Lymphatic bed of layers of the abdominal wall in the human inguinal area]

Lymphatic bed in the layers of the inguinal area was studied in connection with age in 70 human corpses. Polychromic injection of arteries, veins and lymphatic bed, staining of preparations after van Gieson, Weigert, with hematoxylin-eosin and morphometry were the methods applied. It was stated that lymphatic capillaries penetrate through all the layers forming the abdominal wall of the human inguinal area; they arrange interconnected networks in dermis, in external and internal oblique and transversal muscles and in their aponeuroses, as well as in fasciae and in the peritoneum. The lymphatic bed in question changes during ontogenesis. Age transformations of the lymphatic capillaries are in connection with functional loading on the anterior abdominal wall. Intraorganic connections existing between the lymphatic vessels of the anterior abdominal wall and the organs of the small pelvis (urinary bladder, uterus, rectum, etc.) are revealed, they are of a rather great interest for physicians.     

Assay for ADP-ribosyl cyclase by reverse-phase high-performance liquid chromatography.

Cyclic ADP-ribose (cADPR), a natural metabolite of beta-NAD(+), is a second messenger for Ca(2+) signaling in T cells. As a tool for purification and identification of ADP-ribosyl cyclase(s) in T cells, a sensitive and specific enzymatic assay using 1,N(6)-etheno-NAD(+) as substrate was developed. A major problem-the sensitivity of 1,N(6)-etheno-cADPR toward the extraction medium perchloric acid-was solved by replacing the perchloric acid extraction procedure of nucleotides by a filtration step. Standard compounds for the HPLC analysis of ADP-ribosyl cyclases and NAD(+)-glycohydrolases, e.g., 1,N(6)-etheno-cADPR, 1,N(6)-etheno-ADPR, and 1,N(6)-etheno-AMP, were produced by ADP-ribosyl cyclase from Aplysia californica and dinucleotide pyrophosphatase. The assay was applied to subcellular fractions prepared from human Jurkat T cells. As a result ADP-ribosyl cyclase and NAD(+)-glycohydrolase activity could be detected and precisely quantified in different subcellular fractions indicating the presence of different isoenzymes in T cells.     

CD38: a NAADP degrading enzyme

The role of the multifunctional enzyme CD38 in formation of the Ca(2+)-mobilizing second messenger nicotinic acid adenine dinucleotide phosphate (NAADP) was investigated. Gene silencing of CD38 did neither inhibit NAADP synthesis in intact Jurkat T cells nor in thymus or spleen obtained from CD38 knock out mice. In vitro, both NAADP formation by base-exchange and degradation to 2-phospho adenosine diphosphoribose were efficiently decreased. Thus in vivo CD38 appears to be a NAADP degrading rather than a NAADP forming enzyme, perhaps avoiding desensitizing NAADP levels in intact cells.     

Sulfide silver amplification of ferritin iron cores in blood and bone marrow cells. Methods, adaptations to microphysical analyses, and the impact of advanced iron overload

A short exposure of cell suspensions to gaseous hydrogen sulfide, appropriate fixations, and subsequent physical development of silver shells around sulfidated insoluble metals were used to amplify ferritin iron cores in blood and bone marrow cells. The methods described are suitable for both light microscopy and transmission electron microscopy. These techniques made it possible to visualize Prussian Blue stainable ferritin and haemosiderin, as well as a large variety of isoferritin iron and other smaller particles beyond the sensitivity of Prussian Blue staining. Admixtures of sulfidatible zinc and traces of other heavy metals had to be taken into consideration. For further research, adaptations of sulfide silver staining to microphysical analyses were developed. However, conventional energy dispersive X-ray analysis was not sensitive enough to signalize the presence of Fe in sulfide silver amplified iron cores of a single or a few ferritin molecule(s). Proton-induced X-ray emission was used to measure Fe and Zn down to 1 fg/single cell in unstained or sulfide silver stained smears on thin foils. However, multielement analysis of homogeneous cell concentrates was much easier to perform and far more sensitive. In advanced iron overload, highly increased sulfide silver staining was found in peripheral blood cells including lymphocytes, monocytes, eosinophils, basophils, and--in extreme cases--also in neutrophils and platelets.     

