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PubMedDate not supplied

Protoporphyrin derivatives--the use in localization of neoplasmas by titanium laser-induced fluorescence technique.

Examples of in vivo detection of skin neoplasmas (Mercl, melanoma) and breast carcinoma using laser induced fluorescence (LIF) and bis-1[1-alanylo-N]-ethylodeuteroporphyrin (Ala-PP) are described in this study. Photosensitizer, wave titanium laser and standard CCD camera coupled to vision amplifier enable precise neoplasm imaging, Ala-PP retention monitoring and further photodynamic therapy (PDT).

Breast Neoplasms↗
PubMed1983

Fine structure and axonal transport labeling of intraepithelial sensory nerve endings in anterior hard palate of the rat.

The fine structure of intraepithelial free nerve endings, Merkel receptors, and special (possibly chemosensory) corpuscular receptors has been examined in anterior hard palate of the rat. Samples were taken from the two anterior large rugae as well as from the more rostral incisive papilla. Most nerve endings were concentrated in regions in which the epithelium and connective tissue interdigitated extensively. The nerve endings contained relatively pale cytoplasm, a variety of vesicles, and numerous mitochondria. Distinctions could be made among different endings depending on their intraepithelial location, or on the degree of specialization of the neighboring epithelial cells. Simple (free) nerve endings entered the epithelium between epithelial papillae; upon entering, their basal lamina became associated with the epithelial basal lamina in various ways, and their Schwann cell sheath also entered the epithelium for a short length. Free nerve endings were found at various sites among the regular epithelial cells in the basal layers, the stratum granulosum, and stratum spinosum, and next to the stratum corneum border; they did not form specialized junctions with the epithelial cells. Intraepithelial nerve endings at all levels could be labeled by axonal transport from the trigeminal ganglion. Some nerve endings in anterior hard palate were associated with specialized epithelial cells, either in Merkel complexes or in apparent chemosensory corpuscles; both were labeled by axonal transport from the trigeminal ganglion. The specialized epithelial cells in the corpuscular receptors formed possible efferent and afferent synapses with the nerve endings. The nerve endings in the Merkel complexes formed junctions with the specialized Merkel epithelial cells. Both the Merkel complexes and corpuscular receptors were found in their respective characteristic epithelial papillae, which were surrounded by a dense plexus of subepithelial terminal axons and nerve endings.

Animals↗
PubMed1998

Uptake and release of neurotransmitter candidates, [3H]serotonin, [3H]glutamate, and [3H]gamma-aminobutyric acid, in taste buds of the mudpuppy, Necturus maculosus.

Neurotransmitters in vertebrate taste buds have not yet been identified with confidence. Serotonin, glutamate, and gamma-aminobutyric acid (GABA) have been postulated, but the evidence is incomplete. We undertook an autoradiographic study of [3H]serotonin, [3H]glutamate, and [3H]GABA uptake in lingual epithelium from the amphibian, Necturus maculosus, to determine whether taste bud cells would accumulate and release these substances. Lingual epithelium containing taste buds was incubated in low concentrations (0.4-6 microM) of these tritiated transmitter candidates and the tissue was processed for light microscopic autoradiography. Merkel-like basal taste cells accumulated [3H]serotonin. When the tissue was treated with 40 mM K+ after incubating the tissue in [3H]serotonin, cells released the radiolabelled transmitter. Furthermore, depolarization (KCl)-induced release of [3H]serotonin was Ca-dependent: if Ca2+ was reduced to 0.4 mM and 20 mM Mg2+ added to the high K+ bathing solution, Merkel-like basal cells did not release [3H]serotonin. In contrast, [3H]glutamate was taken up by several cell types, including non-sensory epithelial cells, Schwann cells, and some taste bud cells. [3H]glutamate was not released by depolarizing the tissue with 40 mM K+. [3H]GABA uptake was also widespread, but did not occur in taste bud cells. [3H]GABA accumulated in non-sensory epithelial cells and Schwann cells. These data support the hypothesis that serotonin is a neurotransmitter or neuromodulator released by Merkel-like basal cells in Necturus taste buds. The data do not support (nor rule out) a neurotransmitter role for glutamate or GABA in taste buds.

