PubMed Health⌕ Search

Biomedical subjects

P Matousek

Publications and source records attributed to P Matousek.

18 recordsLinked to original sources

Depth profiling in diffusely scattering media using Raman spectroscopy and picosecond Kerr gating.

We demonstrate how pulsed laser Raman excitation (approximately 1 ps) followed by fast optical Kerr gating (approximately 4 ps) can be used to effectively separate Raman signals originating from different depths in heterogeneous diffusely scattering media. The diffuse scattering slows down photon propagation through turbid samples enabling higher depth resolution than would be obtained for a given instrumental time resolution in an optically transparent medium. Two types of experiments on two-layer systems demonstrate the ability to differentiate between surface and sub-surface Raman signals. A Raman spectrum was obtained of stilbene powder buried beneath a 1 mm over-layer of PMMA (poly(methyl methacrylate)) powder. The signal contrasts of the lower stilbene layer and upper PMMA layer were improved by factors >or=5 and >or=180, respectively, by rejecting the Raman component of the counterpart layer. The ability to select the Raman signal of a thin top surface layer in preference to those from an underlying diffusely scattering substrate was demonstrated using a 100 mum thick optically transparent film of PET (poly(ethylene terephthalate)) on top of stilbene powder. The gating resulted in the suppression of the underlying stilbene Raman signal by a factor of 1200. The experiments were performed in back-scattering geometry using 400 nm excitation wavelength. The experimental technique should be well suited to biomedical applications such as disease diagnosis.

Complex Mixtures↗

Subsurface probing in diffusely scattering media using spatially offset Raman spectroscopy.

We describe a simple methodology for the effective retrieval of Raman spectra of subsurface layers in diffusely scattering media. The technique is based on the collection of Raman scattered light from surface regions that are laterally offset away from the excitation laser spot on the sample. The Raman spectra obtained in this way exhibit a variation in relative spectral intensities of the surface and subsurface layers of the sample being investigated. The data set is processed using a multivariate data analysis to yield pure Raman spectra of the individual sample layers, providing a method for the effective elimination of surface Raman scatter. The methodology is applicable to the retrieval of pure Raman spectra from depths well in excess of those accessible with conventional confocal microscopy. In this first feasibility study we have differentiated between surface and subsurface Raman signals within a diffusely scattering sample composed of two layers: trans-stilbene powder beneath a 1 mm thick over-layer of PMMA (poly(methyl methacrylate)) powder. The improvement in contrast of the subsurface trans-stilbene layer without numerical processing was 19 times. The potential applications include biomedical subsurface probing of specific tissues through different overlying tissues such as assessment of bone quality through skin, providing an effective noninvasive means of screening for bone degeneration, other skeletal disease diagnosis, and dermatology studies, as well as materials and catalyst research.

Feasibility Studies↗

Numerical simulations of subsurface probing in diffusely scattering media using spatially offset Raman spectroscopy.

We present the first elementary model predicting how Raman intensities vary for a range of experimental variables for spatially offset Raman spectroscopy (SORS), a recently proposed technique for the effective retrieval of Raman spectra of subsurface layers in diffusely scattering media. The model was able to reproduce the key observations made from the first SORS experiments, namely the dependence of Raman signal intensities on the spatial offset between the illumination and collection points and the relative contributions to the overall spectrum from the top layer and sub-layer. The application of the SORS concept to a three-layer system is also discussed. The model also clearly indicates that an annular geometry, rather than a point-collection geometry, which was used in the earlier experiments, would yield much improved data.

Algorithms↗

Resolution of G(s)alpha and G(q)alpha/G(11)alpha proteins in membrane domains by two-dimensional electrophoresis: the effect of long-term agonist stimulation.

