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At least 235 records · Page 13Linked to original sources
PubMed1997

SOS induction in mycobacteria: analysis of the DNA-binding activity of a LexA-like repressor and its role in DNA damage induction of the recA gene from Mycobacterium smegmatis.

The protein encoded by the lexA gene from Mycobacterium leprae was overproduced in Escherichia coli. The recombinant protein bound to the promoter regions of the M. leprae lexA, M. leprae recA and M. smegmatis recA genes at sites with the sequences 5'-GAACACATGTTT and 5'-GAACAGGTGTTC, which belong to the 'Cheo box' family of binding sites recognized by the SOS repressor from Bacillus subtilis. Gel mobility shift assays were used to confirm that proteins with the same site specificity of DNA binding are also present in Mycobacterium tuberculosis and M. smegmatis. Complex formation was impaired by mutagenic disruption of the dyad symmetry of the M. smegmatis recA Cheo box. LexA binding was also inhibited by preincubation of the M. smegmatis and M. tuberculosis extracts with anti-M. leprae LexA antibodies, suggesting that the mycobacterial LexA proteins are functionally conserved at the level of DNA binding. Finally, exposure of M. smegmatis to DNA-damaging agents resulted in induction of the M. smegmatis recA promoter with concomitant loss of DNA binding of LexA to its Cheo box, confirming that this organism possesses the key regulatory elements of a functional SOS induction system.

Bacterial Proteins↗
PubMed1998

Induction of necrosis by zinc in prostate carcinoma cells and identification of proteins increased in association with this induction.

Zinc exhibits inhibitory effects on apoptosis, and a deficiency in this metal generally causes this type of cell death to occur. In the present study, we found that exposure to zinc results in necrosis of prostate carcinoma cells. When zinc acetate was added to LNCaP or PC-3 cells in monolayer culture, they began to detach from the culture dishes, and viability was lost after 4-8 h. Most of the cell death was found to be due to necrosis as determined by double staining with fluorescein-isothiocyanate-labeled annexin V and ethidium bromide, and by detection of hypodiploid cells. Associated with the induction of necrosis was an increase in low molecular-mass proteins, identified by HPLC analysis to be thymosin beta10, parathymosin and GAGE in LNCaP cells, and thymosin beta4, parathymosin and metallothionein in PC-3. The time course of the increase of thymosin beta10 in LNCaP cells and thymosin beta4 in PC-3 cells was consistent with that of appearance of cell detachment and dead cells. These results indicate that zinc can induce necrosis and suggest that production of proteins including beta-thymosins is involved in induction of processes leading to cell detachment.

Adenocarcinoma↗
PubMed1989

UV induction of coliphage 186: prophage induction as an SOS function.

Our results show that UV induction of the 186 prophage depends upon the phage function Tum, with the mutant phenotype of turbid plaques on mitomycin plates and the expression of which is controlled by the host LexA protein. Tum function, encoded near the right-hand end of the coliphage 186 chromosome, is under the control of promoter p95. This promoter is overlapped by a sequence closely related to the consensus sequence of the LexA-binding site. It is proposed that inactivation of LexA after UV irradiation (or by genetic means) leads to prophage induction by permitting expression of Tum which, by unknown means, induces prophage. This mechanism is basically different from that seen with the UV-inducible lambdoid coliphages, which are not regulated by LexA.

Base Sequence↗
PubMed1999

Induction of hyporesponsiveness to intact foreign protein via retroviral-mediated gene expression: the IgG scaffold is important for induction and maintenance of immune hyporesponsiveness.

IgG molecules can be highly tolerogenic carriers for associated antigens. Previously, we reported that recipients of bone marrow or lipopolysaccharide-stimulated B-cell blasts, both of which were retrovirally gene-transferred with an immunodominant peptide in-frame with the variable region of a murine IgG heavy chain, were rendered profoundly unresponsive to that epitope. To further investigate whether tolerance to larger molecules can be achieved via this approach and whether the IgG scaffold is important for induction and maintenance of immunological tolerance, we engineered two retroviral constructs encoding the cI lambda repressor (MBAE-1-102 and MBAE-1-102-IgG) for gene transfer. Our results show that recipients of bone marrow or peripheral B cells, transduced with the MBAE-1-102-IgG recombinant, are hyporesponsive to p1-102. In addition, the self-IgG scaffold enhanced the induction and maintenance of such an immune hyporesponsiveness. Thus, our studies demonstrate that in vivo-expressed IgG heavy chain fusion protein can be processed and presented on the appropriate MHC class II, resulting in hyporesponsiveness to that antigen and offering an additional therapeutic approach to autoimmune diseases.

