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Biomedical subjects

M Jonsson

Publications and source records attributed to M Jonsson.

At least 37 records · Page 2Linked to original sources

Double bands in DNA gel electrophoresis caused by bis-intercalating dyes.

Many bis-intercalating dyes used for fluorescence detection of DNA in electrophoresis have been reported to give band-splitting and band-broadening, which results in poor resolution and a decreased detection sensitivity. We have studied the dimeric dye YOYO-1, and to some extent also TOTO-1 and EthD-1, and found that in complex with DNA these dyes give rise to two components with different electrophoretic mobilities. Electrophoresis experiments and spectroscopic measurements on the two components show that they differ in that the DNA molecules have different amounts of dye bound. Our results exclude that the extra bands are caused by intermolecular cross-linking. Incubation of the samples for increasing times before electrophoresis makes the bands move closer and closer to each other as the dye molecules become more homogeneously distributed among the DNA molecules. Finally, the two bands merge into one at an intermediate position. This equilibration process is extremely slow at room temperature (days), and is therefore not a practical method to eliminate band-splitting in routine analysis. However, we find that if the temperature is raised to 50 degrees C, the dye-DNA complexes equilibrate completely in only 2 h.

Bacteriophage T7↗

Characterization of the binding of YO to [poly(dA-dT)]2 and [poly(dG-dC)]2, and of the fluorescent properties of YO and YOYO complexed with the polynucleotides and double-stranded DNA.

The interaction between the fluorescent dye YO (oxazole yellow) and the alternating polynucleotides [poly(dA-dT)]2 [the duplex of alternating poly(dA-dT)] and [poly(dG-dC)]2 [the duplex of alternating poly(dG-dC)] has been studied with optical spectroscopic techniques including absorbance, flow linear dichroism, CD, and fluorescence measurements. The principal features of the spectra are very similar for the two polynucleotide solutions, showing that YO binds quite similarly to AT and GC base pairs. From a strongly negative reduced linear dichroism (LDr) in the dye absorption band, an induced negative CD, and transfer of energy from the bases to bound YO, we conclude that at low mixing ratios YO is intercalated in both [poly(dA-dT)]2 and [poly(dG-dC)]2. At higher mixing ratios an external binding mode starts to contribute, evidenced from the appearance of an exciton CD. The conclusion that YO binds in a similar way to AT and GC base pairs should be valid also for the dimer YOYO since its YO units have been found to bind to double-stranded (dsDNA) in the same way as the YO monomer. The fluorescence properties of YO and YOYO complexed with DNA or the polynucleotides have been characterized by studying the dependence of fluorescence intensity on temperature, mixing ratio, and ionic strength. The fluorescence intensity and fluorescence lifetime of YO-DNA decrease strongly with increasing mixing ratio, whereas the fluorescence intensity of YOYO-DNA shows a weaker dependence, indicating that the quantum yield depends on the distance between the YO chromophores on the DNA chain. Further, the fluorescence intensity of YO depends on the base sequence; the quantum yield and fluorescence lifetime for YO complexed with [poly(dG-dC)]2 are about twice as large as for YO complexed with [poly(dA-dT)]2. Measurements of excitation spectra at different mixing ratios and different emission wavelengths indicate that the fluorescence of the externally bound chromophores is negligible compared to the intercalated ones.

Benzoxazoles↗

Dynamic muscle strength training in stroke patients: effects on knee extension torque, electromyographic activity, and motor function.

