Biomedical subjects

L Nilsson

Publications and source records attributed to L Nilsson.

At least 19 recordsLinked to original sources

PubMed2006

Combinations of common chronic paediatric diseases deviate the immune response in diverging directions.

The cytokine pattern of T lymphocytes has not been characterized in children with combinations of paediatric immunological disorders. We describe cytokine secretion in children with type 1 diabetes, coeliac disease and allergy and combinations of two of these diseases after stimulation with 'disease-specific' antigens. Peripheral blood mononuclear cells (PBMC) were collected from 68 children with type 1 diabetes, allergy or coeliac disease, two of these diseases in combination or none of these diseases. Using the enzyme-linked immunospot (ELISPOT) technique, interferon (IFN)-gamma and interleukin (IL)-4 were analysed from fresh PBMC spontaneously and after in vitro stimulation with antigens associated with one or more of these diseases (insulin, gluten, birch and cat extract, beta-lactoglobulin, ovalbumin and phytohaemagglutinin) in order to divide T helper (Th)1- from Th2-like lymphocytes. Stimulation with birch and cat extract caused increased IL-4 secretion in allergic children. A low IFN-gamma response to insulin was found in type 1 diabetic children, whereas allergic children responded to insulin by increased IL-4 secretion. Children suffering from both type 1 diabetes (Th1-prone) and allergy (Th2-prone) reacted distinctly to general mitogen stimulation. Children suffering from two Th1-dominated diseases (type 1 diabetes and coeliac disease) showed hardly any response to either food or inhalation allergens. Our results indicate an important interplay between common immunological diseases in children. The combination of two Th1-deviated diseases is associated with a suppressed immune response, whereas a combination of Th1- and Th2-dominated diseases appears to increase the general immune response.

Adolescent↗

PubMed2005

Measurement of the absolute n p scattering differential cross section at 194 MeV.

We describe a double-scattering experiment with a novel tagged neutron beam to measure differential cross sections for np backscattering to better than +/-2% absolute precision. The measurement focuses on angles and energies where the cross section magnitude and angle dependence constrain the charged pion-nucleon coupling constant, but existing data show serious discrepancies among themselves and with energy-dependent partial-wave analyses. The present results are in good accord with the partial-wave analyses, but deviate systematically from other recent measurements.

Journal Article↗

PubMed2005

Prevalence of self-reported food allergy and IgE antibodies to food allergens in Swedish and Estonian schoolchildren.

OBJECTIVE: To compare the prevalence of self-reported food allergy and IgE antibodies to food allergens in wheezing and non-wheezing Estonian and Swedish schoolchildren, in the light of the disparities in the standard of living, food consumption and prevalence of respiratory allergies that still exist between Estonia and the Scandinavian countries. DESIGN AND SETTING: As a part of the ISAAC Phase II study, children from a random sample of schools in Tallinn in Estonia and Linköping and Ostersund in Sweden participated in skin prick tests to inhalant allergens and the parents replied to questionnaires. IgE antibodies against a panel of food allergens (egg white, milk, soy bean, fish, wheat and peanut) were taken from children with questionnaire-reported wheezing and a random sample of nonwheezing children. SUBJECTS: Children aged 10-11 y. RESULTS: The prevalence of self-reported food allergy was similar in Estonia and Sweden and about twice as high in wheezing children than in nonwheezing children. In Estonia, however, 3% of the children with perceived food allergy reported reactions from at least four different foods, as compared to 31% in Sweden. The prevalence of sensitisation to food allergens was similar in wheezing and nonwheezing children in Estonia (8%) while, in Swedish children, IgE antibodies to food allergens were more likely among wheezing children (Linköping 38 vs 11%, crude OR 5.1, 95% CI 2.2-11.6, and Ostersund 24 vs 7%, crude OR 4.1, 95% CI 1.9-8.5). CONCLUSION: Our study suggests that IgE-mediated food reactions were less likely in Estonian schoolchildren. Moreover, the perception of food allergy and thereby the meaning of self-reported food allergy appears to be different in the two countries.

Allergens↗

PubMed2005

Patient-circumference-adapted dose regulation in body computed tomography. A practical and flexible formula.

