[Formulation and special investigations of innovative intraoral solid dosage forms.] / Not available.

During our work, we summarized the types of solid dosage forms which were in the focus of attention in the last years because of their innovative pharmaceutical technology solution and simple use. The biopharmaceutics of solid dosage forms for intraoral use and the advantages of the use of these dosages forms were presented in general. However, these dosage forms cannot always be prepared with conventional pharmaceutical processes, therefore the special pharmaceutical solutions which can be applied for their preparation were presented. In addition to testing the European Pharmacopoeia dosage forms, the special tests which can be applied for the characterization of innovative solid dosage forms were highlighted.

Chicken dendritic cells and type II pneumocytes express a common intracellular epitope.

1. CVI-ChNL 74·3, a dendritic cell-specific monoclonal antibody (mAb) also identifies chicken lung granular pneumocytes (type II pneumocytes), which produce surfactant. 2. The 74·3 mAb does not cross-react with any other avian or mammalian granular pneumocyte, and provides a convenient tool for monitoring the status of type II pneumocytes in the chicken lung.

Origin of the chicken splenic reticular cells influences the effect of the infectious bursal disease virus on the extracellular matrix.

The effects of infectious bursal disease virus (IBDV) (strain F52/70) infection were studied by immunohistochemical methods on the splenic extracellular matrix (ECM). The major fibrillar components of the ECM, the type I and type III collagens and the main ECM organizing glycoproteins (laminin, tenascin and fibronectin) were monitored up to 11 days post-infection (d.p.i.). By 3 d.p.i., the collagens that form the basic scaffold of the antigen-trapping region of the spleen are destroyed, which is followed by deterioration of the glycoproteins. The ECM in the red pulp and the other regions of the white pulp (periarteriolar lymphatic sheath and germinal centre) seem to be normal. The reason for the significantly different pathological alterations in the ECM between the two regions of the spleen may be explained by the origin of the reticular cells. The reticular cells in the antigen-trapping zone and other splenic regions are of haemopoietic and mesenchymal origins, respectively. Possibly, the reticular cells of the haemopoietic origin are more susceptible for the IBDV infection than the mesenchymal ones. Development of the antigen-trapping, B-cell-dependent zone of the splenic white pulp precedes that of the periarteriolar lymphatic sheath and germinal centre, which suggests that this region may contribute to B-cell maturation. Damage of the ECM in the antigen-trapping zones results in impairment of tissue organization, which may contribute to the permanent immunosuppression.

The elevated interferon gamma production is an important immunological marker in HAM/TSP pathogenesis.

Human T-lymphotropic virus type 1 (HTLV-1) is the agent of the HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP), which may occur in >5% of patients during their lifetime. HTLV-1-infection causes disturbances in the immune system, and the viral load may also play an important role in the pathogenesis of HAM/TSP. Some cytokines are involved in the pathogenesis of this disorder. We have determined IL-2, IL-4, IL-10, IL-12 p70, IFN-gamma and TNF-alpha production among HTLV-1-infected subjects from our HTLV-out Clinic in Institute of Infectious 'Emílio Ribas' in Sao Paulo city, Brazil. PBMC obtained from healthy controls (n = 32), asymptomatic HTLV-1 carriers (n = 68) and HAM/TSP patients (n = 44) were grown in the absence and in the presence of phytohaemagglutinin (PHA), and the supernatants' fluids were measured for cytokines production. IL-2 levels were increased in the asymptomatic HTLV-1 carriers, and IFN-gamma was increased in both groups of patients (asymptomatic HTLV-1 carriers and more significantly among HAM/TSP patients). IL-4, IL-10, TNF-alpha and IL-12 p70 levels were not significantly increased on both groups of patients, as compared with controls. The major finding of this study is that IFN-gamma was an important cytokine for the HAM/TSP pathogenesis. Therefore, immune modulation of IFN-gamma may be critical to treat of HAM/TSP patients.

Identification of the avian B-cell-specific Bu-1 alloantigen by a novel monoclonal antibody.