Animals↗
PubMed2000

Critical energies for SSB and DSB induction in plasmid DNA by low-energy photons: action spectra for strand-break induction in plasmid DNA irradiated in vacuum.

PURPOSE: To measure action spectra for the induction of single-strand breaks (SSB) and double-strand breaks (DSB) in plasmid DNA by low-energy photons and provide estimates for the energy dependence of strand-break formation important for track-structure simulations of DNA damage. MATERIALS AND METHODS: Plasmid pMSG-CAT was irradiated as a monolayer, under vacuum, with 7 150eV photons produced by a synchrotron source. Yields of SSB and DSB were determined by the separation of the three plasmid forms by gel electrophoresis. RESULTS: The yields of SSB per incident photon increased from 1.4x 10(-15) SSB per plasmid per photon/cm2 at 7eV to 7.5 x 10(-14) SSB per plasmid per photon/cm2 at 150 eV. Direct induction of DSB was also detected increasing from 3.4 x 10(-17) DSB per plasmid per photon/cm2 at 7eV to 4.1 x 10(-15) DSB per plasmid per photon/cm2 at 150eV. When the absorption cross-section of the DNA was considered, the quantum efficiency for break formation increased over the energy range studied. Over the entire energy range, the ratio of SSB to DSB remained constant. CONCLUSIONS: These studies provide evidence for the ability of photons as low as 7 eV to induce both SSB and DSB. The common action spectrum for both lesions suggests that they derive from the same initial photoproducts under conditions where the DNA is irradiated in vacuum and a predominantly direct effect is being observed. The spectral and dose-effect behaviour indicates that DSB are induced predominantly by single-event processes in the energy range covered.

DNA↗
PubMed1975

Tolerance induction in B lymphocytes but thymus-dependent antigens. T cells may abrogate B-cell tolerance induction by prevent an antibody response.

Thymus-dependent protein antigens such as fowl gamma globulin (FGG) and dinitrophenylated-human gamma globulin (DNP-HGG), readily induced tolerance of the B cell in the absence of T cells even when these antigens were not deaggregated. However, when the same doses of antigen were given in the presence of T cells, the B-cell population was shown to be protected from tolerance induction, especially when the antigen was not in a deaggregated form. In this case, there was in fact evidence of a priming effect, manifest in both the B-cell and T-cell populations. The priming effect on the B-cell population was demonstrated by an increased response of mice pretreated with DNP-HGG, upon challenge with DNP conjugated to a heterologous carrier. The priming effect on the T-cell population was evident in a helper effect demonstrated in vitro. However, when euthymic mice which had been pretreated with large doses of FGG or DNP-HGG were challenged with the homologous carrier, the results were different. In this case, there was a profound suppression of the response against the carrier or the hapten on that carrier. Suppressor activity was also demonstrated in vitro and was shown to be sensitive to treatment with anti-theta-serum plus complement. Additionally it was shown that the effector phase of the suppression had a definite nonantigen-specific component. Thus, in pretreated euthymic mice, provided the homologous carrier was present, the response to a heterologous carrier was also suppressed. To account for the observation that nondeaggregated antigens can induce B-cell tolerance in athymic mice, but B-cell priming and T-cell-mediated suppression in euthymic mice, it is proposed that B-cell tolerance occurs when antigen at some critical dose interacts with the B cell in the absence of some second signal. This second signal is normally provided by the macrophage, probably with the assistance of the T cell, and its effect is to divert the result of the interaction of the B cell with antigen towards immunization and away from tolerance induction. When a large dose of an antigen that tends to form aggregates is given to an animal possessing functional T cells, both T-dependent helper and T-dependent suppressor activities are generated, thus accounting for a situation where the B-cell population is immunized, but B-cell activation is suppressed in the presence of the original carrier.

Animals↗
PubMed1996

Influence of induction technique on intubating conditions after rocuronium in adults: comparison with rapid-sequence induction using thiopentone and suxamethonium.

