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Biomedical subjects

A Caputo

Publications and source records attributed to A Caputo.

At least 73 records · Page 4Linked to original sources

Pyridoxol L,2-pyrrolidon-5 carboxylate prevents active fibroplasia in CCl4-treated rats.

In the present study we evaluated the protective activity of pyridoxol L,2-pyrrolidon-5 carboxylate (metadoxine) against CCl4 intoxication in rats, especially in relation to liver fibrosis. After 6 consecutive weeks of CCl4 treatment, the animals developed liver fibrosis and inflammation as revealed by histological analysis which also included semiquantitative scoring of these features. In addition the serum levels of the immunoreactive prolyl hydroxylase (SIRPH), an enzyme involved in the hydroxylation of the procollagen molecule, were significantly higher (44.2 +/- 16.3 micrograms/ml; P less than 0.005) in this group of animals than in controls (26.1 +/- 8.06). On the contrary, animals treated with CCl4 + metadoxine (200 mg/kg i.p.) had less severe liver fibrosis and normal SIRPH levels (21.5 +/- 14.6). These data suggest that metadoxine may be an effective pharmacological tool for preventing the progression of liver disease in rats exposed to CCl4 to cirrhosis.

Animals↗

Reciprocal in vitro interactions between human herpesvirus-6 and HIV-1 Tat.

The transactivation induced by human herpesvirus 6 (HHV-6) on the HIV-1 promoter was studied both in the presence and in the absence of the retroviral transactivator protein (Tat) constitutively expressed in Jurkat cells. Jurkat-tat cells were infected with HHV-6, transfected with a plasmid containing HIV-1 long terminal repeat (LTR)/chloramphenicol acetyltransferase (CAT), and CAT assays were performed. HHV-6 infection in the presence of Tat resulted in significantly higher LTR activation than that observed by Tat or HHV-6 alone, indicating that HHV-6 and Tat interact synergically. Furthermore, it was demonstrated that expression of HIV-1 tat inhibits HHV-6 replication, as shown by a 3.6-15.4-fold reduction in infectious yield. We suggest that HHV-6 could trigger HIV reactivation in HIV-seropositive patients which, in turn, could inhibit HHV-6 production.

Cell Line, Transformed↗

Transcellular transactivation by the human immunodeficiency virus type 1 tat protein.

The human immunodeficiency virus type 1 (HIV-1) transactivator (tat) protein produced in one cell activated HIV-1 promoter-directed gene expression in a second cell, provided the cells were in direct contact with one another. This observation suggests that the tat protein produced in HIV-1-infected cells has a physiological effect on neighboring cells.

Cell Line↗

High expression of exogenous cDNAs directed by HIV-1 long terminal repeat in human cells constitutively producing HIV-1 tat and adenovirus E1A/E1B.

A eukaryotic vector-host cell system is described where the additive transactivating effects of HIV-1 tat and adenovirus E1A on HIV-1 long terminal repeat (LTR) are exploited to increase expression of exogenous cDNAs. Human 143B and 293 cells, the latter constitutively producing E1A, were used as host cell lines. The bacterial gene chloramphenicol acetyltransferase (CAT) and the hepatitis B surface antigen (HBs-Ag) gene were employed as reporter genes inserted in pRPneoU3R, an episomal vector containing BK virus replication origin and early region, where cDNAs are expressed under control of HIV-1 LTR. The 293 cells were transformed by tat expression vectors to constitutively express tat. Stable cell clones of 293tat cells, constitutively expressing CAT after transformation with pRPneoU3R-CAT, show a CAT activity 600-fold higher than normal 293 transformed cells. CAT expression obtained in normal 293 cells can be transiently increased 10-fold by transfection by vectors expressing tat. The 293tat cells transformed by pRPneoU3R-HBs, an episomal vector expressing HBs-Ag from HIV LTR, yielded stable cell clones secreting HBs-Ag in the culture medium at a concentration up to 744 ng/ml or 44 ng/10(6) cells/24 h, 48-fold more than normal 293 cells. The use of this system for constitutive or inducible expression of sequences under control of HIV-1 LTR is discussed in view of possible applications for diagnostic, vaccinal and therapeutic purposes.

