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Biomedical subjects

R Gatti

Publications and source records attributed to R Gatti.

At least 145 records · Page 8Linked to original sources

[Seasonal changes in seminiferous tubules and Leydig cells of hibernating bats].

We studied the seasonal course of Leydig cells combined with spermatogenesis and blood testosterone concentrations. The reproductive cycle of Pipistrellus kuhli kuhli and Eptesicus serotinus is characterized by seminiferous tubules which reactivate after a period of complete quiescence extending for the whole winter hibernating sleep and spring (March-June) periods. During hibernation, Leydig cells show ultrastructural features which stress a low secretory activity with corresponding low concentration of blood testosterone. With the beginning of summer, Leydig cells increase their metabolic activity reaching a top in the mating season (fall) when also blood testosterone and spermatogenesis reach the highest levels. The authors, after some comparative morphological considerations with different hibernating animals, assert that the reproductive processes of these two bats are not strictly species-linked, but rather dependent, besides seasonal thermal changes, upon habitat and feeding sources.

Animals↗

Determination of salicilic acid in saliva by high-performance liquid chromatography.

A sensitive reversed-phase HPLC method with fluorimetric detection was developed for the determination of salicilic acid in saliva samples. The method, involving a preliminary solid-phase extraction procedure, was found to be suitable for the determination of salivary salicilic acid concentrations following the administration of a single oral dose of some commercial acetylsalicilic acid formulations.

Aspirin↗

The migration of lymphocytes and polymorphonuclear leukocytes across the endothelial wall of the absorbing peripheral lymphatic vessel.

We studied the process of transendothelial migration of lymphocytes and polymorphonuclear leukocytes in the absorbing peripheral lymphatic vessels of small intestine and urinary bladder, under normal conditions, lymphatic stasis and prolonged fast. By ultrathin serial sections and three-dimensional models we found that lymphocytes and leukocytes share a common migration pathway from the interstitium into the lumen of the lymphatic vessel, namely 'the intraendothelial channel'. The migratory process of these cells occurs in several steps: approach to the endothelial wall, entry and run along the channel and finally coming out into lymphatic lumen. Our findings are compared with those concerning lymphatics in pathological conditions and blood vessels. Further considerations concern the possible mechanisms underlying this kind of transendothelial migration.

Animals↗

Macrophage migration through the endothelium in the absorbing peripheral lymphatic vessel of the small intestine.

TEM and three-dimensional models of the absorbing peripheral lymphatic vessel (ALPA) of the small intestine demonstrate that macrophages migrate from the interstitium into the lumen through the endothelial cell (transcellular pathway). Macrophages cross through a 'migration pore' which arises by an 'erosive process' in the cytoplasmic expansion of the endothelial cell. The pore is completely independent of the intercellular junction which is always sealed by junctional complexes. Furthermore several sites of contact between macrophage and endothelial cell membranes stress a possible functional role played by cellular interactions. Transcellular migration processes under normal conditions are compared with pathological or experimental ones.

Animals↗

Determination of minoxidil in pharmaceutical formulations by difference spectrophotometry and liquid chromatography (HPLC).

A simple difference spectrophotometric method has been developed for the selective and rapid determination of minoxidil in pharmaceutical formulations. The method is based on the characteristic modifications of the minoxidil UV spectrum induced by pH changes; a difference spectrum can be obtained and the absorbance A282.8 or the absorbance difference delta A = (A282.8-A259.2) can be used for the selective drug quantitation. As an alternative, a derivative second-order spectrophotometric method was also developed. The spectrophotometric methods were applied to the quality control of commercial minoxidil dosage forms and the results were comparable with those obtained by a reference HPLC procedure.

Chromatography, High Pressure Liquid↗

Immunization against bovine rotaviral infection.

Calves fed with colostrum from cows vaccinated with an inactivated rotavirus vaccine were refractory to experimental infection with strain 81/36F of bovine rotavirus. In the field study, 458 pregnant cows from 26 herds were involved. In each herd, cows were selected and randomly subdivided in two groups. Cows in one group (248 head in total) were vaccinated, whereas cows in the other group (210 head in total) were left as unvaccinated controls. At calving, colostrum was collected from each cow and stored at -30 degrees C until used for feeding calves. The newborn calves, beginning the second day of life and for the next 7-10 days, were each fed a daily supplement of 400 ml of colostrum from its dam. The diarrhea occurred in 86 (40.9%) calves that had received colostrum from unvaccinated dams (normal colostrum), and in 7 (2.8%) calves which were fed colostrum from vaccinated dams (immune colostrum). The disease was very severe in the normal colostrum-fed calves and 52 of them died. By contrast, the 7 immune colostrum-fed calves displayed a rather mild enteric condition, and all recovered without any sequela being observed.