The effects of training with isokinetic maximal voluntary knee extensions were studied in stroke patients. Two groups of 10 patients each trained twice a week for 6 weeks. One group trained exclusively eccentric movements and the other exclusively concentric movements. The effects were evaluated from the following tests before and after the training period. The maximal voluntary strength in concentric and eccentric actions of the knee extensor and flexor muscles was recorded together with surface electromyography at constant velocities of 60, 120, and 180 deg.s-1 on three different days. The body weight distribution on the legs while rising and sitting down was measured with two force plates. The self-selected and maximal walking speeds and the swing to stride ratio of the paretic leg were measured. After the training period, the knee extensor strength had increased in eccentric and concentric actions in both groups (p < .05). The eccentric and the concentric strength in the paretic leg relative to that of the nonparetic leg increased in the eccentrically trained group (p < .05) but not in the concentrically trained group. The restraint of the antagonistic muscles in concentric movements increased after concentric (p < .05) but not eccentric training. A nearly symmetrical body weight distribution on the legs in rising from a sitting position was noted after eccentric (p < .05) but not concentric training. Changes in walking variables were not significantly different between the groups. Eccentric knee extensor training was thus found to have some advantages as compared to concentric training in stroke patients.

Aged↗

Subcutaneous implanted chambers in different mouse strains as an animal model to study genetic stability during infection with Lyme disease Borrelia.

Tissue metal net cages were implanted subcutaneously in BALB/cJ and C3H/Tif mice as an experimental model of Borrelia burgdorferi infection. B. burgdorferi sensu stricto strain Sh2-82 could be isolated up to 14 weeks after the inoculation. However, a significant difference in infectivity between the two mice strains was observed. C3H/Tif mice were more susceptible to developing chronic B. burgdorferi s.s. infections than BALB/cJ mice. Although a B. burgdorferi infection was established, no rearrangements in the ospA and ospB genes were observed in any of the infected mice.

Animals↗

Human papillomavirus infection is transient in young women: a population-based cohort study.

The prevalence of human papillomavirus (HPV) infection in cervical cell scrapes from a cohort of 276 young women was determined by a general two-step polymerase chain reaction. HPV infection fluctuated among young women during a 2-year interval. The total prevalence of HPV infection decreased from 21% to 8.3%. The most prevalent HPV types at enrollment were HPV-16 (3.3%) and HPV-6 (2.9%). At follow-up, the most common type was HPV-16 (2.9%), while no HPV-6 was detected. In 2 women only, the same HPV type persisted. Regression of HPV infection was found in 80% of the women. A new HPV type-specific infection was detected in 7.2% of the women and was independently associated with a new sex partner or an abnormal smear since enrollment.

Adult↗

The silent suffering women--a population based study on the association between reported symptoms and past and present infections of the lower genital tract.

OBJECTIVES: To assess the prevalence of lower genital tract symptoms and the association between reported symptoms and past and present signs of sexually transmitted diseases (STD) in young women. DESIGN: All women belonging to the 19-, 21-, 23- and 25-year age cohorts and living in the catchment area of the community health centre, were invited by mail to take part in a population-based study. The participants answered a structured questionnaire and a gynaecologic examination was performed. Samples for wet smear, cervical Pap smear, HPV DNA determination and Chlamydia trachomatis culture were taken at the gynaecologic examination. The presence of genital warts was noted. A blood sample was analysed for antibodies against C trachomatis and HSV-2. SETTING: The community health care centre was located in Umeå, a city in Northern Sweden. RESULTS: Of the 886 women who were eligible, 611 (70%) participated in the investigation. One out of four women reported symptoms from the lower genital tract. The most commonly reported symptoms were itching, followed by discharge, and soreness. The most commonly reported STD was C trachomatis (15%). The most prevalent present STD was HPV infection (20%) whereas C trachomatis infection could be isolated from 2.7% of the women. Antibodies against C trachomatis and HSV-2 were present among 22% and 6% of the women, respectively. There was a significant correlation between the women's complaint of vaginal discharge and previous C trachomatis infection, lack of lactobacilli and presence of leucocytosis in wet smear. CONCLUSIONS: We have in a population-based study of young healthy women found that one out of four women had some kind of lower genital tract complaint. Itching was the most commonly reported symptom and was associated with pseudohyphae and acetowhite patches. Reported vaginal discharge and soreness were associated with the history of a past C trachomatis infection and signs of a disturbed vaginal flora.

Adult↗

A bactericidal antibody to Borrelia burgdorferi is directed against a variable region of the OspB protein.

