Biomedical subjects

T Ahola

Publications and source records attributed to T Ahola.

At least 37 records · Page 2Linked to original sources

PubMed1997

Non-invasive measurement of cardiac output: whole-body impedance cardiography in simultaneous comparison with thermodilution and direct oxygen Fick methods.

OBJECTIVE: To determine the reliability of whole-body impedance cardiography (ICGWB), with electrodes attached to wrists and ankles, in the measurement of cardiac output (CO) on the basis of simultaneous comparison with thermodilution (TD) and direct oxygen Fick (Fick) methods. DESIGN: Prospective clinical study. SETTING: A surgical intensive care unit at a university hospital. PATIENTS: Thirty consecutive subjects undergoing a coronary artery bypass surgery were investigated preoperatively. MEASUREMENTS: ICGWB derived CO was measured simultaneously with the TD and Fick methods to establish the biases and limits of agreement (LA) between the methods. RESULTS: The results obtained by ICGWB and the invasive methods showed good agreement. The bias and LA between COTD and COICG were 0.00 l/min: 1.37 and 1.37 l/min, respectively, and were close to those obtained between COTD and COFICK, 0.32 l/min; 1.74 and -1.10 l/min. The bias and LA between the COFICK and COICG were -0.32 l/min; -2.24 and 1.60 l/min respectively. The repeatability value of consecutive single measurements for ICGWB (RVICG = 0.57 l/min) was much better than for the TD method (RVTD = 1.10 l/min). CONCLUSION: There was close agreement between the results of the three methods in the measurement of CO. In sedated preoperative patients the accuracy of ICGWB is within clinically acceptable limits and its repeatability is excellent. ICGWB provides a useful alternative to the TD and Fick methods in cases where the pressures supplied by the pulmonary artery catheter are not essential.

Adult↗

PubMed1997

Monoclonal antibodies specific for Semliki Forest virus replicase protein nsP2.

A panel of monoclonal antibodies (MAbs) was raised against Semliki Forest virus (SFV) nonstructural protein nsP2, which is a protease, an NTPase, a putative RNA helicase, and a regulator of the synthesis of the subgenomic 26S mRNA encoding the structural proteins. nsP2, used for immunization, was expressed as a histidine fusion protein in Escherichia coli and purified by metal affinity chromatography. Dot-blot assay, using a membrane fraction from SFV-infected cells as antigen, gave 33 positive clones. Of these, 30 MAbs recognized nsP2 in Western immunoblotting, and 25 showed positive indirect immunofluorescence (IFAT) in SFV-infected cells; 15 MAbs stained the cytoplasmic vacuoles (CPVI), which are the sites of viral RNA synthesis in alphavirus-infected cells. MAb 3B5 recognized only CPVIs, as shown by double immunofluorescence staining with polyclonal anti-nsP3 antiserum. Most of the MAbs (20/33) recognized the nuclear form of nsP2, which may be associated with SFV neurovirulence. Immunoprecipitation with MAbs revealed that the SFV nonstructural proteins are associated with each other. None of the MAbs recognized Sindbis virus nsP2 in immunoblotting, indicating that they were directed to non-conserved sequences specific for SFV. Interestingly, these epitopes were located mostly within the N-terminal half of nsP2. Unexpectedly, the anti-nsP2 MAb 1E9 cross-reacted strongly with a host protein of 78 kDa from uninfected human, murine, avian and insect cells. This protein was identified as the immunoglobulin binding protein, BiP, by 2-D gel mapping and reaction with anti-BiP antiserum.

Animals↗

PubMed1997

Critical residues of Semliki Forest virus RNA capping enzyme involved in methyltransferase and guanylyltransferase-like activities.

