Endovascular stent-graft placement for treatment of traumatic penetrating subclavian artery injury.
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Acute Disease↗
Biomedical subjects
A Meier
Publications and source records attributed to A Meier.
Use of a mechanical dissociation device to improve standardization of flow cytometric cytokeratin DNA measurements of colon carcinomas.
In order to standardize dual-fluorescence DNA flow cytometry using cytokeratin (CK) antibodies, normal colonic mucosa and tumor tissue were sampled from 308 colorectal surgical specimens. Fresh colon specimens were processed directly and stored frozen until dissociation. The samples were divided into aliquots for manual dissociation with tweezers and scalpel, and parallel dissociation with an automated disaggregation device (Medimachine, DAKO Diagnostika GmbH, Hamburg, Germany). An indirect immunofluorescence method with anti-cytokeratin antibodies and propidiumiodide was applied and measured on a single-laser flow cytometer (FACScan, Becton Dickinson [BDI], Heidelberg, Germany). Evaluation with CellFit (BDI) or MultiPlus (Phoenix Flow Systems, San Diego, CA) showed that dual-parameter fluorescence propidiumiodide (DNA staining) and fluorescein-isothiocyanate (cytokeratin labeling) provides a reasonable staining method for DNA analysis of epithelial cells. No significant differences in coefficient of variation in CK-gated versus ungated cells could be observed. Normal colon mucosa served as a reliable internal, diploid DNA control. Medimachine dissociation led to a significantly higher gain of cytokeratin-positive cells compared to percentage of cytokeratin-positive cells after manual tissue disaggregation. Cytokeratin gating led to a clear-cut separation of S-phase fractions within the respective ploidy groups, irrespective of manual or automated dissociation. The S-phase fraction increased significantly from normal tissue to diploid and nondiploid tumors. In general, automated tissue preparation with the Medimachine allows simple cell-isolation for dual DNA/CK-flow cytometric measurement, improving the gain of CK-positive cells, and facilitating a standardized DNA analysis.
Carcinoma↗
Smad1 and Smad5 have distinct roles during dorsoventral patterning of the zebrafish embryo.
Smad1 and smad5 encode transcription factors that have been implicated in the transduction of signaling by the bone morphogenetic proteins Bmp2 and/or Bmp4. Here we report the characterization of Smad1 and Smad5 from the zebrafish, Danio rerio. Although smad1, smad5, bmp2b, and bmp4 are all expressed during gastrulation and although all four proteins have ventralizing activities, they appear to play distinct roles during dorsoventral pattern formation. smad1 expression starts shortly before the onset of gastrulation. It is expressed on the ventral side of the embryo, whereas smad5 transcripts are both maternally and zygotically provided and ubiquitously distributed. Injection studies and mutant analyses suggest that the ventral smad1 expression is positively regulated by Bmp2b, but not by Bmp4 signaling, whereas smad5 expression is independent of Bmp2b. Also, the dorsalized phenotype of bmp2b-mutant embryos can be rescued by exogenous Smad1, but not by Smad5. Together, these data suggest that smad1 acts later than smad5 and is itself a transcriptional target of Smad5-mediated Bmp2b signaling. During later stages of development, smad1 is expressed in eyes, dorsal cells of rhombomeres 1, 3, and 5, and somites, with highest mRNA levels in the presumptive sclerotome and adaxial regions near the notochord. Injection experiments indicate that this somitic smad1 expression is positively regulated by hedgehog signaling from the dorsal midline, thus perhaps accounting for the recently reported sonic hedgehog-induced competence of sclerotomal cells to Bmp2/4 signals.
Animals↗
Comparison of four 90Sr groundwater analytical methods.