Borrelia burgdorferi, an agent of Lyme disease, is killed by some monoclonal antibodies in the absence of complement or phagocytes. In the present study, the bactericidal action of monoclonal antibodies against B. burgdorferi and B. hermsii, a cause of relapsing fever, was further characterized. H6831, an antibody recognizing the OspB proteins of some B. burgdorferi strains, and H4825, an antibody specific for one serotype of B. hermsii, were purified, and Fab fragments of the antibodies were prepared. In time-kill studies, more than 99.9% of strain B31 B. burgdorferi cells were killed after 30 min of exposure to H6831 Fab fragments. The MBC of the Fab fragments was 10 micrograms/ml. Electron microscopy revealed that the bactericidal Fab fragments produced numerous blebs and cell lysis of the borrelias for which they were specific. To identify the epitope for H6831, the OspB sequences of H6831-susceptible and -resistant strains and mutants were determined. The deduced OspB proteins of all H6831-resistant strains and mutants differed from the strain B31 OspB at residue 253. Murine antisera raised against a 21-mer synthetic peptide representing the region around residue 253 were specific for strain B31 by Western blot (immunoblot) and growth inhibition assays. Furthermore, the antipeptide serum inhibited the binding of H6831 to whole borrelias. These findings indicated that the linear component of the bactericidal antibody's epitope was located at or near residue 253.

Amino Acid Sequence↗

Identification of a highly cross-reactive outer surface protein B epitope among diverse geographic isolates of Borrelia spp. causing Lyme disease.

The outer surface lipoprotein B (OspB) of Borrelia burgdorferi is a major component of the borrelial protein profile and has been shown to be highly immunogenic in experimentally immunized and infected mammals. However, the ospB loci of different strains show considerable heterology at the nucleic acid sequence level, and the progeny of a clonal strain of B. burgdorferi exhibited OspB polymorphisms with respect to apparent molecular weights and reactivities with monoclonal antibodies. These data suggest that OspB is not a good candidate for vaccination or diagnostic purposes. The present study describes a monoclonal antibody, designated 84C, directed against a very highly conserved domain of the OspB lipoprotein. Western immunoblot analysis with 84C demonstrated reactivity in 84.2% of human, tick, and other vertebrate isolate strains examined from widely diverse geographic regions, including strains of B. burgdorferi sensu stricto and two closely related species, B. garinii and B. afzelii. The 84C-binding region was delimited to a highly conserved 11-amino-acid region in the carboxyl terminus of OspB as demonstrated by (i) DNA sequence analysis of wild-type and 84C-resistant mutant ospB alleles and (ii) deletion mutagenesis of a recombinant ospB gene in Escherichia coli. Finally, the 84C epitope was demonstrated to be exposed on the borrelial surface in situ as (i) the monoclonal antibody 84C was able to agglutinate borrelias in culture and (ii) 84C-resistant escape variants were isolated. These data suggest that the potential value of OspB as a vaccine candidate or diagnostic tool be examined more closely, in the context of the 84C-reactive domain.

Amino Acid Sequence↗

Cancer morbidity in blood recipients--results of a cohort study.

Blood transfusions involve the transfer of relatively large volumes of body fluids and cellular material between individuals. A variety of pathogens like viruses, some of which are associated with development of certain tumours, are known to be transmitted by this route. Blood recipients were identified during 1981-1982 in the register of the hospital blood centre, and in-patients by the in-patient and discharge register of the hospital. Tumour occurrence and vital status were determined by means of the population-based regional tumour register. Age, gender and calendar-year specific rates from the general population were used to calculate expected values. In a cohort study of 3177 blood recipients, increased numbers of malignant lymphomas [13 vs. 4.8 expected, standard morbidity ratio (SMR) 2.70 95% confidence interval (CI) 1.44-4.62] and skin cancers [12 vs. 5.2 expected, SMR 2.29, 95% CI 1.19-4.01] were seen 3 to 9 years after transfusion. In a second cohort study of 29,910 hospitalised patients, a total of 37 (29.8 expected) malignant lymphomas was found in 28,338 patients with no transfusion and 10 (2.73 expected) in 1572 patients with a transfusion, 3 to 9 years after the hospitalisation. The incidence rate ratio between these groups was 3.11 (95% CI 1.56-6.20) using a Mantel-Haenszel estimator with age stratification. Non-melanomatous skin cancers had an incidence ratio of 2.74 (95% CI 1.25-6.00). We conclude that, in the cohorts discussed here, malignant lymphomas and skin cancer occur more often in blood recipients than in controls. It remains to be established whether this is due to factors covariating with transfusion or by the transfusion itself. Further studies on these putative associations are warranted, as are analytical studies of the epidemiology of malignant lymphomas, especially non-Hodgkin's lymphoma, whose aetiology is still poorly understood.