PURPOSE: To illustrate that the attenuation formula based on monochromatic radiation in homogeneous objects may be used for dose regulation in body computed tomography (CT) based on patient circumference and using a simple cloth measuring tape. MATERIAL AND METHODS: Based on the attenuation formula for monochromatic radiation the following Microsoft Excel equation was derived: mAs(x) = mAs(n)*EXP((0.693/ HVT)*(O(x)-O(n))/PI()), where mAs(x) (milliampere second) in a patient with circumference O(x) is calculated based on the nominal mAs(n) set for a reference patient with the circumference O(n) with regard to indication, scan protocol, and available CT scanner. The HVT = half-value thickness (object thickness change in cm affecting mAs setting by a factor of 2) resulting in the least mAs difference compared with published studies investigating the mAs needed for constant image noise in abdominal CT phantoms at 80-140 kVp was evaluated. Clinically recommended HVT values were applied to 20 patients undergoing abdominal CT using 130 effective mAs and 94 cm circumference as nominal settings, and an HVT of 9 cm. RESULTS: The object-sized dependent mAs for constant image noise at 80-140 kVp in 10-47 cm diameter abdominal phantoms (31-148 cm in circumference) differed, with few exceptions, by no more than 10% from those obtained with our formula using an HVT of 3.2-3.8 cm. An HVT of 9 cm in the patient study resulted in the same image noise-patient circumference relation as a phantom study using a "clinically adapted mAs" resulting in an acceptable noise according to diagnostic requirements. Clinical experiences recommend an HVT of about 8 cm for abdominal CT and 12 cm in thoracic CT. Changing the kVp from 120 to 80, 100, or 140 requires a mAs change roughly by factors of 4, 2, and 0.6, respectively, for constant image noise. CONCLUSION: Until fully automatic automatic exposure control systems have been introduced, applying the formula in a computer program provides the radiologist with an easy, quick, flexible, and practical instrument for reasonably good patient-sized adjusted exposure levels in clinical practice.

Adult↗

PubMed2005

Growth inhibition of Listeria monocytogenes by a nonbacteriocinogenic Carnobacterium piscicola.

AIMS: This study elucidates the mechanisms by which a nonbacteriocinogenic Carnobacterium piscicola inhibits growth of Listeria monocytogenes. METHODS AND RESULTS: Listeria monocytogenes was exposed to live cultures of a bacteriocin-negative variant of C. piscicola A9b in co-culture, in a diffusion chamber system, and to a cell-free supernatant. Suppression of maximum cell density (0-3.5 log units) of L. monocytogenes was proportional to initial levels of C. pisciola (10(3)-10(7) CFU ml(-1)). Cell-to-cell contact was not required to cause inhibition. The cell-free C. piscicola supernatant caused a decrease in L. monocytogenes maximum cell density, which was abolished by glucose addition but not by amino acid, vitamin or mineral addition. The fermentate also gave rise to a longer lag phase and a reduction in growth rate. These effects were independent of glucose and may have been caused by acetate production by C. piscicola. 2D gel-electrophoretic patterns of L. monocytogenes exposed to C. piscicola or to L. monocytogenes fermentate did not differ. Treatment with C. piscicola fermentate resulted in down-regulation (twofold) of genes involved in purine- or pyrimidine metabolism, and up-regulation (twofold) of genes from the regulon for vitamin B12 biosynthesis and propanediol and ethanolamine utilization. CONCLUSIONS: A nonbacteriocinogenic C. piscicola reduced growth of L. monocytogenes partly by glucose depletion. SIGNIFICANCE AND IMPACT OF THE STUDY: Understanding the mechanism of microbial interaction enhances prediction of growth in mixed communities as well as use of bioprotective principles for food preservation.

Bacteriological Techniques↗

PubMed2005

Urinary inflammatory mediators and inhalation of hypertonic saline in children.

BACKGROUND: The inflammatory mechanisms of hypertonic saline-induced bronchoconstriction are not well understood. METHODS: Seventeen asthmatics with (n=11) and without bronchial hyperresponsiveness (BHR) (n=6) and 18 randomly selected nonatopic nonasthmatic controls without BHR were evaluated by urine samples collected before and 1 h after hypertonic saline provocation test. Histamine, 11beta-PGF2alpha, and LTE4 were analysed by enzyme immunoassay (EIA) and eosinophil protein X (EPX) by radioimmunoassay (RIA). RESULTS: The levels of leukotriene E4 (LTE4) increased significantly after the challenge tests, both in the asthmatics (median: 354 pg/mg pre-challenge vs. 628 pg/mg post-challenge; P=0.05) and in the controls (median: 294 pg/mg pre-challenge vs. 460 pg/mg post-challenge; P <0.01). The levels of histamine also increased significantly in the latter (median: 299 micromol/mg pre-challenge vs. 569 micromol/mg post-challenge; P=0.03). However, the levels of 11beta-PGF2alpha and EPX did not change significantly after the challenge tests either in the asthmatics or in the controls. CONCLUSIONS: The inhalation of hypertonic saline increased urinary excretion of LTE4 both in the asthmatics and in the controls. The slight increase of leukotrienes was enough to induce airway obstruction in some of the asthmatics, because of the hyperresponsiveness in their airways.

