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Biomedical subjects

J Merke

Publications and source records attributed to J Merke.

27 records · Page 2Linked to original sources

No 1,25-dihydroxyvitamin D3 receptors on osteoclasts of calcium-deficient chicken despite demonstrable receptors on circulating monocytes.

1,25-dihydroxyvitamin D3 (1,25(OH)2D3) is known to stimulate osteoclastic bone resorption in vivo and whole organ bone culture systems in vitro. It has not been established whether 1,25(OH)2D3 acts directly on osteoclasts or whether its action on osteoclasts is mediated via other bone cells (e.g., osteoblasts) or recruitment of osteoclast precursor cells. Circulating monocytes have been characterized as osteoclast precursors. In the present study, vitamin D3-replete chicken on a calcium-deficient diet were studied. Circulating monocytes, whole bone cell preparations, and isolated osteoclasts (differential sedimentation) were examined for presence of 1,25(OH)2D3 receptors. Reversible, specific, and saturable binding of [3H]-1,25(OH)2D3 to a 3.5 S macromolecule was demonstrated in nuclear fractions of monocytes (maximal binding capacity, 48 fmol/mg protein; dissociation constant, 1.3 X 10(-10) M) and of whole bone cell preparations. 1,25(OH)2D3 receptors were not demonstrable in osteoclast preparations (70% pure; detection threshold, 2 fmol/mg protein). Data are consistent with indirect action of 1,25(OH)2D3 on osteoclastic bone resorption.

Animals↗

Nuclear testicular 1,25-dihydroxyvitamin D3 receptors in Sertoli cells and seminiferous tubules of adult rodents.

1,25(OH)2D3 receptors were studied in whole testes, Sertoli cells, seminiferous tubules, Leydig cells and spermatogonia of adult NMRI mice and SD rats. Specific reversible high affinity binding (KD 1.4 x 10(-10)M; Nmax 72 fmol/mg protein) by a 3.5 S macromolecule was demonstrated in whole testes, Sertoli cells and seminiferous tubules. With identical techniques, no receptors were found in Leydig cells despite previous reports of 1,25(OH)2D3 actions on Leydig cell function.

Animals↗

Demonstration and characterization of 1,25-dihydroxyvitamin D3 receptors in basal cells of epidermis of neonatal and adult mice.

Nuclear and cytosolic receptors for 1,25-dihydroxycholecalciferol [1,25(OH)2D3] were demonstrated in the epidermis of neonatal and adult mice. The macromolecular binding protein sedimented at 3.5 S (sucrose density gradient) and was distinct from the 6.0 S binding protein for 25-hydroxycholecalciferol [25(OH)D3]. Analysis at different ionic strengths suggested the presence of unoccupied nuclear receptors. Digestion with proteases or nucleases, respectively, and inactivation with alkylating agents demonstrated that the binding macromolecule is a protein with SH groups at the active site. Binding of 1,25(OH)2D3 was specific and reversible. In neonatal mice KD was 1.6 X 10(-10) M for both cytosolic and nuclear fractions, binding capacity was 54 fmol/mg protein in the cytosolic and 108 in the nuclear fractions, respectively. The phenotypic expression of the 1,25(OH)2D3 receptor (dissociation constant, binding capacity) was identical in neonatal and adult epidermis. Half maximal displacement of 1,25(OH)2D3 was achieved with an 80-fold and 200-fold molar excess of 25(OH)D3 and 1-alpha-hydroxycholecalciferol [1(OH)D3], respectively. Using Percoll density gradient centrifugation, 1,25(OH)2D3 receptors could be localized in the basal cell fraction. DNA cellulose chromatography with 1,25(OH)2D3 receptor elution from DNA at 0.25 M KCl (linear gradient) points to a possible role in gene transcription. In mouse primary epidermal cell cultures, 1,25(OH)2D3, but not 25(OH)D3, 24,25(OH)2D3, and 1(OH)D3 influenced [3H]thymidine incorporation (at physiological concentrations); the magnitude of change depending on the concentration of 1,25(OH)2D3 and the time of incubation. These data demonstrate that skin is a target organ for the active vitamin D secosterol.

