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PubMed1989

A comparison of anaesthetic breathing systems during spontaneous ventilation. An in-vitro study using a lung model.

Five anaesthetic breathing systems (Magill, Lack, Humphrey ADE, enclosed Magill and Bain) were compared using spontaneous ventilation in a simple lung model. The fresh gas flow at which rebreathing occurred was determined for each system by the application of four modified definitions of rebreathing. Two were based on the measurement of minimum inspired and two on end-expired carbon dioxide. The four A systems performed similarly with each individual definition. The rebreathing points found for each individual breathing system differed markedly between definitions, with those determined by the minimum inspired CO2 occurring at low, and probably misleading, FGF/VE ratio. The Bain system demonstrated rebreathing at considerably higher fresh gas flows whichever definition was used.

Anesthesiology↗

PubMed1993

The additional work of breathing imposed by Mapleson A systems.

The additional work attributable to breathing through five Mapleson A anaesthetic breathing systems (Magill, Lack, Parallel Lack, Humphrey ADE and Enclosed Magill) was studied using a lung model. With all five systems, the additional work was found to be a function of fresh gas flow, respiratory flow as well as system geometry. Within the range of fresh gas flow and respiratory flow studied, the additional work ranged between 80 mJ.l-1 and 182 mJ.l-1. Expiratory work was always greater than the inspiratory workload. Increasing fresh gas inflow into the system increases expiratory work, both resistive and elastic components. The Magill system posed the least work expenditure. The values for the additional work obtained with the lung model were of the same order of magnitude when measurements were taken in volunteers.

Adult↗

PubMed1989

Isolation of Erwinia chrysanthemi mutants altered in pectinolytic enzyme production.

Various mutations in the pectin catabolic pathway of Erwinia chrysanthemi were isolated by selection of Mu-lac insertions, resulting in expression of the lac genes inducible by pectin degradation products. This approach allowed us to isolate lacZ fusions with the genes pelC, pelD, ogl and pem, encoding pectate lyases PLc and PLd, oligogalacturonate lyase and pectin methylesterase, respectively. Moreover, we obtained mutations affecting the regulation of pectinolytic enzymes; a locus called pecl appeared to be involved in induction of pectate lyases and pectin methylesterase. A second locus, called pecL, may encode an activator protein acting on pectate lyase production. Both pecl and pecL expression are induced in the presence of pectic polymers. The expression of the pem gene was studied in more detail by analysis of the pem-lacZ fusions. The expression of pem appears to be controlled by the negative regulatory gene kdgR, which controls all the genes involved in pectin degradation (pem, pel, ogl, kduD, kdul, kdgK, kdgA). This study confirmed that 2-keto-3-deoxygluconate is a key intermediate for the induction of the pectin catabolic pathway. The three genes pem, pelD and pecl were localized in the same region, near the ade-377 marker on the genetic map of the E. chrysanthemi strain 3937. The pem gene was located more precisely on an 18kb DNA fragment containing the pelADE cluster. However, this 18kb DNA fragment did not complement the pecl mutation. The pecL mutations were located near the ile-2 marker on the genetic map of E. chrysanthemi strain 3937.

Chromosome Mapping↗

PubMed1989

Molecular cloning of the outJ gene involved in pectate lyase secretion by Erwinia chrysanthemi.

The phytopathogenic enterobacterium Erwinia chrysanthemi secretes a number of enzymes involved in plant-tissue degradation, notably the five isoenzymes of pectate lyase. We have cloned a region involved in pectate lyase and cellulase secretion by complementation of non-secretory outJ mutants of E. chrysanthemi strain 3937 using the RP4::miniMu plasmid pULB110. The cloned region maps near the ade-22 marker on the E. chrysanthemi 3937 chromosome. An R-prime containing a chromosomal DNA insert of about 30kb was first obtained; subcloning into pBR325 permitted the isolation of a 4kb ClaI/SspI fragment able to complement outJ mutations in E. chrysanthemi. The isolation of phoA fusions in this fragment allowed us to determine the direction of transcription of the encoding region, which extends over about 2.5kb, and demonstrate that this region encodes exported protein(s). When the TnphoA insertions were transferred back into E. chrysanthemi chromosome, the recombined strains no longer secreted pectate lyases or cellulases. Identification of the products encoded by the ClaI/SspI fragment demonstrated that outJ encodes an 83 kD polypeptide which is processed to an 81 kD polypeptide by cleavage of a signal sequence. The cloned DNA fragment did not endow Escherichia coli with the ability to secrete pectate lyases.