Asthma↗

PubMed2005

Respiration can be monitored by photoplethysmography with high sensitivity and specificity regardless of anaesthesia and ventilatory mode.

BACKGROUND: Photoplethysmography (PPG) is a non-invasive optical technique used, for instance, in pulse oximetry. Beside the pulse synchronous component, PPG has a respiratory synchronous variation (PPGr). Efforts have been made to utilize this component for indirect monitoring of respiratory rate and volume. Assessment of the clinical usefulness as well as of the physiological background of PPGr is required. We evaluated if anaesthesia and positive-pressure ventilation would affect PPGr. METHODS: We recorded reflection mode PPGr, at the forearm, and the respiratory synchronous changes in central venous pressure (CVP), peripheral venous pressure (PVP) and arterial blood pressure (ABP) in 12 patients. Recordings for each patient were made on three occasions: awake with spontaneous breathing; anaesthetized with spontaneous breathing; and anaesthetized with positive-pressure ventilation. We analyzed the sensitivity, specificity, coherence and time relationship between the signals. RESULTS: PPGr sensitivity for breath detection was [mean (SD)] >86(21)% and specificity >96(12)%. Respiratory detection in the macrocirculation (CVP, PVP and ABP) showed a sensitivity >83(29)% and specificity >93(12)%. The coherence between signals was high (0.75-0.99). The three measurement situations did not significantly influence sensitivity, specificity or time shifts between the PPGr, PVP, ABP, and the reference CVP signal despite changes in physiological data between measurements. CONCLUSION: A respiratory synchronous variation in PPG and all invasive pressure signals was detected. The reflection mode PPGr signal seemed to be a constant phenomenon related to respiration regardless of whether or not the subject was awake, anaesthetized or ventilated, which increases its clinical usefulness in respiratory monitoring.

Adult↗

PubMed2004

The contribution of bacteriocin to inhibition of Listeria monocytogenes by Carnobacterium piscicola strains in cold-smoked salmon systems.

AIMS: To study the importance of bacteriocin production for the antilisterial effect of a bacteriocinogenic Carnobacterium piscicola strain A9b on growth of Listeria monocytogenes in broth and cold-smoked salmon systems. METHODS AND RESULTS: Acriflavin treatment of strain A9b resulted in loss of bacteriocin production and of immunity to carnobacteriocin B2. Two plasmids present in the wild-type were lost in the variant that was also more sensitive to bavaricin and leucocin A than the wild-type indicating cross-resistance to class IIa bacteriocins. The growth rate of the bac- mutant was higher than that of the wild-type at 5 and 37 degrees C but not at 25 or 30 degrees C. In salmon juice the maximum cell density of L. monocytogenes was suppressed 3 and 6 log by co-culture with C. piscicola A9b bac- and bac+, respectively, as compared with the control. Sterile filtered cultures of C. piscicola A9b bac- caused a limited suppression of the maximum cell density of L. monocytogenes similar to that observed when sterile buffer was added in equal amounts. Semi-purified carnobacteriocin B2 caused a 3.5 log decline in viable cell count after 6 day of incubation in cold-smoked salmon juice at 5 degrees C. High resistance level to carnobacteriocin B2 was observed for L. monocytogenes cells exposed to semi-purified and in situ produced carnobacteriocin B2. CONCLUSIONS: The presence of bacteriocin production in C. piscicola enhances its inhibition of L. monocytogenes. SIGNIFICANCE AND IMPACT OF THE STUDY: Due to the emergence of resistance, a bacteriocin negative lactic acid bacteria may be more suited for practical use as a bioprotective agent against L. monocytogenes in ready-to-eat foods.

Animals↗

PubMed2003

Respiratory variations in the reflection mode photoplethysmographic signal. Relationships to peripheral venous pressure.

