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Biomedical subjects

J Pawlowski

Publications and source records attributed to J Pawlowski.

At least 37 records · Page 2Linked to original sources

Propofol versus propofol-ketamine sedation for retrobulbar nerve block: comparison of sedation quality, intraocular pressure changes, and recovery profiles.

UNLABELLED: We compared sedation quality, intraocular pressure (IOP) changes, and recovery profiles in patients who received propofol or propofol-ketamine sedation during placement of the retrobulbar nerve block (RBB). Seventy elderly patients undergoing cataract extraction according to a prospective, randomized, double-blinded protocol were preoperatively evaluated with a Mini-Mental State examination and baseline IOP. A hypnotic dose was provided with either propofol (Group P) or a propofol-ketamine (Group PK) combination. The IOP measurement was repeated, and the surgeon initiated the RBB. Supplemental study drug was given if needed. The level of sedation was considered acceptable if the patient exhibited minimal or no movement and grimacing with needle insertion. Patients were evaluated in terms of quality of sedation, cardiopulmonary stability, and recovery profile. Compared with patients in Group P, patients in Group PK had a significantly faster onset of acceptable sedation (Group P 235 +/- 137 s versus Group PK 164 +/- 67 s) and required significantly less supplemental sedation (Group P 1.1 +/- 1.9 mL versus Group PK 0.15 +/- 0.3 mL). Additionally, none of the Group PK patients required ventilatory assistance, but two patients in Group P required assisted mask ventilation. In conclusion, the addition of ketamine (13.2 +/- 3.3 mg) to propofol (44 +/- 11 mg) decreased the hypnotic requirement and improved the quality of sedation without prolonging recovery. IMPLICATIONS: Anesthesiologists frequently perform retrobulbar blocks while simultaneously providing sedation. Using ketamine to supplement propofol sedation provided a faster onset and improved the quality of sedation during the retrobulbar block procedure.

Aged↗

Mycobacterium tusciae sp. nov.

A new, slow-growing, scotochromogenic mycobacterium was isolated from a lymph node of an immunocompromised child and subsequently from tap water and from a respiratory specimen of a patient with chronic fibrosis. Alcohol-acid-fastness, lipid patterns and the G + C content clearly support the placement of this organism in the genus Mycobacterium. The isolates grew very slowly at temperatures ranging from 25 to 32 degrees C and showed activities of nitrate reductase, catalase, urease, arylsulfatase and Tween 80 hydrolysis. The organism was susceptible to all antimycobacterial drugs tested. The 16S rDNA sequence was unique and phylogenetic analysis placed the organism close to fast-growing species such as Mycobacterium farcinogenes, Mycobacterium komossense and Mycobacterium aichiense. These data support the conclusion that the isolates represent a new mycobacterial species, for which the name Mycobacterium tusciae sp. nov. is proposed. The type strain is strain FI-25796T; a culture of this strain has been deposited in the DSMZ as strain DSM 44338T.

Bacterial Typing Techniques↗

Molecular versus taxonomic rates of evolution in planktonic foraminifera.

Neogene planktonic foraminifera are among the most widely used microfossils in the study of tempo and mode of evolution. Comparisons of taxonomic rates between the two major clades in this group have shown that the nonspinose globorotaliids have undergone a significantly more rapid evolutionary turnover than the spinose globigerinids (S. M. Stanley et al., 1988, Paleobiology 14, 235-249). In order to test if similar fluctuations are observed in molecular data, we have used different methods to calculate absolute and relative rates of substitutions based on 16 partial SSU rDNA sequences from representatives of both groups. According to our data, rates of substitution are relatively constant within the globigerinids with a mean value of 4.3 subst./site/10(9) years, but vary in the globorotaliid clade with three species having a rate of about 1 subst./site/10(9) years and two species evolving much faster with rates of more than 7 subst./site/10(9) years. Assuming that the fast rates result from recent accelerations, the globorotaliids have basically much slower molecular evolutionary rates than the globigerinids, in opposition to the fossil data.

Animals↗

Use of the ITS rDNA for elucidation of some life-cycles of Mesometridae (Trematoda, Digenea).

