Mapping of estradiol binding sites through receptor micro-autoradiography in the endometrial stroma of early pregnant mice.

Estradiol triggers key biological responses in the endometrium, which rely on the presence and levels of its cognate receptors on target cells. Employing the receptor micro-autoradiography (RMAR) technique, we aimed to provide a temporal and spatial map of the functional binding sites for estradiol in the mouse endometrial stroma during early pregnancy. Uterine samples from days 1.

Pulse capture without carrier absorption in dynamic Q photonic crystal nanocavities.

We develop a gallium arsenide (GaAs) photonic crystal nanocavity device capable of capturing and releasing a pulse of light by dynamic control of the Q factor through free carrier photoexcitation. Unlike silicon-based devices where the performance of this dynamic optical control is limited by absorption from free carriers with nanosecond-order lifetimes, the short carrier lifetime (∼ 7 ps) of our equivalent GaAs devices enables dynamic control with negligible absorption losses. We capture a 4 ps optical pulse by briefly cycling the Q factor from 40,000 to 7900 and back just as the light couples to the nanocavity and confirm that the captured energy can be subsequently released on demand by a second injection of free carriers.

Whole-body and microscopic autoradiography to determine tissue distribution of biopharmaceuticals -- target discoveries with receptor micro-autoradiography engendered new concepts and therapies for vitamin D.

Information about the distribution of biopharmaceuticals is basic for understanding their actions. Tissue and cellular localization is a key to function. Autoradiography with radiolabeled compounds has provided valuable information with both low resolution whole-body macro-autoradiography and high resolution microscopic autoradiography (micro-autoradiography).

The world epidemic of sleep disorders is linked to vitamin D deficiency.

An observation of sleep improvement with vitamin D supplementation led to a 2 year uncontrolled trial of vitamin D supplementation in 1500 patients with neurologic complaints who also had evidence of abnormal sleep. Most patients had improvement in neurologic symptoms and sleep but only through maintaining a narrow range of 25(OH) vitamin D3 blood levels of 60-80 ng/ml. Comparisons of brain regions associated with sleep-wake regulation and vitamin D target neurons in the diencephalon and several brainstem nuclei suggest direct central effects of vitamin D on sleep.

Drugs in the brain--cellular imaging with receptor microscopic autoradiography.

For cell and tissue localization of drugs, receptor microscopic autoradiography is reviewed, including its development history, multiple testing, extensive applications and significant discoveries. This sensitive high-resolution imaging method is based on the use of radiolabeled compounds (esp. tagged with (3)H or (125)I), preservation through freezing of in vivo localization of tissue constituents, cutting thin frozen sections, and close contact with the recording nuclear emulsion.

In vivo target recognition with high-resolution imaging: significance for drug development.

In vivo target identification is basic for understanding mechanisms of drug action. Target identification requires cellular resolution. Extrapolation from blood bioavailability, low-resolution scans, radioassays, or in vitro tests regularly produce false-negatives and false-positives.

Vitamin D and the digestive system.

Target tissues of in vivo receptor binding and deposition of 1,25(OH)2 vitamin D3 and its oxygen analog OCT are reviewed in rats, mice, hamsters and zebra finch, identified with high-resolution microscopic autoradiography. Throughout the digestive system numerous sites with nuclear receptor binding of 3H-1,25(OH)2 vitamin D3 and 3H-OCT exist: in the oral region, epithelial cells of the oral cavity, tongue and gingiva, teeth odontoblast and ameloblast precursor pulp and stratum intermedium cells; in the parotid, submandibular and sublingual salivary glands, epithelial cells of striated ducts and granular convoluted tubules, intercalated ducts and acinar cells, as well as myoepithelial cells; in the stomach, neck mucous cells of gastric glands, endocrine cells of the antrum, and muscle cells of the pyloric sphincter; in the small and large intestine, absorptive and crypt epithelial cells; in the pancreas, predominantly islet B-cells. Perisinusoidal stellate (Ito) cells in the liver concentrate and retain variable amounts of radiolabeled compound in regions of their cytoplasm after administration of 3H-I,25(OH)2 vitamin D3 and 3H-25(OH) vitamin D3, probably sites of specific storage, similar to vitamin A.

Salivary glands epithelial and myoepithelial cells are major vitamin D targets.

Receptor binding with 3H-1,25(OH)2 vitamin D3 (vitamin D) and its oxygen analog 3H-OCT is demonstrated in rat, hamster, and mice submandibular, sublingual and parotid glands, using receptor microautoradiography high-resolution imaging. Nuclear uptake and retention of radiolabeled compound exist strongest in epithelial cells of striated ducts, granular convoluted tubules and in myoepithelial cells throughout, scattered in epithelial cells of intercalated ducts and relatively low in cells of serous and mucous acini. Deposition and retention of radiolabeled compound is also observed in interstitial spaces.

Skin research and drug localization with receptor microscopic autoradiography.

For the localization of drugs and related functional characterization, cellular-subcellular resolution can be achieved with radiolabelled compounds of high specific activity and receptor microscopic autoradiography, which is the method of choice for high-resolution qualitative and quantitative imaging. Detailed information together with integrative surveys can be obtained which is impossible with other methods. The history of discoveries of drug targets testifies to the utility and potential of receptor microscopic autoradiography that was designed to preserve in vivo conditions by excluding any liquid treatment during tissue preparation and to avoid translocation and loss of diffusible compounds.

Memo to the FDA and ICH: appeal for in vivo drug target identification and target pharmacokinetics Recommendations for improved procedures and requirements.

