Biomedical subjects

A Diaz

Publications and source records attributed to A Diaz.

At least 91 records · Page 5Linked to original sources

PubMed1993

Transcriptional modulation of cartilage-specific collagen gene expression by interferon gamma and tumour necrosis factor alpha in cultured human chondrocytes.

To examine the possibility that cytokines produced in inflamed joint tissues may contribute to the loss of articular cartilage by causing inhibition of synthesis of cartilage-specific matrix macromolecules, we studied the effects of interferon gamma (IFN gamma) and tumour necrosis factor alpha (TNF alpha), alone and in combination, on the expression of the genes for types-II, -IX and -XI collagens in cultured human chondrocytes. Chondrocytes isolated from human fetal epiphyseal cartilage by sequential enzymic digestions were cultured in the presence of IFN gamma (30 pM), TNF alpha (15 pM) or a combination of suboptimal concentrations of both cytokines (1.5 pM IFN gamma plus 0.3 pM TNF alpha). IFN gamma caused a maximal decrease of 23.3-32.6% in the biosynthesis of collagen by chondrocytes. TNF alpha was a more potent inhibitor causing a 42.8-45.3% decrease at one-half the concentration of IFN gamma. A synergistic inhibitory effect of 58.2% was observed with the combination of 1.5 pM IFN gamma plus 0.3 pM TNF alpha. Electrophoretic analysis of the biosynthesized proteins showed a co-ordinate decrease in the production of the three cartilage-specific collagen types II, IX and XI. These effects were accompanied by parallel changes in the steady-state levels of their corresponding mRNAs. In vitro transcription assays showed that the collagen inhibitory effects of the cytokines occurred largely at the transcriptional level. Similar effects of the cytokines were observed on biosynthesis of types-II, -IX and -XI collagens and steady-state mRNA levels for type-II collagen by chondrocytes obtained from adult articular cartilage. These observations indicate that IFN gamma and TNF alpha can induce a synergistic inhibition of the synthesis of cartilage-specific collagens by fetal and adult human chondrocytes and suggest that these effects may contribute to the articular cartilage loss that occurs in inflammatory joint diseases.

Cartilage, Articular↗

PubMed1993

Regulation of human lung fibroblast alpha 1(I) procollagen gene expression by tumor necrosis factor alpha, interleukin-1 beta, and prostaglandin E2.

We investigated the participation of prostaglandin (PG) E2 in the regulation of the alpha 1(I) procollagen gene expression by tumor necrosis factor alpha (TNF alpha), and interleukin-1 beta (IL-1 beta) in normal adult human lung fibroblasts. TNF alpha (100 units/ml) and IL-1 beta (100 units/ml) stimulated the production of PGE2 and caused a dose-dependent inhibition of up to 54 and 66%, respectively, of the production of type I procollagen. Preincubation of cultures with indomethacin partially reversed the inhibition of procollagen production induced by the cytokines. Cytokine-stimulated endogenous fibroblast PG accounted for 35 and 68% of the inhibition induced by TNF alpha and IL-1 beta, respectively. Steady-state mRNA levels for alpha 1(I) procollagen paralleled the changes in collagen production. The transcription rate of the alpha 1(I) procollagen gene was reduced by 58% by TNF alpha and by 43% by IL-1 beta. Cytokine-stimulated endogenous PG production accounted for half of these effects. These results indicate that TNF alpha and IL-1 beta inhibit the expression of the alpha 1(I) procollagen gene in human lung fibroblasts at the transcriptional level by a PGE2-independent effect as well as through the effect of endogenous fibroblast PGE2 released under the stimulus of the cytokines.

Adult↗

PubMed1993

Effect of anti-gamma-interferon and anti-interleukin-4 administration on the resistance of mice against infection with reticulotropic and myotropic strains of Trypanosoma cruzi.

