Practical value of a new serum digitoxin assay

The purpose of this study was to evaluate a new fluorescence polarization immunoassay, TDx, for digitoxin by comparing the results of this assay with those of a radioimmunoassay (RIA). Thirty-three serum samples were obtained from 15 patients during, and for 4 weeks after, a 4-week course of digitoxin therapy. Each sample was separated by centrifugation, coded, and frozen until analysis. At the time of analysis, each sample was divided and analyzed simultaneously by TDx and RIA. Nine samples yielded results less than 2 ng/ml (limit of assay sensitivity) by one or both methods and were excluded from further data analysis. Linear regression analysis of the results of the remaining 24 paired samples (x = TDx, y = RIA) revealed a strong correlation coefficient of r2 = 0.95, slope = 0.95, and a y intercept of -0.99 (y = -0.99 + 0.95x). Additionally, the TDx results were lower than the RIA values in only five of 33 paired samples; and these occurred in four patients who had a significantly lower mean estimated creatinine clearance than that of the other 11 patients (39.0 +/- 9.1 ml/min/1.73 m2 vs. 63.3 +/- 11.8 ml/min/1.73 m2, p less than 0.01). The TDx system is a comparable alternative to the RIA method, but differences in specificity and sensitivity may exist and should be evaluated more thoroughly.     

Ultrastructural observations of organelle accumulation in the equine recurrent laryngeal nerve

Summary The left recurrent laryngeal nerves from five horses with sub-clinical neuropathy were examined by light and electron microscopy in a study designed to examine accumulation of axonal organelles at paranodal and internodal locations. Transverse sections of the nerve showed scattered fibres with split myelin sheaths and axonal accumulation of organelles. On longitudinal sections these collections were seen to result from an axonal outpouching in which dense lamellar bodies and mitochondria had accumulated. These paranodal collections, which could be found on both sides of the node, were often associated with infoldings of the terminal loops of myelin and with occasional paranodal demyelination. The fact that many of the organelles in the outpouches were lysosomal in nature was confirmed by their positive staining for cathepsin D activity. Longitudinal sections demonstrated a number of axons which were swollen over a long distance and which contained focal accumulations of similar organelles. In places, however, there was a clear separation between these organelles and the cytoskeletal proteins. In each case these swollen axons were surrounded by Schwann cell nuclei and their processes, forming well-ordered onion bulbs. The possibility that these two types of changes, i.e. the paranodal accumulations and the axonal swellings could result from a disturbance in axonal transport in this distal axonopathy is discussed.     

The use of laser-light scattering and controlled shear in platelet aggregometry

Laser-light scattering was used to observe and quantify the dynamics of human blood platelet aggregation in platelet-rich plasma (PRP). Aggregation was performed in a controlled shear environment by placing the PRP in the annular space between a rotating cylindrical rod and a stationary cylindrical tube. The instrument was capable of very sensitive continuous semi-quantitative measurements of chemically-induced microaggregation. As a demonstration of the technique, results are presented for ADP-induced aggregation at doses of 10, 1, and 0.1 microM and collagen-induced aggregation at a dose of 5 micrograms/ml, each at shear rates of 1,000 s-1 and 500 s-1. Extensive aggregation was observed in response to ADP at even the low dose of 0.1 microM, indicating a high sensitivity to microaggregates. The sensitivity of the ultimate size of the ADP-induced aggregates to ADP concentration was shear dependent. The formation of microaggregates by collagen stimulation was shown to be almost immediate, as contrasted with a 10-20 s typical lag when observed turbidometrically. Disaggregation was observed with 1 microM ADP, but this was only partial, as contrasted with the complete recovery of transmittance observed in the turbidometric technique. Electronic particle sizing and counting was employed to semiquantitatively verify the aggregate size distributions found from mathematical conversion of the laser-light scattering data.     

Functional activity of the neutrophils in the bronchoalveolar space in chronic bronchitis and bronchiectasis]

Bronchoalveolar lavage fluid neutrophils were studied by the cytologic methods in 58 patients with chronic bronchitis, 63 ones with bronchiectasis, and 8 normal controls. The study included cytospectrophotometry of myeloperoxidase and alkaline phosphatase activity and estimation of active oxygen-producing cells in the NBT test. Neutrophilic functional activity was different in the patients with chronic bronchitis and bronchiectasis. Neutrophilic myeloperoxidase and alkaline phosphatase activities were lower in the patients with chronic bronchitis than in those with bronchiectasis, whereas the counts of cells active in the NBT test were low in both patient populations.     

Central nervous control of hepatic circulation.

This study was undertaken to assess the role of the hypothalamus and medulla oblongata in regulation of liver circulation in anesthetized dogs. Blood pressure, flow in hepatic artery and portal vein, and shifts of blood volume in the liver were recorded. Stimulation of the anterior hypothalamus produced changes in arterial pressure which were followed by passive changes in hepatic arterial blood flow; changes in hepatic artery resistance were rather small. Stimulation of the medial and posterior hypothalamus increased hepatic arterial resistance by 65-170%. Liver portal blood flow during stimulation of most of the hypothalamic sites decreased, hepatic portal pressure rose and vascular portal venous resistance increased 2.5-3 times. Three areas only (sympatho-inhibitory area, paraventricular and lateral hypothalamic nuclei) when stimulated produced dilatation of hepatic portal and splanchnic vascular beds, thus increasing portal blood flow. All cases of stimulation led to the decrease of blood volume in the liver by 10-36%. Stimulation of medullary structures (n. tractus solitarii, reticular nn.) caused similar changes in hepatic circulation, however the amplitude of reaction was 1.5-6 times smaller than upon hypothalamic stimulation. Central impulses to the hepatic vessels are transmitted by sympathetic adrenergic nerve fibers through vascular alpha-adrenoreceptors. It is concluded that the hypothalamic level of the central nervous system, unlike the bulbar one, exerts considerable, differentiated, well coordinated and to some extent specific influences on hepatic circulation.     

The hepatotropic properties of nonsteroidal anti-inflammatory agents

The hepatotropic effect of nonsteroidal anti-inflammatory drugs (NSAID) such as indomethacin, voltaren, piroxicam, phenylbutazon, mefenamic acid was studied. It was found that according to their level of the pharmacological protection of the liver against tetrachlormethan these agents may be arranged in the following sequence: mefenamic acid, phenylbutazon, voltaren, piroxicam. The hepatoprotective effect of NSAID correlates with the antioxidant properties and fails to correlate with the antioxidant ones. The hepatotoxic effect of NSAID was determined by their ability to suppress synthesis of prostaglandins.