Data are presented for 45 Long Island groundwater samples each measured for 90Sr using four different analytical methods. 90Sr levels were first established by two New York State certified laboratories, one of which used the U.S. Environmental Protection Agency Radioactive Strontium in Drinking Water Method 905.0. Three of the 90Sr methods evaluated at Brookhaven National Laboratory can reduce analysis time by more than 50%. They were (a) an Environmental Measurements Laboratory Cerenkov technique and (b) two commercially available products that utilize strontium-specific crown-ethers supported on either a resin or membrane disk. Method independent inter-laboratory bias was <12% based on 90Sr results obtained using both U.S. Department of Energy/Environmental Measurements Laboratory and U.S. EPA/National Environmental Radiation Laboratory samples of known activity concentration. Brookhaven National Laboratory prepared a National Institute of Standards and Technology traceable 90Sr tap-water sample used to quantify test method biases. With gas proportional or liquid scintillation counting, minimum detectable levels (MDLs) of 37 Bq m(-3) (1 pCi L(-1)) were achievable for both crown-ether methods using a 1-L processed sample beta counted for 1 h. The primary radiological contaminants of the Brookhaven National Laboratory groundwater samples were 3H (14.8-518 kBq m(-3)) and 90Sr/Y (37 Bq m(-3)-18.5 kBq m(-3)). Thirty samples were above the U.S. EPA drinking water standard of 300 Bq m(-3) for beta emitting 90Sr. Regression of paired data sets showed that both crown-ether methods quantified 90Sr levels to within 5% of the "EPA method results" for samples greater than the drinking water standard and within 30-40% for samples less than the drinking water standard. The Cerenkov method, with a minimum detectable level of 150 Bq m(-3), was only useful for samples greater than the drinking water standard and underestimated 90Sr levels by 13%. Precision, as measured by the relative percent difference and mean difference statistics, was acceptable for both crown-ether methods when compared to the paired "EPA method" results. Cerenkov method precision exceeded pre-defined acceptance criteria for 12 of 33 samples analyzed.
New York↗
Development and field performance of a broad-spectrum nonviable asporogenic recombinant strain of Bacillus thuringiensis with greater potency and UV resistance.
The main problems with Bacillus thuringiensis products for pest control are their often narrow activity spectrum, high sensitivity to UV degradation, and low cost effectiveness (high potency required). We constructed a sporulation-deficient SigK(-) B. thuringiensis strain that expressed a chimeric cry1C/Ab gene, the product of which had high activity against various lepidopteran pests, including Spodoptera littoralis (Egyptian cotton leaf worm) and Spodoptera exigua (lesser [beet] armyworm), which are not readily controlled by other Cry delta-endotoxins. The SigK(-) host strain carried the cry1Ac gene, the product of which is highly active against the larvae of the major pests Ostrinia nubilalis (European corn borer) and Heliothis virescens (tobacco budworm). This new strain had greater potency and a broader activity spectrum than the parent strain. The crystals produced by the asporogenic strain remained encapsulated within the cells, which protected them from UV degradation. The cry1C/Ab gene was introduced into the B. thuringiensis host via a site-specific recombination vector so that unwanted DNA was eliminated. Therefore, the final construct contained no sequences of non-B. thuringiensis origin. As the recombinant strain is a mutant blocked at late sporulation, it does not produce viable spores and therefore cannot compete with wild-type B. thuringiensis strains in the environment. It is thus a very safe biopesticide. In field trials, this new recombinant strain protected cabbage and broccoli against a pest complex under natural infestation conditions.
Animals↗
The smad5 mutation somitabun blocks Bmp2b signaling during early dorsoventral patterning of the zebrafish embryo.
Signaling by members of the TGFbeta superfamily is thought to be transduced by Smad proteins. Here, we describe a zebrafish mutant in smad5, designated somitabun (sbn). The dominant maternal and zygotic effect of the sbntc24 mutation is caused by a change in a single amino acid in the L3 loop of Smad5 protein which transforms Smad5 into an antimorphic version, inhibiting wild-type Smad5 and related Smad proteins. sbn mutant embryos are strongly dorsalized, similarly to mutants in Bmp2b, its putative upstream signal. Double mutant analyses and RNA injection experiments show that sbn and bmp2b interact and that sbn acts downstream of Bmp2b signaling to mediate Bmp2b autoregulation during early dorsoventral (D-V) pattern formation. Comparison of early marker gene expression patterns, chimera analyses and rescue experiments involving temporally controlled misexpression of bmp or smad in mutant embryos reveal three phases of D-V patterning: an early sbn- and bmp2b-independent phase when a coarse initial D-V pattern is set up, an intermediate sbn- and bmp2b-dependent phase during which the putative morphogenetic Bmp2/4 gradient is established, and a later sbn-independent phase during gastrulation when the Bmp2/4 gradient is interpreted and cell fates are specified.
