Biomedical subjects
Publications and source records attributed to M Lindgren.
The telithromycin susceptibility of 210 erythromycin-resistant pneumococci was tested with the agar diffusion method. Twenty-six erm(B)-positive isolates showed heterogeneous resistance to telithromycin, which was manifested by the presence of colonies inside the inhibition zone. When these cells were cultured and tested, they showed stable, homogeneous, and high-level resistance to telithromycin.
Pressure sores are a current problem in hospitals and care of the elderly, leading to protracted hospital stays and a high care burden. The trauma for the patients is severe, and the cost of pressure sore prevention and treatment, is considerable. Antidecubitus mattresses are used for prevention and in treatment, but they also contribute to the cost of treating pressure sores. The problem highlighted in the review is that the mattresses' effectiveness in preventing and treating pressure sores has not been sufficiently evaluated. When antidecubitus mattresses are evaluated, it is often only with regard to aspects of the interface pressure and the mattresses' ability to redistribute the pressure. The review points out the important observation that, to be able to evaluate the efficacy of the antidecubitus mattress, the mattress's effect on tissue viability needs to be studied. The parameters that ought to be considered when evaluating a support surface are: interface pressure, pressure and blood flow distribution, temperature and humidity in the skin-support surface interface. The authors propose that the effect on tissue viability of external loading can be assessed by simultaneous measurement of the interface pressure and tissue perfusion.
Event-related potentials (ERPs) and behavioural measures were used to investigate recognition memory and source-monitoring judgements about previously perceived and imagined pictures. At study, word labels of common objects were presented. Half of these were followed by a corresponding picture and the other half by an empty frame, signalling to the participants to mentally visualise an image. At test, participants in a source-monitoring task made a three-way discrimination between new words and words corresponding to previously perceived and imagined pictures. Participants in an old/new-recognition task indicated whether test words were previously presented or not. In both tasks, correctly identified old items elicited more positive-going ERPs than correctly judged new items. This widely distributed old/new effect was found to have an earlier onset and to be of a greater magnitude for imagined than for perceived items. Task (source versus item-memory) affected the old/new effects over prefrontal areas and the reaction times to remembered old items. The present findings are consistent with the view that a greater amount, or a different type, of information is necessary for accurate source-memory judgements than for correct recognition, and moreover, that different types of source-specifying information revive at different rates. In addition, the results add weight to the view that the late widespread ERP-old/new effect is sensitive to the quality or the amount of information retrieved from memory.
A mobile screen (0.5 x 0.4 m) producing ultra-clean exponential LAF (air-flow central zone 0.6 m/s and peripheral zone 0.4 m/s) was investigated as an addition to conventional turbulent/mixing operating room ventilation. The evaluation was performed during strictly standardized sham operations reflecting conditions during major surgery. The study consisted of a pilot experiment designed to give high counts of sedimenting aerobic colony forming units (cfu). In a second main study, recording dust particles, air-borne and sedimenting aerobic cfu, the screen was associated with optimal operating room clothing. In the pilot experiment the use of the screen resulted in a substantial reduction of sedimenting bacteria from 3835-4940 to 0-390 cfu/m(2)/h. In the main study, the use of the additional LAF reduced the surface contamination from 416-329 to 7-78 cfu/m(2)/h up to 1.6 m from the screen (P=0.001-0.0001). Measured in the wound area the screen reduced the air counts of bacteria from 9-14 to 0.2-0.4 cfu/m(3) (P=0.008-0.0001) and a marked reduction of air-borne dust particles was recorded (P=0.007-0.009). In conclusion, the additional mobile LAF screen reduced the counts of aerobic air-borne and sedimenting bacteria-carrying particles as well as dust particles to the levels gained with complete ultra-clean LAF room ventilation. Thus, the screen might prove a valuable addition to operating room ventilation as well as in other areas where asepsis is essential.
Phase memory relaxation times (T(M) or T(2)) of spin labels in human carbonic anhydrase II (HCA II) are reported. Spin labels (N-(1-oxyl-2,2,5,5-tetramethyl-3-pyrrolidinyl)iodoacetamide, IPSL) were introduced at cysteines, by site-directed mutagenesis at seven different positions in the protein. By two pulse electron paramagnetic resonance (EPR), electron spin echo decays at 45 K are measured and fitted by stretched exponentials, resulting in relaxation parameters T(M) and x. T(M) values of seven positions are between 1.6 micros for the most buried residue (L79C) and 4.7 micros for a residue at the protein surface (W245C). In deuteriated buffer, longer T(M) are found for all but the most buried residues (L79C and W97C), and electron spin echo envelop modulation (ESEEM) of deuterium nuclei is observed. Different deuterium ESEEM patterns for W95C and W16C (surface residue) indicate differences in the local water concentration, or accessibility, of the spin label by deuterium. We propose T(M) as a parameter to determine the spin label location in proteins. Furthermore, these systems are interesting for studying the pertaining relaxation mechanism.