The Semliki Forest virus (SFV) replicase protein nsP1 has methyltransferase (MT) and guanylyltransferase-like (GT) activities, which are involved in the capping of viral mRNAs. MT catalyzes the transfer of the methyl group from S-adenosylmethionine (AdoMet) to position 7 of GTP, and this reaction is followed by GT-catalyzed formation of the covalent complex m7GMP-nsP1. These reactions are virus specific and thus potential targets for inhibitors of virus replication. We have mutated residues of SFV nsP1, which are conserved in related proteins of the large alphavirus-like superfamily. Mutations of D64, D90, R93, C135, C142, and Y249 to alanine destroyed or greatly reduced the MT activity of nsP1. All MT-negative mutants lost also the GT activity, confirming that methylation of GTP is an essential prerequisite for the synthesis of the covalent guanylate complex. Mutation of H38 prevented the GT reaction without destroying MT activity. Conservation of residues essential for both reactions in the alphavirus-like superfamily implies that they use a capping mechanism similar to that for the alphaviruses. Residues D64 and D90 were necessary for AdoMet binding, as measured by UV cross-linking. Secondary structure predictions of nsP1 and other proteins of the superfamily place these residues in positions corresponding to AdoMet-binding sites of cellular methyltransferases, suggesting that they all may be structurally related.

Amino Acid Sequence↗

PubMed1996

The effects of palmitoylation on membrane association of Semliki forest virus RNA capping enzyme.

The nonstructural protein Nsp1 of Semliki Forest virus has guanine-7-methyltransferase and guanylyltransferase-like activities, required in the capping of viral mRNAs. It is palmitoylated and tightly associated with the cytoplasmic surface of the plasma membrane, endosomes, and lysosomes. To localize the acylation site(s) and the putative membrane-targeting domain, a number of deletions were made in the nsp1 gene. Most deletions resulted in the expression of nonpalmitoylated, enzymatically inactive, cytoplasmic protein. Palmitate could be released from Nsp1 with neutral hydroxylamine, indicating a thioester linkage to a cysteine residue. Therefore we mutated the conserved cysteine residues of Nsp1 to alanine. Triple mutation of Cys418, Cys419, and Cys420 resulted in nonpalmitoylated Nsp1, which was enzymatically active and still associated with membranes. However, it could be released from the membranes with 1 M NaCl, whereas 50 mM sodium carbonate (pH 12) was required to release wild type Nsp1, suggesting a conversion from an integral to a peripheral membrane protein. Indirect confocal immunofluorescence microscopy showed that the nonpalmitoylated Nsp1 colocalized with the plasma membrane marker, concanavalin A. However, it was not detected in filopodia, which were heavily stained in cells expressing wild type Nsp1. These results indicate that the acylation of Nsp1 was not needed for its targeting to the plasma membrane, but it was necessary for the migration to the filopodial extensions of the plasma membrane.

Acylation↗

PubMed1996

Synthesis of Semliki Forest virus RNA polymerase components nsP1 through nsP4 in Saccharomyces cerevisiae by expression of cDNA encoding the nonstructural polyprotein.

A Semliki Forest virus nonstructural polyprotein, P1234, expressed in the yeast Saccharomyces cerevisiae in the absence of a replicative RNA template appeared to be properly cleaved into nsP1 to nsP4. All nsPs were membrane associated, and nsP2 was also transported to the nucleus. The membrane fraction containing nsPs showed guanine-7-methyltransferase and guanylyltransferase-like activities, typical for Semliki Forest virus nsP1.

DNA-Directed RNA Polymerases↗

PubMed1995

Reaction in alphavirus mRNA capping: formation of a covalent complex of nonstructural protein nsP1 with 7-methyl-GMP.

After the start of transcription, the 5' ends of eukaryotic mRNA molecules are modified by the addition of a guanylyl residue to form a cap structure, G(5')ppp(5')N. The guanylyltransferase (GTP:mRNA guanylyltransferase, EC 2.7.7.50) reaction responsible for cap formation usually proceeds via a covalent enzyme-GMP intermediate. We have studied the alphavirus-specific guanylyltransferase by incubating lysates from Semliki Forest and Sindbis virus-infected cells with [alpha-32P]GTP, using vaccinia virus and mock-infected cells as controls. One additional 32P-labeled protein was detected in alphavirus-infected cells but only in the presence of S-adenosylmethionine. This protein was identified as the nonstructural protein nsP1. The properties of the covalent enzyme-guanylate complex were studied with Semliki Forest virus nsP1 expressed in recombinant baculovirus-infected cells. S-Adenosylmethionine and divalent cations were required for the complex formation. The reaction was specific for guanylate nucleotides (GTP, dGTP) and was inhibited by pyrophosphate. nsP1 could be labeled with S-adenosyl[methyl-3H]methionine but only under conditions in which the nsP1-guanylate complex was formed. 7-Methyl-GMP was released from the nsP1-guanylate complex by treatment with acid or acidic hydroxylamine. Similar treatment of vaccinia virus capping enzyme released GMP. These findings suggest that in the capping of alphavirus mRNAs the guanine is methylated before linkage to the mRNA molecule.