Amino Acid Sequence↗
Effects of weight loss on regional fat distribution and insulin sensitivity in obesity.
Weight loss (WL) decreases regional depots of adipose tissue and improves insulin sensitivity, two parameters that correlate before WL. To examine the potential relation of WL-induced change in regional adiposity to improvement in insulin sensitivity, 32 obese sedentary women and men completed a 4-month WL program and had repeat determinations of body composition (dual-energy X-ray absorptiometry and computed tomography) and insulin sensitivity (euglycemic insulin infusion). There were 15 lean men and women who served as control subjects. VO2max was unaltered with WL (39.2 +/- 0.8 vs. 39.8 +/- 1.1 ml x fat-free mass [FFM](-1) x min(-1)). The WL intervention achieved significant decreases in weight (100.2 +/- 2.6 to 85.5 +/- 2.1 kg), BMI (34.3 +/- 0.6 to 29.3 +/- 0.6 kg/m2), total fat mass (FM) (36.9 +/- 1.5 to 26.1 +/- 1.3 kg), percent body fat (37.7 +/- 1.3 to 31.0 +/- 1.5%), and FFM (59.2 +/- 2.3 to 55.8 +/- 2.0 kg). Abdominal subcutaneous and visceral adipose tissue (SAT and VAT) were reduced (494 +/- 19 to 357 +/- 18 cm2 and 157 +/- 12 to 96 +/- 7 cm2, respectively). Cross-sectional area of low-density muscle (LDM) at the mid-thigh decreased from 67 +/- 5 to 55 +/- 4 cm2 after WL. Insulin sensitivity improved from 5.9 +/- 0.4 to 7.3 +/- 0.5 mg x FFM(-1) x min(-1) with WL. Rates of insulin-stimulated nonoxidative glucose disposal accounted for the majority of this improvement (3.00 +/- 0.3 to 4.3 +/- 0.4 mg x FFM(-1) x min(-1)). Serum leptin, triglycerides, cholesterol, and insulin all decreased after WL (P < 0.01). After WL, insulin sensitivity continued to correlate with generalized and regional adiposity but, with the exception of the percent decrease in VAT, the magnitude of improvement in insulin sensitivity was not predicted by the various changes in body composition. These interventional weight loss data underscore the potential importance of visceral adiposity in relation to insulin resistance and otherwise suggest that above a certain threshold of weight loss, improvement in insulin sensitivity does not bear a linear relationship to the magnitude of weight loss.
Abdomen↗
Follistatin and noggin are excluded from the zebrafish organizer.