Identification of larval stages is crucial for elucidating the life-cycles of various Digenea. However, in many digenean species, the larvae lack distinctive morphological features and it is impossible to establish the affiliation between the larval and adult stages by using morphological criteria. Molecular methods, based on DNA sequencing or PCR-restriction fragment length polymorphism analysis, can offer a new tool for larval-stage identification. In this study, the sequences of internal transcribed spacer of the ribosomal DNA were used to identify the cercariae of three out of five species of the family Mesometridae (Centroderma spinosissima, Elstia stossichianum and Wardula capitellata). The three species differ from one another by number of repeats in the region of internal transcribed spacer 1. The phylogeny of Mesometridae was inferred from their internal transcribed spacer ribosomal DNA sequences. The PCR-linked restriction fragment length polymorphism approach was developed for future life-cycle and ecological studies of this family.

Animals↗

Molecular phylogeny of Mesometridae (Trematoda, Digenea) with its relation to morphological changes in parasites.

Complete ITS (Internal Transcribed Spacer) ribosomal DNA sequences were obtained for the six species know at present time within the Mesometridae Poche, 1926. The adult stages are intestinal parasites of herbivorous sparid teleosts. Aligned sequences were analysed with Maximum Parsimony, Maximum Likelihood and Neighbor-Joining phylogenetic methods to infer evolutionary relationships among mesometrid species. The ITS-based phylogeny obtained showed the two Wardula species as a sister group to other Mesometridae, and as compared to morphological data, suggest some general tendencies in the morphological evolution of this group. It consists mainly in changes from elongated to subcircular forms, regression of the pharynx, and the development of a strong accessory attachment organ.

Animals↗

Phylogeny and rates of molecular evolution of planktonic foraminifera: SSU rDNA sequences compared to the fossil record.

Planktonic foraminifera are marine protists, whose calcareous shells form oceanic sediments and are widely used for stratigraphic and paleoenvironmental analyses. The fossil record of planktonic foraminifera is compared here to their molecular phylogeny inferred from ribosomal DNA sequences. Eighteen partial SSU rDNA sequences from species representing all modern planktonic families (Globigerinidae, Hastigerinidae, Globorotaliidae, Candeinidae) were obtained and compared to seven sequences representing the major groups of benthic foraminifera. The phylogenetic analyses indicate a polyphyletic origin for the planktonic foraminifera. The Candeinidae, the Globorotaliidae, and the clade Globigerinidae + Hastigerinidae seem to have originated independently, at different epochs in the evolution of foraminifera. Inference of their relationships, however, is limited by substitution rates of heterogeneity. Rates of SSU rDNA evolution vary from 4.0 x 10(-9) substitutions/site/year in the Globigerinidae to less than 1.0 x 10(-9) substitutions/site/year in the Globorotaliidae. These variations may be related to different levels of adaptation to the planktonic mode of life. A clock-like evolution is observed among the Globigerinidae, for which molecular and paleontological data are congruent. Phylogeny of the Globorotaliidae is clearly biased by rapid rates of substitution in two species (G. truncatulinoides and G. menardii). Our study reveals differences in absolute rates of evolution at all taxonomic levels in planktonic foraminifera and demonstrates their effect on phylogenetic reconstructions.

Animals↗

Extreme differences in rates of molecular evolution of foraminifera revealed by comparison of ribosomal DNA sequences and the fossil record.

Foraminifera have one of the best known fossil records among the unicellular eukaryotes. However, the origin and phylogenetic relationships of the extant foraminiferal lineages are poorly understood. To test the current paleontological hypotheses on evolution of foraminifera, we sequenced about 1,000 base pairs from the 3' end of the small subunit rRNA gene (SSU rDNA) in 22 species representing all major taxonomic groups. Phylogenies were derived using neighbor-joining, maximum-parsimony, and maximum-likelihood methods. All analyses confirm the monophyletic origin of foraminifera. Evolutionary relationships within foraminifera inferred from rDNA sequences, however, depend on the method of tree building and on the choice of analyzed sites. In particular, the position of planktonic foraminifera shows important variations. We have shown that these changes result from the extremely high rate of rDNA evolution in this group. By comparing the number of substitutions with the divergence times inferred from the fossil record, we have estimated that the rate of rDNA evolution in planktonic foraminifera is 50 to 100 times faster than in some benthic foraminifera. The use of the maximum-likelihood method and limitation of analyzed sites to the most conserved parts of the SSU rRNA molecule render molecular and paleontological data generally congruent.