Bacterial Proteins↗

PubMed2005

The Hunter-McAlpine syndrome results from duplication 5q35-qter.

In 1977 Hunter et al. J Med Genet 1977: 14 (6): 430-437, reported a family with six affected members, connected over three generations through unaffected individuals. Subsequently, several other patients purported to have the condition were reported. The condition became known as the Hunter-McAlpine syndrome, and there was debate as to whether or not it was identical to the Ruvalcaba syndrome or a type of tricho-rhino-phalangeal syndrome. In this article we confirm that the original family and a patient reported by Ades et al. Clin Dysmorphol 1993: 2 (2): 123-130 have cryptic translocations resulting in duplication of 5q35-qter. Similarities are noted between our patients and others in the literature with duplication of this chromosome segment.

Abnormalities, Multiple↗

PubMed2005

A randomized, controlled trial comparing IV iron sucrose to oral iron in anemic patients with nondialysis-dependent CKD.

BACKGROUND: Although iron deficiency frequently complicates anemia in patients with nondialysis-dependent CKD (ND-CKD), the comparative treatment value of IV iron infusion and oral iron supplementation has not been established. METHODS: In a randomized, controlled multicenter trial, we compared the efficacy of iron sucrose, given as 1 g in divided IV doses over 14 days, with that of ferrous sulfate, given 325 mg orally thrice daily for 56 days in patients with ND-CKD stages 3 to 5, Hb < or =11 g/dL, TSAT < or =25%, and ferritin < or =300 ng/mL. Epoetin/darbepoetin therapy, if any, was not changed for eight weeks prior to or during the study. RESULTS: The proportion of patients achieving the primary outcome (Hb increase > or =1 g/dL) was greater in the IV iron treatment group than in the oral iron treatment group (44.3% vs. 28.0%, P= 0.0344), as was the mean increase in Hb by day 42 (0.7 vs. 0.4 g/dL, P= 0.0298). Compared to those in the IV iron group, patients in the oral iron treatment group showed a greater decline in GFR during the study (-4.40 vs. -1.45 mL/min/1.73m2, P= 0.0100). No serious adverse drug events (ADE) were seen in patients administered IV iron sucrose as 200 mg IV over two to five minutes, but drug-related hypotension, including one event considered serious, occurred in two females weighing less than 65 kg after 500 mg doses were given over four hours. CONCLUSION: IV iron administration using 1000 mg iron sucrose in divided doses is superior to oral iron therapy in the management of ND-CKD patients with anemia and low iron indices.

Administration, Oral↗

PubMed2006

Undergraduate clinical orthodontic experience: a discussion paper.

Undergraduate clinical orthodontic experience may be expressed in a variety of ways. The most common way (used, inter alia, in the DentEd school visits) is by number of curriculum hours. Other ways include the number of patients seen, or number of procedures carried out. Whilst any of these methods may allow comparison between cohorts of students within a school or between schools, none of them are suitable to determine how much experience is 'satisfactory', nor do they indicate what the student should know, understand, or be able to do, to be considered 'satisfactory', and hence fit to graduate. This situation may be addressed by the use of competences and/or learning outcomes, and in 2004 the Association for Dental Education in Europe (ADEE) adopted a document that contained one major competence and five supporting competences in orthodontics. This paper considers the shortcomings of conventional methods of recording orthodontic experience in relation to the acquisition of these competences, and some ways in which staff and students may assess competence.

Clinical Competence↗

PubMed1982

Somatic cell genetic approaches to Down's syndrome.

Somatic cell genetic analysis of mutants of Chinese hamster ovary cells with deficient purine synthesis and of hybrids between these mutants and human cells is described. Data are presented substantiating that two genes for enzymes of purine synthesis, AdeC and AdeG, can be coordinately regulated in mammalian cells. Analysis of a human-hamster hybrid cell, Ade C/21, which contains a normal complement of hamster chromosomes and human chromosome 21 as its only human genetic component recognizable by electrophoretic and immunogenetic techniques demonstrates that genes associated with the presence of human chromosome 21 and required for the synthesis of specific polypeptides and specific human lethal cell surface antigens can be detected in these hybrids.

Animals↗

PubMed1995

Methionine recycling pathways and antimalarial drug design.