We studied the effect of in vivo administration of anti-gamma-IFN and anti-IL-4 monoclonal antibodies on the resistance of mice against myotropic and reticulotropic strains of Trypanosoma cruzi. Anti-gamma-IFN treatment augmented the susceptibility of mice when infected with the reticulotropic RA and Tulahuén strains of T. cruzi but did not alter the course of infection with the myotropic CA-I strain of the parasite. In vivo administration of anti-IL-4 enhanced the resistance of mice when infected with either Tulahuén or RA strains but did not affect the course of parasitemia when infected with CA-I. The possible biological relevance of these observations is discussed.

Animals↗

PubMed1993

Pigmented paravenous retinochoroidal atrophy.

Pigmented paravenous retinochoroidal atrophy is a rare disorder that is not well understood or classified. We examined a patient who had chorioretinal atrophy with pigment clusters located in the paravenous areas without macular involvement. Other conditions which may also present retinochoroidal atrophy and/or pigmentary degeneration are discussed. The electroretinography results were subnormal but the evoked potentials were not totally extinguished. Fluorescein angiography and retinography confirmed the relative and absolute scotomas corresponding to the atrophic paravenous areas.

Aged↗

PubMed1993

Human immunodeficiency virus seropositivity in adolescents with syphilis.

OBJECTIVE: The purpose of this study was to assess the relationship between syphilis and human immunodeficiency virus (HIV) infection among inner-city, minority group adolescents. METHODS: From August 1989 through June 1990, serum from all positive serologic tests for syphilis, obtained from patients attending a comprehensive adolescent health center in an acquired immunodeficiency syndrome epicenter and its two school-based clinics, were frozen without patient identifiers and were subsequently screened for HIV by enzyme-linked immunosorbent assay with confirmatory Western blot for positives. In addition, a retrospective chart review was performed for all patients with a positive serologic test for syphilis during the study period. RESULTS: Of the 59 specimens with a positive syphilis serologic test, 9 (15.3%) were HIV seropositive. Of the patients with syphilis, 57.4% were black and 42.6% were Hispanic; 16.4% were male (mean age 18.1) and 83.6% were female (mean age 17.8). Only 1 subject (female) was an injection drug user; 4 of the male subjects self-identified as having had sex with other males. Of the subjects, 27.8% had primary, 19.7% had secondary, and 52.5% had latent syphilis at the time of diagnosis. A prior or concurrent sexually transmitted disease was present in 90% of the males and 80% of the females; gonorrhea was the most prevalent sexually transmitted disease in the males (89%) and chlamydia was most prevalent in the females (35%). A history of chancroid and/or herpes was present in 16.4% of the subjects. CONCLUSIONS: It is concluded that the diagnosis of syphilis in an adolescent is a risk factor for HIV infection. All sexually active adolescents should be routinely screened for syphilis, regardless of sexual practices. Those with syphilis should be specifically counseled about their increased risk for HIV infection and the importance of consistent condom use, and they should be referred for formal HIV pretest counseling.

Adolescent↗

PubMed1992

Time-resolved fluorescence study of human recombinant interferon alpha 2. Association state of the protein, spatial proximity of the two tryptophan residues.

Human recombinant interferon alpha 2 belongs a to family of proteins active against a wide range of viruses. It contains two tryptophan residues located at positions 77 and 141 in the peptide sequence. The fluorescence emission spectrum of these tryptophan residues displays a maximum at 335 nm. The fluorescence intensity decay is described by one broad excited-state-lifetime population centered around a value of 1.7 ns (full width at half maximum, 1.5 ns). These observations suggest that in the native protein, both tryptophan residues emit from similar environments, not directly exposed to the surrounding solvent. The anisotropy decay is essentially biexponential. The correlation-time value characterizing the Brownian rotation of the protein varies linearly with the viscosity/temperature ratio. The calculated hydrodynamic volumes are compatible with the existence of a dimer and a tetramer, at pH 5.5 and 9.4, respectively. Addition of urea at pH 5.5 disrupts the dimer and modifies to some extent the excited-state-lifetime distribution which becomes more heterogeneous. Disulfide-bond reduction also dissociates the dimer and leads to a highly heterogeneous fluorescence-intensity decay with four excited-state-lifetime populations. An opening of the local structure in the Trp region of the protein is likely to occur in these conditions. The fast-anisotropy-decay components can be due to either fast rotation or energy transfer between the indoles. Close proximity of the two Trp residues (less than 1 nm) is suggested from steady-state and time-resolved fluorescence-anisotropy measurements in vitrified medium [95% (by mass) glycerol at -38 degrees C]. This suggestion is in agreement with the recently published three-dimensional structure of the homologous protein murine interferon beta [Senda, T., Shimazu, T., Matsuda, S. Kawano, G., Shimizu, H., Nakamura, K. T. & Mitsui, Y. (1992) EMBO J. 11, 3193-3201].

