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Biomedical subjects

M Ott

Publications and source records attributed to M Ott.

At least 73 records · Page 4Linked to original sources

A negatively powered lens in the chameleon.

Chameleons are arboral lizards that spot their prey visually and catch it by highly precise shots with their long sticky tongue. They scan their environment by large-amplitude independent saccadic eye movements; once an insect is detected, the head axis is aligned towards the target ('head tracking', both eyes come forward to fixate the insect and, in a phase called 'initial protrusion', the sticky tongue is loaded with tension by a special hyoid apparatus and subsequently shot out of the mouth with great precision. Lenses placed in front of the eyes produce predictable errors in distance estimation, suggesting that chameleons rely on accommodation cues when measuring the distance to their prey, but focusing has never been measured directly. Using a new technique to measure accommodation, we now show that accommodation is precise enough to serve as the major distance cue. Because accurate focusing requires large retinal images, we have tested image magnification and find that it is higher than in any other vertebrate eye scaled to the same size. This is a result of a unique optical design: unlike other vertebrate eyes, the crystalline lens of the chameleon has negative refractive power. Although there is a trend among vertebrates to increase corneal power and to decrease lens power with higher visual acuity, only in the chameleon eye has this tendency led to a reversal of the sign of the power of the lens.

Accommodation, Ocular↗

Tremor as a factor in prolonged reaction times of parkinsonian patients.

Prolonged reaction times and tremor are symptoms of Parkinson's disease (PD). Recently, we showed the existence of a systematic phase relationship between tremor-at-rest and the onset of voluntary motor responses in PD patients. In the present study we investigated whether this phase relationship contributes to the prolongation of reaction time in PD. Eight PD patients with prominent tremor-at-rest and five age-matched controls performed rapid stimulus-evoked index finger abductions under isometric conditions. Force and surface electromyogram (EMG) signals from first dorsal interosseus muscle were recorded and analyzed off-line by means of automatic routines. Reaction times of PD patients exhibited a significant dependence of mean values and variability on the current tremor phase at the onset of the voluntary motor response. Responses with an onset of contraction during the beginning of an EMG tremor burst were substantially delayed (on average 50 ms) and showed more variability in comparison to responses initiated at later times in the tremor cycle. This effect can be modeled by a simple gating process splitting the tremor cycle into two different system states that supports and inhibits the initiation of voluntary motor responses, respectively. We conclude that attraction of voluntary motor initiation to the tremor oscillator clearly contributes to prolongation of reaction time in PD.

Aged↗

Isolation of a Legionella pneumophila strain serologically distinguishable from all known serogroups.

A Legionella pneumophila strain (Jena-1) was isolated from a water sample collected from the hot water system of a scientific institution in Jena, Germany. Protein profile, ubiquinone and fatty acid content of the outer membrane were in accordance with those described for other Legionella pneumophila serogroups. DNA extracted from strain Jena-1 gave a positive amplification by using an L. pneumophila (mip)-specific commercially available PCR-kit confirming that the isolate belonged to the species L. pneumophila. Strain Jena-1 reacted with a monoclonal antibody specific for the major outer membrane protein of the species L. pneumophila and another one recognizing a lipopolysaccharide epitope of L. pneumophila serogroups 2-6, 8-10, and 12-15. Cross-absorption studies using absorbed and unabsorbed rabbit antisera to serogroups 1-15 and the newly isolated strain showed that strain Jena-1 cross-reacted mainly with serogroup 4, and to a lesser extent, with serogroups 5, 8, and 10. These cross-reactions could be removed by cross-absorption without significant effects on the homologous titres. It is concluded that strain Jena-1 represents a new serogroup of Legionella pneumophila.

Animals↗

Measurement of human chorionic gonadotropin-related immunoreactivity in serum, ascites and tumour cysts of patients with gynaecologic malignancies.