Cell-penetrating peptides, CPPs, have been shown to translocate into living cells by a receptor-independent mechanism and to carry macromolecules over the plasma membrane. This article reports studies of the internalization of pVEC, an 18-amino acid-long peptide derived from the murine sequence of the cell adhesion molecule vascular endothelial cadherin, amino acids 615-632. Fluorophore-labeled pVEC entered four different cell lines tested: human aortic endothelial cells, brain capillary endothelial cells, Bowes melanoma cells, and murine brain endothelial cells. In order to evaluate the translocation efficiency of pVEC, we performed a side-by-side comparison with penetratin, a well-characterized CPP. The cellular uptake of pVEC was highest for murine brain endothelial cells. All cell lines tested contained equal or slightly higher concentrations of pVEC than penetratin. pVEC mainly accumulated in nuclear structures but was also found throughout the cells. Furthermore, pVEC functioned as a transporter of both a hexameric peptide nucleic acid molecule of 1.7 kDa and a 67-kDa protein, streptavidin-FITC, and cellular uptake of fluorophore-labeled pVEC took place at 4 degrees C, suggesting a nonendocytotic mechanism of translocation. In conclusion, our results indicate that pVEC is efficiently and rapidly taken up into cells and functions as a potent carrier peptide.
Upon injury of a blood vessel, activated factor VII (FVIIa) forms a high-affinity complex with its allosteric regulator, tissue factor (TF), and initiates blood clotting. Active site-inhibited factor VIIa (FVIIai) binds to TF with even higher affinity. We compared the interactions of FVIIai and FVIIa with soluble TF (sTF). Six residues in sTF were individually selected for mutagenesis and site-directed labeling. The residues are distributed along the extensive binding interface, and were chosen because they are known to interact with the different domains of FVIIa. Fluorescent and spin probes were attached to engineered Cys residues to monitor local changes in hydrophobicity, accessibility, and rigidity in the sTF--FVIIa complex upon occupation of the active site of FVIIa. The results show that inhibition of FVIIa caused the structures around the positions in sTF that interact with the protease domain of FVIIa to become more rigid and less accessible to solvent. Thus, the presence of an active site inhibitor renders the interface in this region less flexible and more compact, whereas the interface between sTF and the light chain of FVIIa is unaffected by active site occupancy.
The use of projective and psychometric psychological tests at the Department of Forensic Psychiatry in Stockholm (Huddinge), Sweden, was studied for a population of 60 men, including many patients with neuropsychological disabilities and multiple psychiatric disorders. The results showed that the use of projective tests like Rorschach, Object Relations Test, and House-Tree-Person was more frequent than the use of objective psychometric tests. Neuropsychological test batteries like the Halstead-Reitan Neuropsychological Test Battery or Luria-Nebraska Neuropsychological Battery were not used. The majority of patients were, however, assessed by intelligence scales like the WAIS-R. The questionable reliability and validity of the projective tests, and the risk of subjective interpretations, raise a problem when used in a forensic setting, since the courts' decisions about a sentence to prison or psychiatric care is based on the forensic psychiatric assessment. The use of objective psychometric neuropsychological tests and personality tests is recommended.
Vascular endothelial cadherin (VE-cadherin) is an endothelial cell-specific cadherin that plays an important role in the control of vascular organization. Blocking VE-cadherin antibodies strongly inhibit angiogenesis, and inactivation of VE-cadherin gene causes embryonic lethality due to a lack of correct organization and remodeling of the vasculature. Hence, inhibitors of VE-cadherin adhesive properties may constitute a tool to prevent tumor neovascularization. In this paper, we tested different monoclonal antibodies (mAbs) directed to human VE-cadherin ectodomain for their functional activity. Three mAbs (Cad 5, BV6, BV9) were able to increase paracellular permeability, inhibit VE-cadherin reorganization, and block angiogenesis in vitro. These mAbs could also induce endothelial cell apoptosis in vitro. Two additional mAbs, TEA 1.31 and Hec 1.2, had an intermediate or undetectable activity, respectively, in these assays. Epitope mapping studies show that BV6, BV9, TEA 1.31, and Hec 1.2 bound to a recombinant fragment spanning the extracellular juxtamembrane domains EC3 through EC4. In contrast, Cad 5 bound to the aminoterminal domain EC1. By peptide scanning analysis and competition experiments, we defined the sequences TIDLRY located on EC3 and KVFRVDAETGDVFAI on EC1 as the binding domain of BV6 and Cad 5, respectively. Overall, these results support the concept that VE-cadherin plays a relevant role on human endothelial cell properties. Antibodies directed to the extracellular domains EC1 but also EC3-EC4 affect VE-cadherin adhesion and clustering and alter endothelial cell permeability, apoptosis, and vascular structure formation.