This memorandum is addressed to members of regulatory agencies, as well as managers of pharmaceutical companies. Pharmacokineticists and toxicologists may consider this proposal, weigh its merits, and provide input for implementation. Experience from academic research and ADME experiments during drug development has prompted this appeal for improved drug target recognition.

Sweat gland epithelial and myoepithelial cells are vitamin D targets.

Nuclear receptor binding of 1,25(OH)(2)-vitamin D(3) (vitamin D) in skin keratinocytes of epidermis, hair sheaths and sebaceous glands was discovered through receptor microscopic autoradiography. Extended experiments with (3)H-1,25(OH)(2)-vitamin D(3) and its analog (3)H-oxacalcitriol (OCT) now demonstrate nuclear receptor binding in sweat gland epithelium of secretory coils and ducts as well as in myoepithelial cells, as studied in paws of nude mice after i.v.

The dose makes the medicine.

Dose and time considerations in the development and use of a drug are important for assessing actions and side effects, as well as predictions of safety and toxicity. This article deals with epistemological aspects of dose selection by probing into the linguistic and cultural roots for the measure of medicine mediated by the medical doctor. Because toxicity is related to dose, historic and recent views suggest that less can be more.

[Micro-autoradiography: in vivo nuclear receptor binding of OCT].

Maxacalcitol, 1alpha,25-dihydroxy-22-oxavitamin D(3) (OCT), is a new synthetic analogue of 1alpha,25 (OH)(2)vitamin D(3) (1alpha,25 (OH)(2)D(3)), to be used for the treatment of secondary hyperparathyroidism. The side effect of hyper-calcemia can be prevented by short plasma half life of OCT, while there is a possibility of short retention in the target site. Micro-autoradiography is a powerful method to demonstrate the direct cellular distribution of drugs, especially nuclear receptor-binding materials as vitamin D.

Drug localization and targeting with receptor microscopic autoradiography.

This review is an argument in favor of better drug target identification. It presents the many merits and feasibilities of drug localization and target identification through the use of a suitable technique: receptor microautoradiography. Studies of drug targets and target bioavailability require methods with high resolution and sensitivity to gain information for understanding mechanisms of action, sound modeling, prediction of effects, and toxicity.

Receptor microscopic autoradiography for the study of percutaneous absorption, in vivo skin penetration, and cellular-intercellular deposition.

Introduction: Microscopic autoradiography with cellular resolution and preservation of in vivo conditions is potentially the method of choice to gain detailed information about sites of deposition and retention in the epidermis and of penetration to the dermis after topical application of drugs. We tested this using (3)H-Maxacalcitol.

Methods: Dorsal skin of adult rats was treated in vivo with ointment containing 1 or 40 microg/kg body weight of the vitamin D analogue (3)H-Maxacalcitol for periods of 0.

Estradiol receptor binding to the epithelium of uterine lumen and glands: region- and time-related changes during preimplantation and periimplantation periods studied by autoradiography.

The presence and changes of estradiol nuclear binding and related functions in uterine luminal and glandular epithelium were studied before and after blastocyst implantation using receptor autoradiography with (3)H-estradiol-17beta in association with (3)H-thymidine incorporation and immunocytochemical binding of antibody to estrogen receptor ER-alpha. (3)H-estradiol nuclear binding is present but variable during days 1.5-7.

In vivo time-course of receptor binding in the parathyroid gland of the vitamin D analogue [(3)H]1,25-dihydroxy-22-oxavitamin D(3) compared with [(3)H]1,25-dihydroxyvitamin D(3), determined by micro-autoradiography.

1,25-Dihydroxy-22-oxavitamin D(3) (22-oxacalcitriol, OCT), is a new synthetic analogue of 1,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3), calcitriol), to be used in the treatment of secondary hyperparathyroidism. This study used receptor micro-autoradiography in the parathyroid gland to determine and compare the time-course of receptor binding between OCT and 1,25(OH)(2)D(3). Mice were injected with 4 microg/kg of [26-(3)H]OCT or [26,27-methyl-(3)H]1,25(OH)(2)D(3), and killed at 5, 15, 30 min, 1, 2, 4, 8, 12, and 24 h afterwards.

Synthesis and pharmacokinetics of 1 alpha-hydroxyvitamin D3 tritiated at 22 and 23 positions showing high specific radioactivity.

A novel synthesis of a radioactive compound of 1 alpha-hydroxyvitamin D3 (1 alpha OHD3) (1) and its pharmacokinetics are described. Radioactive 1 alpha OHD3 tritiated at 22 and 23 positions ([22,23-(3)H4]1 alpha OHD3) (5) was prepared via key reactions of the reduction of acetylenic side chain in the ketone (12) with tritium gas in the presence of palladium-charcoal and the subsequent Wittig reaction with the A-ring synthon (16). [22,23-(3)H4]1 alpha OHD3 (5) showed high specific radioactivity (111.

In vivo nuclear uptake of a vitamin D analog (OCT) in different tumor cell populations of FA-6 cancer xenograft in nude mice by receptor autoradiography.

1 alpha, 25-dihydroxyvitamin D3 [1 alpha, 25(OH)2D3] and its analogs have been shown to repress the production of parathyroid hormone-related peptide (PTHrP) in tumors, which is a major factor causing humoral hypercalcemia associated with various cancers. Since vitamin D analogs may be applicable to the treatment of cancer patients, the present study was undertaken to examine whether OCT, an analog with little calcemic activity, is incorporated into tumor tissues, and to identify cellular and subcellular sites of its specific uptake and retention. [26-3H]OCT was injected i.