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J P Germain

Publications and source records attributed to J P Germain.

14 recordsLinked to original sources

Zonation of gluconeogenesis, ketogenesis and intracellular pH in livers from normal and diabetic ketoacidotic rats: evidence for intralobular redistribution of metabolic events in ketoacidosis.

The intralobular distribution of metabolism was examined in the livers from rats with severe diabetic ketoacidosis (DKA), perfused at pH 6.8, and compared with that in livers from normal starved animals perfused at either pH 7.4 or 6.8. With lactate and palmitate as substrates, the perivenous uptake of periportally synthesized glucose seen in normal livers at pH 7.4 was abolished during DKA; indeed, gluconeogenesis was most active in the perivenous region. Whereas in normal livers perfused at pH 7.4 the periportal region showed a markedly elevated intracellular pH (pH(i)) compared with the perivenous zone, this distribution of pH(i) was reversed in DKA, with an intermediate distribution in normal livers perfused at pH 6. 8. 3-Hydroxybutyrate was generated throughout the lobule. Some acetoacetate generated periportally was converted to 3-hydroxybutyrate more perivenously. A steep gradient of oxygen uptake along the radius of the lobule was apparent in all three groups; oxygen uptake was greatly decreased perivenously despite adequate oxygen supply. These findings are consistent with our previous observations of the lobular co-location of high pH(i) and gluconeogenesis, and might offer an explanation of how high gluconeogenic rates can continue in spite of severe systemic acidosis in DKA. The findings provide direct evidence for a marked redistribution of intralobular metabolism in DKA.

Animals↗

Gluconeogenesis, glucose handling, and structural changes in livers of the adult offspring of rats partially deprived of protein during pregnancy and lactation.

Maternal protein restriction is a model of fetal programming of adult glucose intolerance. Perfused livers of 48-h- starved adult offspring of rat dams fed 8% protein diets during pregnancy and lactation produced more glucose from 6 mM lactate than did control livers from rats whose dams were fed 20% protein. In control livers, a mean of 24% of the glucose formed from lactate in the periportal region of the lobule was taken up by the most distal perivenous cells; this distal perivenous uptake was greatly diminished in maternal low protein (MLP) livers, accounting for a major fraction of the increased glucose output of MLP livers. In control livers, the distal perivenous cells contained 40% of the total glucokinase of the liver; this perivenous concentration of glucokinase was greatly reduced in MLP livers. Intralobular distribution of phosphenolpyruvate carboxykinase was unaltered, though overall increased activity could have contributed to the elevated glucose output. Hepatic lobular volume in MLP livers was twice that in control livers, indicating that MLP livers had half the normal number of lobules. Fetal programming of adult glucose metabolism may operate partly through structural alterations and changes in glucokinase expression in the immediate perivenous region.

Animals↗

A method for determination in situ of variations within the hepatic lobule of hepatocyte function and metabolite concentrations.

A method is described for the production of detailed maps of intralobular variations of hepatocyte function and metabolite concentrations, based on variable destruction by digitonin of the lobule from the centrilobular direction. Instead of the conventional approach, in which isolated hepatocytes are then prepared and studied in suspension, perfusion is continued after digitonin treatment and the function of the unaffected lobular remnants is determined, or mean metabolite concentrations are measured by 31P-NMR. These measurements are plotted against the degree of destruction, determined precisely after each study by automated quantitative histomorphometry. These plots are transformed into curves of the function or metabolite concentration of nominal single cells at any point along the radius of the lobule. Gluconeogenesis from lactate remained stable, although reduced, even after 85-90% lobular destruction, predominated periportally and disappeared by 50% along the radius of the lobule. In 31P-NMR studies, employing 1.5 mM lactate as substrate, narrowing of the intracellular P1 resonance was observed as digitonin destruction increased; this was attributed to a decrease in the intralobular heterogeneity of the intracellular pH, which fell from approx. 7.9 to < 7.4 along the first 16% of the lobular radius (from the periportal end) and to < 7.3 in the remainder of the lobule. The ATP concentration rose, and then fell, along the radius of the lobule in a centripetal direction. The method is potentially generally applicable to a wide range of hepatocellular functions and to the measurement of metabolite concentrations, most conveniently those susceptible to estimation by NMR.

Animals↗

Histochemical studies of acid and alkaline phosphatases in rat tooth germs with undecalcified resin-embedded specimens.

A novel technique for the histochemical demonstration of acid phosphatase (AcPase) and alkaline phosphatase (AkPase) in hard tissues has been proposed. Fresh, unfixed, undecalcified samples of rat tooth germs and surrounding structures were embedded in LR Gold resin at -20 degrees C. Sections of 2 microns were taken and subsequently processed for enzyme histochemistry. AkPase reaction product appeared as strong linear staining outlining cell boundaries and was present in the enamel organ, dental pulp, and osteoblast cells. Tartrate-resistant AcPase staining was seen exclusively in the osteoclasts of developing alveolar bone. Our results demonstrated that the use of unfixed, undecalcified LR Gold resin-embedded specimens for histochemistry is a novel technique which may be of value for certain studies when decalcification of specimens is undesirable. The technique appears to give good preservation of enzyme activity combined with the ability to prepare sections with excellent morphological detail.