Fluorescence Polarization↗

PubMed1992

Activation of expression of genes coding for extracellular matrix proteins in Tat-producing glioblastoma cells.

The Tat protein of human immunodeficiency virus type 1 has been increasingly implicated in directly contributing to the disease AIDS by altering the expression of strategic cellular genes. In this study we demonstrate that the presence of the human immunodeficiency virus type 1 regulatory protein Tat is associated with a significant induction in the expression of certain protein components of the extracellular matrix in glial-derived cells. Northern blot analysis reveals that in cells expressing Tat there is a marked elevation in the steady-state RNA levels for fibronectin and types I and III collagen. Metabolic labeling of the Tat-producing cells demonstrates that this induction is also reflected at the level of protein synthesis. Transient transfection experiments indicate that the presence of Tat results in increased transcription of fibronectin and alpha I type I collagen promoters. Possible mechanisms for this phenomenon and their significance with regard to AIDS are discussed.

Collagen↗

PubMed1992

Alternative splicing of human prostaglandin G/H synthase mRNA and evidence of differential regulation of the resulting transcripts by transforming growth factor beta 1, interleukin 1 beta, and tumor necrosis factor alpha.

Prostaglandin G/H synthase (PGG/HS) is the rate-limiting enzyme in the conversion of arachidonic acid to prostaglandins and thromboxanes. We screened a human lung fibroblast cDNA library with an ovine PGG/HS cDNA and isolated a 2.3-kilobase clone (HCO-T9). Sequence analysis of this clone showed that (a) it contained the entire translated region of PGG/HS and (b) it displayed an in-frame splicing of the last 111 base pairs encoded by exon 9, which resulted in the elimination of the N-glycosylation site at residue 409. Polymerase chain reaction amplification with specific oligonucleotides of reverse-transcribed mRNA from diverse human tissues and cultured cells yielded 400- and 300-base pair fragments that corresponded, respectively, to the intact and spliced transcripts. The expression of these two transcripts in cultured human lung fibroblasts was differentially regulated by serum, transforming growth factor beta 1, interleukin 1 beta, tumor necrosis factor alpha, and phorbol 12-myristate 13-acetate, as each of these conditions stimulated preferentially the expression of the unspliced transcripts. The elimination of one of the four N-glycosylation sites by the alternative splicing of exon 9 and the differential regulation of this process by relevant cytokines and growth factors may represent a mechanism for the regulation of PGG/HS enzymatic activity under physiological or pathological conditions.

Amino Acid Sequence↗

PubMed1992

Predictors of repeat pregnancy outcome among black and Puerto Rican adolescent mothers.

This prospective study investigated predictors of repeat pregnancies by 12 months after the delivery of a first child and their outcomes in inner-city adolescent mothers. The sample included four groups: those who had therapeutic abortions, miscarriages, full-term deliveries, and no repeat pregnancy. The therapeutic abortion group had more pregnancies before their first delivery (41%) than did full term (20%) and no repeat (15%), p less than .01. More delayed grade placement was found in therapeutic abortion -1.6 years (1.3) and full term -1.8 years (.9) than in no repeat pregnancy -.6 years (1.1), p less .001. Reading achievement scores were higher in no repeat 86.3 (17.1) than in full term 75.0 (16.5), p less than .05. School attendance was higher in no repeat (65%) than in therapeutic abortion (35%) and full-term (24%) p less than .01 groups. Depressive symptoms at baseline were higher among therapeutic aborters 18.9 (9.9) than among full term 10.2 (8.2) and no repeat pregnancy groups 12.2 (6.2). Logistic regression analyses identified delayed grade placement as the most important predictor of pregnancy outcome.

