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PubMed1994

Epithelial transport and bioavailability of intranasally administered human growth hormone formulated with the absorption enhancers didecanoyl-L-alpha-phosphatidylcholine and alpha-cyclodextrin in rabbits.

The transepithelial transport of biosynthetic human growth hormone (hGH) formulated with the absorption enhancers didecanoyl-L-alpha-phosphatidylcholine (DDPC) and alpha-cyclodextrin (alpha-CD) was studied after intranasal administration to rabbits. Plasma concentrations of the hormone were determined until 240 min post administration by ELISA, and the absolute bioavailability was estimated to be in the vicinity of 20%. The localization of hGH was studied 15 min after application of the powder formulation in the initial absorptive phase. To visualize the hormone, a two-step indirect immuno-gold technique was used on semithin and ultrathin cryosections and Epon sections. Polyclonal rabbit anti-hGH was used as primary antibody and gold-conjugated goat anti-rabbit IgG as secondary antibody, succeeded by silver enhancement. Growth hormone was mainly found in the cytoplasm and nuclei of ciliated cells, showing distinct morphological signs of early necrosis, and in lamina propria, including the venules. Minute amounts of hGH were found in endocytotic vesicles in morphologically normal epithelial cells and in the intercellular compartment. We conclude that the major transport route of hGH formulated with absorption enhancers DDPC and alpha-CD was transcellular through lethally damaged ciliated cells.

Absorption↗
PubMed2005

The role of liquid-based cytology in the investigation of breast lesions using fine-needle aspiration: a cytohistopathological evaluation.

INTRODUCTION: Liquid-based cytology processing of breast fine-needle aspiration (FNA) specimens has been used in our institution since 1999. The aim of this study was to investigate the role of liquid-based cytology (Thin-Prep(R)) in the evaluation of breast FNA specimens. PATIENTS AND METHODS: This study was carried out on 352 cases corresponding to equal number of women (134 malignant and 218 benign) examined from September 1999 to June 2003. A direct to vial study was planned. Aspirated material was directly immersed to cytolytR vial (Cytyc Corporation, Boxborough, MA). The specimens were transported to the laboratory non-refrigerated within 24 hr. Thin-Prep slides were made using the Thin-Prep 2000 processor. The remaining material was embedded in paraffin. RESULTS: Altogether, 352 cases were examined and unsatisfactory smears were observed in 14 (3.9 %) out of all cases. The histologic examination verified that 128 out of 131 total malignant cases and 203 out of 207 total benign cases were correctly diagnosed with the cytological technique. False positives were found in four cases and false negative in three cases. Liquid-based technique presented a sensitivity of 97.7%, a specificity of 98%, a positive predictive value (PPV) of 96.9%, a PNV of 98.5%, and an over all accuracy (OA) of 97.9%. CONCLUSIONS: The processing of FNA breast specimens using liquid-based cytology offers an accurate diagnostic tool. Liquid-based cytology could reduce the uncertainty based in the cytological evaluation of FNA, because it offers the possibility to embed material in paraffin and take tissue sections with histological appearance identical to those observed in routine histological sections. Moreover it offers the potential advantage of retraining extra material for the purpose of ploidy analysis and immunohistochemical study in order to offer information concerning the expression of prognostic markers such as p53, MIB-1, steroid hormone receptors, and EGFR.

Biopsy, Fine-Needle↗
PubMed1976

Frozen section- frozen attitudes.

Patients with breast cancer who have clinically negative axillae, tumors 2 cm in diameter or less, and tumors with a moderate or low degree of anaplasticity are candidates for treatment by local excision and irradiation. A definitive histologic diagnosis should be made in all cases of breast cancer before a final treatment decision is made. This can be accomplished by needle or excisional biopsy. After a histologic diagnosis is established, the decision regarding treatment can be arrived at with the patient enering into the process. Those patients requiring mastectomy will be better able to cope with this, knowing why the operation was considered best for them. The elimination of frozen section diagnosis will free the patient from one of the great fears of the current treatment of breast cancer- subjecting herself to biopsy without knowing what the outcome will be. It will free the surgeon from the rigid sequence of frozen section followed by mastectomy and allow him to begin to rationally assign patients with proper criteria to conservative treatment, reserving mastectomy for those who do not meet these criteria.