Adenovirus Early Proteins↗

Nucleotide sequence and genomic organization of a human T lymphocyte serine protease gene.

We have determined the nucleotide sequence of 4508 base pairs of human genomic DNA which contain the human serine esterase gene from cytotoxic T lymphocytes (SECT) (equivalent to the 1-3E cDNA clone) and include 879 bp of 5' flanking DNA and 393 bp of 3' flanking DNA. The gene consists of five exons of 88, 148, 136, 261, and 257 nucleotides separated by four introns of 1043, 455, 205, and 643 nucleotides. The location of introns with respect to protein coding sequences in the SECT gene is identical to that of the human cathepsin G and murine granzyme B genes. Comparison of SECT gene exonic sequences to murine granzyme B-F cDNA sequences indicates similarities of 75 and 72% for granzymes B and C and 61, 59, and 61% for granzymes D, E, and F, respectively. The 5' flanking sequence of the SECT gene showed similarity only to the 5' flanking sequence of the murine granzyme B gene, indicating that these genes are homologous. Comparison of the SECT gene sequence to the human cathepsin G sequence indicated no similarity in the 5' flanking DNA although the exonic sequences show 64% sequence similarity overall and 45% sequence similarity in the respective 3' untranslated regions. These similarities suggest that the SECT and cathepsin G genes are members of the same family of serine protease genes. Evidence from high and low stringency Southern transfer analysis of human genomic DNA indicates the presence of another gene of at least 85% sequence similarity to the SECT gene.

Base Sequence↗

Relative flexural strength of dental restorative ceramics.

The relative flexural strengths of ten brands of dental restorative ceramics were evaluated by a three-point bending test. The materials consisted of three low-fusing and one high-fusing feldspathic porcelains and six reinforced dental restorative ceramics which are currently in clinical use. The reinforced ceramic materials investigated utilized a number of different strengthening processes, including alumina and ceramic fiber reinforcement and controlled crystallization. The results of the investigation indicate that significant differences exist among the measured breaking strengths of the various materials. The processes of controlled crystallization and alumina reinforcement appear to be adequate means of improving the bending strength of restorative dental ceramics.

Aluminum Oxide↗

The potential role of lonidamine (LND) in the treatment of malignant glioma. Phase II study.

Up-to-date unsatisfactory results obtained in multimodality treatments of malignant glioma have prompted the research of new therapeutic modalities with 'unconventional' modes of action. Lonidamine (LND) is a drug which reduces aerobic glycolytic activity in both human and experimental tumors. This effect mainly depends on the inhibition of mitochondrially-bound hexokinase (HK) which is present in large amounts in malignant cells. A Phase II study was conducted on patients with recurrent glioma; 12 patients were admitted to the study. Clinical side effects were moderate, necessitating a reduction of the dosage in only 1 case. The objective results were evaluated according to the indications of Levin. 2 responders and 3 cases of stable disease were observed out of 10 evaluable patients. The potential value of this new drug is discussed.

Adolescent↗

[A comparative analysis of the characteristics of META-DENT denture resin system].

A newly developed denture resin based on a 4-META containing monomer became recently commercially available. It has been claimed that this resin system has the unique ability of creating bond to Cr-Co alloys without any mechanical retention. The objective of this study was to determine the chemical composition of the new resin, to study the bonding mechanism with a Cr-Co alloy and to compare its physical and mechanical properties to a commonly used acrylic denture resin. The analysis revealed that the new system contains 5% by weight 4-META in MMA liquid and 28% poly (ethyl methacrylate) in PMMA. No practical differences were noted in the stress-strain characteristics and hardness values between the resin systems. The META-DENT resin was more abrasion resistant. The bond of META-DENT to the Cr-Co alloy was greatly superior than that of the conventional resin. The bonding mechanism may be related to enhanced physical wetting due to reduced META-DENT viscosity and probably complexation of the anhydride groups with either Cr or Co.