Low-density membrane-domain fractions were prepared from S49 lymphoma cells and clone e2m11 of HEK293 cells expressing a large number of thyrotropin-releasing hormone receptor (TRH-R) and G(11)alpha by flotation on sucrose density gradients. The intact cell structure was broken by detergent-extraction, alkaline-treatment or drastic homogenization. Three types of low-density membranes were resolved by two-dimensional electrophoresis and analyzed for G(s)alpha (S49) or G(q)alpha/G11) (e2m11) content. Four individual immunoblot signals of Gsalpha protein were identified in S49 lymphoma cells indicating complete resolution of the long G(s)alpha L+/-ser and short G(s)alpha S+/-ser variants of G(s)alpha. All these were diminished by prolonged agonist (isoprenaline) stimulation. In e2m11-HEK cells, five different immunoblot signals were detected indicating post-translational modification of G proteins of G(q)alpha/G(11)alpha family. The two major spots corresponding to exogenously (over)expressed G(11)alpha and endogenous G(q)alpha were reduced; the minor spots diminished by hormonal stimulation. Parallel analysis by silver staining of the total protein content indicated that no major changes in protein composition occurred under these conditions. Our data thus indicate that agonist-stimulation of target cells results in down-regulation of all different members of G(s) and G(q)/G(11) families. This agonist-specific effect may be demonstrated in crude membrane as well as domain/raft preparations and it is not accompanied by changes in overall protein composition.

Cell Line, Tumor↗

[Personal experience with laser photocoagulation of varicose veins].

Based on the published information on the use of operation lasers for endoluminal photocoagulation to seal varices the authors began to use KTP laser in 2001. The cohort included 200 patients operated on in 2002. The cohort was composed of three groups of patients: patients with reticular varices, those with non-saphenous varices and patients with the stem saphenous varices. The intervention was made by a KTP laser, wavelength 532 nm, light-conducting wires of 0.2, 0.3 or 0.6 mm diameter.

Adolescent↗

Ultrafast dynamics of photochemical radical formation from

The excited-state dynamics and photochemistry of [Re(R)(CO)3(dmb)] (R=Me, Et); dmb=4,4'-dimethyl-2,2'-bipyridine) in CH2Cl2 have been studied by time-resolved visible absorption spectroscopy on a broad time scale ranging from approximately 400 fs to a few microseconds, with emphasis on the femtosecond and picosecond dynamics. It was found that the optically prepared Franck-Condon 1MLCT (singlet metal-to-ligand charge transfer) excited state of [Re(R)(CO)3(dmb)] undergoes femtosecond branching between two pathways (< or =400 fs for R=Me; approximately 800 fs for R=Et). For both methyl and ethyl complexes, evolution along one pathway leads to homolysis of the Re-R bond via a 3SBLCT (triplet sigma-bond-to-ligand charge transfer) excited state, from which [Re(S)(CO)3(dmb)]* and R* radicals are formed. The other pathway leads to an inherently unreactive 3MLCT state. For [Re(Me)(CO)3(dmb)], the 3MLCT state lies lowest in energy and decays exclusively to the ground state with a lifetime of approximately 35 ns, thereby acting as an excitation energy trap. The reactive 3SBLCT state is higher in energy. The quantum yield (0.4 at 293 K) of the radical formation is determined by the branching ratio between the two pathways. [Re(Et)(CO)3(dmb)] behaves differently: branching of the Franck-Condon state between two pathways still occurs, but the 3MLCT excited state lies above the dissociative 3SBLCT state and can decay into it. This shortens the 3MLCT lifetime to 213 ps in CH2Cl2 or 83 ps in CH3CN. Once populated, the 3SBLCT state evolves toward radical photoproducts [Re(S)(CO)3(dmb)]* and Et*. Thus, population of the 3MLCT excited state of [Re(Et)(CO)3(dmb)] provides a second, delayed pathway to homolysis. Hence, the quantum yield is unity. The photochemistry and excited-state dynamics of [Re(R)(CO)3(dmb)] (R=Me, Et) complexes are explained in terms of the relative ordering of the Franck-Condon, 3MLCT, and 3SBLCT states in the region of vertical excitation and along the Re-R reaction coordinate. A qualitative potential energy diagram is proposed.

Journal Article↗

Low interleukin-12 activity in severe Plasmodium falciparum malaria.

