Polarized light microscopy of weakly birefringent biological specimens

A simple, but little known technique for producing marked colour contrast in images of weakly birefringent biological samples obtained with a polarized light microscope is investigated. The technique involves inserting a full wave plate at a small angle of horizontal rotation with respect to the analyser or polarizer, rather than at the conventional angle of 45° commonly employed in the field of mineralogy for examining strongly birefringent specimens. Inserting the full wave plate at a small angle (typically less than 10°) enhances the contrast between regions of tissue having different optical path differences and, in particular, different orientations of optic axes; improving the detection of structures of interest with the polarized light microscope.     

HIV-1 envelope proteins gp120 and gp160 potentiate NMDA-induced [Ca2+]i increase, alter [Ca2+]i homeostasis and induce neurotoxicity in human embryonic neurons

The envelope glycoprotein gp120 of the human immunodeficiency virus HIV-1 has been proposed to cause neuron death in developing murine hippocampal cultures and rat retinal ganglion cells. In the present study, cultured human embryonic cerebral and spinal neurons from 8- to 10-week-old embryos were used to study the neurotoxic effect of gp120 and gp160. Electrophysiological properties as well as N-methyl-D-aspartate (NMDA)-induced current were recorded from neurons maintained in culture for 10-30 days. Neither voltage-activated sodium or calcium currents nor NMDA-induced currents were affected by exposure of neurons to 250 pM gp120 or gp160. In contrast, when neurons were subjected to photometric measurements using the calcium dye indo-1 to monitor the intracellular free Ca2+ concentration ([Ca2+])i, gp120 and gp160 (20-250 pM) potentiated the large rises in [Ca2+]i induced by 50 microM NMDA. The potentiation of NMDA-induced Ca2+ responses required the presence of Ca2+ in the medium, and was abolished by the NMDA antagonist D-2-amino-5-phosphonovalerate (AP5) and the voltage-gated Ca2+ channel inhibitor nifedipine. Moreover, exposure of a subpopulation of spinal neurons (25% of the cells tested) to 20-250 pM gp120 or gp160 resulted in an increase in [Ca2+]i that followed three patterns: fluctuations not affected by AP5, a single peak, and the progressive and irreversible rise of [Ca2+]i. The neurotoxicity of picomolar doses of gp120 and gp160 cultures was estimated by immunofluorescence and colorimetric assay. Treatment of cultures with AP5 or nifedipine reduced gp120-induced toxicity by 70 and     

Gonadoblastoma: an ovarian tumor with characteristic pelvic calcifications

Typical circumscribed mottled calcifications in the pelvis are a frequent finding in gonadoblastomas. Six patients with this tumor are reviewed. Radiologically visible calcifying gonadoblastomas were found in three cases.     

Changes in Flavor Components of Garlic Resulting from Gamma Irradiation

Formation of diallyl disulfide, identified as a main flavor component of garlic, was gradually decreased by irradiation of 0.05-0.5 kGy, but increased slightly at higher doses of 1.0-3.0 kGy. In garlic stored for 5 months, a dose of 0.1 kGy had little influence on the amount of diallyl disulfide, but higher dose irradiation caused an apparent reduction in the amount of the component.     

The Wenckebach phenomenon between electric pacemaker and ventricle

Two cases which exhibited a Wenckebach form of exit conduction between an electrical pacemaker and the ventricular myocardium are presented. This manifests with increasing latency (stimulus to QRS intervals) and ultimate stimulus failure (block). The cycle then repeats itself. The QRS duration also increases progressively within each cycle. The phenomenon connotes an adverse prognosis. Several postulates regarding the mechanism are discussed.     

