Serum alkaline phosphatase isoenzymes as markers of liver disease

A perspective on serum alkaline phosphatase isoenzymes in liver disease is provided with a brief discussion of the location of the enzyme in liver and its presumed function. Mechanisms of entry of alkaline phosphatase into serum in liver disease are discussed. Characterization of high molecular weight alkaline phosphatase in obstructive jaundice is reviewed. The relationship between blood group O and the appearance of the intestinal enzyme in sera of such subjects with cirrhosis of liver is discussed. Properties of hepatoma alkaline phosphatase and the genesis of liver alkaline phosphatase in diseases not related to the liver are explored. Methods for detection of serum alkaline phosphatase isoenzymes in liver disease are discussed from the standpoint of the limitations of electrophoretic procedures, and the promise of procedures such as isoelectric focusing and high performance liquid chromatography that are currently non-routine.     

Neuronal maturation and synaptogenesis in the rat ventrobasal complex: Alignment with developmental changes in rate and severity of axon reaction

Developmental alterations in the rate and severity of axon reaction are thought to be influenced by the level of neuronal maturation and synaptogenesis. As an initial step in more carefully examining this issue, two concurrent series of studies were performed using the ventrobasal complex as a model. Sprague-Dawley albino rats, ranging in age from neonate through 60 days postnatal (dpn) were subjected to unilateral aspiration lesions encompassing somato-sensory cortex. Two to four animals were sacrificed at various postoperative periods of from one through 40 days. The reactive ventrobasal (VB) complex was qualitatively and quantitatively analysed employing electron microscopy and the Cajal-DeCastro reduced silver technique or a hematoxylin-eosin-Orange-G method for light microscopic study. Neuronal degenerative changes are prominent within 48 hours following ablation of neonate through 9 dpn animals. These alterations include a loss of cytoplasmic basophilia together with vacuolization, and rapid disappearance of the cell. Swelling and vacuolization of axons and dendrites is seen throughout the zone of degeneration accompanied by the rapid accumulation of numerous reactive non-neuronal elements, some of which resemble those mesodermal elements designated as “M” cells. Following equivalent lesions of animals 12 through 60 dpn, neurons within the VB undergo degenerative changes which are qualitatively similar to those seen in younger animals but require between 6 and 20 days postoperative to appear. While most such cells will disappear the degenerative process is temporally extended by as much as one order of magnitude. The normal ventrobasal complex was examined in 7,12,15,20, and 60 dpn rats. Golgi-Cox impregnations of thalamic neurons reveal such cells at 7 dpn to be characterized by short, stubby, irregular dendrites with distinct growth cones and filopodia, whereas by 12 dpn Golgi analysis demonstrates prominent morphological changes indicative of considerable progress in thalamic neuronal differentiation including dendritic elongation with secondary and tertiary branching, and the appearance of spinous protrusions. The use of the Rasmussen stain for synapses, as well as electron microscopy, shows distinctive trends of synaptogenesis typified by a paucity of synaptic complexes but large numbers of dendritic and axonal growth cones at 7 dpn. However, by 12 dpn a 3-fold increase in the number of synapses has taken place together with an enhanced morphological complexity and variety of individual synaptic profiles, some of which are known to represent lemniscal input. Further prominent increases in synaptic population density are observed at 15dpn but the rate of synaptogenesis appears to become attenuated after this period. Finally, patterns of distribution of perikaryal RNA become altered from a predominance of polyribosomes at 7 dpn to characteristic multilamellate aggregations of granular endoplasmic reticulum initially observed at 12 dpn. With subsequent neuronal enlargement this component proportionally increases in amount but is qualitatively unchanged with respect to morphological arrangement. The close temporal correlation between the specific developmental events documented in this study and the postnatal age during which VB neurons become less susceptible to injury suggests that important changes in maturation of intrinsic neuronal metabolism involving rates of RNA and protein synthesis may be, in part, initiated and regulated by extrinsic factors related to the establishment of pre-synaptic input upon VB neurons. A number of putative molecular substrates for these influences are discussed.     