Photoplethysmography (PPG) is a non-invasive optical way of measuring variations in blood volume and perfusion in the tissue, used in pulse oximetry for instance. Respiratory-induced intensity variations (RIIVs) in the PPG signal exist, but the physiological background is not fully understood. Respiration causes variations in the blood volume in the peripheral vascular bed. It was hypothesised that the filling of peripheral veins is one of the important factors involved. In 16 healthy subjects, the respiratory synchronous variations from a PPG reflection mode signal and the peripheral venous pressure (PVP) were recorded. Variations of tidal volume, respiratory rate and contribution from abdominal and thoracic muscles gave significant and similar amplitude changes in both RIIV and the respiratory variation of PVP (p<0.01). The highest amplitudes of both signals were found at the largest tidal volume, lowest respiratory rate and during mainly thoracic breathing, respectively. The coherence between PVP and RIIV signals was high, the median (quartile range) being 0.78 (0.42). Phase analysis showed that RIIV was usually leading PVP, but variations between subjects were large. Although respiratory-induced variations in PVP and PPG showed a close correlation in amplitude variation, a causal relationship between the signals could not be demonstrated.

Adult↗

PubMed2003

An unusual case of tumor thrombus in the inferior vena cava. A case report.

Adrenal cortical carcinoma (ACC) is a rare malignancy. Patients present either with a functional tumor or secondary to mass effect. In non-functioning tumors, the tumor size often exceeds 5 cm by the time of diagnosis, and tumor thrombus can occur. We report on a case of a small non-functioning ACC causing a large tumor thrombus in the inferior vena cava.

Adrenal Cortex Neoplasms↗

PubMed2003

Macrocirculation is not the sole determinant of respiratory induced variations in the reflection mode photoplethysmographic signal.

Photoplethysmography (PPG) is a non-invasive optical technique sensitive to variations in blood volume and perfusion in the tissue. Reflection mode PPG may have clinical advantages over transmission mode PPG. To improve clinical usefulness and further development of the reflection mode PPG, studies on factors that modify the signal are warranted. We studied the coherence between the respiratory induced intensity variations (RIIV) of the PPG signal and respiratory synchronous pressure variations in central venous pressure (CVP), peripheral venous pressure (PVP) and arterial blood pressure (ABP) during positive pressure ventilation on 12 patients under anaesthesia and on 12 patients with spontaneous breathing. During positive pressure ventilation the coherence between all signals was high. Inspiration was followed first by an increase in CVP, then by increases in ABP and PVP and lastly by RIIV indicating less back-scattered light. In spontaneously breathing patients the coherence was high, but the phases between the signals were changed. During inspiration, ABP decreased slightly before CVP, followed by a decrease in RIIV and PVP. The phase relation between RIIV and respiratory induced variation in macrocirculation changed with ventilatory mode, but not in a uniform way, indicating the influence of mechanisms other than macrocirculation involved in generating the RIIV signal.

Adolescent↗

PubMed2003

High body mass index, asthma and allergy in Swedish schoolchildren participating in the International Study of Asthma and Allergies in Childhood: Phase II.

AIM: To assess the relationship between high body mass index (BMI) and asthma and atopic manifestations in 12-y-old children. METHODS: The relationship between high BMI and asthma symptoms was studied in 457 sixth-grade children, with (n = 161) and without (n = 296) current wheeze. High BMI was defined as > or = 75th percentile of gender-specific BMI reference values for Swedish children at 12 y of age; overweight as a subgroup of high BMI was defined as > or = 95th percentile. Children with a BMI < 75th percentile served as controls. Questionnaires were used to assess asthmatic and allergic symptoms, and bronchial hyperresponsiveness was assessed by hypertonic saline provocation tests. RESULTS: Current wheeze was associated with high BMI after adjustment for confounding factors (adjusted OR 1.7, 95% CI 1.0-2.5) and overweight had an even more pronounced effect (adjusted OR 1.9, 95% CI 1.0-3.6). In addition, asthma severity was associated with high BMI, as evaluated by the number of wheezing episodes during the previous 12 mo among the wheezing children (adjusted OR 2.0, 95% CI 1.0-4.0). There was also an association between high BMI and the presence of eczema in wheezing children (adjusted OR 2.2, 95% CI 1.0-4.6). However, high BMI was not significantly associated with hay fever, positive skin prick tests or bronchial hyperresponsiveness. CONCLUSION: The study confirms and extends a previously observed relationship between BMI and the presence of wheezing and asthma.

Adolescent↗

PubMed2002

Antibiotic prescription practices, consumption and bacterial resistance in a cross section of Swedish intensive care units.