Adult↗

Pharmacokinetics of ceftazidime in acutely ill hospitalised elderly patients.

The i.v. and i.m. dose pharmacokinetics of ceftazidime 1 g b.i.d. were investigated in steady state in 52 consecutive cases of hospitalised patients with underlying diseases and serious supervening bacterial infections. Following i.v. bolus injection, serum concentrations of greater than 4 mg/l persisted for 8 h in all patients. Compared to values found in younger patients studied previously, clearance was markedly reduced in the elderly, t1/2 beta more than doubled, the area under the curve four times enlarged and the apparent volume of distribution reduced. The values obtained in serum after i.m. injection were similar, though the peak concentration was delayed by about 2 h. The serum uptake was approximately 70%, and serum concentration was greater than 4 mg/l for 8 h. The concentrations in tissue fluid after i.v. administration were similar to those in serum, but the penetration into tissue fluid was slower and only 20% of that reported previously for young people. The elimination was slow, with greater than 4 mg/l maintained for 7 h. Tissue concentrations after i.m. injection were almost comparable with those after i.v. injection, but lagged behind by about 1 h. In conclusion, suitable concentrations in blood and tissue are attained with 1 g ceftazidime b.i.d., whether administered by the i.v. or i.m. route.

Aged↗

Heterogeneity of outer membrane proteins in Borrelia burgdorferi: comparison of osp operons of three isolates of different geographic origins.

Biochemical and immunochemical studies of the outer membrane proteins of Borrelia burgdorferi have shown that the OspA and OspB proteins from strains of different geographic origins may differ considerably in their reactivities with monoclonal antibodies and in their apparent molecular weights. To further characterize this variation in Osp proteins between strains, the osp operons and deduced translation products from two strains, one from Sweden (ACAI) and one from eastern Russia (Ip90), were studied. Polyacrylamide gel electrophoresis and Western blot (immunoblot) analyses confirmed differences between ACAI, Ip90, and the North American strain B31 in their Osp proteins. The sequences of the ospA and ospB genes of ACAI and Ip90 were compared with that of the previously studied osp operon of B31 (S. Bergström, V. G. Bundoc, and A. G. Barbour, Mol. Microbiol. 3:479-486, 1989). The osp genes of ACAI and Ip90, like the corresponding genes of B31, were found on plasmids with apparent sizes of about 50 kb and are cotranscribed as a single unit. Pairwise comparisons of the nucleotide sequences revealed that the ospA genes of ACAI and Ip90 were 85 and 86% identical, respectively, to the ospA gene of strain B31 and 86% identical to each other. The ospB sequences of these two strains were 79% identical to the ospB gene of B31 and 81% identical to each other. There was significantly greater similarity between the ospA genes of the three different strains than there was between the ospA and ospB genes within each strain. These studies suggest that the duplication of osp genes in B. burgdorferi occurred before the geographical dispersion of strains represented by ACAI, Ip90, and B31.

Amino Acid Sequence↗

Comparison of a one-step and a two-step polymerase chain reaction with degenerate general primers in a population-based study of human papillomavirus infection in young Swedish women.