We have assessed the effect of anaesthetic technique on intubating conditions after rocuronium 0.6 mg kg-1 in four groups (n = 25 each) of unpremedicated patients in whom anaesthesia was induced with either thiopentone 5 mg kg-1 or propofol 2.5 mg kg-1 alone, or supplemented with alfentanil 20 micrograms kg-1. Fifty control patients were anaesthetized with thiopentone followed by suxamethonium. Laryngoscopy was commenced at 45 s. Overall intubating conditions after rocuronium were similar to those after suxamethonium (good and excellent > or = 96%) only when alfentanil was part of the induction regimen. However, intubation time was similar in all five groups and averaged 55 (SD 3.2) s, and the tube could be passed through open vocal cords within 70 s. After rocuronium the response of the diaphragm to intubation was more pronounced in the two groups of patients not receiving alfentanil (P < 0.0001) and in patients anaesthetized using propofol with alfentanil (P < 0.01) than in the control group. Opioids (in doses equivalent to alfentanil 20 micrograms kg-1) constitute an integral part of an induction regimen containing rocuronium 0.6 mg kg-1, regardless of whether or not thiopentone or propofol is used, in order to achieve overall intubating conditions similar to those after suxamethonium.

Adolescent↗
PubMed1997

Aneuploidy induction by benzo[a]pyrene and polyploidy induction by 7,12-dimethylbenz[a]anthracene in Chinese hamster cell lines V79-MZ and V79.

We found that different ploidy effects were induced in four Chinese hamster-derived cell lines (V79-MZ, V79, CHL and CHO-K1) treated through two cell cycles with polycyclic aromatic hydrocarbons in the absence of a metabolic activation system. 5-Bromodeoxyuridine was used to investigate cell cycle delay and sister chromatid exchanges (SCE) induced by the chemicals. Benzo[a]pyrene (BP) induced aneuploidy at 2.5-10 micrograms/ml in V79-MZ cells. 7,12-Dimethylbenz[a]anthracene (DMBA) induced polyploidy at 3.125-6.25 and 6.25-1.25 micrograms/ml in V79-MZ and V79 cells respectively. Higher concentrations caused cell cycle delay and, therefore, did not affect ploidy. BP and DMBA did not induce a significant increase in SCE frequency at the above doses. 3-Methylcholanthrene tested up to its solubility limit (10 micrograms/ml) did not induce numerical aberrations in any cell line. The clastogen mitomycin C, tested up to 0.01 microgram/ml, did not produce numerical aberrations but did significantly increase SCE frequency in all cell lines. The spindle poison colchicine, tested up to 0.1 microgram/ml, induced ploidy changes in the four cell lines that showed different sensitivities. Four cell lines showed no arylhydrocarbon hydroxylase activity, and V79-MZ, but not the other cells lines, showed high glutathione S-transferase activity. Aneuploidy induction by BP and polyploidy induction by DMBA in the absence of S9 mix in vitro have not been described before, and the finding might be due to the effect on tubulin. Due to their specificity and high sensitivity, the V79-MZ and V79 cell lines might be good systems for detecting aneuploidogens.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1987

Induction of transplantation tolerance in rats by spleen allografts. III. The role of T suppressor cells in the induction of specific unresponsiveness.

Heterotopic (WAG x AGUS)F1 spleen allografts survive indefinitely when transplanted to normal AGUS recipients and induce long-term donor-specific unresponsiveness. In this report, we have examined the immune reactivity of spleen graft recipients soon after transplantation, in an attempt to define the immunological mechanisms responsible for the induction of donor-specific unresponsiveness. Unresponsiveness develops as early as one week after splenic transplantation. T cells obtained from the recipient lymph node and spleen exhibit reduced mixed lymphocyte reaction responses to donor (WAG) but respond normally to third-party (PVG) stimulators. In contrast, T cells obtained from the spleen graft are unresponsive to both donor and third-party stimulators. Donor specific T suppressor cells (Ts) appear in the recipient's lymph node and spleen by one week posttransplantation--however, at this time antigen nonspecific suppressor cells predominate in the spleen graft. Only minimal cytotoxic T cell activity could be detected in the spleen graft, with the host spleen and lymph nodes being devoid of cytotoxic T lymphocytes. Sera obtained one or two weeks following splenic transplantation did not contain cytotoxic alloantibodies, and only a very weak response could be detected at one month. These data demonstrate that the unresponsiveness associated with the spontaneous acceptance of spleen allografts is correlated with the early induction of antigen specific Ts in recipient lymphoid tissue and the presence of nonspecific suppressor cells at the graft site.