Animals↗
PubMed2003

Multi-photon microscopy of cell types in the viable taste disk of the frog.

The morphology of viable taste disks of the frog was explored with multi-photon microscopy. In order to identify single sensory or supporting cells within the tissue, we searched for fluorescent dyes that stained subsets of the cell population or possibly cell types. Some cell types indeed stained preferentially with certain fluorescent dyes. A subset of glia-like cells (type Ic) stained with BCECF, a H+-sensitive dye, and indo-1, a Ca2+-sensitive dye, both presented in the membrane-permeant ester form. BCECF-ester also stained the dendrites of type III receptor cells, but indo-1 ester did not. Receptor cells of type II stained with MQAE, a positively charged Cl- -sensitive dye. A subset of type II cells accumulated amiloride, a positively charged fluorescent diuretic. Certain supporting cells, i.e., wing cells (type Ib) and glia-like cells (type Ic), were labeled by negatively charged dyes, e.g., calcium green-1 dextran. Mucus cells (type Ia) were stained with only two of the 19 dyes examined, and Merkel-like basal cells (type IV) were stained only with a membrane-labeling voltage-sensitive dye, presumably by endocytosis. No dye was found which would stain all types of cells or all receptor cells. This finding reveals a potential problem for future functional imaging aiming at population responses, as the responses of unstained cells then would remain unobserved. Specificity of dyes with respect to cell types was sufficient to identify supporting cells and receptor cells. Cell shape could then be reconstructed, using optical slicing and rendering techniques. Thus populations of dye-loaded elongated cells, especially types Ic, II and III, could for the first time be visualized in three dimensions.

Acridines↗
PubMed1988

[DNA synthesis and content in the nuclei of epidermal cells of mice during their differentiation and specialization].

Synthesis and content of DNA in the nuclei of differentiating cells of mouse skin epidermis was studied by using cytomorphometric, autoradiographic and cytophotometric methods. It has been shown that the cells of the keratinoid series divide only in the basal layer and contain 2-4c DNA. Keratinocytes of the thorny layer are mostly tetraploid, 2c cells are lacking. H4c and 8c cells comprise 12% of the population. In the keratinocytes of the granular layer DNA content is somewhat lower due to nuclei break down and conversion of cells into anucleate scale. Part of the melanocytes of the basal layer also contain 4c DNA. Highly specialized element of the basal layer Merkel and Langerhans cells are polyploid. Conclusion is drawn that DNA hyper-replication by multiplication of the whole genome is part of the development program of the population.

Animals↗
PubMed2000

A micromechanic study of cell polarity and plasma membrane cell body coupling in Dictyostelium.

We used micropipettes to aspirate leading and trailing edges of wild-type and mutant cells of Dictyostelium discoideum. Mutants were lacking either myosin II or talin, or both proteins simultaneously. Talin is a plasma membrane-associated protein important for the coupling between membrane and actin cortex, whereas myosin II is a cytoplasmic motor protein essential for the locomotion of Dictyostelium cells. Aspiration into the pipette occurred above a threshold pressure only. For all cells containing talin this threshold was significantly lower at the leading edge of an advancing cell as compared to its rear end, whereas we found no such difference in cells lacking talin. Wild-type and talin-deficient cells were able to retract from the pipette against an applied suction pressure. In these cells, retraction was preceded by an accumulation of myosin II in the tip of the aspirated cell lobe. Mutants lacking myosin II could not retract, even if the suction pressures were removed after aspiration. We interpreted the initial instability and the subsequent plastic deformation of the cell surface during aspiration in terms of a fracture between the cell plasma membrane and the cell body, which may involve destruction of part of the cortex. Models are presented that characterize the coupling strength between membrane and cell body by a surface energy sigma. We find sigma approximately 0.6(1.6) mJ/m(2) at the leading (trailing) edge of wild-type cells.

Animals↗
PubMed1992

Conditions for isolation and culture of porcine myogenic satellite cells.