Adult↗
PubMed1990

Accuracy and pitfalls of frozen section during thyroid surgery.

The purpose of this study was to assess the accuracy of frozen sections performed during thyroid surgery and to define any pitfalls they may have. The material includes a series of 190 frozen section examinations of thyroid pathology. The overall accuracy of frozen section in this series was 95%. There were no false positives in our series and false negatives related to only follicular adenomas. In two patients, the report was deferred for permanent slide examination. Nine patients had lesions reported as malignant; however, the distinct cell type was not designated on frozen section. Eight specimens initially reported to have follicular adenoma on frozen section turned out to have angioinvasion or capsular invasion indicative of follicular carcinoma. Minor discrepancies between frozen and permanent sections in the types of benign disease were noted occasionally, but were not clinically significant. There was a discrepancy in the diagnosis of Hashimoto's thyroiditis on three occasions. The major discrepancy in patients with malignant pathology was related to the cell type. The diagnosis of anaplastic thyroid cancer, though suspected, was deferred for permanent sections in all cases. The diagnosis of medullary cancer of thyroid was difficult to make on frozen section. The major problem with frozen section was the diagnosis of follicular adenoma versus carcinoma. The benign diagnosis was changed in eight instances (out of 24 frozen sections) from follicular ademona to follicular carcinoma. Because of this experience, we do not provide the frozen section diagnosis to the patient and we wait until the final diagnosis is available.

Adenocarcinoma↗
PubMed1993

Transmission of corneal collagen during ArF excimer laser ablation.

The time-resolved transmission of collagen films and 10-microns sections of bovine cornea during ArF laser ablation has been investigated. The film studies were performed on thin layers of extracted bovine corneal collagen, the principal chromophore in 193 nm photoablation. Transmission measurements were made on both dry and water-saturated films to assess the sensitivity of the ablation process to hydration. Distinct transient optical changes were observed in both fully desiccated and rehydrated films. Dehydrated films exhibit rapid reduction in film absorption over the time-course of the ablating laser pulse, presumably due to chromophore bleaching or annihilation. In contrast, rehydrated films demonstrate a short-lived enhancement of the attenuation. In either case, a single ablative laser pulse increased the long-term transmission of the film, although this increase was a factor of five greater for dehydrated films than for rehydrated samples. Results obtained from corneal tissue sections were essentially identical to those derived from hydrated collagen films.

Absorption↗
PubMed1989

Chromatin arrangement in mouse sperm nuclei: an ultrastructural cytochemical study.

The arrangement of mouse sperm nuclei chromatin and, in particular, of DNA has been studied by electron microscopic cytochemistry. It had been previously shown that, after a Feulgen-type reaction using an osmium ammine complex (OAC), the OAC-stained DNA was distributed in a spotted pattern in the nucleus (Biggiogera: Basic Appl Histochem 30:501-504, 1986). The present chapter shows that this pattern is characteristic of mouse spermatozoa from testis to vas deferens, with the exception of some testicular spermatozoa, in which DNA was homogeneously stained. DNase digestion of thin-sectioned nuclei resulted in a distribution of residual material complementary to the pattern of the unstained zones after the OAC reaction. These findings are discussed considering the role of -S-S- crosslinks, characteristics of this extremely condensed chromatin, in limiting the availability of DNA to acid hydrolysis.

Animals↗
PubMed1997

In vivo observation of anisotropic motion of brain water using 2H double quantum filtered NMR spectroscopy.

The 2H DQF NMR spectra of deuterated water molecules were measured for the first time in in vivo rat brain. The observation of the DQF signal indicates that there is a water population that exhibits anisotropic motion. The characteristics of the DQF spectra premortem and postmortem are very similar (lineshape and relaxation times). In the 1st h there is a 10-15% decrease in the signal intensity of the DQF spectra followed by a gradual but a much slower decrease in the DQF signal intensity that reaches 65-70% of its initial value after only 12 h. When the brains were kept at 4 degrees C, a 40% decrease in the DQ signal intensity was observed only after 7 days. Mechanical chopping of the brain tissues causes an immediate loss of more that 97% of the DQ signals. The slow, temperature-sensitive decay of the signal, and its sensitivity to mechanical treatment point out that these signals originate from water molecules that interact with structural components in the brain. The characteristics of the DQF spectra depend on the amount of bulk water as exemplified by increased residual quadrupolar interaction and relaxation rates obtained when dehydrating the brain tissue.

