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Four monoclonal antibodies were raised against crude gap junction fractions of rat liver to clarify the distribution of gap junctions during animal development and to analyze gap junction expression in vivo and the polarity of hepatocytes in vitro. Among the monoclonal antibodies obtained, HAM8 antibody recognized the 27-kDa rat liver gap junction protein connexin 32. This antibody recognized gap junctions at the contiguous faces of hepatocytes, and the antigen was also observed in exocrine pancreas and salivary gland but not in kidney, heart, esophagus, or thymus. HAM8 did not react with amphibian or fish liver, heart, esophagus, stomach, or intestine as assessed via the immunofluorescence method on frozen sections. A few hepatocytes and many hemopoietic cells were seen in rat fetal liver at 15 days of gestation. HAM8 antigen was expressed on some hepatocytes but not on any hemopoietic cells. As the fetus grew, the number of hepatocytes in the liver increased gradually, together with the amount of HAM8 antigen. The distribution of HAM8 antigen at 25 days after birth was similar to that in adult liver. When the expression of HAM8 antigen was examined in primary cultured hepatocytes using the immunofluorescence method, the antigen was observed clearly between the hepatocytes. However, most of the HAM8 antigen on the free surface of hepatocytes disappeared within 4 hr. HAM8 antigen was not expressed on AH-7974 rat hepatoma cells when they formed small islets in the rat peritoneal cavity or within the liver. When HAM8 IgG antibody was injected intravenously, the HAM8 signal was expressed in the liver.
Anat Rec 1993 Mar
PMID:Immunohistochemical analysis of rat liver using a monoclonal antibody (HAM8) against gap junction. 838 23

The T cell receptor (TCR)-alpha and -delta loci are contained on the same chromosomal region, and yet are developmentally and genetically independent. The first element of the J alpha cluster (psi J alpha) is the site of an active rearrangement in the human thymus (delta Rec-psi J alpha rearrangement) and is localized downstream of a region expressed as a germ-line sterile transcript (TEA) in the human developing thymus. We hypothesized that the transcription of TEA could be indicative of (or responsible for) the opening of the J alpha to the V(D)J recombinase and undertook to analyze cis-acting sequences controlling the TEA transcription. The promoter of TEA was characterized. It was part of a region that is highly conserved between human and mouse and contained many sites for the putative binding of T cell-specific transcription factors. The in vitro activity of this promoter was dependent on the association with an enhancer. A strong DNase I hypersensitive site was found in the vicinity of this promoter again suggesting the possible presence of protein-DNA interactions in this region. The implications of these results in the general perspective of TCR-alpha/delta gene regulation is discussed.
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PMID:Functional characterization of the promoter for the human germ-line T cell receptor J alpha (TEA) transcript. 838 96

The process of T-lymphocyte differentiation within the thymus involves a series of molecular interactions. In this work we have carried out an analysis of the chick thymus microenvironment in order to evaluate its heterogeneity during development. We have produced 11 monoclonal antibodies whose staining patterns detected by the immunoperoxidase technique allowed us to divide them into five groups. A first group (E19-E2, P0-E5, and P15-T1) binds to thymic medullary stroma showing a reticular pattern on medullary epithelial cells and whose significance would be related to thymic stromal secretion. The second group of monoclonal antibodies (P15-T3) stains thymic corpuscles of 10- and 15-day chicks. The third group of antibodies includes P0-E1, P0-E3, P5-A6, and P15-T2 whose staining pattern is both medullary and cortical. The fourth group (P10-HB1 and P10-HB2) binds to thymic stromal and cortical thymocytes, and the fifth group (P5-A1) is characterized by the staining of medullary vessels of 5-day chicks. These five groups of monoclonal antibodies corroborate the existence of an antigenic diversity of the chick thymus microenvironment. Their possible relationships with T-cell differentiation and stromal-thymocyte interactions are discussed.
Anat Rec 1993 Feb
PMID:A study of the chick thymus microenvironment during development: analysis by monoclonal antibodies against thymic epithelium. 842 Mar 97