Some epidemiological studies indicate an association between extremely low frequency electromagnetic field (ELF-EMF) exposure and cancer risks. These studies have mainly taken residential and occupational exposure into consideration. Outdoor environments are often considered as low level areas, but in this paper we show that this is not true in a city environment. We have mapped the ELF magnetic flux densities along certain stretches of sidewalk in central Göteborg City, Sweden. About 50% of the investigated street length shows flux densities of the same order of magnitude (0.2 microT and above) as those associated with increased risks of cancer in epidemiological studies. We conclude that the outdoor exposures in a city environment also should be considered in exposure assessments and risk evaluations. These elevated flux densities are probably due to stray currents. We also found strong magnetic flux densities (> 1.0 microT) close to ordinary distribution pillars, power substations, shoplifting alarms, and other electrical devices.
Children with specific language impairment (SLI) often have a family history of language disorder. In this study, ERPs in response to a visual semantic priming task were recorded in parents of children with SLI. Despite equal performance, the ERPs displayed differences in language processing: larger N400 amplitudes indicated that the parents, especially the fathers, were less primed by the preceding context. Difference waveforms showed that the fathers of SLI children, contrary to controls, had less differentiated responses to congruent versus incongruent sentences. We propose that the N400 observations may be residual markers of past language deficiencies in the fathers. No differences in the N400 effect were found in the mothers of SLI children.
The specific complex between the extracellular part of tissue factor (sTF) and factor VIIa (FVIIa) was chosen as a model for studies of the binding interface between two interacting proteins. Six surface-exposed positions in sTF, residues known to contribute to the sTF-FVIIa interaction, were selected for cysteine mutation and site-directed labeling with spin and fluorescent probes. The binding interface was characterized by spectral data from electron paramagnetic resonance (EPR) and steady-state and time-domain fluorescence spectroscopy. The labels reported on compact local environments at positions 158 and 207 in the interface region between sTF and the gamma-carboxyglutamic acid (Gla) domain of FVIIa, and at positions 22 and 140 in the interface region between sTF and the first epidermal growth factor-like (EGF1) domain of FVIIa. The tightness of the local interactions in these parts of the interface is similar to that seen in the interior of globular proteins. This was further emphasized by the reduced local polarity detected by the fluorescent label upon FVIIa binding, especially in the sTF-Gla region. There were indications of structural rigidity also at positions 45 and 94 in the interface region between sTF and the protease domain (PD) of FVIIa, despite the perturbed cofactor function of these sTF variants. The results of the present study indicate that the multi-probing approach enables comparison of the tightness and characteristics of interaction along the binding interface of a protein complex. This approach also increases the probability of acquiring reliable structural data that are descriptive of the wild-type proteins.
Two different spin labels, N-(1-oxyl-2,2,5,5-tetramethyl-3-pyrrolidinyl)iodoacetamide (IPSL) and (1-oxyl-2,2,5,5-tetramethylpyrroline-3-methyl) methanethiosulfonate (MTSSL), and two different fluorescent labels 5-((((2-iodoacetyl)amino)-ethyl)amino)naphtalene-1-sulfonic acid (IAEDANS) and 6-bromoacetyl-2-dimetylaminonaphtalene (BADAN), were attached to the introduced C79 in human carbonic anhydrase (HCA II) to probe local structural changes upon unfolding and aggregation. HCA II unfolds in a multi-step manner with an intermediate state populated between the native and unfolded states. The spin label IPSL and the fluorescent label IAEDANS reported on a substantial change in mobility and polarity at both unfolding transitions at a distance of 7.4-11.2 A from the backbone of position 79. The shorter and less flexible labels BADAN and MTSSL revealed less pronounced spectroscopic changes in the native-to-intermediate transition, 6.6-9.0 A from the backbone. At intermediate guanidine (Gu)-HCl concentrations the occurrence of soluble but irreversibly aggregated oligomeric protein was identified from refolding experiments. At approximately 1 M Gu-HCl the aggregation was found to be essentially complete. The size and structure of the aggregates could be varied by changing the protein concentration. EPR measurements and line-shape simulations together with fluorescence lifetime and anisotropy measurements provided a picture of the self-assembled protein as a disordered protein structure with a representation of both compact as well as dynamic and polar environments at the site of the molecular labels. This suggests that a partially folded intermediate of HCA II self-assembles by both local unfolding and intermolecular docking of the intermediates vicinal to position 79. The aggregates were determined to be 40-90 A in diameter depending on the experimental conditions and spectroscopic technique used.
