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At least 595 records · Page 33Linked to original sources
PubMed2005

Proteomic characterization of the acid tolerance response in Lactococcus lactis MG1363.

Exponentially growing cells of Lactococcus lactis MG1363 are able to develop an Acid Tolerance Response (ATR) when incubated at pH 5, in both rich (M17)--and chemically defined (SA)--culture media. Physiological and proteomic characterization of this adaptive response indicated that L. lactis reorganizes its metabolism in response to acid stress to a great extent and quite differently in the two media. The development of ATR was fully dependent on protein de novo synthesis in SA and only partly dependent in M17. 2D gel electrophoresis revealed a total of 90 spots induced by acidity, 80 of which were identified by mass spectrometry. Only 10 proteins (BglA, PycA, GlmS, HasC, ArgS, GatA, AtpA, ArcB, Cfa, and SodA) were overproduced in the two media. A transcriptional analysis of the corresponding genes suggested that for half of them the mode of regulation may differ in the two media. Among the protein spots upregulated during the ATR in SA but not in M17, 13 already displayed an elevated rate of synthesis in M17 at neutral pH. These proteins could play an important role in the development of the protein de novo synthesis-independent ATR observed in M17.

Adaptation, Physiological↗
PubMed1997

Reconstitution of a functional bovine papillomavirus type 1 origin of replication reveals a modular tripartite replicon with an essential AT-rich element.

A functional replication origin was reconstituted using oligonucleotide cassettes corresponding to three sequence subelements within the Bovine Papillomavirus Type 1 (BPV-1) replication origin: the 23-bp AT-rich region (ATR), the 18-bp binding site for the viral replication initiator protein E1 (E1BS), and a binding site for the viral transcriptional transactivator and replication enhancer protein E2 (E2BS). Replication of the reconstituted origin depended on heterologous expression of both the E1 and E2 proteins and on the presence of both the E1BS and E2BS, indicating that it is functionally analogous to the authentic BPV-1 origin. In addition, pairwise testing of subelement combinations revealed that the ATR was also essential and that a functional origin required at least one copy of all three subelements. While the E1BS and E2BS are sequence-specific elements, the function of the BPV-1 ATR could be at least partially substituted with heterologous AT-rich sequences, suggesting that the role of this element is primarily AT content-dependent rather than sequence-dependent. A stringent requirement for the ATR was also observed in the context of an authentic minimal origin sequence confirming that it is an intrinsic property of the BPV-1 origin and not simply an artifact of the reconstitution system. This study indicates that the minimal functional BPV-1 origin shares the tripartite modular organization characteristic of other simple eukaryotic replication origins. The reconstitution system described now provides a convenient approach to define the physical and functional interrelationships between the three subelements in a systematic fashion.

Adenovirus E2 Proteins↗
PubMed1981

Oxygen deficit during incremental exercise.

The oxygen deficit and debt have conventionally been determined during exercise at constant work rates. During this study these were calculated during and after exercise at progressively incremented work rates. Five men performed two successive incremental exercise tests to exhaustion on an electronically braked cycle ergometer. The two tests were separated by a 5 min rest period. The oxygen deficit was defined as the sum of the minute differences between the measured oxygen uptake and the oxygen uptake occurring during steady state work at that same rate. The oxygen deficit was quantified for the work periods before and after the anaerobic threshold (AT) as determined from respiratory gas analysis (ATR). The measured deficit for the period before the ATR was smaller than the deficit measured in the same subjects during steady state work at low intensity (below the ATR) and was also less than the rapid component of the oxygen repayment as determined after the second incremental test. It was concluded that this test could be used for the determination of anaerobic capacity as represented by the total oxygen deficit (within motivational limits), but that the lactacid and alactacid components of the deficit could not be differentiated. A considerable portion of the alactacid component of the deficit was incurred after the onset of the ATR.

Adolescent↗
PubMed1981

The anaerobic threshold as determined before and during lactic acidosis.