Animals↗

Sequence variations in the large-subunit ribosomal RNA gene of Ammonia (Foraminifera, Protozoa) and their evolutionary implications.

An unusually high divergence was observed in the ribosomal RNA genes of a free-living population of foraminifera belonging to the genus Ammonia. The sequences of a large-subunit (LSU) rDNA expansion segment D1 and flanking regions were obtained from 20 specimens named Ammonia sp. 1 and Ammonia sp. 2. The sequence divergence between the two species averages 14%. Within each species it ranges from 0.2% to 7.1% in Ammonia sp. 1 and from 0.7% to 2.3% in Ammonia sp. 2. We did not find two specimens having identical sequences. Moreover, in opposition to the generally accepted view, rDNA sequence variations were also found within a single individual. The variations among several rDNA copies in a single specimen of Ammonia may reach up to 4.9%. Most of the observed variations result from multiplication of CA or TA serial repeats occurring in two particularly variable regions. For single base changes, C-T transitions are most frequently observed. We discuss the evolution of expansion segments and their use for phylogenetic studies.

Animals↗

Early origin of foraminifera suggested by SSU rRNA gene sequences.

Foraminifera are one of the largest groups of unicellular eukaryotes with probably the best known fossil record. However, the origin of foraminifera and their phylogenetic relationships with other eukaryotes are not well established. In particular, two recent reports, based on ribosomal RNA gene sequences, have reached strikingly different conclusions about foraminifera's evolutionary position within eukaryotes. Here, we present the complete small subunit (SSU) rRNA gene sequences of three species of foraminifera. Phylogenetic analysis of these sequences indicates that they branch very deeply in the eukaryotic evolutionary tree: later than those of the amitochondrial Archezoa, but earlier than those of the Euglenozoa and other mitochondria-bearing phyla. Foraminifera are clearly among the earliest eukaryotes with mitochondria, but because of the peculiar nature of their SSU genes we cannot be certain that they diverged first, as our data suggest.

Animals↗

Origin of the Mesozoa inferred from 18S rRNA gene sequences.

The phylum Mesozoa comprises small, simply organized wormlike parasites of marine invertebrates and is composed of two classes, the Rhombozoa and the Orthonectida. The origin of Mesozoa is uncertain; they are classically considered either as degenerate turbellarians or as primitive multicellular animals related to ciliated protists. In order to precisely determine the phylogenetic position of this group we sequenced the complete 18S rRNA gene of one rhombozoid, Dicyema sp., and one orthonectid, Rhopalura ophiocomae. The sequence analysis shows that the Mesozoa branch early in the animal evolution, closely to nematodes and myxozoans. Our data indicate probably separate origins of rhombozoids and orthonectids, suggesting that their placement in the same phylum needs to be revised.

Animals↗

Two different 16S rRNA genes in a mycobacterial strain.

Sequencing of the gene coding for 16S rRNA (16S rDNA) is a well-established method used to identify bacteria, particularly mycobacteria. Unique sequences allow identification of a particular genus and species. If more than one 16S rDNA is present on one mycobacterial genome, their sequences are assumed to be strictly or almost identical. We have isolated a slowly growing Mycobacterium strain, "X", identified by conventional biochemical tests as Mycobacterium terrae. Identification by amplification and direct sequencing of 16S rDNA yielded ambiguous results in two variable regions, suggesting the presence of different copies of the sequenced gene. Total DNA was digested by restriction enzymes and hybridized after Southern blotting to a probe representing about two-thirds of the 16S rDNA. Two copies of 16S rDNA were identified and cloned. By sequencing, the clones were of two different types, A and B, differing in 18 positions. Oligonucleotides specific to each copy of the 16S rDNA were used to distinguish the positions of the two genes observed in the Southern blot. We conclude that Mycobacterium strain "X" has two different copies of 16S rDNA. Variations in the sequence between two copies of 16S rDNA gene have been described in archaeobacteria, but not in mycobacteria. When placed in a phylogenetic tree together with other slowly growing mycobacteria gene A shows a common root with M. terrae, whereas gene B is placed separately.