Human chorionic gonadotropin (hCG)-like molecules have been reported to be elevated in a substantial fraction of serum samples from patients with various gynaecologic tumours and have been discussed as possible markers in these malignancies. Employing highly sensitive and specific immunoradiometric assays, we determined total hCG-related immunoreactivity (hCG/hCG beta), as well as free alpha-subunit (alpha-SU), common to all glycoprotein hormones, in serum (n = 106) and malignant effusions (n = 26) of women with gynaecologic malignancies. For comparison, we also measured hCG/hCG beta in nonmalignant ascitic fluids (n = 21). HCG/hCG beta serum levels were elevated (> 5 IU L-1) in 39 of 106 patients (37%) with gynaecologic malignancies, whereas free alpha-SU was above normal range only in seven (6.6%). Frequencies of hCG/hCG beta elevations were similar in women with endometrial, (n = 39), cervical (n = 40) and ovarian (n = 27) cancer, being 30%, 35% and 41%, respectively. In malignant ascites (n = 15) and tumour cyst fluids (n = 11) of patients with ovarian cancer, hCG/hCG beta concentrations were significantly higher than in the corresponding serum samples and benign ascitic samples. Free alpha-SU, on the other hand, was increased in only one of 26 malignant effusions. In conclusion, hCG/hCG beta is frequently elevated in serum of patients with endometrial, cervical and ovarian cancer and may serve as a tumour marker in these malignancies, particularly in patients where other markers are negative. In this respect, analysis of ascitic or tumour cyst fluids may be of higher diagnostic value as serum measurements.

Adult↗

Cloning and genetic characterization of the flagellum subunit gene (flaA) of Legionella pneumophila serogroup 1.

The gene flaA, encoding the flagellum subunit protein of Legionella pneumophila serogroup 1, has been isolated from an expression library of L. pneumophila isolate Corby in Escherichia coli K-12 by using an antiflagellin specific polyclonal antiserum. DNA sequence analysis of the flaA gene revealed the presence of a 1,428-bp open reading frame encoding a protein of 475 amino acids with an apparent molecular mass of 48 kDa that is expressed independently of an E. coli vector promoter. Peptide sequencing of the N terminus of the isolated flagellum subunit protein confirmed that this open reading frame encodes the flagellin. By comparing the FlaA amino acid sequence with those of flagellins of various other bacteria, high degrees of homology in the N-terminal and C-terminal amino acids could be observed. The flaA-specific mRNA was determined to be 1.6 kb in size, the expected size of a monocistronic mRNA. Temperature-dependent expression of flagellin was found to be regulated at the transcriptional level. Sequence analysis and primer extension experiments indicated that the transcription of the gene flaA is directed by a sigma 28-like RpoF-FliA factor. By using fliA and fliA+ E. coli K-12 mutants, it was shown that flaA expression in E. coli required the sigma 28 factor. A flaA-specific DNA probe hybridizes with genomic DNA isolated from L. pneumophila and with most of the genomic DNAs from non-L. pneumophila Legionella strains. Two L. pneumophila strains and isolates of Legionella bozemanii and Legionella feeleii (serogroup 1) carry flaA-specific sequences but were not able to produce flagella.

Amino Acid Sequence↗

[Local therapy and adjuvant interferon therapy in genital papilloma virus infections].

OBJECTIVE: Is it possible to reduce the recurrence rates of HPV-positive genital tract lesions by systemic interferon alfa-2a in addition to local therapy? METHODS: Thirty-three of 63 patients with first manifestation of papillomavirus infection or monolocal manifestation were treated by local therapy. The other 30 patients with recurrent or multiorgan infections received 3 courses with 12 x 10(6) IU interferon alfa-2a subcutaneously. RESULTS: For the remaining 47 patients (16 were lost to follow-up) we found a significantly lower recurrence rate of 21% (5 of 24) in the group of interferon-treated patients compared to 52% (12 of 23) of patients treated without interferon. CONCLUSIONS: The systemic treatment of HPV-positive genital tract lesions with interferon alfa-2a in addition to CO2 laser surgery or cone biopsy seems to reduce the recurrence rates of HPV-positive lesions.