Measurement of lung expansion with computed tomography and comparison with quantitative histology

The total and regional lung volumes were estimated from computed tomography (CT), and the pleural pressure gradient was determined by using the milliliters of gas per gram of tissue estimated from the X-ray attenuation values and the pressure-volume curve of the lung. The data show that CT accurately estimated the volume of the resected lobe but overestimated its weight by 24 +/- 19%. The volume of gas per gram of tissue was less in the gravity-dependent regions due to a pleural pressure gradient of 0.24 +/- 0.08 cmH2O/cm of descent in the thorax. The proportion of tissue to air obtained with CT was similar to that obtained by quantitative histology. We conclude that the CT scan can be used to estimate total and regional lung volumes and that measurements of the proportions of tissue and air within the thorax by CT can be used in conjunction with quantitative histology to evaluate lung structure.     

PURIFICATION AND PARTIAL CHARACTERIZATION OF WHITE KIDNEY BEAN (PHASEOLUS VULGARIS)β-AMYLASE INHIBITORS FROM TWO EXPERIMENTAL CULTIVARS

β-Amylase inhibitors WKB 858B and WKB 858B were purified to homogeneity from different cultivars of white kidney beans by extraction from the ground beans and by sequential heat treatment, ethanol fractionation, DEAE-cellulose chromatography, Sephadex G-75 gel chromatography and CM-cellulose chromatography. The inhibitors were homogeneous by 7.5% polyacrylamide gel electrophoresis; no isoinhibitors were found. Inhibitors WKB 858A and WKB 858B had isoelectric points of 5.0 and 4.65, respectively, and molecular weights of 42,000 and 20,000, respectively, by FPLC Superose 12 gel filtration chromatography. Inhibitor WKB 858A had molecular weights of 40,000 and 38,000 by Sephadex G-75 gel filtration chromatography and by native gel electrophoresis, respectively. Inhibitor WKB 858A contained 11.0% carbohydrate, N-linked to asparagine residues, with a composition of 1 fucose, 1 xylose, 4 galactose, 8 N-acetylglycosamine and 13 mannose residues per mol of inhibitor. Amino acid analysis of Inhibitor WKB 858A gave a high content of Asx, Glx, Ser, Thr and Val (combined total of 60% molar ratio) and low content of sulfur amino acids (0.8% molar ratio of Met and no 1/2 cystine). No-SH groups were found. The amino acid composition was similar to that of eight other a-amylase inhibitors from beans. Inhibitor WKB 858A formed a 1:1 stoichiometric complex with porcine pancreatic a-amylase with a Ki of 1.0 × 10^?11 M at pH 5.4 and 30C; it had no trypsin inhibitory activity. At pH 6.90 and 30C, the rate of complex formation between Inhibitor WKB 858A and porcine pancreatic β-amylase was 2.76 times faster at 1.385 vs 0.035 ionic strength (with Na₂SO₄), indicating hydrophobic bonds are most important in complex formation.     

Protein concentration of subcutaneous interstitial fluid in the human leg. A comparison between the wick technique and the blister suction technique

The wick technique and the blister suction technique are the most common methods for sampling of subcutaneous interstitial tissue fluid in man. The blister suction technique has the advantage of being less invasive than the wick technique, but the reliability of this method is still controversial. The aim of this study was to evaluate whether the simpler blister suction technique using large (8 mm) blisters could replace the wick technique in the investigation of patients with postreconstructive leg edema. Fifteen patients with ipsilateral leg edema following infrainguinal bypass surgery for lower limb atherosclerosis were investigated. The two different fluid sampling techniques were applied simultaneously on both legs. The concentration of total protein and albumin as well as colloid osmotic pressure of the subcutaneous interstitial tissue fluid in the leg were measured in all fluid samples. Agreement analysis was applied to compare the two methods, while the correspondence between the methods was estimated with linear regression analysis. The agreement index was found to be positive for all variables from the operated as well as from the contralateral control limb. Furthermore, all values were within the agreement limit. The best agreement between the two methods was found for colloid osmotic pressure on the operated side. According to the equation of linear regression, there was a slight overestimation of the wick values compared to the observed blister values. In conclusion, there was a good methodological agreement between the blister suction technique and the wick technique. The less invasive blister suction technique should be regarded as the method of choice for the investigation of subcutaneous interstitial tissue fluid in patients with postreconstructive leg edema.