, p=0.0001), there was no correlation in the older group (r=-0.48, p=0.832). The influence of age caused a significant increase of ocular aberrations of the 3rd and 4th order, in particular a tenfold extension of coma (C07) (p=0.002), a twofold extension of spherical aberration (C12) (p=0.0001), and an increase of the 3rd and 4th order RMS values (p=0.001). CONCLUSIONS: Increased age induced an increase in optical aberrations of the eye, which demonstrates the influence of the lens on ocular aberrations. The combination of corneal and ocular diagnostic methods is recommendable for a better understanding of visual performance.

Racial group differences in plasma concentrations of antioxidant vitamins and carotenoids in hemodialysis patients.

Approximately 50% of the mortality in hemodialysis patients is due to cardiovascular disease. Antioxidant vitamins and carotenoids may be protective because oxidation of low-density lipoproteins appears to be a necessary prerequisite for the development of atherogenesis, and hemodialysis itself may stimulate the generation of oxygen reactive species. African Americans comprise a substantial proportion of dialysis patients because they have higher rates of hypertension, glomerulonephritis, and diabetic end-stage renal disease than do whites. The purpose of this cross-sectional study was to determine the plasma concentrations of antioxidant vitamins and carotenoids in hemodialysis patients and to investigate whether differences in these concentrations in the major racial or ethnic groups exist. Plasma concentrations of alpha- and gamma-tocopherol, carotenoids, and retinol were measured with HPLC and plasma vitamin C was measured with a spectrophotometric method in 109 white and African American hemodialysis patients. Dietary intakes of selected micronutrients were also compared by using data from a food-frequency questionnaire. Overall, plasma vitamin C and alpha-tocopherol concentrations were comparable but plasma carotenoid concentrations were lower than those reported for other populations. African American patients had significantly higher mean plasma concentrations of retinol (P < 0.04), lutein (P < 0.02), and total carotenoids minus lycopene (P < 0.04); whites had significantly higher mean plasma concentrations of alpha-tocopherol (P < 0.02), independent of age and plasma lipid concentrations. Diabetes comorbidity had an independent negative association with plasma beta-carotene concentration but was not associated with other measures.

Aging of liver: morphological and biochemical changes.

Light-microscopically there are no significant age-dependent changes in the liver, due to the slight increase in biochemically determined connective tissue components. Electron-microscopical comparison of young and old animals show a larger size and decreased numbers of liver cell mitochondria in older animals. Collagen synthesis in liver is accelerated during growth, as demonstrated by assay of the [3H] proline incorporation rates and of the specific activity of hydroxyproline, corresponding to a decrease in the prolyl hydroxylase activity with age. The neutral salt-soluble collagen fraction may be regarded as a parameter for collagen synthesis, which is activated during the growth period and then remains constant without age and sex differences. Not much is known about the significance of these changes with increasing age or about their initiating factors, although there are parallel findings for liver intoxication and inflammation.

Properties of a microcoaxial electrode designed for unit recording from human peripheral nerves.

A new microcoaxial electrode is presented, which has been used successfully for recording single- and multiunit activity from myelinated and unmyelinated human nerve fibres. The main advantages (as compared with conventional needle electrodes) are electrical stability, mechanical durability, capability of being re-used and an insulation not easily scraped off, resulting in a low leakage capacitance. The AC polarization impedance and the potential transducing properties of the microelectrode have also been studied.

Responses of human muscle spindle afferents during isotonic position holding and active movements.

Discharge characteristics of muscle spindle primary afferents from human finger flexors were investigated during load-bearing isotonic position holding, and intentional shortening or lengthening contractions. During position holding, units mostly showed a negative relationship between muscle length and impulse rate, particularly when the load was small. When the load was increased, Ia-discharge rates increased more in the elongated than in the shortened muscle, and the inverse position response became less pronounced. During active lengthening contractions, a stretch response pattern was sometimes absent or could be abolished by increasing the load.

Response of human muscle spindle afferents to abrupt load perturbations.

Response patterns of single muscle spindle primary afferents to abrupt load perturbations inflicted upon the contracting finger flexors were investigated in awake human subjects using the technique of microneurography. Units showed a transient high-frequency discharge in the rising phase of the stretch. No repeated burst activity attributable to irregularities of the stretch velocity or to mechanical oscillations of the muscle could be detected.

Measurements of muscle stiffness, the electromyogram and activity in single muscle spindles of human flexor muscles following conditioning by passive stretch or contraction.

In experiments on adult human subjects we examined the effect on passive mechanical properties of a muscle by conditioning it with either an isometric contraction or passive muscle extension. The test measurement was the amount of muscle displacement (stiffness) and the accompanying EMG in response to a brief torque pulse. Two muscles were tested, flexor digitorum profundus (FDP) and brachialis. In FDP the discharge of single muscle spindles was recorded as well. After muscle extension and return to the initial length, passive stiffness was less than after an isometric contraction. The changes in stiffness were accompanied by changes in pattern of EMG and in the responses of muscle spindles. It is suggested that in resting muscle there are stable cross bridges between actin and myosin filaments of muscle fibres which largely determine the passive stiffness. Muscle extension leads to detachment of these cross bridges which then re-form at the longer length. Return of the muscle to its starting length leads to development of slack in muscle fibres because, stiffened by the presence of the stable cross bridges, they are unable to shorten. Slack in muscle fibres lowers their measured stiffness. Muscle contraction, on the other hand, will result in any preexisting slack being taken up by the actively shortening muscle fibres, thereby raising muscle stiffness. Stiffness in intrafusal fibres is likely to follow a similar pattern to that in extrafusal fibres, leading to changes in stretch responsiveness of muscle spindles and consequently in the reflex EMG. It is concluded that the changes in stiffness and accompanying reflexes observed in this study are likely to be seen, at least under some conditions, in normal movements.