Adolescent↗

[Genomic DNA fingerprints of Legionella pneumophila serogroup 2 strains as an epidemiologic marker].

Using pulsed-field gel electrophoresis, DNA fingerprints of eleven Legionella pneumophila isolates of serogroup 2 were generated. It was shown that two strains from a patient suffering from pneumonia as well as three environmental strains isolated from the shower in the hotel where the patient stayed 5 days before his illness were identical. Six strains of the same serogroup isolated from other sources were clearly separated. Thus, DNA fingerprints by pulsed-field gel electrophoresis are excellent epidemiological markers for the rarely occurring serogroup 2 of Legionella pneumophila.

Antibodies, Monoclonal↗

Analysis of Legionella pneumophila strains associated with nosocomial pneumonia in a neonatal intensive care unit.

A premature child received continuous mechanical ventilation in a neonatal intensive care unit. On day 10 of his life he developed pneumonia due to Legionella pneumophila serogroup 1, monoclonal subtype Bellingham. The strain was cultured from a tracheal secretion taken on day 10 and detected by immunofluorescence using monoclonal antibodies on days 10, 12 and 17. Legionella pneumophila serogroups 1 and 6 (10(2)-4 x 10(4) cfu/l) were cultured from both central and peripheral hot water systems. Monoclonal antibody testing, macrorestriction analysis of the genomic DNA using pulse-field electrophoresis, and electrophoretic alloenzyme typing showed the isolate from the child to be identical to the serogroup 1 strains from the hot water system. Four unrelated Legionella strains of the same monoclonal subgroup Bellingham were studied for comparison. Legionellae were also isolated from two other incubators, but no clinical or microbiological indications of legionellosis were found in the neonates hospitalised there. Serogroup 1 strains isolated from the patient and from the hot water system and serogroup 6 isolates from the hot water supply were able to multiply in cultured Acanthamoeba castellanii cells and in guinea pigs. The serogroup 6 strain, although prevalent in the incubators, was not found in any of the clinical specimens by either culture of immunofluorescence.

Animals↗

Nutritional and metabolic variables correlate with amino acid forearm flux in patients with severe head injury.

OBJECTIVE: To measure the arterial-venous amino acid flux across the forearm muscle in patients with severe head injury. DESIGN: Prospective, interventional study. SETTING: Level I trauma hospital in the neurosurgery intensive care unit (ICU) at a university medical center. PATIENTS: Eight nonsteroid-treated patients with severe head injury. INTERVENTIONS: Patients were prospectively randomized to receive either standard or supplemental intravenous zinc therapy. MEASUREMENTS AND MAIN RESULTS: Net forearm alanine, glutamine, tyrosine, phenylalanine, and branch-chain amino acid forearm flux were measured and compared with metabolic markers of energy expenditure and nitrogen excretion. There was a significant inverse relationship between the measured energy expenditure/predicted energy expenditure ratio and glutamine flux (r2 = .62; p < .05). The patients with the highest measured energy expenditure/predicted energy expenditure ratio had the greatest release of glutamine from forearm muscle. Nitrogen balance was significantly correlated with leucine flux (r2 = .53; p < .05) and with isoleucine flux (r2 = .67; p < .05). The patients with the most positive nitrogen balance had the least release of branch-chain amino acids from skeletal muscle. Tyrosine flux was highly correlated with net amino acid flux (r2 = .76; p < .01). Tyrosine flux was therefore indicative of overall muscle catabolism. Four patients had an overall negative flux of amino acids from skeletal muscle. Three patients had an overall negative flux of branch-chain amino acids. CONCLUSIONS: This preliminary descriptive report suggests that increased skeletal muscle efflux of amino acids correlates significantly with metabolic variables of hypermetabolism and hypercatabolism in nonsteroid-treated, head-injured patients.