Myogenic satellite cells were isolated from semimembranosus muscles of 4-8 week-old pigs. Muscles were ground and incubated in 0.8 mg/ml Pronase solution for 40 min at 37 degrees C. Following enzymatic digestion, cells were separated from muscle debris by differential centrifugation and sequential filtering through 500 and 53 microns nylon mesh. Primary cultures grown in 16 mm diameter cell culture wells were used to evaluate five sera, media, and substrata for their ability to promote satellite cell proliferation and differentiation. Porcine satellite cell proliferation and myotube formation were optimized in cultures grown on gelatin-coated substratum in the presence of Minimum Essential Medium-alpha supplemented with 10% fetal bovine serum (FBS) (P less than 0.01). Maximum fusion was induced by 48 hr exposure to 2% FBS, horse serum, or lamb serum. These data 1) document the first evidence that myogenic satellite cells can be isolated from porcine skeletal muscle, and 2) identify culture conditions which optimize proliferation and myotube formation of porcine satellite cells.

Animals↗
PubMed2005

Regulation of activity in vitro and in vivo of three phospholipases B from Saccharomyces cerevisiae.

The genome of the yeast, Saccharomyces cerevisiae, contains three highly similar genes coding for phospholipases B/lysophospholipases. These enzymes behave differently with respect to substrate preferences in vitro and relative contributions to phospholipid catabolism in vivo [Merkel, Fido, Mayr, Pruger, Raab, Zandonella, Kohlwein and Paltauf (1999) J. Biol. Chem. 274, 28121-28127]. It is shown in the present study that, in vitro, pH markedly affects the substrate preference of Plb1p and Plb2p, but not of Plb3p. At the pH optimum of 2.5-3.5, the order of substrate preference of Plb1p and Plb2p is PtdSer (phosphatidylserine)>PtdIns>PtdCho (phosphatidylcholine>PtdEtn (phosphatidylethanolamine). At pH values of 5 and above, the substrate preferences change to PtdCho=PtdEtn for Plb1p and PtdSer=PtdEtn for Plb2p. Accordingly, with cultured cells the ratio of PtdIns/PtdCho breakdown, as reflected in the ratio of GroPIns (glycerophosphoinositol)/GroPCho (glycerophosphocholine) released into the culture medium, is inversely related to the pH of the growth medium. This effect is ascribed to the pH response of Plb1p, because Plb2p does not contribute to the degradation of PtdIns and PtdCho in vivo. Bivalent and tervalent cations activate phospholipases B at pH 5.5, but are inhibitory at pH 2.5. Al3+ at a concentration of 20 mM increases Plb1p activity in vitro by 8-fold and leads to a 9-fold increase in GroPCho release by whole cells. In vivo, cycloheximide strongly inhibits the breakdown of PtdIns, and to a lesser extent PtdCho. However, Al3+-stimulated GroPCho release is almost completely inhibited by cycloheximide. Deletion of PLB3 leads to increased sensitivity to toxic Al3+. Addition of SDS or melittin to cultured cells leads to a significant increase in phospholipid degradation, which is insensitive to inhibition by cycloheximide. Deletion mutants defective in the PLB1 gene are significantly more resistant to SDS than are wild-type cells.

Aluminum↗
PubMed1996

Testosterone up-regulates androgen receptors and decreases differentiation of porcine myogenic satellite cells in vitro.