5'-Deoxy-5'-(methylthio)adenosine (MTA) is an S-adenosylmethionine metabolite that is generated as a by-product of polyamine biosynthesis. In mammalian cells, MTA undergoes a phosphorolytic cleavage catalyzed by MTA phosphorylase to produce adenine and 5-deoxy-5-(methylthio)ribose-1-phosphate (MTRP). Adenine is utilized in purine salvage pathways, and MTRP is subsequently recycled to methionine. Whereas some microorganisms metabolize MTA to MTRP via MTA phosphorylase, others metabolize MTA to MTRP in two steps via initial cleavage by MTA nucleosidase to adenine and 5-deoxy-5-(methylthio)ribose (MTR) followed by conversion of MTR to MTRP by MTR kinase. In order to assess the extent to which these pathways may be operative in Plasmodium falciparum, we have examined a series of 5'-alkyl-substituted analogs of MTA and the related MTR analogs and compared their abilities to inhibit in vitro growth of this malarial parasite. The MTR analogs 5-deoxy-5-(ethylthio)ribose and 5-deoxy-5-(hydroxyethylthio)ribose were inactive at concentrations up to 1 mM, and 5-deoxy-5-(monofluoroethylthio)ribose was weakly active (50% inhibitory concentration = 700 microM). In comparison, the MTA analogs, 5'-deoxy-5'-(ethylthio)adenosine,5'-deoxy-5'-(hydroxyethylthio)ade nosine (HETA), and 5'-deoxy-5'-(monofluoroethylthio)adenosine, had 50% inhibitory concentrations of 80, 46, and 61 microM, respectively. Extracts of P. falciparum were found to have substantial MTA phosphorylase activity. Coadministration of MTA with HETA partially protected the parasites against the growth-inhibitory effects of HETA. Results of this study indicate that P. falciparum has an active MTA phosphorylase that can be targeted by analogs of MTA.

Animals↗

PubMed2004

Novel resistance-nodulation-cell division efflux system AdeDE in Acinetobacter genomic DNA group 3.

Resistance-nodulation-cell division type efflux pump AdeDE was identified in acinetobacters belonging to genomic DNA group 3. Inactivation of adeE showed that it may be responsible for reduced susceptibility to amikacin, ceftazidime, chloramphenicol, ciprofloxacin, erythromycin, ethidium bromide, meropenem, rifampin, and tetracycline. However, unlike what was found for other similar efflux systems, the open reading frame for the outer membrane component was not found downstream of the adeDE gene cluster.

Acinetobacter↗

PubMed1989

Transformation by Complementation of an Adenine Auxotroph of the Lignin-Degrading Basidiomycete Phanerochaete chrysosporium.

Swollen basidiospores of an adenine auxotroph of Phanerochaete chrysosporium were protoplasted with Novozyme 234 and transformed to prototrophy by using a plasmid containing the gene for an adenine biosynthetic enzyme from Schizophyllum commune. Transformation frequencies of 100 transformants per mug of DNA were obtained. Southern blot analysis of DNA extracted from transformants demonstrated that plasmid DNA was integrated into the chromosomal DNA in multiple tandem copies. Analysis of conidia and basidiospores from transformants demonstrated that the transforming character was mitotically and meiotically stable on both selective and nonselective media. Genetic crosses between double mutants transformed for adenine prototrophy and other auxotrophic strains yielded Ade progeny, which indicated that integration occurred at a site(s) other than the resident adenine biosynthetic gene.

Journal Article↗

PubMed1991

Autoregulatory response of Phaseolus vulgaris L. to symbiotic mutants of Rhizobium leguminosarum bv. phaseoli.

In Rhizobium-legume symbiosis, the plant host controls and optimizes the nodulation process by autoregulation. Tn5 mutants of Rhizobium leguminosarum bv. phaseoli TAL 182 which are impaired at various stages of symbiotic development, were used to examine autoregulation in the common bean (Phaseolus vulgaris L.). Class I mutants were nonnodulating, class II mutants induced small, distinct swellings on the roots, and a class III mutant formed pink, bacterium-containing, but ineffective nodules. A purine mutant (Ade-) was nonnodulating, while a pyrimidine mutant (Ura-) formed small swellings on the roots. Amino acid mutants (Leu-, Phe-, and Cys-) formed mostly empty white nodules. Each of the mutants was used as a primary inoculant on one side of a split-root system to assess its ability to suppress secondary nodulation by the wild type on the other side. All mutants with defects in nodulation ability, regardless of the particular stage of blockage, failed to induce a suppression response from the host. Only the nodulation-competent, bacterium-containing, but ineffective class III mutant induced a suppression response similar to that induced by the wild type. Suppression was correlated with the ability of the microsymbiont to proliferate inside the nodules but not with the ability to initiate nodule formation or the ability to fix nitrogen. Thus, the presence of bacteria inside the nodules may be required for the induction of nodulation suppression in the common bean.