A mutagenic steroidal derivative (3 beta-Acetoxy-5 alpha-Cholestano[6 alpha,5-d']1'-3'oxathiolane-2' thione) structurally related to cholesterol caused strand scission and induced nicks in calf thymus, supercoiled pBR322 and single stranded M13 mp8 phage DNAs. S1 nuclease hydrolysis, reaction with pBR322 and M13 phage DNA as well as treatment of E. coil mutant strains and phage was used to evaluate the effect of test steroid on the DNA molecule. The strand scission/nicking of DNA by the test steroid was enhanced by some metal ions, especially the Cu(II). Scavengers of active oxygen radical species significantly inhibited the S1 nuclease hydrolysis by the test steroid indicating the major role of active oxygen species in DNA strand scission and nicking. The steroid brought about the DNA degradation even in the absence of S1 nuclease. There was an appreciable reduction in the survival of steroid treated polA and lig mutants of E. coli K12 compared to the wild type strain. Phage on steroid treatment also lost its plaque forming units (P.F.U.) which was more pronounced in the polA and rec A background.
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PMID:Steroid induced single strand breaks in DNA mediated by active oxygen species and its biological consequences. 848 66

Localization of eosinophil granule major basic protein by immunofluorescence permits recognition of both eosinophil infiltration and degranulation. Over the past decade and a half, our laboratory has shown that eosinophil infiltration and degranulation occur in many diseased tissues in humans; among normal tissues studied as controls, only the gut showed striking eosinophil infiltration and degranulation. Using an indirect immunofluorescence procedure for the detection of major basic protein, we extended our analyses of normal human tissues to include tissues from essentially all body organs; a total of 117 biopsy/autopsy specimens were analyzed. To determine whether the method of tissue procurement affected the level of eosinophil degranulation in the normal gastrointestinal tract, normal proximal jejunum from six patients was biopsied using either an endoscopic forceps or a scalpel at the time of elective surgery and examined by immunofluorescence. Spleen, lymph node, and thymus tissues showed eosinophil infiltration with scant evidence of degranulation, but the only organ showing both eosinophil infiltration and remarkable degranulation was the gastrointestinal tract. Eosinophil degranulation was significantly increased in specimens obtained by endoscopic forceps compared to those obtained by scalpel (P = 0.021). These results indicate that tissue procurement methods affect the degree of eosinophil degranulation in the gut. Thus, among normal human body organs, both eosinophil infiltration and appreciable degranulation consistently occur only in the gut.
Anat Rec 1998 11
PMID:Eosinophil infiltration and degranulation in normal human tissue. 981 Dec 20

Only scant information is available in the scientific literature on the parathyroids and ultimobranchial bodies in the primitive mammals, the echidna (Tachyglossus aculeatus) and platypus (Ornithorhynchus anatinus). The major aim of this paper is to describe the morphology of the monotreme parathyroid gland and to compare it with parathyroids in mammals and reptiles. The gross anatomy and light microscopic structure of the ultimobranchial body, thymus, and thyroid are also given. Animals were dissected and routine light and electron microscopic techniques used to examine the microscopic morphology. The locations of parathyroid hormone, calcitonin and calcitonin gene-related peptide in tissue sections were identified by immunostaining. Monotremes have one pair of parathyroid glands located in the thorax and they are often associated with thymic tissue but never with the thyroid which is also present in the mediastinum. Ultimobranchial bodies are ventrolateral to the commencement of the trachea. Thymic lobules with Hassall's corpuscles are scattered in the fibrofatty tissue of the mediastinum and the ventral surface of the pericardium. Histologically, principal cells, water-clear cells, and non-secretory cells were identified in the parathyroid glands. Principal cells showed polarity and had microlamellar projections that formed intercellular canaliculi. Non-secretory cells had features similar to those of thymic epithelial reticular cells. Immunostaining of parathyroid hormone showed a diffuse distribution in parathyroid principal cells and none in ultimobranchial bodies. Identification of the ultimobranchial bodies was confirmed by immunostaining. The monotreme parathyroid gland, ultimobranchial bodies and thyroid show reptilian as well as mammalian features.
Anat Rec 1999 02 01
PMID:Parathyroids and ultimobranchial bodies in monotremes. 997 12

The ultrastructure of the thymus in the chick (Gallus domesticus) was studied after unilateral vagotomy at survival times of 3, 7 and 10 days. Ultrastructural changes in the ipsilateral thymus were observed in axon boutons as well as in myoid and cystic cells in the medulla, especially those situated near the corticomedullary junction. Structural changes in axon boutons ranged from granular degeneration of the axonal cytoskeleton to vacuolation of the axoplasm. Myelin figures of different sizes and configurations and clumping of small agranular vesicles were commonly observed in the axon terminals. Degeneration of myoid cells appeared to peak at 7 days post-vagotomy. Changes ranged from oedematous appearance and intense vacuolation of the peripheral cytoplasm to disorganisation and clumping of myofibrils. In some myoid cells the sarcomeres showed granular degeneration at the I-bands and in others, the myofibrils were completely degenerated such that amorphous material and partially degenerated organelles filled the entire cell. The majority of cystic cells at 3 days post-vagotomy showed a uniform increase in electron density. Numerous electron dense bodies, some displaying concentric lamellation, were observed throughout the expanse of the cytoplasm. At 7 days post-vagotomy, the cytoplasm of some cells gave a "moth-eaten" appearance. Dying cystic cells were encountered at 10 days after vagotomy. Degeneration in the myoid and cystic cells suggests that these cellular components may be the putative targets of the vagal fibres in the chick thymus. The changes in these cells reflect a disturbance in the cell metabolism presumably brought about by the removal of vagal influence.
Anat Rec 1999 07 01
PMID:Ultrastructural changes in the chick thymus following unilateral vagotomy. 1041 94