During this study the relationships between venous lactate concentration and accociated changes in respiratory gas exchange were investigated. Five men performed two successive incremental exercise tests to exhaustion on an electronically braked cycle ergometer. These tests were separated by a 5 min rest period. During the initial test venous lactate concentrations showed a characteristic curvilinear increase and the anaerobic threshold (AT1) was determined conventionally. During the second test lactate concentrations were still decreasing at higher work rates than the AT1, and a second anaerobic threshold (AT2) was determined as the point where lactate concentrations again increased. The departure from linearity of the ventilatory response to both exercise tests occurred at a similar work rate, irrespective of whether venous lactate concentrations were increasing or decreasing. Carbon dioxide production was similar during the two exercise tests. The anaerobic thresholds as determined by respiratory gas analysis (ATR) were therefore similar for both tests. Results of this study indicate that changing venous lactate concentrations were not responsible for the ventilatory drive which occurred at the ATR. The venous lactate response to work at a constant rate determined within the range AT1-AT2 was also investigated. It was concluded that the lactate response to constant work rate will vary predictably at work rates falling within the AT1 to AT2 range. At AT1 no increase in venous lactate concentrations occurred, while at AT2 these increased progressively, and the test was terminated at varying times (12-15 min) due to subject exhaustion. At work rates determined from the ATR venous lactate concentrations varied according to the placement of the ATR within the AT1 AT2 range.

Acidosis↗
PubMed2001

On the interactions between antimuscarinic atropine and NMDA receptor antagonists in anticholinesterase-treated mice.

Both organophosphate (OP) and carbamate pesticides may produce seizures and death commonly attributed to the inhibition of acetylcholinesterase (AChE) and subsequent excess of acetylcholine (ACh). The anticonvulsant and neuroprotective properties of N-methyl-D-aspartate (NMDA) receptor antagonists in animals encouraged us to investigate their effects on the toxic and convulsant properties of OP and carbamate pesticides. Adult Swiss mice were systemically injected with the OP pesticide, chlorfenvinphos (CVP), or the carbamate pesticide, methomyl (MET). Both CVP and MET induced dose-dependent seizure activity and death in mice. Pretreatment with the muscarinic antagonist, atropine (ATR), at a dose of 1.8 mg/kg did not prevent seizures but decreased the lethal effects of CVP and MET. Pretreatment with the NMDA antagonists, dizocilpine (MK-801) at a dose of 1 mg/kg or 3-((R,S)-2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid (CPP) at a dose of 10 mg/kg, influenced neither MET-induced seizures nor CVP- or MET-induced death. However, both MK-801 and CPP blocked CVP-induced seizures. Concurrent administration of ATR and the NMDA antagonists prevented seizures produced by CVP, but not those produced by MET. Nevertheless, both MK801 and CPP coadministered with ATR markedly enhanced its antilethal effects in CVP- and MET-intoxicated mice. The antidotes had no influence upon brain AChE activities in mice treated with saline or CVP or MET. It seems that combined treatment with ATR and NMDA receptor antagonists might be of clinical relevance.

Acetylcholinesterase↗
PubMed2006

Enhancement of recombinant streptokinase production in Lactococcus lactis by suppression of acid tolerance response.

Lactococcus lactis is a potential host for production of recombinant proteins, especially of therapeutic importance. However, in glucose-grown cultures, lowering of pH due to accumulation of lactic acid and the concomitant induction of acid tolerance response (ATR) may affect the recombinant protein produced. In this work, we have analyzed the effect of culture pH and the associated ATR on production of recombinant streptokinase. Streptokinase gene was cloned and expressed as a secretory protein in L. lactis under the control of P170 promoter. It was found to undergo degradation to form inactive products leading to low productivity. The extent of degradation and productivity of streptokinase was greatly influenced by the development of ATR, which was dependent on the pH of the culture and initial phosphate concentration of the medium. It was found that high pH and high initial phosphate concentration leads to suppression of ATR and this results in at least 2.5-fold increase in streptokinase productivity and significant decrease in degradation of streptokinase.

Adaptation, Physiological↗
PubMed1997

Molecular linkage of the human alpha 1-antitrypsin and corticosteroid-binding globulin genes on chromosome 14q32.1.