Adult↗

Genetic approaches to study Legionella pneumophila pathogenicity.

Legionella pneumophila is an intracellular pathogen replicating in human macrophages during the course of infection of the lungs. Infection by legionellae often leads to severe pneumonia, termed Legionnaires' disease. Genetic approaches to identify the factors responsible for L. pneumophila pathogenicity started with the construction of genomic libraries in Escherichia coli. Various L. pneumophila-specific genes were cloned in E. coli K-12 by identification using functional assays, antibody screening and hybridization ('reverse genetics'). By disrupting the genes via allelic exchange, mutants have been created to assess the influence of the factors on pathogenicity. Among the cloned genes, only for the gene product of the mip gene, encoding a 24-kDa surface-associated protein (macrophage infectivity potentiator) unequivocal evidence for its contribution to pathogenicity could be provided. Two hemolytic factors that have been cloned do not seem to play a role in L. pneumophila pathogenicity. Genetic systems for transposon mutagenesis of the L. pneumophila genome (Tn5, Tn903dIIlacZ, MudphoA), including Tn phoA shuttle mutagenesis, have been established and specifically adapted to identify mutants which displayed an impaired capability to multiply inside macrophages and with a reduced in vivo virulence. Furthermore, by complementation of avirulent mutants, genetic loci could be identified which restored the virulence.

Gene Expression Regulation, Bacterial↗

Excision of large DNA regions termed pathogenicity islands from tRNA-specific loci in the chromosome of an Escherichia coli wild-type pathogen.

Uropathogenic Escherichia coli 536 (O6:K15:H31) carries two unstable DNA regions, which were shown to be responsible for virulence. These regions, on which the genes for hemolysin production (hly) and P-related fimbriae (prf) are located, are termed pathogenicity islands (PAI) I and II, and were mapped to positions 82 and 97, respectively, on the E. coli K-12 linkage map. Sequence analysis of the PAI region junction sites revealed sequences of the leuX and selC loci specific for leucine and selenocysteine tRNAs. The tRNA loci function as the targets for excision events. Northern (RNA) blot analysis revealed that the sites of excision are transcriptionally active in the wild-type strain and that no tRNA-specific transcripts were found in the deletion mutant. The analysis of deletion mutants revealed that the excision of these regions is specific and involves direct repeats of 16 and 18 nucleotides, respectively, on both sides of the deletions. By using DNA long-range mapping techniques, the size of PAI I, located at position 82, was calculated to be 70 kb, while PAI II, mapped at position 97, comprises 190 kb. The excision events described here reflect the dynamics of the E. coli chromosome.

Base Sequence↗

Sequence determination and mutational analysis of the lly locus of Legionella pneumophila.

The lly (legiolysin) locus codes for a 39-kDa protein which confers hemolysis, pigment production, and fluorescence on recombinant Escherichia coli K-12 clones carrying the lly gene. The nucleotide sequences of the lly genes from two Legionella pneumophila isolates were determined. The lly loci exhibited identical nucleotide sequences. They contained open reading frames of 348 amino acid residues, encoding proteins with a deduced molecular mass of 38.9 kDa. N-terminal amino acid sequencing further confirmed that the Lly protein corresponds to the open reading frame sequenced. The amino acid sequence of the Lly protein exhibits a high degree of homology with the sequences of the MelA protein responsible for melanin production in the freshwater bacterium Shewanella colwelliana and the 4-hydroxyphenylpyruvate dioxygenase of Pseudomonas spp. 4-Hydroxyphenylpyruvate dioxygenase is involved in the degradation of aromatic amino acids in various organisms. An Lly-negative mutant of L. pneumophila Philadelphia I derivative JR32 and an Lly-positive transcomplementant were constructed. The Lly-negative mutant lost the ability to produce brown pigment and to confer fluorescence but retained hemolysis. Introduction of a plasmid carrying the lly locus restored pigment production and fluorescence. Intracellular survival of L. pneumophila in U937 macrophage-like cells and in Acanthamoeba castellanii was not affected by mutagenization of the lly locus.