The numbers and distribution of T and B cells in the thoracic thymus, spleen and intestinal tissue and the proliferation of T lymphocytes were examined during pouch life and in the adult to determine when the developing brushtail possum reaches immunological maturity. CD3-positive cells were observed in the thoracic thymus at day 2 post-partum indicating that the thymus produces T lymphocytes at or soon after birth. By day 25 the thymus was fully populated with CD3-positive T lymphocytes and they were observed in distinct regions of the cortex and medulla. By day 48 post-partum, B and T lymphocytes were identified in the follicles and parafollicular areas of the spleen. Although the numbers of T and B cells in the spleen increased significantly from day 25 to day 100 post-partum (P < 0.005), fewer cells were present at day 150 post-partum than in the adult (P < 0.05). Peyer's patches were not observed in the intestines up to day 73 post-partum. However, both T and B cells were observed in the intestinal lymph nodes. Although the T lymphocytes at weaning showed a proliferative response, the response was not as great as that observed in the adult possum. Thus, the immune system of the possum is not fully developed at weaning but continues its development after pouch life.
Anat Rec 1999 12 01
PMID:Ontogeny of the immune system of the brushtail possum, Trichosurus vulpecula. 1058 22

Sixteen European badgers (Meles meles) from three statutory removal operations were studied. Samples of tracheal aspirate, pooled lymph nodes and urine were cultured for mycobacteria. Seven of the badgers were infected with Mycobacterium bovis and had tuberculous pulmonary lesions which varied in severity from extensive granulomatous consolidation to microgranulomas which were not detectable grossly. Tuberculous lesions were also observed in the upper respiratory airways, intestines, kidneys, spleen, liver, thymus, pleura and lymph nodes. One badger had tuberculous bite wounds. The histopathological characteristics of the tuberculous reactions and the associated tissue damage in various organs, together with the gross pathology, indicate that both mildly and severely infected badgers have the potential to excrete M. bovis by several routes.
Vet Rec 2001 Mar 10
PMID:Pathology of natural Mycobacterium bovis infection in European badgers (Meles meles) and its relationship with bacterial excretion. 1131 35

Lymphocytes in the bronchoalveolar space are routinely obtained and examined in lung diseases such as asthma or sarcoidosis. In a pig model, labeled lymphocytes were found in regional lymph nodes after intrabronchial instillation, indicating that reentry of lymphocytes from the bronchoalveolar space into the body is possible. In the present study, the route and kinetics of the reentry of bronchoalveolar lymphocytes were investigated in a congenic rat model using immunohistochemistry on cryostat and semithin sections and confocal laser scanning microscopy. As early as 15 min after intratracheal instillation lymphocytes were found to leave the bronchoalveolar space by transmigration through alveolar but not bronchial epithelium and were observed in interstitial alveolar tissue. At 6 hr after intratracheal instillation, T and B lymphocytes appeared in the draining lymph nodes of the lung with an increase after 24 and 48 hr. The kinetic pattern clearly differed in nondraining lymph nodes and other organs. After 6 hr, only single cells were found in nondraining lymph nodes, spleen, and blood with a slight increase after 24 hr, and only occasionally were single cells seen in the liver, thymus, or Peyer's patches 24 and 48 hr after instillation. In conclusion, T and B lymphocytes can leave the alveolar space by reentry into the lung tissue through alveolar epithelium. They reach regional lymph nodes by means of lymphatic vessels and are then distributed all over the body to rejoin the systemic immune system. Coming into contact with environmental antigens, these lymphocytes could perform an important function in the lung immune system and might be a target for inhalative therapy.
Anat Rec 2001 11 01
PMID:Lymphocytes in the bronchoalveolar space reenter the lung tissue by means of the alveolar epithelium, migrate to regional lymph nodes, and subsequently rejoin the systemic immune system. 1159 5


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