Analysis of spatial correlation between 99mTc-Sestamibi uptake and radiographic breast density

Breast scintigraphy is a technique by which the biological properties of breast lesions can be assessed using an injected radiopharmaceutical. It may be particularly useful for women with radiographically dense breasts, in whose mammograms, lesions are often obscured by breast tissue. We are evaluating a dual modality breast scanner developed at the University of Virginia for its ability to distinguish between benign and malignant lesions. The scanner obtains a digital mammogram and a gamma ray emission image in quick succession with the breast held under mild compression, resulting in a fused image in which structures in the digital mammogram can be directly correlated with those in the scintigram. Our experience has shown that radiopharmaceutical uptake by normal breast tissue can sometimes obscure uptake by small lesions. It would therefore be advantageous to correct for this background uptake if possible. One potential way of accomplishing this is to use the information from the digital mammogram to help predict the background radiopharmaceutical distribution. With this in mind, we retrospectively investigated the degree of spatial correlation between the distribution of background activity and the distribution of radiodense breast tissue in normal breasts. Using a histogram-based analysis, we have quantified the degree of correlation in 16 images obtained from a total of 8 patients. We also used the mammographic images to quantify the radiographic density of each breast. Our results suggest that spatial correlation between areas of high radiopharmaceutical uptake and parenchymal density exists in the most dense regions of the breast for either extremely dense or heterogeneously dense breasts. High correlation was also observed for some homogeneously fatty breasts. In the latter case however, variation in breast thickness appeared to be the cause of the increased correlation. Correlation properties are approximately equal in both right and left breasts for a particular patient, except in cases exhibiting focal radiotracer uptake in a lesion. Although our preliminary results suggest that correlation between radiopharmaceutical uptake and parenchymal density exists, the number of cases thus far is too small for definitive conclusions. In addition, the planar nature of the dual modality scans imposes an inherent limitation on our ability to take into account attenuation of the emitted gamma radiation, which thus constitutes an uncontrolled variable in the correlation analysis. In principle, this problem can be eliminated by 3-dimensional imaging.     

Real Time-PCR HBV-DNA Analysis: Significance and First Experience in Armed Forces

Background

HBV DNA quantitation is used extensively world wide for the diagnosis and monitoring of treatment of Hepatitis B virus (HBV) infection. However, it has still to be popular in India. The aim of this study was to quantitate HBV – DNA by Real time – PCR method in Hepatitis B and in immuno-compromised patients, to compare the results with HBeAg detection and to monitor the response to therapy of chronic Hepatitis B patients to antivirals.

Methods

Ninety one serum samples of Hepatitis group of patients (all HBsAg positive), 41 samples from immuno-compromised patients (all HBsAg negative) and 49 patients of Chronic Hepatitis B group (all HBsAg positive) were the subjects of this first ever study in Armed Forces. Twenty serum samples from healthy volunteers and non-hepatitis B patients served as negative controls. The amplification detection was carried out in a Rotor-Gene 2000-sequence detector

Results

Amongst Hepatitis B group, 33% (30/91) of the samples were positive for HBV-DNA and 26% (24/91) of samples were positive for HBeAg. In the immuno-compromised group of patients 14.6% (6/11) of samples were positive for HIV-DNA and 9.7% (4/41) were positive for HBeAg. Of the Chronic Hepatitis B patients on treatment, all (100%) were positive by HBV-DNA, whereas 29/49 (59.2%) were positive by HBeAg before treatment. After treatment with antivirals, 06/49 (12.2%) were positive by both tests and 11/49 (22.5%) were positive only by HBV-DNA. 32/49 (65.3%) patients became negative serologically after therapy.

Conclusion

HBeAg status did not necessarily reflect HBV-DNA level in the serum, as 10/91 (11%) in the Hepatitis B group, 2/41 (4.9%) in the immuno compromised group and 20/49 (40.8%) patients in the Chronic Hepatitis B group were positive for HBV-DNA but negative for HBeAg. HBV-DNA was not found to be positive amongst any of the negative controls. Real time – PCR is a sensitive and reproducible assay for HBV-DNA quantitation and may be started in Armed Forces referral centers in the near future.Key Words: Real time – PCR, Chronic Hepatitis B, HBV – DNA, Antivirals     

A simple and inexpensive device for microvascular training

A simple and inexpensive device is described that is easy to assemble and provides an excellent model for developing sound microvascular technique.