The genes encoding alpha 1-antitrypsin (alpha 1AT; gene symbol PI) and corticosteroid-binding globulin (CBG) are part of a cluster of structurally related serine protease inhibitor (serpin) genes on human Chromosome (Chr) 14q32.1. This cluster also includes the genes encoding alpha 1-antichymotrypsin (AACT) and protein C inhibitor (PCI), as well as an alpha 1-antitrypsin-related sequence (ATR; gene symbol PIL). In this report we present a detailed restriction map of a 110-kb region of genomic DNA that includes the alpha 1AT, ATR, and CBG genes. Gene order in this interval is tel-alpha 1AT-ATR-CBG-cen, and all three genes are transcribed in a distal-to-proximal orientation. Within the gene cluster, ATR is approximately 12 kb downstream of alpha 1AT, and CBG is about 57 kb downstream of alpha 1AT. Repetitive DNA sequences have been mapped throughout the interval, and several new restriction site polymorphisms in the region are described.

Chromosome Mapping↗
PubMed1991

Cholinolytic antagonism to the disruptive effects of oral low doses of pyridostigmine on simple discrimination performance in rats.

We have previously reported that acute oral administration of low doses (less than or equal to 12 mg/kg) of pyridostigmine bromide (PYR) to rats resulted in a dose-dependent decrement in reinforcement rate under two different multiple schedules of response-produced water presentation, which involved motivational dysfunction rather than motor impairment and alterations in visual perception. The purpose of the present investigation was to examine further if the anticipated operant behavioral deficits of PYR are mediated by central and/or peripheral cholinergic mechanisms. Lever-press responses of male Sprague-Dawley rats were maintained under a multiple fixed-ratio GO/differential-reinforcement-of-low-rate NO GO, brightness discrimination, schedule of water reinforcement. The effects of the muscarinic antagonists atropine (ATR) and methylatropine (MAT), both at doses of 0.25, 0.5 and 1.0 mg/kg (SC), against a single oral low dose of PYR (12 mg/kg)-induced behavioral disruption were compared. ATR partially antagonized the reinforcement loss of PYR with concomitant dose-related increases in nonreinforced responses, whereas MAT completely antagonized the reinforcement loss without affecting the frequency of nonreinforced responses. These results suggest that in rats, the debilitating effects of oral PYR on operant behavior are primarily due to the stimulation of peripheral muscarinic receptors via its anticholinesterase activity. The increments of nonreinforced responses observed after coadministration of PYR with ATR may reflect a central, excitatory action of ATR which could affect the discrimination performance. The present results have practical implications for the clinical utilization of PYR in combination with the peripherally active muscarinic antagonist in situations where optimal performance is required.

Administration, Oral↗
PubMed2006

Bongkrekic acid and atractyloside inhibits chloride channels from mitochondrial membranes of rat heart.

The aim of this work was to characterize the effect of bongkrekic acid (BKA), atractyloside (ATR) and carboxyatractyloside (CAT) on single channel properties of chloride channels from mitochondria. Mitochondrial membranes isolated from a rat heart muscle were incorporated into a bilayer lipid membrane (BLM) and single chloride channel currents were measured in 250/50 mM KCl cis/trans solutions. BKA (1-100 microM), ATR and CAT (5-100 microM) inhibited the chloride channels in dose-dependent manner. The inhibitory effect of the BKA, ATR and CAT was pronounced from the trans side of a BLM and it increased with time and at negative voltages (trans-cis). These compounds did not influence the single channel amplitude, but decreased open dwell time of channels. The inhibitory effect of BKA, ATR and CAT on the mitochondrial chloride channel may help to explain some of their cellular and/or subcellular effects.

Animals↗
PubMed2005

Acid response of exponentially growing Escherichia coli K-12.