Fabaceae↗

PubMed1994

Homologous expression of recombinant manganese peroxidase in Phanerochaete chrysosporium.

The promoter region of the glyceraldehyde-3-phosphate dehydrogenase gene (gpd) was used to drive expression of mnp1, the gene encoding Mn peroxidase isozyme 1, in primary metabolic cultures of Phanerochaete chrysosporium. A 1,100-bp fragment of the P. chrysosporium gpd promoter region was fused upstream of the mnp1 gene to construct plasmid pAGM1, which contained the Schizophyllum commune ade5 gene as a selectable marker. pAGM1 was used to transform a P. chrysosporium ade1 auxotroph to prototrophy. Ade+ transformants were screened for peroxidase activity on a solid medium containing high carbon and high nitrogen (2% glucose and 24 mM NH4 tartrate) and o-anisidine as the peroxidase substrate. Several transformants that expressed high peroxidase activities were purified and analyzed further in liquid cultures. Recombinant Mn peroxidase (rMnP) was expressed and secreted by transformant cultures on day 2 under primary metabolic growth conditions (high carbon and high nitrogen), whereas endogenous wild-type mnp genes were not expressed under these conditions. Expression of rMnP was not influenced by the level of Mn in the culture medium, as previously observed for the wild-type Mn peroxidase (wtMnP). The amount of active rMnP expressed and secreted in this system was comparable to the amount of enzyme expressed by the wild-type strain under ligninolytic conditions. rMnP was purified to homogeneity by using DEAE-Sepharose chromatography, Blue Agarose chromatography, and Mono Q column chromatography. The M(r) and absorption spectrum of rMnP were essentially identical to the M(r) and absorption spectrum of wtMnP, indicating that heme insertion, folding, and secretion were normal.(ABSTRACT TRUNCATED AT 250 WORDS)

Base Sequence↗

PubMed1987

Ploidy and DNA content of Candida stellatoidea cells.

Three isolates of Candida stellatoidea contained approximately the same amount of DNA per blastospore (38.3 to 41.9 fg) as did a known diploid isolate of Candida albicans and about twice as much as did a haploid isolate of Saccharomyces cerevisiae. The diploidy of C. stellatoidea was supported by demonstration of mitotic segregation of an ade marker.

Candida↗

PubMed1998

Integrated physical and genetic mapping of Bacillus cereus and other gram-positive bacteria based on IS231A transposition vectors.

The genome structure of Bacillus cereus is relatively complex, its DNA being modulated between a size-varying chromosome and large plasmids. To study the genetic organization of the B. cereus type strain ATCC 14579, thermosensitive transposition vectors were designed on the basis of IS231A-derived cassettes containing uncommon restriction sites. A highly preferred insertion site for IS231A was detected in the chromosome by Southern blotting and pulsed-field gel electrophoresis (PFGE) analyses of independent insertion mutants. However, once this insertional hot spot was occupied, secondary IS231A insertions occurred randomly, as demonstrated by isolation of independent B. cereus auxotrophs at a frequency of approximately 0.6%. The hot-spot site, as well as several auxotrophic mutations, were mapped by using NotI, SfiI, and AscI PFGE restriction profiles. It was confirmed by sequencing that one of the insertions, generating an Ade- phenotype, had disrupted a gene of the purine synthesis pathway. These results showed that combined PFGE and sequencing analyses of mini-IS231A insertions enable the construction of integrated physical and genetic maps of B. cereus type strain. Moreover, the presence of the ultrarare I-SceI restriction site in the mini-IS231A allowed the isolation, in double-insertion mutants, of contiguous and nonoverlapping large chromosomal fragments, convenient for direct sequencing. The system detailed in this report is therefore a powerful tool for comparative genetic studies among members of the B. cereus group (i.e., B. cereus, B. thuringiensis, B. mycoides, and B. anthracis) and could also be applied to more distantly related gram-positive bacteria.

