Computed tomography osteoabsorptiometry-based investigation on subchondral bone plate alterations in sacroiliac joint dysfunction.

Sacroiliac joint dysfunction (SIJD) is an underappreciated source of back pain. Mineralization patterns of the sacroiliac (SIJ) subchondral bone plate (SCB) may reflect long-term adaptations to the loading of the joint. Mineralization densitograms of 27 SIJD patients and 39 controls, were obtained using CT osteoabsorptiometry. Hounsfield unit (HU) values of the SCB mineralization of superior, anterior and inferior regions on the iliac and sacral auricular surfaces were derived and statistically compared between SIJD-affected and control cohorts. Healthy controls showed higher HU values in the iliac; 868 ± 211 (superior), 825 ± 121 (anterior), 509 ± 114 (inferior), than in the sacral side; 541 ± 136 (superior), 618 ± 159 (anterior), 447 ± 91 (inferior), of all regions (p < 0.01). This was similar in SIJD; ilium 908 ± 170 (superior), 799 ± 166 (anterior), 560 ± 135 (inferior), sacrum 518 ± 150 (superior), 667 ± 151 (anterior), 524 ± 94 (inferior). In SIJD, no significant HU differences were found when comparing inferior sacral and iliac regions. Furthermore, HU values in the inferior sacral region were significantly higher when compared to the same region of the healthy controls (524 ± 94 vs. 447 ± 91, p < 0.01). Region mineralization correlated negatively with age (p < 0.01). SIJD-affected joints reflect a high mineralization of the sacral inferior region, suggesting increased SIJD-related mechanical stresses. Age-related SCB demineralization is present in all individuals, regardless of dysfunction.

Intrahepatic distribution of human glutathione-dependent formaldehyde dehydrogenase.

By means of a microelectrophoretic separation technique the different forms of alcohol dehydrogenase can be detected in microdissected liver tissue samples of the nanogram range. Alcohol dehydrogenase 3 (the glutathione-dependent formaldehyde dehydrogenase) was demonstrated to be zonally distributed in the human liver parenchyma. A periportal/perivenous gradient is evident in both sexes, however, the periportal/perivenous ratio is higher in males. The biological functions of this enzymatic form and the possible role of the periportal maximum are discussed.

Distribution of alcohol dehydrogenase isoenzymes in the human liver acinus.

By the use of a newly developed technique of ultrathin-layer electrophoresis, class I and class II alcohol dehydrogenase activity could be demonstrated in microdissected samples of the periportal, intermediate, and perivenous zones of the liver acinus in men and women. It could be demonstrated that both classes exhibit low activity in the periportal zone. From there, a rising gradient in the direction of the perivenous end was apparent. This increase, however, was found to be significant only in women. The analysis of class I alcohol dehydrogenase isoenzymes showed that the expression of alpha-, beta-, and gamma-containing isoforms did not differ in relation to the intraacinar position. The constant proportions of the isoenzymes to the maxima and minima of the total alcohol dehydrogenase activity support the view that the adult liver-specific isoenzyme pattern is determined during postnatal development.

Ultrathin-layer zone electrophoresis of alcohol dehydrogenase in partly rehydrated polyacrylamide gels: an alternative to starch gel electrophoresis.

A highly sensitive electrophoretic technique for the separation of alcohol dehydrogenase isoenzymes by zone electrophoresis in partly rehydrated polyacrylamide gels is described. Five hundred microm thin polyacrylamide gels are polymerized under standardized conditions. After polymerization the gels are washed thoroughly with distilled water to remove any unreacted monomers, catalysts or still soluble polymers. The washed gels are then impregnated with 0.5% Tween 20 and dried. Before electrophoresis the dry gels are rehydrated to a thickness of 250 microm, which makes up 50% of the original gel volume. Rehydration is carried out by use of a degassed buffer solution. This method permits the demonstration of the isoenzymes of alcohol-dehydrogenase class I and II in man and allows quantitative determination.

Intra-acinar profiles of cytosolic and mitochondrial malate dehydrogenase isoenzymes in rat liver.

A new method for electrophoretic separation of the isoforms of malate dehydrogenase in microdissected tissue samples was applied to rat liver. The intra-acinar activity profiles of cytosolic (cMDH) and mitochondrial (mMDH) malate dehydrogenase were determined in male and female control animals and in animals fasted for 84 hr. Measurements were carried out on lyophilized liver tissue samples of 50-100 ng from the whole length of the sinusoid. The results showed that both in fed and fasted animals, mMDH activity was almost evenly distributed throughout the acinus in livers of both sexes. cMDH showed higher activity in the periportal area compared with the perivenous area by a factor of approximately 1.35 in all animals studied. Our results favor a slightly higher capacity of the malate-aspartate shuttle in the periportal area in both fed and fasted animals. Furthermore, the distribution pattern of mMDH suggests that this isoenzyme is not a marker for the zonation of the oxidative metabolism in the liver acinus.

Ultrathin-layer microelectrophoretic determination of lactate dehydrogenase isoenzymes in corneal and conjunctival epithelium of the cow.

In order to evaluate the impact of tissue oxygenation on the distribution pattern of lactate dehydrogenase isoenzymes, activities of the isoenzymes were measured in microdissected samples of bovine tissue. A highly sensitive ultrathin-layer electrophoretic technique was used to determine the distribution pattern of lactate dehydrogenase isoenzymes in basal, intermediate and superficial layers of the epithelium of central and peripheral cornea and in the epithelium of the bulbar conjunctiva. Measurements revealed almost homogeneous intraepithelial distribution patterns of lactate dehydrogenase isoenzymes in both tissues. In the cornea the lactate dehydrogenase isoenzymes 4 and 5, which are regarded to be specialized for anaerobic glucose metabolism, were found to predominate. In the well-oxygenated conjunctival epithelium most of the activity could be ascribed to the lactate dehydrogenase isoenzyme 3. In contrast to the isoenzymatic activities, total activity of lactate dehydrogenase was inhomogeneously distributed; maximum activities were found in the basal layer of corneal epithelium and in the intermediate layer of conjunctival epithelium. The results indicate that oxygen supply is relevant rather for the intraepithelial distribution of total enzyme activity than for the expression of lactate dehydrogenase isoenzymes.

Ultrathin-layer zone electrophoresis of lactate dehydrogenase isoenzymes in microdissected liver samples.

A highly sensitive electrophoretic separation of lactate dehydrogenase isoenzymes in 150-microns ultrathin-layer polyacrylamide gels is described. By means of the incorporation of miniature-sized wells into the gel and by executing all the analytical steps under paraffin oil, this technique allows the exact and sharp separation of the lactate dehydrogenase isoenzymes in nanogram-sized microdissected liver tissue samples. With this method the heterogeneous distribution pattern of lactate dehydrogenase isoenzymes in the liver acinus of various mammals could be demonstrated, and a new interpretation of their functional roles is offered.