Animals↗
PubMedDate not supplied

Actomyosin ATPase. I. Quantitative measurement of activity in cryostat sections.

A method for the quantitative study of the actomyosin ATPase activity (Ca,MG-ATPase) in thin sections cut in a cryostat is presented. This method is based on the liberation of 32P from [gamma 32P]ATP or 45Ca phosphate precipitation. The advantage of this method lies in the requirement for only a small muscle sample and the availability of serially cut sections for other assays including Ca uptake by sarcoplasmic retciulum and histochemical tests for oxidative and glycolytic enzymes. The actomyosin ATPase activity for various types of muscles determined by this method showed the same sequence found in isolated protein, that is, fast-twitch skeletal greater than slow-twitch skeletal greater than cardiac. The Ca,Mg-ATPase of cryostat sections showed Ca sensitivity. The fact that the sections retained Ca sensitivity at 37 degrees C, in contrast to myofibrils, which have been reported to lose Ca sensitivity at this temperature, indicates that the structural integrity of the contractile and regulatory apparatus is preserved to a higher degree in sections than in isolated myofibrils.

Actomyosin↗
PubMed1984

Topographical localization of motor endplates in cryosections of whole human muscles.

The distribution of motor endplates in biceps brachii, tibialis anterior, and sartorius muscles from human adults was studied by staining longitudinal cryosections of whole muscle for cholinesterase. A special freezing technique was used to prevent the muscle from cracking before sectioning on a heavy cryostat microtome. The results from a large number of cryosections from biceps brachii and tibialis anterior muscles were analyzed by a computer and the topographical distributions of endplates in different views of the muscles were reconstructed. In the biceps brachii muscle, the endplates formed a fairly distinct, slightly V-shaped band through the middle of the two heads. In the tibialis anterior muscle, the majority of the endplates were superficially distributed along the whole muscle. In the longitudinal sections from the middle part of the muscle, they gave the pattern of a parabola with its apex at the proximal end of the muscle. In the sartorius muscle, the endplates were scattered throughout the muscle and no endplate band was observed. The findings are in accordance with results obtained 30 years ago in investigations of muscle from small children and stillborn infants.

Adult↗
PubMed1989

Sequential demonstration of antigens and AgNORs in frozen and paraffin sections.

A technique has been designed which enables the sequential demonstration of nucleolar organizer regions (NORs) and various antigens, in both frozen and paraffin wax-embedded sections. The NORs were demonstrated by the standard argyrophil (AgNOR) method and the antigens were shown by either immunoperoxidase (PAP) or immunoalkaline phosphatase (alkaline phosphatase-anti-alkaline phosphatase or avidin-biotin-alkaline phosphatase) methodology. Clear, reproducible results were obtained and AgNOR sites were enumerated with ease. It is suggested that the sequential method may be of great use in the evaluation of AgNOR numbers in neoplasms, where cell populations are heterogeneous. Cell populations may be demarcated with accuracy prior to the counting the AgNORs.

Antigens↗
PubMed1991

The AMeX method: a multipurpose tissue-processing and paraffin-embedding method. III. Extraction and purification of RNA and application to slot-blot hybridization analysis.

RNA was extracted from tissues processed by a new fixation and paraffin-embedding method (the AMeX method) and examined by Northern blot analysis and slot-blot analysis. The RNA extraction method for AMeX-processed tissue sections after the deparaffinization step was the same as that for fresh materials. The total amount of cellular RNA extracted from AMeX-processed mouse liver tissue was slightly less than that extracted from fresh tissue. In tissues of malignant lymphoma, the total amount of cellular RNA extracted from 25 sections each 20 microns thick was about 1.6-1.8 micrograms/mm2, regardless of the histological subtype and period of storage. The extracted RNA was moderately degraded, and usually could not be used for Northern blot hybridization analysis. The intensity of ethidium bromide staining and the hybridization signals of RNA extracted from AMeX-processed tissues were usually reduced in comparison with RNA from fresh material, but specific signals could be detected by slot-blot hybridization analysis. We have demonstrated previously that the AMeX method preserves high-molecular-weight DNA and various antigens. Since the present study showed that information on mRNA can be obtained from AMeX-processed tissue, the versatility and usefulness of this method were further proven.