Acrylic Resins↗

Co-operation in cell transformation between BK virus and the human c-Harvey-ras oncogene.

Early-passage hamster embryo cells were transformed by recombinant DNA molecules containing BK virus (BKV) early-region gene and either the activated c-Ha-ras oncogene (pBK/c-rasA) or the normal c-Ha-ras proto-oncogene (pBK/c-rasN). The recombinant DNAs had a greater transforming ability and converted hamster cells to a more malignant phenotype than the single genes transfected separately. pBK/c-rasA was significantly more powerful than pBK/c-rasN in conferring to cells all the characteristics of transformation. Transfected DNA sequences were integrated mostly as single insertions into cellular DNA. Specific c-Ha-ras and BKV transcripts as well as c-Ha-ras p21 and BKV T antigen were detected in transformed cells. Although stimulation of c-Ha-ras expression by BKV enhancers cannot be excluded in recombinants, super-transfection and co-transfection experiments in hamster embryo cells and pre-neoplastic cell lines showed that BKV early-region and c-Ha-ras co-operate in transformation by contributing separate and independent functions.

Animals↗

Structure and differential mechanisms of regulation of expression of a serine esterase gene in activated human T lymphocytes.

A cDNA clone encoding a polypeptide resembling proteolytic serine esterases (from cytotoxic T-cells; SECT) was isolated from human peripheral blood lymphocytes which had been cultured in the presence of the T-cell mitogen phytohemagglutinin for 72 h. The cDNA encodes a polypeptide of 247 amino acids which show homology of 99% with the protein sequence encoded by a cDNA clone (1-3E) isolated from staphylococcal enterotoxin A-stimulated human peripheral blood lymphocytes and 68% with the protein sequence (cytotoxic cell protease type I) derived from a cDNA clone (C11) encoding a serine esterase isolated from a murine cytotoxic T-cell line. The overall nucleotide sequence homology between the SECT cDNA and 1-3E was 99% and 73% between SECT and C11. Comparing the coding regions of SECT and C11 showed 75% homology, whereas the 5'- and 3'-untranslated regions showed 67% homology. Phytohemagglutinin stimulation results in 30-, 60-, and 370-fold increases in cytoplasmic SECT mRNA with respect to unstimulated cells after 6-, 24-, and 72-h cultures, respectively. At 6 h, the increase in SECT mRNA occurs in the absence of increases in SECT gene transcription and cytoplasmic mRNA stabilization. A 5-fold increase in SECT nuclear RNA seen at this time suggests that stabilization of SECT nuclear RNA transcript is responsible for early increases in SECT mRNA levels. At 24 and 72 h, the increased cytoplasmic SECT mRNA levels can be accounted for by increased transcriptional activity of the SECT gene.

Amino Acid Sequence↗

Effect of lonidamine on human malignant gliomas: biochemical studies.

Lonidamine (LND) has been shown to inhibit tumor aerobic glycolysis. Its effect was evaluated on several human astrocytomas at different degrees of malignancy; a correlation was found between LDN effect on lactate production and tumor malignancy: in grade I and II astrocytomas LND stimulates lactate production, while in grade III, IV and glioblastoma multiforme lactate production is inhibited. In an attempt to explain this different behaviour, hexokinase content and compartmentation was evaluated in astrocytomas from fresh operatory specimens and from cultured cells as well, observing a significative correlation between malignancy, hexokinase activity, percent of mitochondrially-bound hexokinase and LND effect. The results justify from a biochemical point of view the role of LND as a 'non-conventional' agent in multimodality combined treatment for malignant gliomas.

Astrocytoma↗

New BK virus episomal vector for complementary DNA expression in human cells.