Cloning, Molecular↗

Phylogenetic position of foraminifera inferred from LSU rRNA gene sequences.

A 5'-terminal region of 1600-1800 base pairs was amplified, cloned, and sequenced in the large subunit rDNA (LSU rDNA) of four species of foraminifera. These sequences were compared with the homologous regions of 16 eukaryotic taxa in order to establish the phylogenetic position of foraminifera. Analysis of 610 unambiguously aligned bases shows that foraminifera branch closely to plasmodial and cellular slime molds in the middle of the eukaryotic tree--that is, much earlier than suggested by the fossil record. These data, the first DNA sequences reported for foraminifera, will help analyze this class of protists and the early evolution of eukaryotes.

Animals↗

[Diabetes insipidus--a rare complication post partum].

A report is made on the manifestation of diabetes insipidus (neuropituitary syndrome), observed six days after a Caesarean section (29-years old I. Gravida). The progress of pregnancy and delivery as well as the successful treatment with Desmopressin (Minirin) post partum are described. The aetiology is still obscure.

Adult↗

Mechanisms of intrinsic tone in ferret vascular smooth muscle.

1. Circular strips from ferret aorta were used to investigate the mechanism of the intrinsic basal tone. 2. Determinations of stiffness using small sinusoidal length changes showed an abolition of both stiffness and force with cooling, but the temperature dependence of the change in active stiffness did not parallel that of force. At temperatures below 22 degrees C there appeared to be a relatively large population of attached, non-force-generating cross-bridges, indicating that separate mechanisms are involved in regulating cross-bridge attachment and the force per cross-bridge. 3. Active intrinsic tone was not affected by removal of extracellular Ca2+ or removal of endothelium. 4. Intracellular ionized Ca2+ concentrations ([Ca2+]i) as measured with the photoprotein aequorin, did not significantly change when intrinsic tone was abolished by cooling. 5. Myosin light chain phosphorylation, as measured by 2-dimensional polyacrylamide gel electrophoresis, significantly decreased on cooling, but the temperature dependence of phosphorylation did not parallel that of force. The change in phosphorylation in the absence of a change in [Ca2+]i suggests the presence of a constitutively active Ca(2+)-independent form of myosin light chain kinase. 6. Maximal concentrations of staurosporine inhibited but did not eliminate intrinsic tone. 7. Changes in myosin light chain kinase and protein kinase C activities may explain part but not all of the intrinsic tone.

Animals↗

Isolation and partial characterization of a full-length cDNA clone for 3 alpha-hydroxysteroid dehydrogenase: a potential target enzyme for nonsteroidal anti-inflammatory drugs.

Homogeneous 3 alpha-hydroxysteroid dehydrogenase (3 alpha-HSD; EC 1.1.1.50) of rat liver cytosol is a monomeric (Mr 34000) NAD(P)+ dependent oxidoreductase which displays 9-, 11- & 15-hydroxyprostaglandin dehydrogenase activity. The enzyme is potently inhibited by nonsteroidal anti-inflammatory drugs (NSAIDs), suggesting that 3 alpha-HSD may be a target enzyme for NSAIDs. A monospecific, polyclonal anti-sera raised against the purified enzyme was used to screen a lambda gt11 expression library and oligonucleotide probes complementary to the 5' and 3' ends of immunopositive clones were used to isolate a 2.1 kb full-length cDNA. Digestion of the full-length cDNA with Eco RI generated two fragments of 1.1 and 1.0 kb in length. Both fragments were subcloned into pGEM3 and partially sequenced. The 1.1 kb fragment contains the C-terminus of 3 alpha-HSD which was confirmed by an in-frame stop codon and comparison of the predicted amino acid sequence to peptide sequence obtained from two endo lys-C peptides of 3 alpha-HSD. The 1.0 kb fragment is 5' to the 1.1 kb fragment and is sufficient in length to contain the remainder of the entire open reading frame for 3 alpha-HSD. Dideoxysequencing reveals significant sequence homology with bovine lung prostaglandin PGF2 alpha synthase. These findings support the role 3 alpha-HSD in inflammation and suggest that hydroxysteroid dehydrogenases, hydroxyprostaglandin dehydrogenases and prostaglandin F2 alpha synthase may be members of a common gene family.

3-Hydroxysteroid Dehydrogenases↗