Animals↗

Heterogeneity of mRNA expression in Italian fucosidosis patients.

Genomic DNA and mRNA from six unrelated Italian patients affected with fucosidosis were analyzed using two probes (AF3 and AF11B) of partial length cDNA coding for the lysosomal enzyme alpha-L-fucosidase. DNA from patient 2, digested with EcoRI, showed a variant pattern of hybridization caused by the loss of an EcoRI site. The same patient showed a markedly decreased amount of mRNA on Northern blot hybridization. Among the remaining patients, who showed no variation at the DNA level, two apparently lacked mRNA for alpha-L-fucosidase whereas the other three showed a transcript similar in size and amount to that observed in controls. These data confirm the genetic heterogeneity of the molecular defects causing fucosidosis in Italy.

Blotting, Northern↗

Analysis of miconazole and econazole in pharmaceutical formulations by derivative UV spectroscopy and liquid chromatography (HPLC).

Methods based on derivative UV spectrophotometry and high-performance liquid chromatography (HPLC) have been developed for the selective determination of miconazole and econazole in pharmaceutical dosage forms. A solid-phase extraction (SPE) procedure using a diol column gave quantitative drug extraction from formulated creams and provided purified sample solutions suitable for assay by the derivative UV spectrophotometric and HPLC methods. The proposed methods gave comparable accurate results, whereas a conventional UV spectrophotometric method was found to be seriously affected by excipients.

Chromatography, High Pressure Liquid↗

Determination of ethacrynic acid in pharmaceutical formulations by difference ultraviolet spectrophotometry after derivatisation with N-acetylcysteine.

A selective difference spectrophotometric procedure is described for the assay of ethacrynic acid (a diuretic drug) in pharmaceutical dosage forms. The method is based on the reaction of the drug with N-acetylcysteine at pH 7.4 and ambient temperature, affording thiol adducts having different spectral properties, and involves the measurement of the absorbance of the reaction mixture relative to an equimolar solution of unreacted ethacrynic acid. The absorbance at 270 nm (A(270)) and the absorbance difference, delta A=A(270-A(244), obtained from the difference spectrum, were found to be linearly correlated with the drug concentration. The proposed spectrophotometric procedure was applied successfully to the determination of ethacrynic acid in pharmaceutical formulations using a reversed-phase high-performance liquid chromatographic procedure as a reference method.

Acetylcysteine↗

Molecular analysis of a t(14;14) translocation in leukemic T-cells of an ataxia telangiectasia patient.

We have detected and cloned two rearrangements in the T-cell receptor alpha locus from a clone of somatic cell hybrids carrying a t(14;14)(q11;q32) chromosomal translocation derived from an ataxia telangiectasia patient with T-cell chronic lymphocytic leukemia. The T-cell clone carrying the t(14;14) chromosomal translocation was known to be present for greater than 10 years before the onset of overt leukemia. One molecular rearrangement of the T-cell receptor alpha locus corresponded to a functional variable-joining region (V-J) joining, whereas the other derived from the breakpoint of the t(14;14)(q11;q32) translocation. Chromosomal in situ hybridization of the probe derived from the t(14;14) breakpoint localized the breakpoint region to 14q32.1, apparently the same region that is involved in another ataxia telangiectasia characteristic chromosome translocation, t(7;14)(q35;q32). The 14q32.1 breakpoint is at least 10,000 kilobase pairs (kbp) centromeric to the immunoglobulin heavy chain locus. Sequence analysis of the breakpoint indicates the involvement of a J alpha sequence during the translocation. Comigration of high-molecular weight DNA fragments involved with t(7;14) and t(14;14) translocations suggests the presence of a cluster of breakpoints in the 14q32.1 region, the site of a putative oncogene, TCL1.

Alleles↗

Fine structure of the excretory system of the deep posterior (Ebner's) salivary glands of the human tongue.