Amino Acid Sequence↗

Comparison of BACTEC Plus 26 and 27 media with and without fastidious organism supplement with conventional methods for culture of sterile body fluids.

We compared the BACTEC Plus 26/27 culture system (Becton Dickinson Diagnostic Instrument Systems, Sparks, Md.) with and without fastidious organism supplement with conventional centrifugation preparation and plating for the recovery and speed of detection of microorganisms. A total of 1,101 sterile body fluid specimens were collected and processed at five hospital laboratories, yielding 234 (21%) positive cultures. Of the 176 isolates considered clinically significant, 133 (76%) were recovered by both the BACTEC system and conventional culture, while 28 (16% [P < 0.005]) were recovered by BACTEC only and 11 (6%) were recovered by conventional culture alone. There were no statistically significant differences in the speed of detection of microbial growth. It was found that BACTEC, with or without the addition of fastidious organism supplement, exhibited improved sensitivity for the recovery of microorganisms, including fastidious bacteria.

Bacteria↗

Clonal differentiation of uropathogenic Escherichia coli isolates of serotype O6:K5 by fimbrial antigen typing and DNA long-range mapping techniques.

Escherichia coli isolates of serotype O6:K5 are the most common causative agents of cystitis and pyelonephritis in adults. To answer the question, as to whether strains of this particular serotype represent one special clonal group, out of a collection of 34 serotype O6:K5 isolates [Zingler et al. (1990) Zentralbl. Bakteriol Mikrobiol Hyg [A] 274:372-381] 15 strains were selected and analyzed in detail. The flagellar (H) antigen and the outer membrane protein (OMP) pattern were determined. Further serum resistance properties and the genetic presence and expression of other virulence factors, including hemolysin, aerobactin, P fimbriae, S/F1C fimbriae and type 1 fimbriae was evaluated. In addition the XbaI-macrorestriction pattern of ten representative isolates was elaborated and the fimbrial (F) antigen type of the P fimbriae was determined, to obtain the complete O:K:H:F pattern. These analyses could clearly show that the O6:K5 isolates do not represent one clonal group. The XbaI-macrorestriction profiles were heterogeneous and marked differences in the hybridization patterns, using virulence-associated gene probes in Southern hybridization of long-range-separated genomic DNA, were observed among the strains. However, some of strains showed similarities in the genomic profiles, arguing for clonal groupings among the O6:K5 isolates. Interestingly the strains grouped together exhibited the same fimbrial F type that many indicate a coincidence of this phenotypic trait with clonality.

Adhesins, Escherichia coli↗

Interleukin-2, soluble interleukin-2-receptor, neopterin, L-tryptophan and beta 2-microglobulin levels in CSF and serum of patients with relapsing-remitting or chronic-progressive multiple sclerosis.

Cerebrospinal fluid (CSF) and serum levels of interleukin-2 (IL-2), soluble IL-2 receptors (sIL-2R), neopterin, L-tryptophan (L-TRP) and beta 2-microglobulin (beta 2-M) were measured in 31 untreated multiple sclerosis patients in acute exacerbation and 27 normal controls. Twenty-six patients showed the relapsing-remitting type of disease (RRMS); 5 had a chronic-progressive course (CPMS). No changes in serum IL-2 and sIL-2R were found between RRMS patients and controls, whereas serum and CSF levels as well as the CSF/serum ratio of neopterin were significantly elevated in the RRMS group. IL-2 was not detectable in CSF of patients or controls and sIL-2R levels were at the level of the lower detection (LD) sensitivity of the ELISA method. Four of 23 RRMS patients versus 1 of 25 controls showed CSF sIL-2R levels above the LD sensitivity, indicating a trend towards elevation in acute relapse. No difference was found in serum and CSF L-TRP and beta 2-M of patients and controls. In CSF of RRMS patients neopterin and L-TRP correlated negatively, reflecting the interferon-gamma mediated activation of macrophages in acute relapse. A significant positive correlation (Spearman rank 0.57, P = 0.001) between serum IL-2 levels and duration of acute relapse (mean 30 days) warrants further evaluation.