Acid Phosphatase↗

Stable acid phosphatase: I. Demonstration and distribution.

After a pH-dependent reactivation a highly stable form of acid phosphatase (SAPhase) could be demonstrated in active cells of the macrophage/giant cell/osteoclast series and also in epiphyseal chondrocytes, in cells lining bone undergoing resorption and in hamster eosinophils. Because acid phosphatases of epithelial cells in rat, hamster and Macaca sp. tissues did not possess this stability, SAPhase served as a useful cell marker for the above mesenchymal cell types in paraffin and glycol-methacylate sections even after rapid demineralization in acidic buffers. Conformational alterations appear to occur in the enzyme during formaldehyde fixation, embedding, and reactivation. The granular staining of SAPhase and the successful use of a non-aqueous fixative suggest an association of SAPhase with lysosomes and their membranes. Cells of mesencymal origin that are actively engaged in intra- and/or extracellular digestion contain high levels of SAPhase. The distribution and properties of SAPhase indicate an interrelationship between mononuclear and mutinuclear cell types actively engaged in such digestive processes.

Acid Phosphatase↗

The gingival epithelium of rodent molars with limited eruption.

We have re-examined the arrangement of epithelia surrounding molar teeth with limited eruption in the rat, mouse and hamster. Our methods permitted enamel to be retained in paraffin and glycolmethacrylate sections, so providing optimal preservation of epithelial relationships. Three basic patterns of epithelial arrangement were observed on the various aspects of molars. Non-keratinized junctional epithelium covers the interdental septum and the base of all gingival crevices even in areas of gingival recession. In all areas other than the interdental septum, a fold of keratinized gingival epithelium lies beneath the lateral aspects of junctional epithelium. The buccal and lingual aspects of interdental septa show a zone of transition between these two basic types of epithelial arrangement. In intact gingiva the junctional epithelium can be seen to extend a considerable distance on to enamel surfaces, with the result that actual gingival crevice depths are even less than previously assumed. Artefacts resulting from loss of enamel during processing are discussed in the light of previous attempts to explain epithelial arrangements in rodent gingivae.

Animals↗

Rapid demineralization in acidic buffers.

The demineralization of routine histological specimens in buffers of weakly ionized organic acids, unbuffered formic acid, and EDTA was investigated. The rate of demineralization was measured by a chemical method and from radiographs. Lactate-containing buffers and buffers of formic acid with its potassium salt were more rapid in effect than any other agent. Acidic buffers and unbuffered formic acid produced rapid diffuse demineralization with secondary precipitation of calcium salts. Preservation of dental enamel in such buffers resulted from the significantly slower rate of enamel demineralization than that for bone and dentine. In rapid demineralizing agents the secondary salts were quickly redissolved while in slow buffers these salts persisted. Multivalent ions such as citrate and maleate slowed the rate of demineralization, and a citrate-containing buffer was the slowest of all the agents tested. Demineralization in EDTA exhibited a different pattern with the establishment of a well-defined front of demineralization without apparent reprecipitation. EDTA attacked enamel, bone and dentine at the same rate. An attempt was made to relate the observed rates of demineralization to current theories of the demineralization process.

Animals↗

Dermatologic problems in amputees.

Improvements in surgical techniques and prosthetic devices and the establishment of prosthetic clinics have altered the outlook for the amputee. A cooperative effect on the part of professionals looking after amputee patients, as carried out in these clinics, offers the best means of recognition and treatment of difficulties as they arise. Until the "bionic man" becomes a reality, amputees will continue to have skin problems. In describing a number of illustrative cases, we have attempted to: (a) renew interest in these problems; (b) demonstrate the value of the group approach; and (c) encourage the participation of interested dermatologists in the prosthetic clinic team to facilitate earlier recognition and treatment of troublesome skin disorders in the amputee.

Adolescent↗

Osteopetrosis.

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Adult↗

The reversal of tissue differentiation around screws.

Mechanical factors influence the morphology and function of differentiated as well as undifferentiated cells. Observations of tissues surrounding screws, during a 4-week period of stability followed by a 4-week period of instability, demonstrate resorption of the newly formed callus and subsequently to screw lossening. Four weeks of instability followed by 4 weeks of stability produced not only a cessation of the osteoclastic activity, but also the appearance of an osteoclastic activity. Thus, the holding power of screws is not finally determined at the moment of their insertion nor after cellular differentiation has occurred around the screws. Local mechanical conditions throughout the period of fracture consolidation are of eqaul importance.

Animals↗