A panel of monoclonal antibodies was generated against the guinea fowl's bursal cells. One of the antibodies, designated BoA1, recognized both cortical and medullary B cells of bursal follicles and B cell dependent regions of peripheral lymphoid organs, like germinal centers and splenic periellipsoidal regions. The staining pattern of this monoclonal antibody is similar to other antibodies (L22, 11G2, AV20), which also identify the Bu-1 antigens. Under reducing conditions, the molecular weight of the BoA1 antigen is 70 to 73 kDa, and after immunoprecipitation it proved to be identical with the antigen recognized by the AV20 antibody. It is unique for this novel monoclonal antibody that it shows wide range cross-reactivity with different avian species, like chicken, quail, guinea fowl, and turkey. Therefore, this Bu-1-specific monoclonal antibody could be a versatile tool for studying the B cell development in different domesticated birds.

Identification of the gene product recognized by monoclonal antibody GIIF3.

The chicken as a research model has a disadvantage compared with the mouse and the human because of the low number of available antibodies against gene products of interest. The goal of this study was to identify the antigen recognized by monoclonal antibody (mAb) GIIF3, which is a 42 kDa protein that appears in follicle-associated epithelium of the guinea hen as well as in different muscle types during chicken embryonic development. The 42 kDa protein, immunoprecipitated from chicken gizzard protein lysates, was evaluated by mass spectrometry. Mass spectrometry analysis revealed peptides specific for the chicken ß- or γ-actin isoforms. The mAb GIIF3 can be used as a new research tool for smooth muscle cell and bursa of Fabricius developmental studies.

Effect of soluble antigen on the ellipsoid-associated cells of the chicken's spleen.

The ellipsoid-associated cells (EAC) localize on the surface of the ellipsoid. They bind the IV injected horseradish peroxidase (HRP) and bovine serum albumin (BSA), migrate to the red pulp and then enter the systemic circulation. After protein injection, the number of white blood cells increased above the normal level. The highest number of white blood cells occurred between 4 and 6 hr after HRP injection. In this period, the cell enhancement resulted in HRP-positive mononuclear cells which might come from the spleen. This cell population may vary in size and number in the blood and may be identical with the dendritic cell of the chicken. The other type of blood mononuclear cell is histologically identical with the classical monocyte endogenous peroxidase positive type and their number is lower than that of the former. This type of monocytic (MN) cell could be of bone marrow origin.

Effect of surgical bursectomy on the ellipsoid, ellipsoid-associated cells, and periellipsoid region of the chicken's spleen.

Surgical bursectomy resulted in cellular depletion of the periellipsoid white pulp, confirming its bursa dependency. Also, in bursectomized birds, the ellipsoid could not be identified, although a small number of abnormal ellipsoid-associated cells (EAC) were observed in the periellipsoid region. The most characteristic finding was the degeneration of the EAC. Degeneration of EAC indicated that the intact bursa was mandatory for normal differentiation of cells of the periellipsoid white pulp into EAC. The promoting effect of the bursa might take place by a bursal hormone. The histological impairment of the EAC was followed by reduced carbon binding and migrating capabilities. Bursectomy resulted in a shift in bacterial phagocytosis in that many cells of the periellipsoid phagocytosed Salmonella. The reduced heterophil infiltration of the ellipsoid in bursectomized birds might be explained by the impaired granular content of the EAC. The impaired migration capability of the EAC might contribute to the low number of germinal centers in bursectomized birds.

Oesophageal tonsil of the chicken.

The oesophageal tonsil of the chicken is a novel member of the mucosal-associated lymphoid tissue (MALT), which is located around the entrance of the proventriculus. It consists of 6 to 8 single units, which are surrounded by a thin fibrous capsule. Each one is organised around the bottom of the longitudinal folds of the oesophagus, and serves as a 'tonsillar crypt'. Stratified squamous epithelium is infiltrated by lymphoid cells, i.e. T cells, plasma cells, macrophages, and dendritic cells, but not B cells, to form lymphoepithelium (LE). In the LE vimentin-, MHC II- and ATPase-positive cells possibly represent Langerhans' cells, but the appearance of 74.3 positive cells in the LE is unusual, because the 74.3 monoclonal antibody (mAb) recognises chicken follicular dendritic cells in the germinal centre and medulla of the bursal follicles. The subepithelial lymphoid tissue is organised into T- and B-dependent regions, which are the interfollicular areas and the germinal centres, respectively. Existence of high-endothelial venules in the interfollicular region suggests an extensive cellular connection between the oesophageal tonsil and the other lymphoid organs. In the resting oesophagus the lumen is closed, but during swallowing a bolus the crypt opens and the lymphoepithelium can be exposed to undigested food, antigens, infectious agents and vaccines. The location of the oesophageal tonsil, cranial to the stomach, may provide this organ with a unique role as compared to the other parts of the MALT; namely, it may contribute to the replication of infectious bursal disease virus (IBDV) and/or the pathogenesis of infectious bursal disease.