Animals↗
PubMed2006

Induction with rabbit antithymocyte globulin versus induction with corticosteroids in liver transplantation: impact on recurrent hepatitis C virus infection.

This study compares the clinical course of recurrent hepatitis C virus (HCV) infection between 64 patients, who were randomized to receive either rabbit antithymocyte globulin (RATG) or steroids as induction therapy with tacrolimus for maintenance. The HCV recurrence was assessed by HCV RNA levels, peak ALT at 3-6 months, the grade of inflammation at biopsy at 3-6 months posttransplant, progression of fibrosis, and survival. All patients had also received antiviral therapy with interferon alpha 2b and ribavirin, if there were no contraindications. There was no statistically significant difference between the two groups in terms of inflammation at 3 months, peak ALT, or HCV RNA. The survival between the two groups of patients was similar. It appears that steroid-free liver transplantation with RATG induction does not have any negative influence on HCV recurrence in hepatitis C patients after liver transplantation.

Adrenal Cortex Hormones↗
PubMed1993

Generalized Induction of Defense Responses in Bean Is Not Correlated with the Induction of the Hypersensitive Reaction.

Transcripts for phenylalanine ammonia-lyase, chalcone synthase, chalcone isomerase, and chitinase accumulated in common bean after infiltration with the Pseudomonas syringae pv tabaci Hrp- mutant Pt11528::Hrp1, even though a hypersensitive reaction did not occur. The temporal pattern of this transcript accumulation was similar to that seen after infiltration with wild-type P. s. tabaci Pt11528, which resulted in a hypersensitive reaction. Escherichia coli DH5[alpha], P. fluorescens Pf101, heat-killed Pt11528 cells, and Pt11528 cells treated with protein synthesis inhibitors also induced accumulation of defense transcripts but not a hypersensitive reaction. In contrast, these transcripts were not detected in plants infiltrated with water or P.s. pv phaseolicola NPS3121, a compatible pathogen that causes halo blight. Phytoalexins were produced in bean after infiltration with Pt11528, Pt11528::Hrp1, Pt11528 cells treated with neomycin, or Pf101, but not in plants infiltrated with NPS3121 or water. These results suggest that there are unique biochemical events associated with the expression of a hypersensitive reaction which are distinct from other plant defense responses such as phytoalexin biosynthesis. In addition, our results support the hypothesis that there is a general, nonspecific mechanism for the induction of defense transcripts and phytoalexins by pathogenic and saprophytic bacteria that is distinct from the more specific mechanism associated with the induction of the hypersensitive reaction.

Journal Article↗
PubMed2005

Mycophenolate mofetil in pediatric renal transplantation: non-induction vs. induction with basiliximab.

North American Pediatric Renal Transplant Cooperative Study (NAPRTCS) reports have shown anti-T cell antibody, OKT3, to be deleterious in pediatric renal transplant recipients treated with mycophenolate mofetil (MMF). Unlike OKT3, basiliximab is a chimeric monoclonal antibody to the alpha subunit of the interleukin-2 receptor on activated T-lymphocytes. We sought to examine the outcome of MMF with or without basiliximab induction therapy in pediatric renal transplantation. Between January 1998, and June 2001, 49 pediatric renal transplants were performed at our center and 41 met the criteria for this study. We retrospectively analyzed the records of 25 patients who received MMF, Prednisone, CSA or TAC, alone (group I) and 16 patients who received MMF, CSA or TAC, and Prednisone in combination with basiliximab (group II). The two groups were similar with respect to recipient or donor age, gender, ethnicity, donor source (LD vs. CAD), cold ischemia time, and primary diagnosis. The basiliximab group had a shorter follow up period because of its more recent addition to our pediatric immunosuppression protocol, 12.9 +/- 5.9 months vs. 35.5 +/- 7.2 months for group I (p < 0.0001). At 6 months, the acute rejection rate was 16% (group I) compared with 25% (group II) (p = 0.689). The patient and graft survival at 6 and 12 months were 100% respectively for both groups. Basiliximab was well tolerated without significant adverse events. At 6 months, there was no significant difference between the groups in the incidence of urinary tract infection or cytomegalovirus infection. These data suggest that in the short-term, MMF with or without basiliximab induction therapy appears to yield excellent and statistically similar outcomes. However, further controlled studies are necessary to verify these findings as well as to define the role of basiliximab in MMF-treated pediatric renal transplant recipients.