Bacillus cereus↗

PubMed1971

Genetic mapping of genes concerned with glutamyl polypeptide production by Bacillus licheniformis and a study of their relationship to the development of competence for transformation.

By transduction with SP-15, we have mapped some of the genetic sites involved in glutamyl polypeptide (Pep) production by Bacillus licheniformis 9945A. Eleven peptide mutations were assigned to group 1 which lies between the gly-42 and his-6 markers. Three other mutations were assigned to group 2 which lies between the ade-2 and str-1 sites. A series of 28 additional Pep(-) mutants did not belong to either group 1 or 2; the chromosomal location of those peptide mutations is unknown. All three mutants of group 2 were highly transformable, but only two of group 1 transformed well under the conditions employed. Knowing the chromosomal markers that were linked to peptide mutations made it possible to investigate the effect peptide mutations might have on the development of competence for transformation. Nontransformable organisms were rendered transformable upon the introduction, by transduction, of Pep(-) mutations from transformable strains. These results supported the conclusion that the ability of cells to develop competence for transformation when grown under appropriate conditions was related to the physiological effects of particular peptide mutations.

Bacillus↗

PubMed1974

Active transport of exogenous S-adenosylmethionine and related compounds into cells and vacuoles of Saccharomyces cerevisiae.

Saccharomyces cerevisiae 4094-B (alpha, ade-2, ura-1) in potassium phosphate buffer with glucose under aerobic conditions took up (-)S-adenosyl-l-methionine from the medium in sufficient quantity to permit the demonstration of its accumulation in the vacuole by ultraviolet micrography. The same result was obtained with (+/-)S-adenosyl-l-methionine, (+/-)S-adenosyl-d-methionine, and (-)S-adenosyl-l-ethionine. The rate of uptake was slow with (-)S-adenosyl-S(n-propyl)-l-homocysteine and S-adenosyl-d-homocysteine. S-Adenosyl-l-homocysteine was assimilated rapidly, but intracellular degradation precluded accumulation and ultraviolet micrographic studies. The uptake of 5'-methyl-, 5'-ethyl-, 5'-n-propylthioadenosine, and 5'-dimethylsulfonium adenosine was minimal.

Aerobiosis↗

PubMed1975

Intrageneric transformation of neisseria gonorrhoeae and neisseria perflava to streptomycin resistance and nutritional independence.

Auxotrophic mutants of Neisseria gonorrhoeae and Neisseria perflava were transformed to prototrophy using homologous and heterologous deoxyribonucleic acid (DNA). Within either species the efficiencies of transformation for nutritional markers were found to be very similar to the values obtained for transformation to streptomycin resistance. The number of transformants in the interspecific N. perflava (donor) - - leads to N. gonorrhoeae (recipient) cross was 100-fold lower than the number obtained in the intraspecific N. gonorrhoeae - - leads to N. gonorrhoeae cross for streptomycin resistance, as well as for several nutritional markers. In the reciprocal experiment the difference in the number of transformants in the interspecific N. gonorrhoeae - - leads to N. perflava cross and the number obtained in the intraspecific N. perflava - - leads to N. perflava cross varied from 600 to 1,000-fold for the streptomycin resistance marker. Of greater interest was the finding that N. perflava auxotrophs, although transformable to prototrophy with wild-type N. perflava DNA, were not transformed to nutritional independence by gnoncoccal DNA. These same mutants were transformable to streptomycin resistance using the heterologous gonococcal DNA. When the DNAs of N. meningitidis, N. flava, and N. lactamicus were used to transform N. gonorrhoeae to prototrophy or streptomycin resistance, the transformation frequencies obtained fell along a gradient that in general reflected taxonomic relationships. On the other hand, with N. perflava as the recipient for these same DNAs, only N. flava DNA could transform auxotrophs to prototrophy, although transformation to streptomycin resistance occurred in all cases. DNA from N. perflava - - leads to N. gonorrheae streptomycin-resistant or Ade+ intergenotic transformants transformed N. gonorrhoeae cells at a 100-fold-higher efficiency than did DNA from N. perflava. Our findings suggest that (i) N. gonorrhoeae and N. perflava are more closely related than hitherto suspected and (ii) N. perflava is more selective with respect to heterologous DNA than is N. gonorrhoeae.

Amino Acids↗