Induction of acid tolerance response (ATR) of exponential-phase Escherichia coli K-12 cells grown and adapted at different conditions was examined. The highest level of protection against pH 2.5 challenges was obtained after adaptation at pH 4.5-4.9 for 60 min. To study the genetic systems, which could be involved in the development of log-phase ATR, we investigated the acid response of E. coli acid resistance (AR) mutants. The activity of the glutamate-dependent system was observed in exponential cells grown at pH 7.0 and acid adapted at pH 4.5 in minimal medium. Importantly, log-phase cells exhibited significant AR when grown in minimal medium pH 7.0 and challenged at pH 2.5 for 2 h without adaptation. This AR required the glutamate-dependent AR system. Acid protection was largely dependent on RpoS in unadapted and adapted cells grown in minimal medium. RpoS-dependent oxidative, glutamate and arginine-dependent decarboxylase AR systems were not involved in triggering log-phase ATR in cells grown in rich medium. Cells adapted at pH 4.5 in rich medium showed a higher proton accumulation rate than unadapted cells as determined by proton flux assay. It is clear from our study that highly efficient mechanisms of protection are induced, operate and play the main role during log-phase ATR.

Adaptation, Physiological↗
PubMed2003

Myopic astigmatism and presbyopia trial.

PURPOSE: No prospective double-masked study has evaluated whether low astigmatism benefits or harms patients with presbyopia, whose intermediate and near vision might theoretically benefit from enhanced depth of focus provided by astigmatism. The purpose of the first Myopic Astigmatism and Presbyopia (MAP I) study was to determine whether low myopic astigmatism enhances or harms the visual acuity, stereopsis, or quality of life in patients with presbyopia. DESIGN: Prospective, randomized, double-masked, crossover design clinical trial. METHODS: Fifteen patients with presbyopia aged 45 to 68 years were recruited from an academic center population. These patients were given a baseline eye examination, including manifest refraction, Early Treatment of Diabetic Retinopathy Study (ETDRS) logarithm of minimal angle of resolution (logMAR) visual acuity at distance, intermediate, and near, accommodative amplitudes, and stereo vision. Each patient was then cycled in random order through three masked pairs of soft contact lenses. The power of each contact lens pair was calculated by the subtraction method to maintain a spherical equivalent of -0.5 diopters, while providing either no astigmatism (spherical arm, SPH), 1 diopter of with-the-rule (WTR) astigmatism, or 1 diopter of against-the-rule (ATR) astigmatism. Actual refractive errors produced were measured by masked examiner. Outcomes measured at the end of 1 week of usage of each contact lens arm were binocular (ETDRS) logMAR visual acuity at three distances (far [4 m], intermediate [1 m], and near [33cm]); near stereoacuity, using the quantitative Titmus Stereotest; and quality of life, measured using the Refractive Status and Vision Profile (RSVP), a standardized questionnaire. RESULTS: Visual acuity results across the three arms were similar. However, 1-m logMAR visual acuity was better for the spherical arm than either astigmatic arm (-0.06 SPH, +0.01 WTR, +0.02 ATR). Near (33 cm) and distance (4 m) acuities were similar across arms. Stereoacuity was better in ATR than WTR (50 vs 102 seconds, P =.01). Subjects preferred SPH slightly over the WTR astigmatic arm by the RSVP quality-of-life survey instrument (101 vs 104, P =.05). Other intergroup comparisons showed no difference in RSVP scores. CONCLUSIONS: This study has demonstrated that intermediate distance acuity and refractive quality of life are slightly better with spherical low myopic refractive error vs either astigmatic arm. Near and far distance acuity were unaffected by low myopic astigmatism compared with spherical low myopia. Near stereopsis was best in the ATR arms, but this did not produce better near visual acuity or RSVP quality of life.

Aged↗
PubMed2003

Anthrax toxin receptor proteins.

Anthrax toxin is a key virulence factor for Bacillus anthracis, the causative agent of anthrax. Here we discuss what is known about the anthrax toxin receptor (ATR), the cellular receptor for anthrax toxin, and how this information is being used to develop treatments for anthrax as well as to understand aspects of cancer. ATR was identified recently as a type I transmembrane protein with unknown function that contains an extracellular integrin-like inserted (I) domain. The ATR I domain contains the toxin binding site, and a soluble form of this domain was shown to serve as an effective antitoxin to protect cultured cells from toxin action. ATR is encoded by the tumor endothelial marker 8 (TEM8) gene, which is selectively up-regulated during blood vessel formation and in tumor vasculature, raising the possibility that this protein normally functions in angiogenesis. Therefore, identification of the cellular receptor for anthrax toxin has made possible new avenues of research in the areas of anthrax pathogenesis, antitoxin development, and cancer biology.