Plasma cell proliferation in the chicken harderian gland.

Studies to examine the percentages of proliferating plasma cells (PPC) in the Harderian gland (HG) were carried out in chicks between 5 and 12 weeks of age. Two methods, 5-bromo-2'-deoxyuridine (BrdUrd) incorporation into DNA and flow cytometric analysis of propidium iodide (PI) stained cells, were employed in control and emetine dihydrochloride treated birds. Flow cytometric analysis of PI stained cells showed the percentages of plasma cells in S phase were highest between 6 and 8 weeks of age. After this period of time, the number of S phase plasma cells decreased and remained low through 12 weeks of age. The lowest percentages of plasma cells in G0 + G1 were found at 6 and 8 weeks of age, and all ages had equal percentages of plasma cells in G2 + M phase. After administration of the protein synthesis inhibitor emetine dihydrochloride a common pattern of plasma cell depletion and repopulation in the HG was observed. At 3 and 5 days post-treatment the plasma cell population in the gland decreased and by 7 days post-treatment repopulation of the gland with plasma cells had taken place. Anti-BrdUrd staining of frozen sections revealed that the number of PPC were decreased at 3 days after emetine treatment but were as high as, or higher than, controls at 5 and 7 days post-treatment. Flow cytometric analysis indicated that some birds were more severely affected by emetine. Namely, the percentages of plasma cells in S phase were lower at 3 and 5 days post-treatment. Even though most birds were severely affected by emetine treatment during the experiments, they possessed a cell population with the proliferative capacity to quickly repopulate the HG by 7 days post-emetine treatment.

Angiotensin II analogues. 13. Role of the hydroxyl group of position 4 tyrosine in pressor activity.

In order to determine the features of the phenolic ring in position 4 of [Asn1,Ile5]angiotensin II that contribute to pressor activity, analogues with selected aromatic substituents were synthesized by the solid-phase method. They showed pressor activities in the rat: [Asn1,Phe(4-NH2)4]AII, 24%; [Asn1,Phe(4-NO2)4AII, 0.1%; [Asn1,Tyr(3,5-Me2)4]AII, 2.2%; [Asn1,D-Tyr(3,5-Me2)4]AII, 1.4%. These results indicate that the activity contributed by the aromatic character of the phenyl ring in the side chain of position 4 is enhanced by a group in the para position that may function as a proton donor in hydrogen-bond formation. Bulky substituents ortho to this hydrogen-bonding group decrease activity by steric interference with hydrogen-bond formation. Bulky groups than cannot act as hydrogen donors in the para position of the aromatic ring drastically decrease the activating effect of the aromatic ring on pressor activity.

Lymphopineal tissue in the chicken.

We have studied the chicken's pineal gland by light and electron microscopy at 3 and 4 weeks of age. The results indicated that lymphocytes, plasma cells, secretory cells, basophil and eosinophil granulocytes enter the parenchyma and a new histologically defined tissue is formed which we call lymphopineal tissue. Inside the pineal parenchyma, the number of thymidine labeled cells is almost three times higher than in the interstitium suggesting that the pineal's products might be blastogenic for the cells and/or exert an influence on post-thymic T-cell differentiation.

Anti-vimentin monoclonal antibodies differentiate two resident cell populations in chicken spleen.

Splenic stromal cells were studied with anti-vimentin monoclonal antibody clones V9 and 3B4. V9 positive cells were widely distributed both in the red and white pulp, specifically the ellipsoid, but were absent from the germinal centers. These observations suggested that this clone recognized splenic reticular cells. The original name given to the splenic ellipsoid region by Schweigger-Seidel, "Kapillarhülse" or capillary sleeve, would be a more appropriate term than ellipsoid, at least in the chicken because the V9 positive ellipsoid covered the entire length of the penicilliform capillary. The capillary sleeve was decorated by large, 3B4 positive cells that were identical to the ellipsoid-associated cells, a dendritic-like cell. The 3B4 monoclonal antibody recognized large stellate-shaped cells in germinal centers. These data emphasized the cellular difference between ellipsoid-associated cells and the cells of the ellipsoid; the cytoskeletal similarities of the ellipsoid-associated cells and dendritic cells of germinal centers advocate their common cell lineage.