The prevalence of human papillomavirus (HPV) infection in cervical cell scrapes from young women was determined by polymerase chain reaction (PCR) by using general primer pairs localized within the L1 region. With a one-step general PCR, 5.9% (35 of 590) of young women in a population-based study were found to contain HPV DNA. The proportion of HPV-positive women increased with age, from 1.4% (1 of 69) among women aged 19 years to 9.2% (13 of 142) among women aged 25 years. Among the cervical scrapes from women with normal cytology, 5.6% (30 of 539) harbored HPV DNA. A total of 5 of 19 (26.3%) of the women with pathological signs were positive for HPV DNA. By a two-step PCR, using nested general primers, 20.3% (118 of 581) of all women were shown to contain HPV DNA. The proportion of HPV-positive women also increased with age, from 17.4% (12 of 69) among women aged 19 years to 31.9% (43 of 135) among women aged 25 years, when the two-step PCR was used. Some 19.2% (102 of 530) of cervical scrapes from women with normal cytology contained HPV DNA. Among the women with pathological signs, 16 of 19 (84.2%) were positive for HPV DNA. The HPV DNA-positive specimens were demonstrated to contain HPV type 6, 11, 16, 18, 31, 33, 35, 39, 40, 45, 55, or 56. The most prevalent HPV types were 6 (2.0%) and 16 (2.7%). More than one type was found in 16 specimens. Sixty HPV-positive samples could not be typed.

Adult↗

Molecular characterization of Borrelia burgdorferi isolated from Ixodes ricinus in northern Sweden.

Ixodes ricinus ticks, harbouring Borrelia burgdorferi, were found in an area in northern Sweden, not thought to be endemic for Lyme borreliosis. This investigation took place at Norrbyskär, an island situated in the Bothnian Gulf, 63 degrees 33'N/19 degrees 52'E. One of 42 nymphal and 8/43 adult I. ricinus ticks collected carried spirochetes as seen by phase contrast microscopy. Pure bacterial cultures were obtained from 2 of the ticks. Western blot analysis using species-specific monoclonal antibodies showed that the isolated spirochetes were B. burgdorferi. The identity of the isolated spirochetes was confirmed by DNA amplification using B. burgdorferi OspA and flagellin gene specific oligonucleotides as well as partial DNA sequencing of the respective OspA and flagellin genes. The 2 isolated spirochaete populations were different as shown by their protein profiles in sodium dodecyl sulphate polyacrylamide gels. Moreover, the demonstration of Lyme borreliosis in a patient from the island of Norrbyskär indicates the need for clinical consideration of this disease in northern Sweden.

Animals↗

Comparison of European isolates of viruses causing hemorrhagic fever with renal syndrome by a neutralization test.

Different virus isolates causing hemorrhagic fever with renal syndrome (HFRS) were compared using a neutralization test. Patient convalescent sera and antisera prepared in rabbits were used to compare Puumala-related Hantavirus isolates from Finland, Sweden, Belgium, and the USSR. The majority of European isolates were indistinguishable from each other using both homologous rabbit antisera and patient convalescent sera. The European isolates of HFRS were also compared with prototype Hantaan (the etiologic agent of Korean hemorrhagic fever). The one-way cross-reaction between the Hantaan and Puumala viruses, previously described using human convalescent sera tested by indirect immunofluorescence and immunoprecipitation, was also seen by the neutralization test.

Europe↗

Norfloxacin vs. pivmecillinam in the treatment of uncomplicated lower urinary tract infections in hospitalized elderly patients.

In a single-blind, randomized study of multicentre trial design 343 hospitalized geriatric patients (median age 83 years) suffering from symptomatic urinary tract infections (UTI) were treated with either norfloxacin 200 mg twice daily or pivmecillinam 200 mg 3 times daily orally for 7 days to compare the clinical efficacy and safety of the antibiotics in the elderly. The pathogen was Escherichia coli in 56.7%, other gram-negative rods in 37.2% and gram-positive cocci in 6.1%. Up to 18 days after cessation of treatment the original pathogen was eradicated in 60.5% of the 114 who were assessable for the effect of norfloxacin therapy and in 36.5% of the 96 patients in the pivmecillinam group (p less than 0.001). Adverse drug reactions, mostly gastrointestinal disturbances, were noticed in single cases.

Aged↗