Alphavirus↗

PubMed1995

Two alpha-chain hemoglobin variants, Hb Broussais and Hb Cemenelum, characterized by cation-exchange HPLC, isoelectric focusing, and peptide sequencing.

We here report the characteristics of two rare alpha-chain hemoglobin (Hb) variants. The variants were found during quantification of HbA1c by cation-exchange HPLC with the Diamat glycohemoglobin analyzer. They were further characterized by isoelectric focusing and PolyCAT A cation-exchange chromatography. The structure of the abnormal Hbs was established by amino acid analysis after separation of the globin chains by reversed-phase chromatography, digestion with trypsin, separation of the peptides by reversed-phase chromatography, and amino acid sequencing. These studies showed that the two variants were Hb Broussais [alpha 90 (FG2)Lys-->Asn] and Hb Cemenelum [alpha 92 (FG4)Arg-->Trp].

Adolescent↗

PubMed1992

[Breaking the circle of aggression: victim, aggressor and network].

Explore the source record for details and available documents.

Adult↗

PubMed1988

Return of the question "why": advantages of exploring pre-existing explanations.

We suggest that there are many advantages in thoroughly exploring the causal explanations given by clients or members of their social network to account for their problems. Specific interviewing techniques are presented to uncover clients' causal explanations or their impressions about the causal explanations of others. Various advantages of exploring these explanations are discussed. They include improved cooperation, development of "systemic empathy," detachment from the explanations of other professionals, recognition and avoidance of coalitions, loosening of firmly held explanations, dilution of noxious explanations, generation of new and positive explanations toward solutions, and taking a bird's-eye view or meta position about such explanations. We conclude that acceptance and appreciation of the human tendency to believe in causal explanations is a fruitful way to enhance interaction between clinicians and clients.

Communication↗

PubMed1988

Comparison of indirect diagnostic methods for hypolactasia.

The aim of this study was to continue our previously published work and to compare the different indirect diagnostic methods for hypolactasia with the lactase to sucrase ratio obtained by jejunal biopsy. The following tests were performed in 63 adult patients: the breath hydrogen test, the lactose tolerance test with ethanol (serum galactose measurement after oral lactose load with ethanol), the urinary lactose tolerance test (urinary galactose measurement after oral lactose load with ethanol), and the strip test (like the former but using a special test strip for urinary galactose). Specificities of all these tests were good (96-98%). The 3-h breath hydrogen test was less sensitive (69%) than the other methods (81-94%). The strip test is recommended for the general practitioner for the diagnosis of this common cause of abdominal complaints.

Adolescent↗

PubMed1979

Sialic acid secretion of patients with gastric and duodenal ulcers in the dose-response pentastrin test.

The secretion of HCl and sialic acid into the gastric juice was investigated in 13 patients with gastric, 52 with duodenal ulcers and 27 control patients, during the dose-response pentagastrin test. The basal output of sialic acid was clearly highest in patients with gastric ulcers, and the differences between these and other groups of patients were significant. During pentagastrin stimulation a significant initial decrease was found in patients with gastric ulcers with small doses, and the minimum was reached with a dose of 0.1 micrograms/kg/h. After increasing the dose, an increase in sialic acid output occurred in all groups, reaching its maximum with a pentagastrin infusion rate of 1.0 micrograms/kg/h. The basal concentration of sialic acid was also highest in patients with gastric ulcers. In all groups the concentration decreased during the stimulation, which obviously depended on the sialic output not increasing in the same relationship as the volume of gastric secretion.

Adult↗

PubMed1972