Adolescent↗
PubMedDate not supplied

Induction of acrosome reaction by low temperature is comparable to physiological induction by human follicular fluid.

The acrosome reaction was induced by means of low temperature or human follicular fluid (FF) in spermatozoa from 18 patients. Live acrosome-reacted spermatozoa were determined with the triple-stain technique. Both inducers of acrosome reaction had a highly significant correlation with the percentage of acrosome-reacted spermatozoa after induction of the acrosome reaction: (r = 0.9190; P < 0.0001) and the inducibility of AR (r = 0.8472; P < 0.0001). This might indicate that both the non-physiological (low temperature) and the physiological (FF) inducers use the same signal transduction pathway, and demonstrates the usefulness of low temperature induction for diagnostic purposes.

Acrosome↗
PubMed2004

Reproducibility of negative mood induction: a self-referent plus musical mood induction procedure and a controllable/uncontrollable stress paradigm.

Mood induction procedures (MIPs) have been used extensively as a means of inducing a variety of mood states in laboratory settings. Recently, MIPs have been employed in a repeated measures design in combination with such techniques as functional imaging and psychopharmacological investigations. This study attempted to assess the repeatability of two paradigms designed to induce a depressed/negative affect, a self-referent plus musical MIP and a controllable/uncontrollable stress paradigm, to assess the viability of using such procedures in a repeated design. Healthy volunteers were enrolled in to one of two studies. Study 1 administered a self-referent plus musical MIP on three separate occasions: neutral and depressive mood induction (MI) in a balanced order, followed by a further depressive MI. In study 2, volunteers were subjected to sessions of controllable and uncontrollable noise stress in a balanced order on two different occasions. Subjective ratings of mood were obtained prior to and following the interventions. Depressive MI successfully increased ratings of negative affect, although this effect tended to be greater on the first occasion than on the second. Correlation between mood change on the first and second occasions were low, both for the MIP and stress paradigm. As such, caution should be exercised in using these procedures in a repeated measures design. However, the results may vary depending on the type of MIP employed.

Adult↗
PubMed1995

An immunologic approach to induction of epidermal growth factor deficiency: induction and characterization of autoantibodies to epidermal growth factor in rats.

Epidermal growth factor (EGF) in pharmacologic doses is able to induce growth and development in the fetus and the newborn. To investigate the opposite situation, the effects of insufficient amounts of EGF during development, we wanted to establish an in vivo model with a state of EGF deficiency. This was attempted by induction of autoimmunity to EGF in rats. Twenty rats were immunized with EGF. Fifteen of these developed autoantibodies against EGF, which, as judged by Scatchard analysis, had a median apparent affinity constant of 14 x 10(9) L/mol and a median concentration of binding sites of 20 x 10(-9) mol/L. The antibodies recognized purified EGF from the submandibular glands (6 kD) and from urine (45 kD) and further native EGF in saliva and urine. The cross-reactivity toward transforming growth factor-alpha was below 3%. Binding of EGF by antibodies inhibited its binding to the EGF-receptor by approximately 97% in vitro. Investigation of in vivo metabolism of antibody-bound 125I-EGF confirmed these results, that is, the antibodies were able to inactivate EGF. The adult rats were unaffected by the induction and presence of autoantibodies, and the EGF-containing organs did not show any histologic signs of inflammation or tissue damage. Furthermore, as judged by immunohistochemistry, no major changes in the distribution and tissue concentration of EGF were seen in the adult rat. These results show that it is possible to induce homologous antibodies that can inhibit the binding of EGF to its receptor and further suggest that circulatory EGF is of no physiologic importance in the healthy, adult rat.

Animals↗
PubMed1989

Induction of ovarian cysts in progesterone-synchronized immature rats: evidence that suppression of follicular aromatase activity is not a prerequisite for the induction of cystic follicles.