BACKGROUND: The purpose of this work was to study usage of antibiotics, its possible determinants, and patterns of bacterial resistance in Swedish intensive care units (ICUs). METHODS: Prospectively collected data on species and antibiotic resistance of clinical isolates and antibiotic consumption specific to each ICU in 1999 were analyzed together with answers to a questionnaire. Antibiotic usage was measured as defined daily doses per 1000 occupied bed days (DDD1000). RESULTS: Data were obtained for 38 ICUs providing services to a population of approximately 6 million. The median antibiotic consumption was 1257 DDD1000 (range 584-2415) and correlated with the length of stay but not with the illness severity score or the ICU category. Antibiotic consumption was higher in the ICUs lacking bedside devices for hand disinfection (2193 vs. 1214 DDD1000, p=0.05). In the ICUs with a specialist in infectious diseases responsible for antibiotic treatment the consumption pattern was different only for use of glycopeptides (58% lower usage than in other ICUs: 26 vs. 11 DDD1000,P=0.02). Only 21% of the ICUs had a written guideline on the use of antibiotics, 57% received information on antibiotic usage at least every 3 months and 22% received aggregated resistance data annually. Clinically significant antimicrobial resistance was found among Enterbacter spp. to cephalosporins and among Enterococcus spp. to ampicillin. CONCLUSIONS: Availability of hand disinfection equipment at each bed and a specialist in infectious diseases responsible for antibiotic treatment were factors that correlated with lower antibiotic consumption in Swedish ICUs, whereas patient-related factors were not associated with antibiotic usage.

Adult↗

PubMed2002

Living situation, subjective quality of life and social network among individuals with schizophrenia living in community settings.

OBJECTIVE: To investigate the relationships between characteristics of the living situation in the community and subjective quality of life and social network among community-based individuals with schizophrenia. METHOD: A total of 418 individuals with schizophrenia from 10 sites were interviewed with regard to quality of life, psychopathology, social network and needs for care. Characteristics of the living situation investigated were: living alone or not, living with family or not, and having an independent or a sheltered housing situation. RESULTS: An independent housing situation was related to a better quality of life concerning living situation and living with the family to a better quality of life concerning family relations. An independent housing situation was associated with a better social network regarding availability and adequacy of emotional relations. CONCLUSION: People with schizophrenia with an independent housing situation have a better quality of life associated with more favorable perceptions of independence, influence, and privacy. Their social network is better irrespective of whether they live alone or not, or with family or not.

Activities of Daily Living↗

PubMed2002

Green tea extract only affects markers of oxidative status postprandially: lasting antioxidant effect of flavonoid-free diet.

Epidemiological studies suggest that foods rich in flavonoids might reduce the risk of cardiovascular disease and cancer. The objective of the present study was to investigate the effect of green tea extract (GTE) used as a food antioxidant on markers of oxidative status after dietary depletion of flavonoids and catechins. The study was designed as a 2 x 3 weeks blinded human cross-over intervention study (eight smokers, eight non-smokers) with GTE corresponding to a daily intake of 18.6 mg catechins/d. The GTE was incorporated into meat patties and consumed with a strictly controlled diet otherwise low in flavonoids. GTE intervention increased plasma antioxidant capacity from 1.35 to 1.56 (P<0.02) in postprandially collected plasma, most prominently in smokers. The intervention did not significantly affect markers in fasting blood samples, including plasma or haemoglobin protein oxidation, plasma oxidation lagtime, or activities of the erythrocyte superoxide dismutase, glutathione peroxidase, glutathione reductase and catalase. Neither were fasting plasma triacylglycerol, cholesterol, alpha-tocopherol, retinol, beta-carotene, or ascorbic acid affected by intervention. Urinary 8-oxo-deoxyguanosine excretion was also unaffected. Catechins from the extract were excreted into urine with a half-life of less than 2 h in accordance with the short-term effects on plasma antioxidant capacity. Since no long-term effects of GTE were observed, the study essentially served as a fruit and vegetables depletion study. The overall effect of the 10-week period without dietary fruits and vegetables was a decrease in oxidative damage to DNA, blood proteins, and plasma lipids, concomitantly with marked changes in antioxidative defence.

Adult↗

PubMed2001

[Great geographic variations in the prevalence of asthma and allergy. International population studies to explain the associative factors].