T-cell receptor-mediated metabolism of inositol polyphosphates in Jurkat T-lymphocytes. Identification of a D-myo-inositol 1,2,3,4,6-pentakisphosphate-2-phosphomonoesterase activity, a D-myo-inositol 1,3,4,5,6-pentakisphosphate-1/3-phosphatase activity and a D/L-myo-inositol 1,2,4,5,6-pentakisphosphate-1/3-kinase activity.

Stimulation of the human T-lymphocyte cell line Jurkat via the T-cell receptor/CD3 complex by an anti-CD3 antibody (OKT3) induced time-dependent changes in the intracellular concentrations of multiple inositol polyphosphate (InsPn) isomers. Quantitative mass analysis by anion-exchange HPLC and a recently developed postcolumn dye system (Mayr, G. W. (1988) Biochem. J. 254, 585-591) revealed basal intracellular concentrations between less than 5 pmol/10(9) cells for Ins(1,3,4,5)P4 and 6380 +/- 355 pmol/10(9) cells for InsP6. Time course analysis of samples from stimulated Jurkat T-cells showed an increase of Ins(1,3,4,5)P4 to 1125 +/- 125 pmol/10(9) cells within 10 min and remained elevated over more than 30 min. Moreover, increases of the intracellular concentrations of Ins(1,3,4,6)P4, Ins(1,4,5,6)P4, and/or Ins(3,4,5,6)P4 (determined as the enantiomeric mixture), Ins(1,3,4,5,6)P5, Ins(1,2,3,4,6)P5 and InsP6 were observed. In contrast, the concentration of Ins(1,2,4,5,6)P5 and/or Ins(2,3,4,5,6)P5 (determined as the enantiomeric mixture) decreased after stimulation. Using cytosolic extracts from Jurkat T-lymphocytes incubated with purified Ins(1,3,4,5,6)P5, Ins(1,2,3,4,6)P5, or Ins(1,2,4,5,6)P5/Ins(2,3,4,5,6)P5 three enzyme activities were observed. Ins(1,3,4,5,6)P5 was dephosphorylated by a phosphatase removing a phosphate group at the 1 and/or 3 position resulting in the formation of Ins(1,4,5,6)P4 and/or Ins(3,4,5,6)P4 (determined as the enantiomeric mixture). Ins(1,2,3,4,6)P5 was metabolized by a specific phosphatase that cleaved the phosphate group at the 2 position, thereby generating the product Ins(1,3,4,6)P4. On the other hand, Ins(1,2,4,5,6)P5/Ins(2,3,4,5,6)P5 was phosphorylated by a 1/3-kinase activity to InsP6. Together novel receptor-mediated metabolic pathways of inositol polyphosphates were demonstrated in human T-lymphocytes, and corresponding enzyme activities for the inositol pentakisphosphate metabolism were found in cell lysates.     

Die digitiformen Sensillen auf dem Maxillarpalpus von Coleoptera

Summary The digitiform sensilla on the distal segment of the maxillar palps ofAgabus bipustulatus (L.) andHydrobius fuscipes (L.) were investigated by electron microscopic methods. Each sensillum is innervated by a single bipolar sensory cell. The sensilla ofHydrobius are associated with three enveloping cells, which enclose an inner and outer receptor lymph cavity. A single enveloping cell only is found in the completely differentiated sensilla ofAgabus. These sensilla do not form an outer lymph cavity. The area beneath the hair base is filled by the distal process of the enveloping cell and by extensions of epidermal cells. Only one extra-cellular space exists, which seems to be homologous to an inner receptor lymph cavity.The outer dendritic segment surrounded by a dendritic sheath runs to the tip of the hair shaft. In the hair shaft the outer dendritic segment divides into several branches. The poreless hair shaft does not rise over the surface of the cuticle, but it is positioned in a narrow shallow groove. Special socket structures or a tubular body do not exist. The digiti-form sensilla possess neither the typical feature of mechanosensitive, nor gustatory or olfactory sensilla. The functional significance of the structural divergences in the sensilla of both species and the presumed function of the sensilla are discussed referring to hygro- and thermo-receptors.

Unserem verehrten Lehrer, Herrn Prof. Dr. H.Risler, dem wir für vielfache Förderung danken möchten, zum 65. Geburtstag gewidmet.