Animals↗
PubMedDate not supplied

A scanning and transmission electron microscopy study of the parabronchial unit in quail (Coturnix coturnix) and town pigeons (Columba livia).

A combined scanning electron (SEM) and transmission electron microscopy (TEM) investigation was undertaken to gain insight into the complex structural pattern of the atrial compartment and the gas exchange tissue of parabronchial units in quail and town pigeons. The aim was also to depict the changes taking place in the parabronchial unit in the late prehatching and early posthatching periods in quail. The standard SEM and TEM investigation was carried out in 13 mature quail and 8 town pigeons. The developmental study involved embryonic quail (Days 15, 16, 17), newly hatched quail, quail 24 h after hatching, and quail aged 2, 10, 19, and 25 days (3 individuals per group). The luminal relief of the parabronchus is formed by anastomosing interatrial septa delineating the atrial pits, which are thinner and shallower in pigeons. The atrial bottom opens in mature individuals into 3-6 infundibula. The extracellular material represented by trilaminar substance, which does not appear until hatching, veils the surface relief of the parabronchial epithelium, which is consequently hardly accessible to three-dimensional visualization. Only in town pigeons with fewer discontinuous layers of extracellular material was it possible to visualize the surface of the atrial epithelium, that is, of the granular and squamous atrial cells. The SEM analysis has convincingly shown the intricate spatial organization of atria, infundibula, and air and blood capillaries of the gas exchange tissue. The retinacula, that is, parallelly arranged processes of squamous respiratory cells bridging the air-capillary lumina, were evidenced by SEM and TEM. The complex structure of the avian parabronchus has been successfully demonstrated in the present SEM and TEM study.

Animal Structures↗
PubMedDate not supplied

Characterization of an oil-degrading Microcoleus consortium by means of confocal scanning microscopy, scanning electron microscopy and transmission electron microscopy.

A consortium of microorganisms with the capacity to degrade crude oil has been characterized by means of confocal laser scanning microscopy (CLSM), transmission electron microscopy (TEM), and scanning electron microscopy (SEM). The analysis using CLSM shows that Microcoleus chthonoplastes is the dominant organism in the consortium. This cyanobacterium forms long filaments that group together in bundles inside a mucopolysaccharide sheath. Scanning electron microscopy and transmission electron microscopy have allowed us to demonstrate that this cyanobacterium forms a consortium primarily with three morphotypes of the heterotrophic microorganisms found in the Microcoleus chthonoplastes sheath. The optimal growth of Microcoleus consortium was obtained in presence of light and crude oil, and under anaerobic conditions. When grown in agar plate, only one type of colony (green and filamentous) was observed.

Cyanobacteria↗
PubMed1997

Electron microscopic identification of putative liver stem cells and intermediate hepatocytes following periportal necrosis induced in rats by allyl alcohol.

The ultrastructural characteristics of the putative liver stem cells that repopulate the necrotic periportal zones after allyl alcohol induced liver injury are described. Periportal liver cell necrosis was induced in adult female Sprague-Dawley rats by i.p. injection with 0.62 mmol/kg of allyl alcohol. Electron microscopic examination of the livers was carried out at 33, 57, 81 and 129 h after injection. After periportal necrosis small nondescript intraportal cells (putative liver stem cells) as well as three type of "progenitor" cells are seen: type I, immature "precursor" cells; type II, bile duct-like; and type III, hepatocyte-like, with numerous cells of intermediate type between type I and type III. The periportal necrotic zone (zone I) is reconstituted largely by an increase in hepatocyte-like cells containing mitochondria, lysosomes, lipid-filled vacuoles, rare peroxisomes, prominent endoplasmic reticulum and lateral microvilli (type III cells) with a relatively small number of type I (immature) cells participating. The type III cells display different degrees of differentiation; the less mature are termed "restitutive" and the more mature "transitional" hepatocytes to emphasize the probable relationship between these cell types. Immature ductular cells (type II cells) are seen located basally within hyperplastic ducts in the periportal zone. It is postulated that hepatocyte restitution after periportal necrosis is accomplished by proliferation and differentiation of stem cells with both biliary and hepatic potential that specifically differentiate into hepatic cells through "restitutive" and "transitional" intermediates. These postulated liver stem cells may be intraportal cells seen 33-57 h after injury that precede the type I and type III hepatic precursors seen later.

Animals↗