Short-term electrical stimulation enhances the effectiveness of Botulinum toxin in the treatment of lower limb spasticity in hemiparetic patients.

The study tested the spasmolytic effect of Botulinum toxin A in two groups of hemiparetic patients with lower limb spasticity: in the first group (n = 5) 2000 U Dysport were injected into the soleus, tibialis posterior and both heads of gastrocnemius muscles alone; the second (n = 5) received additional repetitive alternating electrical stimulation of M. tibialis anterior and plantar flexors for 30 min six times per day during the 3 days following the injection. Muscle tone, rated by the Ashworth spasticity score, and gait analysis including recording of vertical ground reaction forces, were assessed before and 4 weeks after injection. The combined treatment proved to be more effective with respect to the clinically assessed reduction of muscle tone, gait velocity, stride length, stance- and swing-symmetry (P < 0.05). The result is discussed with reference to animal experiments demonstrating enhanced toxin uptake and accelerated onset of its paralytic effect by electrical stimulation.

Comparative lectin-binding and agglutination properties of the strain-specific, TA-3-St, and the non-strain-specific, TA3-Ha, murine mammary carcinoma ascites sublines. Further studies of receptors in epiglycanin.

The complex carbohydrates at the cell surfaces of two TA3, murine mammary carcinoma ascites sublines (the strain-specific, TA3-St subline and the nonstrain-specific. TA3-Ha line) were compared by binding studies with 125I-labelled concanavalin A (con A), Ricinis communis agglutinin (RCA), and eel-serum agglutinin (ESA). The TA3-Ha cell bound equal amounts of con A, 1.5-fold more RCA, and 4-fold more ESA than the TA3-St cell. Binding-inhibition studies by these lectins and two others [wheat-germ agglutinin (WGA) and potato lectin (STA)] suggest complementary binding-sites between con A and both RCA and ESA. Quantitative agglutination studies with the five lectins, and inhibition determinations by both neuraminidase-treated and untreated epiglycanin revealed that TA3-St, but not TA3-Ha, cells were agglutinated by con A, and that epiglycanin inhibited this agglutination, as well as the agglutination of rabbit erythrocytes by con A. The presence of a con A receptor on epiglycanin was also suggested by the binding of epiglycanin to con A-Sepharose, and its specific elution with methyl alpha-D-mannopyranoside. TA3-St cells were agglutinated at a 10-15-fold lower concentration of either STA or RCA than TA3-Ha cells, but both cells were agglutinated by the same concentration of WGA and ESA. Inhibition by epiglycanin of agglutination of TA3-St cells by either STA or ESA occurred at a concentration lower than that of TA3-Ha cells, but epiglycanin inhibited RCA agglutination of TA3-Hs cells at a concentration lower than that of TA3-St cells. Epiglycanin, but not asialoepiglycanin, inhibited the agglutination of TA3-Ha cells by WGA.

Gait function in spastic hemiparetic patients walking barefoot, with firm shoes, and with ankle-foot orthosis.

The purpose of the study was to investigate the effect of firm shoes and a particular type of a frequently prescribed ankle-foot orthosis in Germany, the Valens calliper, on functional gait parameters in hemiparetic patients with marked extensor spasticity and corresponding heel-strike problems. Cycle times, vertical ground reaction forces, trajectories of the centre of pressure (gait line) under the soles and the mode of initial contact were assessed in 19 hemiparetic patients walking barefoot, with firm shoes and the orthosis. With the shoes, significant improvements (p < 0.006) were observed for: walking velocity, stride length, initial double stance duration and gait line of the non-affected foot. The pattern of the vertical force diagram improved qualitatively in six patients. With the orthosis the following variables showed an even more marked improvement (p < 0.006): walking velocity, cadence, stride length, and gait line of both the affected and non-affected foot. Fourteen patients became rear-foot strikers and the vertical force pattern further improved in six patients. It is concluded that the ankle-foot orthosis (more than the shoes) provide a useful support in the early rehabilitation of hemiparetic patients with marked spasticity.

One-way distribution system for water for injection: process management, microbiological quality control, and meeting regulatory requirements.

The specifications for pharmaceutical water, the qualification and validation of water preparation facilities, strategies to prevent contamination by water-borne bacteria and lastly, the monitoring of microbiological purity are the topics of frequent seminars on Pharma Water Total Quality Management. The same subdivisions are used in the following paper on the process management and microbiological control of a one-way distribution system for Water for Injection. Since 1990, such a system has been in use in the production department of Pharma Hameln GmbH, a contract manufacturer of parenterals. Using this system as an example, the twin needs for flawless microbiological process control and for suitable measures to monitor water quality are discussed, which, together with extensive documentation of the qualification of the production and distribution system, ultimately led to acceptance of the system by regulatory authorities.

[A clinically applicable system for analysis of standing up using measurements of floor reaction forces]. / Klinisch einsetzbares System zur Analyse des Aufstehens mittels Messung der Bodenreaktionskräfte.

A system for clinical analysis of sit-to-stand transfer using two force plates is presented. Distinct time-events and phases are estimated. Illustrative parameters are also determined, such as the distribution of the vertical ground reaction forces, displacement of the center of gravity in the sagittal and frontal planes, and a comparison of the displacement of the center of gravity and the center of pressure. These parameters indicate a symmetrical raising while keeping the middle of the body. Future applications include the validation of therapy, biofeedback and the control of closed-loop FES.