Serum hormone profiles for women and animals with cystic ovaries have led to the hypothesis that elevated serum LH and androgens are involved in the induction of ovarian follicular cysts. To test the ability of LH-like activity to induce cysts, immature rats (bearing progesterone implants to suppress endogenous LH secretion) were assigned to treatment groups that received 0 (control), 0.1, 0.5, or 1.5 IU hCG twice daily for 9 days beginning on day 27 of life. Serum progesterone concentrations were maintained at approximately 156 ng/ml throughout the in vivo treatment period. By the morning of day 36 (day 10 of treatment) the largest follicles in ovaries from control animals were at the small antral stage of development. In contrast, the largest follicles in ovaries from rats receiving 0.1, 0.5, and 1.5 IU hCG were primarily at the large antral, precystic, and cystic stages of development, respectively. On all days tested, progesterone, androstenedione, and estradiol accumulation in medium alone was greater for follicles from rats receiving hCG than for follicles from control rats. Progesterone and androstenedione increased in response to cAMP in vitro in follicular incubations from both control and hCG-treated animals. Only estradiol production by follicles from rats treated with 0.1 or 0.5 IU hCG increased in response to cAMP in vitro. Follicles from all treatment groups produced significantly more estradiol in the presence of a saturating amount of aromatizable substrate than in medium alone. Indeed, on day 36, cystic follicles produced more than 8 ng estradiol when incubated in the presence of either androstenedione or testosterone. In addition, more androstenedione was accumulated in vitro when developing cysts were incubated with exogenous testosterone than when noncystic follicles were incubated under similar conditions. The results of these experiments demonstrate that chronic stimulation by LH-like activity is capable of inducing follicular cysts in a time- and dose-related manner in intact noncycling immature rats. The ability of these cysts to produce 1-2 ng estradiol in medium alone and even greater amounts of estradiol in the presence of exogenous androgen indicates that inhibition of the ability of FSH to induce and stimulate follicular aromatase activity is not a prerequisite for the induction of follicular cysts in these animals. Finally, the increasing ability to accumulate androstenedione in the presence of exogenous testosterone suggests that follicular 17 beta-hydroxysteroid oxidoreductase activity increases in response to chronic stimulation by low doses of LH-like activity during the development of follicular cysts.

Androstenedione↗
PubMedDate not supplied

Development of inductive heating equipment using an inductive aperture-type applicator.

We designed an inductive aperture-type applicator, IATA, consisting of a one-turn, square, column-like coil made of a metal strip. In experiments using a phantom, this applicator was seen to have the following characteristics: (1) it does not excessively heat the fat layer; (2) deep portions can be heated effectively; (3) heat can be applied to the subject, without contact with the applicator; (4) excessive generation of heat does not occur in parts of tissue near the edge of the aperture; (5) size of the aperture can be changed for adaptation to size of the site to be heated; (6) the aperture can be modified to fit the shape of the subject and the depth of penetration is influenced by size of the subject being heated; (7) the applicator can be driven at an arbitrary frequency; (8) efficiency of heat generation to the input power is low. The inductive heating equipment, 'Inductron', which we have developed possesses the necessary requirements for adequate clinical application.

Adipose Tissue↗
PubMed1990

Simultaneous multielement analysis of diet samples by inductively coupled plasma mass spectrometry and inductively coupled plasma atomic emission spectrometry.

Diet samples were collected by a duplicated portion study for an adult male in both Mito, Japan and Los Alamos, U.S.A. The ashed samples were analyzed with inductively coupled plasma mass spectrometry (ICP-MS) and inductively coupled atomic emission spectrometry (ICP-AES) for 22 elements. The trace and ultratrace elements, Ba, Mo, Ni, Co, Cd, Cs, Tl, Pb, Bi, Th, and U were determined by ICP-MS. The major and minor elements, Na, K, P, Ca, Mg, Fe, Zn, Mn, Al, Sr, and Cu were determined by ICP-AES. Accuracy and precision for some elements were examined by analyzing National Institute of Standards and Technology (formerly National Bureau of Standards) Standard Reference Materials, 1577b Bovine Liver, and 1573 Tomato Leaves. Simultaneous multielement analyses of diet samples by using the two methods were found to be very useful.

Adult↗