Animals↗

Demonstration and characterization of 1,25-dihydroxyvitamin D3 receptors in human mononuclear blood cells.

Circulating human mononuclear blood cells were studied for the presence of specific 1,25-dihydroxyvitamin D3 (calcitriol) binding macromolecules. Cells were isolated by density gradient centrifugation and characterized by surface markers. Specific reversible high affinity binding by a 3.5 S macromolecule was demonstrated in malignant B-cells and circulating monocytes. In monocytes specific calcitriol binding was found both in the presence and absence of vitamin D3 to saturate the vitamin D3 binding serum protein. No specific calcitriol binding was found in resting B or T lymphocytes. The data suggest a role of calcitriol in the control of mononuclear blood cell proliferation/differentiation.

B-Lymphocytes↗

Tissue compliance in superficial tissues along body axis in man.

A previously described miniature plethysmograph which allowed the measurement of tissue volumes in superficial tissues was enclosed in a small plexiglass chamber and attached to the frontal area, sternum, dorsum and the tibia. The tissues interposed between bone and skin underneath the chamber were exposed to pressures between +/- 3 and +/- 15 mmHg in order to test tissue deformability. The pressure application induced within the first 5 s a fast component of tissue deformation comprising between 75-90% of the total deformation followed by a slow component which lasted till the end of the pressure application. The highest deformability was found in the tissues of the sternum and dorsum whereas the stiffest tissues were in the pretibial area. Assuming the tissue deformation is due to a translocation of fluid into or out of the pressurized tissue, the tissue compliance was calculated. This calculated tissue compliance was 19.2 ml . 1,000 ml-1 . mmHg-1 in the sternum and 6.4 ml . 1,000 ml-1 . mmHg-1 (P < 0.01) in the pretibial area applying a pressure of +/- 3 mmHg. The differences observed are due to the morphological arrangement of the tissue fibres which in turn have to counteract the gravity forces to which the tissues are usually exposed during upright standing.

Animals↗

A new miniature plethysmograph to measure volume changes in small circumscribed tissue areas.

With an ultrasonic method tissue layer thickness was measured in man in circumscribed superficial tissue areas where the underlaying bone provided good backwall echos. In a 5 mm tissue layer changes of +/- 0.2% could be reliably detected. Knowing the height of the tissue cylinder between the surface of the skin and the bone allowed to calculate the tissue volume. The ultrasonic probes could therefore serve as miniature plethysmograph. Several probes were attached in the frontal region, sternum, along the vertebral column and along the tibia simultaneously. Changes of the volume content of the superficial shell tissues were induced by orthostasis, water immersion and heat exposure. It was possible to assess quantitatively the volume shifts into or out of the superficial tissues. During orthostasis 166 ml of fluid left the superficial tissues of the cephalad parts of the body and 164 ml could be traced in the dependent parts. Heat exposure was followed by a pooling 140 ml in the tissues studied. The most pronounced tissue volume changes were observed in the forehead region during heat exposure.

Electronics↗

Fluid volume distribution within superficial shell tissues along body axis during changes of body posture in man: the application of a new miniature plethysmographic method.

In up to six different sides along the body axis during tilting manoeuvres, volume shifts into or out off superficial tissues were followed with a newly developed miniature plethysmograph. It was possible to localize a region where no or only minor volume changes during the tilt table experiments occurred. This region is identical with the Hydrostatic Indifferent Point (HIP) being localized below the apex of the heart in the upper third of abdominal vena cava. Above the HIP fluid is drained out off the tissues during assumption of upright posture whereas below the HIP fluid volume is pooled. The volume changes occurred in two phases. Within the first 5 s in the cephalad parts of the body a rapid decrease occurred, thereafter the volume remained unchanged or even increased; below the HIP within the first 5 s a large volume increase was followed by a slow continuous volume increment. The functional peculiarities of the low pressure system as a whole were visible studying only superficial shell tissues of the body with the non invasive miniature plethysmographic technique.

Blood Circulation↗