Anti-S-100 antibody recognizes ellipsoid-associated cells and other dendritic cells in the chicken spleen.

The chicken spleen was studied immunohistochemically with anti-S-100 protein polyclonal antibody. S-100-positive cells accumulated around the penicilliform capillaries during the first 3 weeks of life. After 2 weeks posthatch the S-100-positive cells appeared in the red pulp, periarterial lymphatic sheath, and subsequently in the germinal center. Their ontogenetic development and intrasplenic distribution strongly suggested that the S-100-positive cells were identical with ellipsoid-associated cells. The S-100-negative cells of the periellipsoidal white pulp gradually transformed to S-100-positive, functionally active cells on the surface of the ellipsoid. The immunohistological findings support the hypothesis that the interdigitating dendritic cells and follicular dendritic cells were not of monocytic origin but belong to a splenic resident, endocytic cell line located on the surface of the ellipsoid.

Effect of the progestogen, allylestriol, on the bursal development of the chicken.

Fertile eggs were dipped on the fifth day of incubation into varying concentrations of allylestriol (AE), a synthetic progesterone, and testosterone propionate (TP). Concentrations of AE higher than 0.1% inhibited hatchability. The AE inhibited the differentiation of the bursal mesenchyme and follicular development and subsequently resulted in bursectomy. The AE caused bursectomy in 40-50 times lower concentrations than did TP. An unusual lymphocyte accumulation occurred around the postcapillary venules in the bursal mesenchyme of AE-treated birds. This observation suggested that inhibition of mesenchyme differentiation may lead to a modification in bursal function. The AE modification of bursal development was compared to those produced by TP. We demonstrated that AE caused immunosuppression of the B-cell system.

Structural changes in the mouse thymus and lymph node after emetine treatment.

The effect of emetine which is a potent immunosuppresant was studied on the thymus and lymph node. The subcapsular zone of the thymus was depleted and large number of adherent cells accumulated in this thymic region. The medulla enlarged but the cortico-medullary border remained distinct. In the paracortex (T dependent area) of the lymph node many non-lymphoid pyroninophil cells appeared which is followed by an increased cell proliferation 24 hours after emetine injection. 48-60 hours after administration many macrophage-like cells appeared in the medullary sinuses. This macrophage invasion precedes the adherent cell accumulation in the subcapsular zone of the thymus suggesting a possible non-lymphoid (adherent) cell migration from the lymph node's paracortex to the thymus.

Diverse expression of the K-1 antigen by cortico-medullary and reticular epithelial cells of the bursa of Fabricius in chicken and guinea fowl.

The immunocytochemical study of the K-1 monoclonal antibody indicates that the epithelial components of the bursa of Fabricius of the chicken and guinea fowl express the K-1 positive molecule. During embryogenesis, the K-1 antigen expression appears together with the bud-formation. As the number of B cells increases in the developing follicle, the K-1 expression gradually diminishes in the medullary reticular epithelial cells and completely ceases by hatching, which suggests that the molecule is developmentally regulated. After hatching, the expression of the molecule is restricted to the sealing off zone of the lymphoepithelial or medullary region of the follicle: i.e. to the cortico-medullary (CM) epithelial cells and the follicle associated epithelium (FAE) supporting cells in guinea fowl and to the latter ones in the chicken. The expression of the K-1 antigen by these epithelial components may support their structural identity. After hatching, the K-1 molecule is restricted to the CM epithelial cells and/or FAE supporting cells, which suggests that the function of the embryonic epithelial bud is taken over by the CM epithelial cells. The K-1 positive CM epithelial cells form arches, which encompass blast-like cells. The possible relationship of the CM epithelial cells and blast-like cells, which may represent the precursors of bursal secretory dendritic cells is discussed.

The lymphoid substance of the chicken's harderian gland is organized in two histologically distinct compartments.