Animals↗
PubMed2001

Influence of length on force and activation-dependent changes in troponin c structure in skinned cardiac and fast skeletal muscle.

Linear dichroism of 5' tetramethyl-rhodamine (5'ATR) was measured to monitor the effect of sarcomere length (SL) on troponin C (TnC) structure during Ca2+ activation in single glycerinated rabbit psoas fibers and skinned right ventricular trabeculae from rats. Endogenous TnC was extracted, and the preparations were reconstituted with TnC fluorescently labeled with 5'ATR. In skinned psoas fibers reconstituted with sTnC labeled at Cys 98 with 5'ATR, dichroism was maximal during relaxation (pCa 9.2) and was minimal at pCa 4.0. In skinned cardiac trabeculae reconstituted with a mono-cysteine mutant cTnC (cTnC(C84)), dichroism of the 5'ATR probe attached to Cys 84 increased during Ca2+ activation of force. Force and dichroism-[Ca2+] relations were fit with the Hill equation to determine the pCa50 and slope (n). Increasing SL increased the Ca2+ sensitivity of force in both skinned psoas fibers and trabeculae. However, in skinned psoas fibers, neither SL changes or force inhibition had an effect on the Ca2+ sensitivity of dichroism. In contrast, increasing SL increased the Ca2+ sensitivity of both force and dichroism in skinned trabeculae. Furthermore, inhibition of force caused decreased Ca2+ sensitivity of dichroism, decreased dichroism at saturating [Ca2+], and loss of the influence of SL in cardiac muscle. The data indicate that in skeletal fibers SL-dependent shifts in the Ca2+ sensitivity of force are not caused by corresponding changes in Ca2+ binding to TnC and that strong cross-bridge binding has little effect on TnC structure at any SL or level of activation. On the other hand, in cardiac muscle, both force and activation-dependent changes in cTnC structure were influenced by SL. Additionally, the effect of SL on cardiac muscle activation was itself dependent on active, cycling cross-bridges.

Animals↗
PubMed1999

Quantitative orientation measurements in thin lipid films by attenuated total reflection infrared spectroscopy.

Quantitative orientation measurements by attenuated total reflectance (ATR) infrared spectroscopy require the accurate knowledge of the dichroic ratio and of the mean-square electric fields along the three axes of the ATR crystal. In this paper, polarized ATR spectra of single supported bilayers of the phospholipid dimyristoylphosphatidic acid covered by either air or water have been recorded and the dichroic ratio of the bands due to the methylene stretching vibrations has been calculated. The mean-square electric field amplitudes were calculated using three formalisms, namely the Harrick thin film approximation, the two-phase approximation, and the thickness- and absorption-dependent one. The results show that for dry bilayers, the acyl chain tilt angle varies with the formalism used, while no significant variations are observed for the hydrated bilayers. To test the validity of the different formalisms, s- and p-polarized ATR spectra of a 40-A lipid layer were simulated for different acyl chain tilt angles. The results show that the thickness- and absorption-dependent formalism using the mean values of the electric fields over the film thickness gives the most accurate values of acyl chain tilt angle in dry lipid films. However, for lipid monolayers or bilayers, the tilt angle can be determined with an acceptable accuracy using the Harrick thin film approximation. Finally, this study shows clearly that the uncertainty on the determination of the tilt angle comes mostly from the experimental error on the dichroic ratio and from the knowledge of the refractive index.

Biophysical Phenomena↗
PubMed2002

Application of infrared spectroscopy (attenuated total reflection) for monitoring enzymatic activity on substrate films.

Infrared film analysis, a method based on infrared spectroscopy in the mode of attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), is demonstrated as a novel analytical method for monitoring enzymatic activity on surface-attached substrate films in the mid infrared range (400-4000 cm(-1)). The ATR-FTIR technique is sensitive to molecules within a distance of approximately 1 microm from the ATR sampling unit surface (a 7 cm(2) hydrophobic ZnSe crystal). Applying a 0.2-0.3 microm thick film on the ATR unit surface, any chemical changes within this film as well as at the interface can be continuously monitored, even having an aqueous phase on top of the film. Infrared film analysis is considered especially useful for studying detergent enzymes, which act on surface bound films consisting of food component like vegetable oils (triacylglycerols) and carbohydrates (e.g. starch). Experimental data are presented for hydrolysis of a triacylglycerol film (triolein) by use of a triacylglycerol lipase (cutinase), and starch film degradation by use of an alpha-amylase.