The patterning activity of the Spemann organizer in early amphibian embryos has been characterized by a number of organizer-specific secreted proteins including Chordin, Noggin, and Follistatin, which all share the same inductive properties. They can neuralize ectoderm and dorsalize ventral mesoderm by blocking the ventralizing signals Bmp2 and Bmp4. In the zebrafish, null mutations in the chordin gene, named chordino, lead to a severe reduction of organizer activity, indicating that Chordino is an essential, but not the only, inductive signal generated by the zebrafish organizer. A second gene required for zebrafish organizer function is mercedes, but the molecular nature of its product is not known as yet. To investigate whether and how Follistatin and Noggin are involved in dorsoventral (D-V) patterning of the zebrafish embryo, we have now isolated and characterized their zebrafish homologues. Overexpression studies demonstrate that both proteins have the same dorsalizing properties as their Xenopus homologues. However, unlike the Xenopus genes, zebrafish follistatin and noggin are not expressed in the organizer region, nor are they linked to the mercedes mutation. Expression of both genes starts at midgastrula stages. While no patterned noggin expression was detectable by in situ hybridization during gastrulation stages, later expression is confined to presumptive cartilage cells in the branchial arches and the neurocranium and to proximal regions of the pectoral fin buds. follistatin transcripts in gastrulating embryos are confined to anterior paraxial regions, which give rise to head mesoderm and the first five somites. The dorsolateral extent of this expression domain is regulated by Bmp2b, Chordino, and Follistatin itself. In addition, transient expression was observed in a subset of cells in the posterior notochord anlage. Later, follistatin is expressed in brain, eyes, and somites. Comparison of the spatiotemporal expression pattern of follistatin and noggin with those of bmp2b and bmp4 and overexpression studies suggest that Noggin and Follistatin may function as Bmp antagonists in later processes of zebrafish development, including late phases of D-V patterning, to refine the early pattern set up by the interaction of Chordino and Bmp2/4. It thus appears that many, but not all, aspects of early dorsoventral patterning are shared among different vertebrate species.
Amino Acid Sequence↗
Prenatal diagnosis of fetal malformations by ultrafast magnetic resonance imaging.
Prenatal ultrasonography is the primary imaging modality in pregnancy as it allows direct real-time fetal examination. Antenatal magnetic resonance imaging (MRI) has so far been of limited clinical value owing to poor image quality. This was due to the long acquisition times that were needed to achieve a high enough spatial resolution for assessment of the small fetal anatomic structures resulting in severe motion artefacts. This problem has now been overcome by recent technical improvements. We present a case of fetal malformation (MURCS association), where an ultrafast T2-weighted RARE sequence (single shot fast spin echo sequence) provided images of excellent quality, which led to the correct antenatal diagnosis. MRI using this sequence can now be used for prenatal diagnosis and is thus likely to become an important adjunct to ultrasonography, especially in cases in which ultrasound findings are unclear of sonographic images are impaired by maternal obesity or oligohydramnios.
Adult↗
Metal chelating properties of adenylate kinase from Paracoccus denitrificans.
Zinc, a common element of adenylate kinases from Gram-positive bacteria, binds to a structural motif consisting of three or four cysteine residues, Cys-X2-Cys-X16-Cys-X2-Cys/Asp. The enzyme from Paracoccus denitrificans, a Gram-negative bacterium, has structural features much similar to those of adenylate kinases from Gram-positive organisms [Spurgin, P., Tomasselli, A.G., and Schiltz, E. (1989) Eur. J. Biochem., 179, 621-628]. However, adenylate kinase isolated from this bacterium was not reported to bind metal. These findings prompted us to clone the corresponding gene of P. denitrificans, and to characterize the enzyme overproduced in Escherichia coli. The deduced primary structure of adenylate kinase from P. denitrificans revealed two differences from that previously published: Cys was found at position 130 instead of His, and His was found at position 138 instead of Gly. The recombinant enzyme is a dimer which binds either zinc or iron, in a metal/monomer ratio of one. The dissociating sulfhydryl reagent, p-(hydroxy-mercuri)phenylsulfonate, released the metal from the protein, confirming that thiols are involved in zinc- or iron-binding. The iron-chelated form of recombinant P. denitrificans adenylate kinase, which is essentially under reduced form, transfers electrons to the oxidized cytochrome c. In conclusion, the absence of metal in the enzyme isolated from P. denitrificans is not related to the protein structure but most probably due to the physiological properties of the host organism.
Adenylate Kinase↗
Neonatal citrobacter meningitis: neurosonographic observations.
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Citrobacter↗
RNA polymerase II transcription inhibits DNA repair by photolyase in the transcribed strand of active yeast genes.