Human deep posterior lingual glands (von Ebner's glands) are located beneath the circumvallate papillae. They are formed by tubuloalveolar adenomeres, intercalated ducts and excretory ducts coming together in the main excretory duct. The tubuloalveolar cells, pyramid-shaped, show large and dense secretory granules (clear cored) throughout the cytoplasm, rare basal folds and packed cisternae of rough endoplasmic reticulum (RER) at the basal pole. The columnar cells of the intercalated ducts are arranged in a monolayer. They are characterized by dense, clear-core secretory granules (mostly in the apical cytoplasm), a basal nucleus, well-developed RER and Golgi apparatus, and thin filaments distributed in supra- and perinuclear cytoplasm. Striated ducts are absent. Excretory ducts, coming together in the main duct, are lined by a bistratified epithelium. The inner layer consists of columnar cells showing bundles of tonofilaments with scarce secretory activity. The outer layer is composed of basal cells lying on the basal lamina. The main excretory duct, which opens at the bottom of the vallum, shows a stratified epithelium. The outer side is composed of 2-3 layers of malpighian cells lying on the basal lamina. The inner side consists of a single layer of cuboidal-columnar cells with dense apical granules and well-developed organelles synthesizing and condensing secretions. These cells interpolate with goblet cells, rare mitochondria-rich cells, ciliated cells and numerous small globous cells showing a clear matrix and lacking secretory granules. The cilia show a 9 + 2 microtubular structure with basal bodies provided with striated rootlets. Myoepithelial cells surround with their processes the basal portions of the secretory cells and the intercalated ducts. The conclusions concern some comparative aspects and some hypothesis on the functional role of goblet cells, ciliated cells and epithelial cells lining the different ducts, also in relation to the final secretory product.

Aged↗

Use of deoxyribonucleic acid probes in the identification of cell origin and detection of cellular contamination in human lymphoblastoid cell lines.

Established lymphoblastoid cell lines have provided a valuable reference source for studying neoplastic lymphoproliferative disorders in humans. However, two major problems are associated with the establishment and growth of these cell lines: (a) the established cell line may not represent the original neoplastic clone, and (b) contamination of the established cell line with the other cell lines may occur. Lymphoblastoid cell lines "W" and "SP5" were established from splenectomy specimens of two patients with hairy cell leukemia. Both cell lines displayed B cell characteristics by immunophenotypic and Ig gene rearrangement studies. The banding patterns of the rearranged Ig genes (heavy and light chains) in the W cell line were different and in the SP5 cell line were identical with the corresponding untransformed splenic cell lines, indicating that cell line SP5 did and cell line W did not represent the original neoplastic clone. Continuous cultures of some of the subclones derived from cell line W and SP5 led to the growth of the cell lines W15T, W17T, and SP5T which all demonstrated T cell features based on immunophenotypic and T cell receptor rearrangement studies. However, the T cell receptor alpha and beta rearranged bands as well as bands generated by hybridization with highly polymorphic DNA probes p YNH24 and 0-3315-32 in these three lines and a human T cell leukemia line (CEM), were identical indicating that W15T, W17T and SP5T cell lines were contaminated with CEM. Studies of gene establishment patterns and DNA polymorphisms by Southern blotting are effective methods to establish clonal identity and to rule out cellular contamination in lymphoblastoid cell lines.

B-Lymphocytes↗

Fine structure of bat deep posterior lingual glands (von Ebner's)

We studied the morphology and ultrastructure of the bat (Pipistrellus k.k. and Rhinolophus f.e.) deep posterior lingual glands (Ebner's glands) during hibernation, summer and after stimulation with pilocarpine. Ebner's glands are formed by serous tubulo-alveolar adenomeres and by an excretory system organized in intercalated ducts, long excretory ducts and a main excretory duct. The latter opens in the vallum which surrounds the circumvallate papillae and in the groove of the foliate papillae. The secretory cells, which lack basal folds, show abundant and dense granules (PAS+, Alcian blue -), microvilli (scarce during hibernation), a Golgi apparatus (well developed during summer and after stimulation with pilocarpine), a large nucleus and RER cisternae stacked at the basal pole. Centrioles, lipid droplets, heterogeneous bodies (in content and density, probably lipofuscin bodies), lysosomal multivesicular bodies and large, dense granules with a microcrystalline structure were also encountered. The lateral membranes of adjacent cells are joined by desmosomes; their interdigitations are neither numerous nor prominent during summer. Microfilaments, often gathered in small bundles, lie in the lateral, peripheral cytoplasm without any relation with desmosomes. In summer and particularly after stimulation with pilocarpine, the apical pole of the secretory cells is characterized by many long microvilli, pedunculated hyaloplasmic protrusions and secretory granules. During hibernation the lumen is filled with secretory material. Myoepithelial cells are arranged among secretory cells or between them and the basal lamina. The short intercalated ducts show similarities with the analogous ducts of the parotid gland. Striated ducts are absent. Excretory ducts are endowed with: a) an inner layer of cuboidal cells characterized by poorly developed cytoplasmic organelles, rare dense granules and a few small microvilli; b) an outer layer of basal cells lying on the basal lamina. Myoepithelial cells are absent. The main excretory duct is lined by a stratified epithelium with an inner layer of conical-pyramidal cells surrounded by two-three rows of basal cells. The conical-pyramidal cells show poorly developed organelles, an apical border with small short microvilli and a prominent terminal web.