Amylases↗
PubMed2003

Induction of an adaptive tolerance response in the foodborne pathogen, Campylobacter jejuni.

In this study we aimed to determine if Campylobacter had the ability to induce an adaptive tolerance response (ATR) to acid and/or aerobic conditions. Campylobacter jejuni CI 120 was grown to the appropriate phase in Brucella broth under microaerobic conditions. Cells were initially adapted to a mild stress (pH 5.5) for 5 h prior to challenge at pH 4.5, a lethal pH. Survival was examined by determining the numbers of viable cells on Campylobacter blood free selective agar base. Stationary phase cells adapted at pH 5.5 induced an ATR that enabled a 100-fold greater survival compared to an uninduced culture. Aerobic adaptation also protected the cells against acid challenge. The cross protection provided a 500-fold increase in survival when compared to unadapted cells. The incorporation of chloramphenicol during the induction period eliminated the ATR and resulted in death kinetics similar to an uninduced culture. These data suggest that Campylobacter spp. have the ability to induce an ATR to sublethal treatments, which increased their ability to withstand subsequent stresses.

Adaptation, Physiological↗
PubMed2003

Effect of central corneal thickness on surgically induced astigmatism in cataract surgery.

PURPOSE: To evaluate the effect of central corneal thickness (CCT) on surgically induced astigmatism (SIA) in cataract surgery using temporal clear corneal incisions. SETTING: Department of Ophthalmology, Seoul National University Hospital, Seoul, Korea. METHODS: The retrospective nonrandomized clinical study comprised 129 eyes that had cataract surgery performed through a sutureless temporal clear corneal incision. Eyes were categorized according to the axis of the preoperative astigmatism: against the rule (ATR, 0 to 45 degrees; n = 64) or with the rule (WTR, 46 to 90 degrees; n =65). Corneal topography was taken preoperatively and 1 day and 2 months postoperatively to evaluate the change in the cornea. The SIA was calculated using the polar method from simulated keratometric readings obtained with topography. Correlations between SIA and CCT were analyzed by linear regression analysis and compared between the ATR and the WTR groups. RESULTS: One day postoperatively, the SIA was negatively correlated with CCT in the ATR group but not in the WTR group (P =.050 versus P =.92; linear regression analysis). At 2 months, the correlation between CCT and SIA was no longer statistically significant. CONCLUSIONS: Central corneal thickness was negatively correlated with the amount of SIA immediately postoperatively when the preoperative astigmatism was ATR. The correlation was not present 2 months after surgery. These results may be applied to abnormally thin corneas such as those after keratorefractive surgery.

Aged↗
PubMed1992

Effect of suture material on postoperative astigmatism.

Two hundred patients were enrolled in a randomized, prospective clinical trial comparing the use of 10-0 nylon, 10-0 polypropylene (Prolene), 11-0 polyester (Mersilene), and 10-0 polyethylene (Novafil) suture materials on the amount and decay curves of surgically induced astigmatism following intraocular lens (IOL) surgery. Patients with Mersilene and nylon sutures had the highest amounts of induced with-the-rule (WTR) cylinder (significantly more than Prolene) at one day after surgery. However, the WTR cylinder decayed rapidly for nylon during the first three months but more slowly for Mersilene because of its lack of stretchability. The Prolene group had the lowest level of induced WTR cylinder at one day, but against-the-rule (ATR) drift occurred, leaving cases with ATR astigmatism by a year. The nylon group had the second highest amount of induced WTR cylinder at one day, which had decayed to ATR cylinder by five months. Between one and two years postoperatively, the nylon group experienced a significant ATR shift. The amount of early induced WTR cylinder seemed to be related to the knot-tying technique and tissue gripping characteristics, whereas the shape of the decay curve was related to the material characteristics of the suture.

Adult↗