During the 1990's the international variation in the prevalence of asthma and allergy was studied in young adults (European Community Respiratory Health Survey, ECRHS) and in children (International Study of Asthma and Allergy in Children, ISAAC). The prevalence of asthma and allergy was highest in the United Kingdom, North America, Australia and New Zealand, lower in the Baltic countries and very low in some of the poorest republics of the former USSR. Children that lived in the countryside during the first year of life had a lower prevalence of atopic disease as teenager while in adults farmers had the highest prevalence of occupational asthma. In both investigations a negative association was found between having pets as a child and atopic sensitisation. This association could, however, be related to selection mechanisms. In a three year follow-up of the Swedish part of the ECRHS the incidence of asthma was higher in smokers than non-smokers. Passive smoking was associated with respiratory symptoms in both children and adults. Follow-ups of both studies are being planned or performed. Further analyses of the geographic differences may explain the reason for the rapid increase of asthma and allergy in the Western world.

Adolescent↗

PubMed2001

MD simulations of homomorphous PNA, DNA, and RNA single strands: characterization and comparison of conformations and dynamics.

MD simulations of homomorphous single-stranded PNA, DNA, and RNA with the same base sequence have been performed in aqueous solvent. For each strand two separate simulations were performed starting from a (i) helical conformation and (ii) random coiled state. Comparisons of the simulations with the single-stranded helices (case i) show that the differences in the covalent nature of the backbones cause significant differences in the structural and dynamical properties of the strands. It is found that the PNA strand maintains its nice base-stacked initial helical structure throughout the 1.5-ns MD simulation at 300 K, while DNA/RNA show relatively larger fluctuations in the structures with a few local unstacking events during -ns MD simulation each. It seems that the weak physical coupling between the bases and the backbone in PNA causes a loss of correlation between the dynamics of the bases and the backbone compared to the DNA/RNA and helps maintain the base-stacked helical conformation. The global flexibility of a single-stranded PNA helix was also found to be lowest, while RNA appears to be the most flexible single-stranded helix. The sugar pucker of several nucleotides in single-stranded DNA and RNA was found to adopt both C2'-endo and C3'-endo conformations for significant times. This effect is more pronounced for single strands in completely coiled states. The simulations with single-stranded coils as the initial structure also indicate that a PNA can adopt a more compact globular structure, while DNA/RNA of the same size adopts a more extended coil structure. This allows even a short PNA in the coiled state to form a significantly stable nonsequentially base-stacked globular structure in solution. Due to the hydrophobic nature of the PNA backbone, it interacts with surrounding water rather weakly compared to DNA/RNA.

Base Sequence↗

PubMed2001

Structure, dynamics and electrostatics of the active site of glutaredoxin 3 from Escherichia coli: comparison with functionally related proteins.

The chemistry of active-site cysteine residues is central to the activity of thiol-disulfide oxidoreductases of the thioredoxin superfamily. In these reactions, a nucleophilic thiolate is required, but the associated pK(a) values differ vastly in the superfamily, from less than 4 in DsbA to greater than 7 in Trx. The factors that stabilize this thiolate are, however, not clearly established. The glutaredoxins (Grxs), which are members of this superfamily, contain a Cys-Pro-Tyr-Cys motif in their active site. In reduced Grxs, the pK(a) of the N-terminal active-site nucleophilic cysteine residue is lowered significantly, and the stabilization of the corresponding thiolate is expected to influence the redox potential of these enzymes. Here, we use a combination of long molecular dynamics (MD) simulations, pK(a) calculations, and experimental investigations to derive the structure and dynamics of the reduced active site from Escherichia coli Grx3, and investigate the factors that stabilize the thiolate. Several different MD simulations converged toward a consensus conformation for the active-site cysteine residues (Cys11 and Cys14), after a number of local conformational changes. Key features of the model were tested experimentally by measurement of NMR scalar coupling constants, and determination of pK(a) values of selected residues. The pK(a) values of the Grx3 active-site residues were calculated during the MD simulations, and support the underlying structural model. The structure of Grx3, in combination with the pK(a) calculations, indicate that the pK(a) of the N-terminal active-site cysteine residue in Grx3 is intermediate between that of its counterpart in DsbA and Trx. The pK(a) values in best agreement with experiment are obtained with a low (<4) protein dielectric constant. The calculated pK(a) values fluctuate significantly in response to protein dynamics, which underscores the importance of the details of the underlying structures when calculating pK(a) values. The thiolate of Cys11 is stabilized primarily by direct hydrogen bonding with the amide protons of Tyr13 and Cys14 and the thiol proton of Cys14, rather than by long-range interactions from charged groups or from a helix macrodipole. From the comparison of reduced Grx3 with other members of the thioredoxin superfamily, a unifying theme for the structural basis of thiol pK(a) differences in this superfamily begins to emerge.

Bacterial Proteins↗