Animals↗

A herpes simplex virus type 1 mutant resistant to benzhydrazone, an inhibitor of glycoprotein synthesis in herpesvirus-infected cells. Preliminary mapping of benzhydrazone-resistance and of a novel syncytial mutation.

Benzhydrazone (BH) is an inhibitor of glycoprotein biosynthesis. It acts selectively in Herpes simplex virus (HSV)-infected cells and does not significantly affect glycoprotein synthesis in uninfected cells and in cells infected with other viruses. Previously, we reported on a syncytial (syn) mutant, designated HSV-1(13)S11, resistant to BH, and showed that BH-resistance is encoded in the mutant virus DNA and therefore can be transferred into the genome of wild type HSVs. The present paper reports on a preliminary mapping in HSV-1(13)S11 genome of the loci which confer resistance to BH and of three distinct syn mutations present simultaneously in this mutant. Two of them were mapped in previously described syn loci localized in BamHI fragment L (map units 0.707-0.745) (locus syn 1) and BamHI fragment Q (map units 0.296-0.317) (locus syn 5). A third mutation not described before and mapping in BamHI fragment SP (c.a. map units 0.81-0.85) conferred the syn phenotype to both HEp-2 and Vero cells. This novel mutation has been designated herein locus syn 6. Transfer of BH-resistance could be achieved in cotransfection experiments involving two HSV-1(13)S11 fragments, BamHIL and BamHISP.

Animals↗

Comparative study of two strains of Bovid herpesvirus-4.

Two strains of Bovid herpesvirus-4 (BHV-4), i.e. the prototype strain DN-599, obtained from a steer suffering of a respiratory disease, and the strain 85/BH 16TV, originated from a cow with vulvovaginitis, were compared in studies which included restriction endonuclease analysis, experimental infection and reciprocal cross protection tests. The restriction endonuclease analysis revealed that the resultant DNA patterns of the isolates were generally similar with only a difference in one fragment. The two strains were capable of causing respiratory tract infection in calves, even if they displayed a different level of virulence: the strain 85/BH 16TV being the most virulent while the strain DN-599 the least. The two viral strains were mutually protective in that the calves were generally found to be refractory to challenge inoculation with either the homologous or the heterologous virus. Finally, both viral strains failed to evoke the production of neutralizing antibody in the experimental calves.

Animals↗

Determination of amiodarone hydrochloride in pharmaceutical formulations by derivative UV spectrophotometry and high-performance liquid chromatography (HPLC).

Assay procedures based on derivative ultraviolet spectrophotometry and high-performance liquid chromatography (HPLC) have been developed for the specific determination of amiodarone hydrochloride in pharmaceutical dosage forms. The use of first- and second-order derivative spectrophotometry was found to have suppressed the background absorption from the excipients with comparable accuracy and precision to the reversed-phased HPLC reference method. A conventional UV absorption method (lambda = 242 nm) is subject to possible interference by formulation excipients.

Amiodarone↗

The protection of newborn calves against experimental rotavirus infection by feeding mammary secretions from vaccinated cows.

Pregnant cows were given the first injection of an inactivated bovine rotavirus vaccine approximately 4 weeks before calving and a second injection just before calving. This led to the enhancement of rotavirus antibody titers in their colostrum as well as in the milk for at least 5 days after parturition. Thus, when newborn calves were fed with the mammary secretions obtained from the vaccinated cows daily for 5 consecutive days they were fully refractory to experimental infection with 81/36F bovine rotavirus. By contrast, the calves which were given the mammary secretions from unvaccinated cows, had clinical signs consistent with rotavirus infection and viral shedding. Based on these results it is suggested that vaccination of cows according to the scheme followed in this experiment, i.e., two injections within the last month of pregnancy, might be a valid approach which depending on confirmation under field conditions, could help reduce the incidence of rotavirus-induced diarrhea in newborn calves.

Animals↗