In this paper, peak matched multiple windows (PM MW) were used to estimate the spectrogram of the electroencephalogram (EEG). We focussed on the ability to estimate frequency changes, and especially resolving close peaks. A peak of known frequency was evoked in the EEG in a predetermined time interval. The PM MW spectrogram was compared to the commonly used single Hanning window and to weighted overlapped segment averaging in simulations and for real-data. The PM MW were shown to give estimates with good resolution and low variance.
The sound intensity method is usually recommended instead of the pressure method in the presence of strong flanking transmission. Especially when small and/or heavy specimens are tested, the flanking often causes problems in laboratories practicing only the pressure method. The purpose of this study was to determine experimentally the difference between the maximum sound reduction indices obtained by the intensity method, RI,max, and by the pressure method, Rmax. In addition, the influence of adding room absorption to the receiving room was studied. The experiments were carried out in an ordinary two-room test laboratory. The exact value of RI,max was estimated by applying a fitting equation to the measured data points. The fitting equation involved the dependence of the pressure-intensity indicator on measured acoustical parameters. In an empty receiving room, the difference between RI,max and Rmax was 4-15 dB, depending on frequency. When the average reverberation time was reduced from 3.5 to 0.6 s, the values of RI,max increased by 2-10 dB compared to the results in the empty room. Thus, it is possible to measure wall structures having 9-22 dB better sound reduction index using the intensity method than with the pressure method. This facilitates the measurements of small and/or heavy specimens in the presence of flanking. Moreover, when new laboratories are designed, the intensity method is an alternative to the pressure method which presupposes expensive isolation structures between the rooms.
Several shorter analogues of the cell penetrating peptide, transportan, have been synthesized in order to define the regions of the sequence, which are responsible for the membrane translocation property of the peptide. Penetration of the peptides into Bowes melanoma cells and the influence on GTPase activity in Rin m5F cellular membranes have been tested. The experimental data on cell penetration have been compared with molecular modeling of insertion of peptides into biological membranes. Omission of six amino acids from the N-terminus did not significantly impair the cell penetration of the peptide while deletions at the C-terminus or in the middle of the transportan sequence decreased or abolished the cellular uptake. Most transportan analogues exert an inhibitory effect on GTPase activity. Molecular modeling shows that insertion of the transportan analogues into the membrane differs for different peptides. Probably the length of the peptide as well as the location of aromatic and positively charged residues have major impact on the orientation of peptides in the membranes and thereby influence the cellular penetration. In summary, we have designed and characterized several novel short transportan analogues with similar cellular translocation properties to the parent peptide, but with reduced undesired cellular activity.
Chaperonins are molecules that assist proteins during folding and protect them from irreversible aggregation. We studied the chaperonin GroEL and its interaction with the enzyme human carbonic anhydrase II (HCA II), which induces unfolding of the enzyme. We focused on conformational changes that occur in GroEL during formation of the GroEL-HCA II complex. We measured the rate of GroEL cysteine reactivity toward iodo[2-(14)C]acetic acid and found that the cysteines become more accessible during binding of a cysteine free mutant of HCA II. Spin labeling of GroEL with N-(1-oxyl-2,2,5, 5-tetramethyl-3-pyrrolidinyl)iodoacetamide revealed that this additional binding occurred because buried cysteine residues become accessible during HCA II binding. In addition, a GroEL variant labeled with 6-iodoacetamidofluorescein exhibited decreased fluorescence anisotropy upon HCA II binding, which resembles the effect of GroES/ATP binding. Furthermore, by producing cysteine-modified GroEL with the spin label N-(1-oxyl-2,2,5, 5-tetramethyl-3-pyrrolidinyl)iodoacetamide and the fluorescent label 5-((((2-iodoacetyl)amino)ethyl)amino)naphthalene-1-sulfonic acid, we detected increases in spin-label mobility and fluorescence intensity in GroEL upon HCA II binding. Together, these results show that conformational changes occur in the chaperonin as a consequence of protein substrate binding. Together with previous results on the unfoldase activity of GroEL, we suggest that the chaperonin opens up as the substrate protein binds. This opening mechanism may induce stretching of the protein, which would account for reported unfoldase activity of GroEL and might explain how GroEL can actively chaperone proteins larger than HCA II.
A search for antiproton decay has been made at the Fermilab Antiproton Accumulator. Limits are placed on fifteen antiproton decay modes. The results are used to place limits on the characteristic mass scale m(X) that could be associated with CPT violation accompanied by baryon number violation.