Achievement↗

PubMed1992

Streptococcus pneumoniae DNA polymerase I lacks 3'-to-5' exonuclease activity: localization of the 5'-to-3' exonucleolytic domain.

The Streptococcus pneumoniae polA gene was altered at various positions by deletions and insertions. The polypeptides encoded by these mutant polA genes were identified in S. pneumoniae. Three of them were enzymatically active. One was a fused protein containing the first 11 amino acid residues of gene 10 from coliphage T7 and the carboxyl-terminal two-thirds of pneumococcal DNA polymerase I; it possessed only polymerase activity. The other two enzymatically active proteins, which contained 620 and 351 amino acid residues from the amino terminus, respectively, lacked polymerase activity and showed only exonuclease activity. These two polymerase-deficient proteins and the wild-type protein were hyperproduced in Escherichia coli and purified. In contrast to the DNA polymerase I of Escherichia coli but similar to the corresponding enzyme of Thermus aquaticus, the pneumococcal enzyme appeared to lack 3'-to-5' exonuclease activity. The 5'-to-3' exonuclease domain was located in the amino-terminal region of the wild-type pneumococcal protein. This exonuclease activity excised deoxyribonucleoside 5'-monophosphate from both double- and single-stranded DNAs. It degraded oligonucleotide substrates to a decameric final product.

Bacterial Proteins↗

PubMed1992

[Distal aneurysm of the postero-inferior cerebellar artery. Anatomical and surgical considerations].

A patient with a rare peripheral aneurysm of the posterior inferior cerebellar artery (P.I.C.A.) was treated by microsurgery. Anatomical features and surgical procedures are discussed. The aneurysm was located in the retromedullary, tonsillomedullary segment of the right P.I.C.A. and formed a caudal or infratonsillar loop in the cisterna magna close to the inferior part of the tonsil. The operation was performed with the patient in the sitting position using a median suboccipital approach through a cervical 1 (C1) laminectomy. The widened neck of the aneurysm was clipped successfully and the post-operative course was completely asymptomatic. A vertebral angiogram control showed the P.I.C.A. to be patent. The surgical approach should be related to the site of the aneurysm. Those affecting the telovelotonsillar segment of a P.I.C.A., forming cranial or supratonsillar loops, and those involving its cortical segment can be effectively operated using the same median suboccipital approach. Aneurysms arising from the anterior or lateral medullary segment of a P.I.C.A. should be approached through a unilateral suboccipital craniectomy extending down to the foramen magnum. Although clipping the neck of the aneurysm is preferable, trapping is possible for lesions arising from or distal to the telovelotonsillar segment, this preserving the small arteries arising from the lateral and posterior medullary segments.

Adult↗

PubMed1991

Regulation of transforming growth factor-beta 1 gene expression by glucocorticoids in normal human T lymphocytes.

Glucocorticoids (GC) modulate immune function in a number of ways, including suppression of T cell proliferation and other IL-2-mediated T cell functions. These inhibitory effects are similar to those induced by transforming growth factor-beta 1 (TGF-beta 1), a cytokine with potent T cell inhibiting activities. We examined the hypothesis that GC effects may be at least partially achieved through modulation of the expression of the TGF-beta 1 gene in activated T cells. Normal T cells were cultured with or without purified phytohemagglutinin (PHA-p) and 4 beta-phorbol 12-myristate 13-acetate (PMA) in the presence or absence of the synthetic GC, dexamethasone (100-200 micrograms/ml). The production of latent and active forms of TGF beta by these cells were analyzed by immunoblotting and bioassays. The steady-state levels of TGF-beta 1 mRNA were analyzed in total RNA from these cells by Northern hybridizations using a human TGF-beta 1 cDNA. The results showed that dexamethasone caused an increase in TGF beta production and a dose-dependent two to fourfold increase in TGF-beta 1 mRNA in activated as well as in unstimulated T cells, 1 h after exposure of the cultures to the steroid. The increase in TGF-beta 1 mRNA levels by dexamethasone was further potentiated two to threefold by cycloheximide, suggesting that the steroid effect may be due to inhibition of the synthesis of proteins that decrease TGF-beta 1 gene transcription or the stability of its transcripts. Finally, in vitro nuclear transcription studies indicated the dexamethasone effects on TGF-beta 1 gene expression to be largely transcriptional.