Yeast uses nucleotide excision repair (NER) and photolyase (photoreactivation) to repair cyclobutane pyrimidine dimers (CPDs) generated by ultraviolet light. In active genes, NER preferentially repairs the transcribed strand (TS). In contrast, we recently showed that photolyase preferentially repairs the non-transcribed strands (NTS) of the URA3 and HIS3 genes in minichromosomes. To test whether photoreactivation depends on transcription, repair of CPDs was investigated in the transcriptionally regulated GAL10 gene in a yeast strain deficient in NER [AMY3 (rad1Delta)]. In the active gene (cells grown in galactose), photoreactivation was fast in the NTS and slow in the TS demonstrating preferential repair of the NTS. In the inactive gene (cells grown in glucose), both strands were repaired at similar rates. This suggests that RNA polymerases II blocked at CPDs inhibit accessibility of CPDs to photolyase. In a strain in which both pathways are operational [W303-1a (RAD1)], no strand bias was observed either in the active or inactive gene, demonstrating that photoreactivation of the NTS compensates preferential repair of the TS by NER. Moreover, repair of the NTS was more quickly in the active gene than in the repressed gene indicating that transcription dependent disruption of chromatin facilitates repair of an active gene.
Chromatin↗
[The Zurich prospective dementia study 1987-1993].
1987-1993 the Zurich prospective dementia study examined twice yearly 308 nursing-home patients: 192 with senile dementia of Alzheimer-type, 18 with Parkinsonism, 20 with Parkinson dementia and 78 controls, with the Zurich variant of the mini-mental state and rated their dementia state by CDR, activities of daily living with ADL and social behavior with an SVS-scale. At the end of the study or at death the average age was 86.2 years (range 63-103 and 159 autopsies were performed which confirmed the clinical diagnoses at least partly in 70%. Of the 62 cases with autopsies who had been studied more than two years 25 showed M.Alzheimer, 8 M, Parkinson, 7 vascular dementia and 13 a combination of those three diagnoses 40% of these Alzheimer patients exhibited no deterioration in the SVS for two years, 40% respectively 30% during one, resp. two years in the CDR, 20% resp. 40% in the ADL and 23% resp. 5% in the MMS. This demonstrates that good nursing-home care can stabilize behaviour and to a lesser extent even cognitive abilities of a large minority of patients with a pathologically confirmed progressive dementing illness.
Activities of Daily Living↗
The role of ribosomal RNAs in macrolide resistance.
Macrolides are bacteriostatic antibiotics which interfere with the peptidyltransfer function of the ribosome. We have investigated the molecular mechanisms underlying macrolide resistance in Mycobacterium smegmatis, an eubacterium carrying two rRNA operons. Surprisingly, drug resistance was associated not with alterations in ribosomal proteins, but with a single point mutation in the peptidyltransferase region of one of the two 23S RNA genes, i.e. A2058-->G or A2059-->G. This mutation resulted in a heterozygous organism with a mutated and a wild-type rRNA operon respectively. Reverse transcriptase sequencing indicated the expression of both wild-type and mutated rRNAs. The mutated operon was introduced into genetically engineered rrn- strains of M. smegmatis carrying a single functional rRNA operon and into parental M. smegmatis with two chromosomal rRNA operons, using gene transfer as well as gene replacement techniques. The results obtained demonstrate the dominant nature of resistance. As exemplified in our results on macrolide resistance, a complete set of genetic tools is now available, which allows questions of dominance vs. recessivity and gene dosage effects in eubacterial ribosomal nucleic acids to be addressed experimentally in vivo.
Alleles↗
Usefulness of PCR for diagnosis of Pneumocystis carinii pneumonia in different patient groups.