Light and electron microscopical investigations revealed that the lymphoid structure of the chicken Harderian gland is organized in different histological frameworks. In the head the surface epithelium of the central canal can be classified as a lymphoepithelial tissue which covers the dense lymphoid substance. It consists of small and medium-sized lymphocytes, dendritic-like cells, and occasional macrophages. High endothelial venules are associated with intense lymphocyte migration and homing that gives circumstantial evidence for a T-dependent region, as found in a secondary lymphoid organ. The B-dependent germinal centers are also common structural units of the head region's lymphoid substance. The body of the gland is loaded with plasma cells of different maturation stages. They immigrate into the epithelium of the central canal and produce IgM and IgA. Only a few scattered IgG producing plasma cells can be found in the gland of Harder. This plasmocytic region accounts for the immunosurveillance on the conjunctiva and in the upper respiratory tract through antibody production against bacterial or parasitic infections. In both the head and body regions of the gland, anti-B-L (anti-Ia) antibody recognized scattered elongated cells which might represent dendritic cells. The immunological relationship between the two histologically different parts of the Harderian gland is unknown, but we speculate that the dense lymphoid tissue with high endothelial venule receives the blood-borne, immunologically mature, but uncommitted B cells. By the influence of local antigen stimulus, these B cells transform to plasma cells which gradually appear in the body of the gland. The lymphoid structures of the head and the body fulfill the function of secondary and tertiary lymphoid organs, respectively.

A novel monoclonal antibody identifies all avian embryonic myogenic cells and adult smooth muscle cells.

A novel monoclonal antibody, designated GIIF3, recognized prospective and differentiated smooth muscle cells in avian species studied - guinea fowl, chicken and quail. The GIIF3 antigen appeared in the myocardial, and the myotomal cells of the embryos at Hamburger-Hamilton stages 10 and 14, respectively. The expression of the GIIF3 molecule in the vascular smooth muscle cells emerged in the ventral wall of the dorsal aorta at Hamburger-Hamilton stage 16. The visceral smooth muscle cells started to produce the GIIF3 molecule from Hamburger-Hamilton stage 28 onwards. In both cardiac and skeletal muscles the GIIF3 expression gradually diminished, and it was lost by the end of the embryonic period, unlike in the differentiated vascular and visceral smooth muscle cells. In the latter cells the GIIF3 immunoreactive product showed a fine granular pattern that accumulated in the central region of the cytoplasm; it also occurred in the nucleus. A heavily stained discontinuous layer was associated with the cell membrane. The immunoblotting of the GIIF3 antibody recognized protein bands at 50 and 42 kDa in lysates of adult avian gizzard. A detailed comparative immunohistochemical study was made by smooth muscle markers, which confirmed the results of the immunoblotting, namely, that the GIIF3 monoclonal antibody recognized an avian myogenic cell specific molecule. During smooth muscle cell differentiation the GIIF3 molecule appeared as early as the alpha-smooth muscle actin, and in adult birds continued to be expressed; therefore the GIIF3 molecule could be regarded as a novel avian smooth muscle specific marker.

Origin of the bursal secretory dendritic cell.

The origin of vimentin-positive secretory dendritic cells of the bursa of Fabricius was studied by chick-quail chimera, parabiosis and immunohistochemistry using species-specific monoclonal antibodies. Quail bursal primordia of different ages were transferred to coelomic cavity of 3-day-old chicken embryos and further incubated for 18 days. In transplanted quail bursas the secretory dendritic cells of chicken and quail origin were detected by double staining of vimentin plus 74.3 and vimentin plus QCPN monoclonal antibodies, respectively. In bursal primordia of 5- and 6-day-old quail embryos both dendritic cells and B cells were of host, i.e. chicken origin. Mixed dendritic cell population of quail and chick origin emerged in chimeric birds of 6.5 days of age. In quail embryos transplanted at 7 and 8 days of age both dendritic cells and B cells were mixed i.e. of chicken and quail origin. Bursal secretory dendritic cells and medullary epithelial cells create "dendro-epithelial tissue" to receive pre-B cells. Colonization of dendro-epithelial tissue by pre-B cells initiates at day 7, thus the colonization of bursal anlage by blood-borne cells is a two-step process; entering of dendritic cells at day 6.5 is followed by that of B cells at day 7 and afterwards. It is discussed that bursal secretory dendritic cells and their product are key elements of bursal function therefore the mammalian bursa equivalent organ might be represented by a cell, which is analogous with the bursal secretory dendritic cell.