Cells, Cultured↗

PubMed1991

[Secondary penetration of a ventriculo-peritoneal shunt into the intestines. Possible cause of a recurring sepsis?].

We present a 28-year-old-patient with a severe head injury: skull fractures, epidural hematoma, subarachnoid hemorrhage, Glasgow coma score 7, and aspiration following a motorbike accident. A systemic infection with symptoms of shock and Staphylococcus aureus in blood culture specimens developed a few days after admission (later Staphylococcus epidermidis was also cultured). A posttraumatic hydrocephalus was treated by a ventriculo-peritoneal shunt inserted at mini-laparotomy. In multiple microbiological and cytological tests the cerebrospinal fluid (CSF) was always sterile. Enterocolitis occurred with Clostridium difficile and Staphylococcus aureus in stool cultures. After 6 months' intractable sepsis the patient died with multiple-system failure. Autopsy revealed secondary displacement of the shunt catheter into the intestinal lumen. A possible ascending infection was found in the form of a cerebral ventricular empyema. However, prior to death there was no specific clinical sign of peritonitis or encephalitis or a positive microbiological or cytological CSF findings. Despite insertion of a ventriculo-peritoneal shunt under visual control, this case shows that secondary displacement of the peritoneal extremity into the bowel can occur, which may cause a cerebral and eventually a systemic infection. CSF examinations may fail to show contamination; specific clinical signs may be absent or, with multiple-system failure, misleading. An autopsy is generally to be recommended as it contributes to a better understanding of the clinical problems in most cases.

Adult↗

PubMed1990

Frequency of use, knowledge, and attitudes toward the contraceptive sponge among inner-city black and Hispanic adolescent females.

This study assessed the frequency of use, knowledge, and attitudes toward the contraceptive sponge in sexually active, inner-city, black and Hispanic adolescent females. A 54-item, self-administered questionnaire was completed by 208 adolescents (mean age 17.9 years) attending a gynecology/family planning clinic within a comprehensive adolescent health center. Twenty-nine (13.9%) had never used any birth control, and 49 (23.6%) had not used any form of birth control in the 3 months prior to the study. Fifty-three (25.5%) of the adolescents had used a less reliable method such as rhythm and/or withdrawal. The birth control method used least was the sponge. Only four (1.9%) of the adolescents had used the sponge in the 3 months prior to the study, and only ten (4.8%) had ever used it. Knowledge of the correct use of the sponge was poor. However, 69 (44.8%) of the 156 girls who knew about the sponge had a positive attitude toward it, and only 19% said they did not like it. The contraceptive sponge may be a viable option for adolescents, particularly those who are not using birth control or who are using a less reliable method.

Adolescent↗

PubMed1990

Ultrasonic forms of posterior staphyloma.

The most frequent deformations of the globe encountered in pediatric ophthalmology are staphylomas and colobomas. In this paper we reviewed 16 ultrasonographically diagnosed staphylomas and classified the ultrasonographic patterns produced by staphyloma cases into four types: (1) staphylomas involving the posterior pole but not the optic nerve head, (2) staphylomas involving the posterior pole and the optic nerve head, (3) peripapillary staphylomas, and (4) giant peripapillary staphylomas. Morphologic and ultrasonographic characteristics are described.

Eye Abnormalities↗

PubMed1989

The staphylococcal toxic shock syndrome toxin 1 triggers B cell proliferation and differentiation via major histocompatibility complex-unrestricted cognate T/B cell interaction.