We compared interleukin-12 (IL-12) and other cytokine activities during and after an acute clinical episode in a matched-pair case-control study of young African children who presented with either mild or severe Plasmodium falciparum malaria. The acute-phase, pretreatment plasma IL-12 and alpha interferon (IFN-alpha) levels, as well as the acute-phase mitogen-stimulated whole-blood production capacity of IL-12, were significantly lower in children with severe rather than mild malaria. IL-12 levels, in addition, showed strong inverse correlations both with parasitemia and with the numbers of circulating malaria pigment-containing neutrophils. Acute-phase plasma tumor necrosis factor (TNF) and IL-10 levels were significantly higher in those with severe malaria, and the concentrations of both of these cytokines were positively correlated both with parasitemia and with the numbers of pigment-containing phagocytes in the blood. Children with severe anemia had the highest levels of TNF in plasma. In all the children, the levels in plasma and production capacities of all cytokines normalized when they were healthy and parasite free. The results indicate that severe but not mild P. falciparum malaria in young, nonimmune African children is characterized by down-regulated IL-12 activity, contrasting markedly with the up-regulation of both TNF and IL-10 in the same children. A combination of disturbed phagocyte functions resulting from hemozoin consumption, along with reduced IFN-gamma responses, may contribute to these differential effects.

Acute Disease↗

Antibody responses to Plasmodium falciparum: evolution according to the severity of a prior clinical episode and association with subsequent reinfection.

We measured sporozoite- and total parasite antigen-specific IgG and IgM antibodies before and after treatment in matched groups of Gabonese children who presented with either mild or severe Plasmodium falciparum malaria. We investigated the influence of various parameters on these antibody responses, including clinical presentation, age, and post-treatment reinfection profiles. IgG but not IgM responses were strongly influenced by both clinical and parasitological status. IgG responses to the repeat region of the circumsporozoite protein, which were low at admission, particularly so in those with severe anemia, increased after treatment but showed no association with either age or reinfection profiles. Total parasite antigen-specific IgG responses were strongly influenced by parasitological status, and also differed significantly when segregated according to clinical status at admission, age, and reinfection histories. Most notably, anti-parasite IgG responses measured when children were parasite-free were higher and a good indicator of recent reinfections in those who presented with mild rather than with severe malaria. The profile of responses in the latter group suggests some immune system dysfunction, which may reflect the induction of tolerance to parasite antigens.

Animals↗

Interferon-gamma responses are associated with resistance to reinfection with Plasmodium falciparum in young African children.

The contribution of T cell-mediated responses was studied with regard to resistance to reinfection in groups of Gabonese children participating in a prospective study of severe and mild malaria due to infection with Plasmodium falciparum. In those admitted with mild malaria, but not in those with severe malaria, production of IFN-gamma by peripheral blood mononuclear cells (PBMC) in response to either liver-stage or merozoite antigen peptides was associated with significantly delayed first reinfections and with significantly lower rates of reinfection. Proliferative or tumor necrosis factor responses to the same peptides showed no such associations. Production of interferon-gamma by PBMC in response to sporozoite and merozoite antigen peptides was observed in a higher proportion of those presenting with mild malaria. Differences in the Th1/Th2 cytokine balance may be linked to the ability to control parasite multiplication in these young children, helping to explain the marked differences observed in both susceptibility to infection as well as in clinical presentation.

Amino Acid Sequence↗

The role of red blood cell polymorphisms in resistance and susceptibility to malaria.

In regions highly endemic for Plasmodium falciparum malaria, red cell polymorphisms that confer resistance to severe disease are widespread. Sickle cell trait, alpha-thalassemia, glucose-6-phosphate dehydrogenase deficiency, and blood groups were determined in 100 children from Gabon with severe malaria who were matched with 100 children with mild malaria and followed up for evaluation of reinfections. The sickle cell trait was significantly associated with mild malaria and blood group A with severe malaria. During follow-up, the original severe cases had significantly higher rates of reinfection than the original mild cases, with higher parasitemia and lower hematocrit values. Incidence rates did not differ in the context of erythrocyte polymorphisms, but patients with sickle cell trait presented with markedly lower levels of parasitemia than those without. Thus, the severity of malaria is partly determined by the presence of blood group A and the sickle cell trait. The different presentation of reinfections in severe versus mild cases probably reflects different susceptibility to malaria.