The properties of pRP-c, a new vector for complementary DNA (cDNA) expression, are described. The vector contains the early region and replication origin of BK virus (BKV), a human papovavirus. Due to the presence of these BKV sequences, pRP-c replicates in human cells allowing amplification of inserted cDNAs. The promoter, intron and polyadenylation region for cDNA expression are separated by unique restriction sites and can therefore be individually excised and substituted with different transcription signals. Coding sequences of the bacterial genes for chloramphenicol-acetyl transferase (CAT) or neomycin phosphotransferase (neo) were inserted into the cDNA cloning site of pRP-c and expressed in human cells in transient assays or stable clones. In both cases expression of the inserted sequences was significantly more efficient than by using the integration vectors pSV2CAT and pSV2neo, demonstrating the advantages of episomal expression vectors in human cells. Possible uses of pRP-c to express viral and cellular cDNAs in human cells are discussed.

BK Virus↗

Factors affecting amplification of BK virus episomal vectors in human cells. Brief report.

Analysis of factors determining replication of BK virus (BKV) episomal vectors in human cells showed that vector copy number was related to the level of BKV T antigen expression. T antigen was synthesized efficiently, as assessed by indirect immunofluorescence, in vector-transfected primary embryonic fibroblasts undergoing neoplastic transformation. Surprisingly, transfected continuous cell lines (143 B, HeLa and KB), kept under biochemical selection or tested in transient assays, produced negligible amounts or no T antigen, revealed only by a sensitive ELISA test, suggesting that in these cells vector amplification was under the control of cellular factors. Presence or absence of BKV late region sequences, BKV strain, orientation of the inserted genes and presence or absence of selection were not relevant for vector replication. Type of biochemical selection, however, was important, since BKV vectors containing the thymidine kinase gene replicated better than those containing the neo gene. Despite great variability, vector copy number increased in transfected clones of adenovirus 5-transformed 293 cells, in the absence of immunofluorescence detectable T antigen. These cells express adenovirus immediate early proteins E1A and E1B which may directly or indirectly activate BKV origin of replication.

Antigens, Polyomavirus Transforming↗

Modulation of adriamycin uptake by lonidamine in Ehrlich ascites tumor cells.

The effect of Lonidamine, 1-(2,4 dichlorobenzyl)-1-H-indazol-3-carboxylic acid, on the uptake of Adriamycin by Ehrlich ascites tumor cells has been investigated. The uptake of Adriamycin is greatly stimulated by Lonidamine and the increase depends on the energy sources of the cell. In the presence of glucose the intracellular drug content is remarkably lower than that in its absence. This difference lies in the mechanism by which Lonidamine enhances the uptake of Adriamycin. The Adriamycin efflux is via an active transport process and, in the presence of glucose, both aerobic glycolysis and oxidative phosphorylation contribute to ATP synthesis. Although Lonidamine inhibits both these pathways, there is still sufficient ATP to extrude a certain amount of Adriamycin. The elevated intracellular concentration of Adriamycin depends not only on the Lonidamine-inhibited outward transport but also on higher membrane permeability which allows a low concentration of Adriamycin (18 microM) to interfere also with the oxidative metabolism of Ehrlich ascites tumor cells.

Animals↗

Retention magnets in guiding plates of distal-extension removable partial dentures.

This investigation studied vertically placed magnets that act simultaneously as guiding plates and retentive devices. The retention of vertically placed magnets was less than that of horizontally or obliquely placed magnets, but comparable to that of I-bar retainers. Load-induced stresses were lower, transmitted more axially to the abutments and were generally more equitably distributed than stresses produced by magnets in other orientations to the abutment tooth. The force distribution characteristics were less stressful than those of a comparable RPD with distal guiding plates, mesial rests, and I-bar retainers. Although the investigators were initially concerned about the relative capabilities of the vertically placed magnets, in vitro and in vivo studies demonstrated that retention was as good if not better than that of conventional clasp designs.

Dental Stress Analysis↗