The Staphylococcus aureus exotoxin toxic shock syndrome toxin 1 (TSST-1) is a potent activator of T cells and monocytes. We have recently demonstrated that TSST-1 is a superantigen that binds monomorphic determinants on MHC class II molecules. In the present study, we have examined the effect of TSST-1 on the activation and differentiation of high density human tonsillar B cells. TSST-1 bound to tonsilar B cells with high affinity and saturation kinetics. This binding was effectively inhibited by a combination of anti-HLA-DR and anti-HLA-DQ mAbs. Treatment of purified B cells with TSST-1 failed to induce B cell proliferation or Ig production. However, in the presence of irradiated T cells, TSST-1 induced resting B cells to proliferate and differentiate into Ig secretory cells. TSST-1 mimicked nominal antigen in that its induction of B cell responses was strictly dependent on physical contact between T and B cells, and was profoundly inhibited by anti-MHC class II mAbs, anti-CD3 mAbs, and, to a lesser extent, by anti-CD18 mAbs. However, unlike nominal antigen, TSST-1-mediated T/B cell interactions were MHC unrestricted. These results suggest that TSST-1 induces T cell-dependent B cell proliferation and differentiation by virtue of its ability to mediate MHC-unrestricted cognate T/B cell interaction via the TCR/CD3 complex and MHC class II antigens.

Antibodies, Monoclonal↗

PubMed1989

Transforming growth factor-beta stimulation of lung fibroblast prostaglandin E2 production.

Transforming growth factor-beta (TGF beta) stimulated the production of total protein, collagen, and fibronectin by normal human lung fibroblasts. The stimulatory response was maximal at 100 pM TGF beta and reversed toward control at higher concentrations. Inhibition of fibroblast prostaglandin (PG) synthesis enhanced TGF beta-induced stimulation of total protein, collagen, and fibronectin production and reversed the negative slope of the dose-response curve at high concentrations of TGF beta. Determination of the steady-state levels of Types I and III procollagens and fibronectin mRNAs employing specific cDNA probes demonstrated that inhibition of fibroblast PG production increased the stimulatory effect of TGF beta on the levels of these transcripts. Exogenous PGE2 abrogated the stimulatory effects of TGF beta. These findings suggest that fibroblast stimulation by TGF beta may be down-regulated by endogenous PG synthesized in response to TGF beta. This notion was supported by the demonstration that TGF beta markedly stimulated fibroblast PGE2 production. These results indicate that TGF beta-induced stimulation of fibroblast PGE2 production may be an autoregulatory control mechanism to limit the effects of TGF beta on connective tissue protein synthesis.

Blotting, Northern↗

PubMed1989

Characterization of the polA gene of Streptococcus pneumoniae and comparison of the DNA polymerase I it encodes to homologous enzymes from Escherichia coli and phage T7.

The DNA sequence of the polA gene of Streptococcus pneumoniae was determined, and the DNA polymerase I encoded by the gene was purified to homogeneity. Determination of the amino-terminal amino acid sequence of the protein showed it to correspond to the Mr 99,487 polypeptide predicted from the nucleotide sequence. The mRNA transcript was mapped with respect to its sites of initiation and termination in the DNA. Inasmuch as the mRNA begins only two nucleotides before the first codon, it lacks a typical ribosome binding site. Nevertheless, 500 molecules of the protein are produced per cell. Like the Escherichia coli DNA polymerase I, the protein from S. pneumoniae has 5'- and 3'-exonuclease as well as polymerase activities, and it also undergoes a single cleavage on mild proteolysis. Alignment of the two different polymerase I proteins shows 40% of their amino acid residues to be identical. Homology is evident also with the DNA polymerase encoded by phage T7 gene 5. In addition, the amino-terminal regions of the bacterial polymerase I proteins are homologous to the separate 5'-exonuclease protein encoded by phage T7 gene 6. Analysis of the patterns of homology suggests that the bacterial polymerase I may represent the accretion of at least six separate genetic regions.

Base Sequence↗