ABO Blood-Group System↗

Parasite antigen-specific interleukin-10 and antibody reponses predict accelerated parasite clearance in Plasmodium falciparum malaria.

Using strict inclusion criteria, we conducted a hospital-based, case-control study in which 100 Gabonese children with severe Plasmodium falciparum malaria were matched for age, gender and provenance with 100 children presenting with mild malaria. Parasite antigen-specific cellular and humoral immunological responses were measured and compared with post-treatment parasite clearance times in each group. Significantly faster parasite clearance times were associated with in vitro production of IL-10 by acute-phase peripheral blood mononuclear cells (PBMC) in response to both liver and asexual stage parasite antigens, but not with proliferative, IFN-gamma, or TNF responses to the same antigens. In addition, in those children with mild malaria, higher levels of acute-phase antibody responses to liver stage antigen-1 (LSA-1) were associated with faster parasite clearance times, and were correlated with the presence of IL-10 responses to the same antigen. No such associations were found for IL-10 or antibody responses to a range of asexual blood stage antigens. Those with severe malaria had significantly lower levels of anti-LSA-1 antibodies compared to their counterparts with mild malaria. In conclusion, the results of this study suggest that parasite antigen-specific IL-10-mediated antibody responses may play a role in the control of asexual stage parasite multiplication in P. falciparum malaria.

Amino Acid Sequence↗

[The Varix Set--instruments for varices surgery].

The authors describe new, modern instruments for the operation of varicosities, a VARIX set of their own design. The complex of instruments makes sparing and cosmetically effective strisping of the trunk of the large and small saphenous vein possible, as well as extirpation of collateral branches and epifascial severing of perforator veins. Operation with these instruments is easy, safe and sparing for surrounding tissues.

Humans↗

[Perforation of Meckel's diverticulum by a foreign body].

The authors describe two case-histories of acute abdomen which developed as a result of perforation of Meckel's diverticulum by a foreign body. They draw attention to diagnostic difficulties by a foreign body. They draw attention to diagnostic difficulties and the frequent diagnosis of acute appendicitis.

Adolescent↗

No influence of socioeconomic factors on severe malarial anaemia, hyperparasitaemia or reinfection.

Malaria is responsible for nearly 500 million clinical cases per year, only a small proportion of whom will become severely ill. Socioeconomic risk factors may play a role in the development of severe malaria in African children and in their susceptibility to reinfection. In Gabon, 100 children suffering from severe malaria, defined as hyperparasitaemia and/or severe anaemia, were matched for sex, age and provenance to 100 children with mild malaria. Socioeconomic factors were assessed using a standard questionnaire and compared between the 2 groups. The children were followed-up and the time to first reinfection was recorded. No significant influence of socioeconomic factors could be detected on the severity of disease or the time to first reinfection. Socioeconomic factors are not major determinants of severe malarial anaemia and hyperparasitaemia in children in Gabon.

Adult↗

Merozoite surface antigen 1 and 2 genotypes and rosetting of Plasmodium falciparum in severe and mild malaria in Lambaréné, Gabon.

We present a case-control study to investigate the distribution of Plasmodium falciparum genotypes in patients with severe and mild malaria. We compared clinical and parasitological data with the parasites' genotype and rosetting. The study group consisted of 100 children suffering severe malaria, defined as severe anaemia and hyperparasitaemia. These children were matched by age, sex and provenance with 100 children with mild malaria. For characterization of the parasites we used the polymerase chain reaction to determine merozoite surface antigen (MSA) 1 and 2 genotypes and the phenomenon of rosette formation. We found a significant association between rosette formation and disease severity, and a significant association of severe anaemia with the presence of the MSA-1 allele K1. Infections with 2 genotypes in the severely affected group were significantly associated with severe anaemia and the presence of MSA-1 allele K1. Comparison with the findings of other groups led to the conclusion that the occurrence of P. falciparum genotypes seems to differ geographically.

Animals↗