Accumulation of DNA is essential for muscle growth, yet mechanisms of androgen-induced DNA accretion in skeletal muscle are unclear. The purpose of this study was to determine whether androgen receptors (AR) are present in cultured skeletal muscle satellite cells and myotubes and examine the effects of testosterone on satellite cell proliferation and differentiation. Immunoblot analysis using polyclonal AR antibodies (PG-21) revealed an immunoreactive AR protein of approximately 107 kDa in porcine satellite cells and myotubes. Immunocytochemical AR staining was confined to the nuclei of satellite cells, myotubes, and muscle-derived fibroblasts. Administration of 10(-7) M testosterone to satellite cells, myotubes, and muscle-derived fibroblasts increased immunoreactive AR. In satellite cells and myotubes, AR increased incrementally after 6, 12, and 24 h of exposure to testosterone. Testosterone (10(-10) - 10(-6) M), alone or in combination with insulin-like growth factor I, basic fibroblast growth factor, or platelet-derived growth factor-BB, had no effect (P > 0.01) on porcine satellite cell proliferation, and testosterone pretreatment for 24 h did not alter the subsequent responsiveness of cells to these growth factors. Satellite cell differentiation was depressed (20-30%) on days 2-4 of treatment with 10(-7) M testosterone. This effect was not reversible within 48 h after treatment withdrawal and replacement with control medium. These data indicate that satellite cells are direct targets for androgen action, and testosterone administration increases immunoreactive AR protein and reduces differentiation of porcine satellite cells in vitro.

Animals↗
PubMedDate not supplied

[Sentinel lymph node biopsy in cutaneous non-melanoma malignancies].

PURPOSE: To assess the feasibility of the Sentinel lymph node biopsy (SLNB) technique in cutaneous non-melanoma malignancies. MATERIALS AND METHODS: Nine patients were retrospectively evaluated performing a scintigraphy with 99mTc-nanocolloid. On the day of the surgery, an initial dynamic study and static images were obtained. The first drainage station visualized was considered the sentinel node (SLN). The SLN position was marked on the skin and after a correct localization in the surgical field with a gamma probe the SLN was obtained. Patients of this study have been followed up for 8 to 48 months. RESULTS: Lymphoscintigraphy detected the sentinel node in 88,8 % of our studies (the SLN was not observed in a patient with a Merkel's tumour on the back). The SLN was identified intraoperatively in those patients with positive imaging. Those cases without scintigraphic demonstrated migration were also not found intraoperatively. Histopathological analysis of the SLN showed non metastatic disease and none patient developed metastases or local recurrence in the monitoring period. CONCLUSIONS: Sentinel node biopsy can be applied to certain cutaneous non-melanoma malignancies. In patients with unclear drainage and to avoid unnecessary lymphadenectomy, the technique offers clear advantages. In our study the SLN analysis was related to the clinical progress. A large number of patients should be examined to truly assess the benefit of this technique in this kind of malignancies and to determinate when the technique must be performed.

Adult↗
PubMed1995

The frog taste disc: a prototype of the vertebrate gustatory organ.

The frog taste disc (TD) is apparently the largest gustatory organ found in vertebrates and seems to differentiate into a specialized variety of the prototypic scheme of the taste bud. An explanation for this unusual organization is lacking although it is possible to speculate the existence of environmental and nutritional requirements. Up to the present time, the most common model of the TD was based on two main cell types (sensory and sustentacular). This model may oversimplify the morphology of this structure since more numerous cell types have been described. We now propose a new model of the TD, based on comprehensive data on the ultrastructure of the organ obtained in the last 20 years. The main conclusions are the following: (1) the TD is a pluristratified epithelium with a general organization similar to that of the olfactory and vomeronasal epithelium; (2) it has skeleton composed of three different types of epithelial cells; (3) the chemoreceptorial surface is covered by different microenvironments; (4) three different types of neuro-epithelial systems are present; the type II is an 'open' sensory cell with axonal contacts devoid of vesicles; the type III is an 'open' sensory cell with synaptic-like junctions; the type i.v. is a 'closed' sensory cell with a 'Merkel-neurite complex'; (5) the nerve fibers in the basal plexus are mostly cholinergic while the peridiscal nerve fibers are mostly peptidergic. The presence of several cell types in the TD must be considered using these large receptors in electrophysiological studies or as a source of isolated cells, and their complexity must induce caution in the interpretation of the data. Text books of histology usually describe the peripheral structures associated with taste as very simple: an idea that probably must be revised. A taste organ is a highly complex structure composed of several sensory systems and a comparative approach can aid comprehension of its general organization. The study of the 'large taste organs' present in some species of amphibians can provide useful data for knowledge of the gustatory system of vertebrates.

Animals↗