Pneumocystis carinii pneumonia (PCP) is one of the most predominant opportunistic infectious diseases in patients with AIDS. Nested PCR has been described as a sensitive and specific tool for detecting P. carinii DNA in clinical specimens. Little is known about the correlation of positive PCR results and clinical evidence of PCP in patients with different forms of immunosuppression. One hundred and thirty-six sputum samples, 26 tracheal-bronchial aspirate samples, 35 bronchoalveolar lavage samples, and 11 lung biopsy samples from (i) human immunodeficiency virus (HIV)-infected patients with AIDS, (ii) immunocompromised patients with leukemia or lymphoma, and (iii) immunocompetent control patients were investigated by a nested PCR amplifying DNA from the mitochondrial large subunit of P. carinii. All patients suffered from acute episodes of respiratory disease. The resulting data were correlated with clinical evidence of PCP. A high degree of association of positive P. carinii PCR results and clinical evidence of PCP in HIV-infected patients with AIDS was found. When calculated for bronchoalveolar lavage and lung biopsy samples, the positive and the negative predictive values of P. carinii PCR for PCP diagnosis in HIV-infected patients with AIDS were 1 and the specificity and the sensitivity were 100%. In contrast, in the group of patients with leukemia or lymphoma, the positive predictive value of the nested PCR for these materials was found to be as low as 0.09, the negative predictive value was 0.73, the specificity was 44.4%, and the sensitivity was 25.0%. No P. carinii DNA could be detected in specimens from immunocompetent patients. In summary, in contrast to patients with leukemia and lymphoma, nested PCR seems to be a sensitive and specific tool for PCP diagnosis in HIV-infected patients with AIDS.
AIDS-Related Opportunistic Infections↗
Clinical application of capillary isoelectric focusing on fused silica capillary for determination of hemoglobin variants.
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Diagnosis, Differential↗
[Cost reduction in long-term nursing thanks to a memory clinic--results of a case control study].
Between 1991 and 1995, 334 persons from the city of Zurich have been investigated at the gerontologic counselling facility of the memory clinic of the nursing home Entlisberg near Zurich. 49 (14.7%) of these persons suffered from a treatable disease (16 from depression, 13 from chronic intoxication, 20 from other treatable diseases), 10 from other disturbances (debility or postapoplectic state), and 275 from dementia. In 45% of the cases dementia was moderate, in 47% moderate to severe, and in 8% severe. In most cases dementia was of Alzheimer's type (60.7%), others were of multi-infarct type (17.5%), of mixed type (13.1%), or other types (8.7%). Until march 1996, 64 (19.4%) of these 334 on the average 74-year-old persons treated in the memory clinic have been admitted to a city nursing home. They have compared to a random sample, including 312 demented persons of similar ages and similar date of admittance to the same institutions. The proportion of demented persons cared before admittance by a spouse was about 28% in both groups. On an average, the mini mental state (Zurich variant) at admittance to a nursing home was 10.95 points in patients from the memory clinic, compared to 13.43 points in the control group. Thus, patients that have formerly been advised and trained entered the nursing home in a state of dementia more advanced by 2.48 points. Based on an average yearly demelioration by about 2.4 points, this means admittance to a nursing home is delayed by 376 days. Patients for whom admittance to a nursing home could be avoided completely by the treatment proposed by the counselling were not considered in this calculation.
Aged↗
Molecular mechanisms of clarithromycin resistance in Mycobacterium avium: observation of multiple 23S rDNA mutations in a clonal population.
The peptidyltransferase region of the 23S rRNA gene (the probable target site for the macrolides) was investigated in blood isolates of Mycobacterium avium recovered from 38 patients before and after the development of clarithromycin resistance. Point mutations were identified in 100% of the 74 resistant relapse blood isolates but in none of 69 susceptible pretreatment isolates. Multiple mutations were identified in isolates from 23 (61%) of 38 patients. Of the 63 identified mutations, 95% involved adenine at bp 2058. Single-colony clones from cultures that were mixtures of more than one mutation revealed a single mutation within each clone. Pulsed field gel electrophoresis of genomic DNA restriction fragments revealed that 13 (81%) of 16 multiple mutations identified in the same patient were derived from a single infecting strain. In vitro investigation revealed the same point mutations observed in vivo. This study defines the probable mechanism of clarithromycin resistance in M. avium and provides in vivo evidence that mutational resistance is random and selection-directed.
Base Sequence↗