Acute Disease↗

A double-blind, placebo controlled trial of solidified benzyl benzoate applied in dwellings of asthmatic patients sensitive to mites: clinical efficacy and effect on mite allergens.

BACKGROUND: The aim of this double-blind, randomized study was to investigate the effectiveness of an acaricidal cleaning product in modifying both clinical symptoms and mite allergen levels over a period of at least 1 year. METHODS: Twenty-six asthmatic patients with proven Dermatophagoides pteronyssinus (Dp) asthma were selected; three were withdrawn from the trial. The patients' homes were divided into two groups; 11 homes were treated with solidified benzyl benzoate and tenside agents (A), and 12 were treated with a placebo (P). Two applications were performed at the beginning of the trial and at least 6 months later. Patients were examined 1 month before the trial, at the beginning of the trial, and every 3 months over a period of 1 year. Indoor mite exposure was evaluated by three methods: semiquantitative guanine determinations, quantitative guanine determinations, and the measurement of Der p I + Der f 1 (antigen P, of Dp + antigen F1 of D. farinae) levels. RESULTS: The symptom scores established at the beginning of the trial and 12 months later showed a statistically significant improvement only in the A group (p < 0.01). The visual analog scale also showed a statistically significant difference both in the A (p < 0.05) and P groups (p < 0.01). No statistical differences were found between medication scores in the A or P groups. A statistically significant increase was also observed for forced expiratory volume in 1 second and maximal expiratory flow rate 25/75 in the two groups (p < 0.05 for P group; p < 0.01 for A group). The mean decreases in Der p I + Der f 1 in patient mattresses between the beginning of the trial and after 12 months were 20% for the acaricide group and 17% for the placebo group, respectively (NS). For house dust samples with origins other than the patients' mattresses we found significant decreases in Der p I+Der f 1 in the A group (p < 0.01 for carpets and p < 0.05 for upholstery elements).

Adolescent↗

EBV DNA in nodal and extranodal non-Hodgkin's lymphomas: impact of cell lineage, morphology, and site of origin.

The Epstein-Barr virus (EBV) is a transforming herpes virus which is found in high frequencies in lymphoproliferative disorders arising in immunocompromised patients. To address the question of viral involvement in lymphomas of immunocompetent patients, we investigated 299 nodal and extranodal non-Hodgkin's, non-Burkitt's lymphomas of B- and T-cell lineage for the presence of EBV DNA. Epstein-Barr nucleic acid sequences were detected in 23 of 226 (10%) cases of B-cell lymphomas, in 24/72 (33%) T-cell lymphomas and one non-B, non-T lymphoma. Our data imply a possible role for the EBV in lymphoma development or propagation, an influence of viral factors on morphology and an impact of the anatomic site in which lymphoma development occurs.

Cell Transformation, Neoplastic↗

Dynamics of the bacterial genome: deletions and integrations as mechanisms of bacterial virulence modulation.

Bacterial virulence is a multifactorial phenomenon, arising from the coordinate action of special abilities of the infectious agents, termed as virulence factors, which is crucial for the infectious process. The genetic determinants encoding those factors are termed virulence associated genes, which can be located on the bacterial chromosome or on extrachromosomal elements (plasmids). Various examples have repeatedly demonstrated that bacterial genome dynamics contributes to virulence modulation. Strikingly, a reduced in vivo virulence of the pathogens was shown to be due to the spontaneous loss of virulence associated genes. The deletion events can involve chromosomal as well as plasmid regions. Also integration of plasmids into the chromosome are considered as dynamic events. The new genetic location of the formerly plasmid encoded virulence associated genes can result in an alteration of virulence expression.

Bacteria↗