[Noninvasive ventilation of critically ill patients. II: Practical application and procedure].
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Contraindications↗
Biomedical subjects
B Hoffmann
Publications and source records attributed to B Hoffmann.
[Noninvasive ventilation in critically ill patients. I. Mechanism of action and current scientific evidence].
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Critical Illness↗
Testicular endocrine function, seasonality and semen quality of the stallion.
To gain further information on gonadal function of the stallion, concentrations of testicular steroids in blood plasma (bpl) and seminal plasma (spl) and their distribution in the ejaculate were determined. Blood and semen samples from a total of 11 stallions were collected from November to July. Estrone (E1), estrone sulfate (E1S), estradiol-17beta (E2beta) and testosterone (T) were determined in bpl and spl, and in addition androstenedione (A), dehydroepiandrosterone (DHEA) and 5alpha-dihydrotestosterone (5alpha-DHT) were measured in spl. At certain points of time, aliquots of an ejaculate were centrifuged, washed and the distribution of E1, E1S, E2beta and T into seminal plasma and the sperm fraction was assessed. Hormone assay was by RIA, partly after prior separation by HPLC. Mean concentrations (X(g) x DF) were as follows: E2beta (bpl) 31.1 (1.16), (spl) 24.2 (1.42) pg ml(-1); E1 (bpl) 143.3 (1.21), (spl) 117.7 (1.53) pg ml(-1); E1S (bpl) 157.3 (1.44), (spl) 2.92 (1.42) ng ml(-1); T (bpl) 570.6 (1.43), (spl) 23.1 (1.68) pg ml(-1); A (spl) 17.9 (1.39) pg ml(-1); DHEH (spl) 12.4 (1.51) pg ml(-1); 5alpha-DHT (spl) 9.7 (1.29) pg ml(-1). Except for E2beta and A in seminal plasma, a seasonal pattern was established for all other steroids with lowest mean values occurring from November to April. From the semen parameters determined, only motility was correlated to season. There was a higher correlation among oestrogen in blp than in spl and the only correlation identified between oestrogenic and androgenic steroids was between T and E2beta in blp. In spl, T was correlated with A and 5alpha-DHT. T was the dominant free steroid in bpl while it was E1 in spl; T and E1S concentrations were about 23- and 54-fold lower in spl compared to bpl with E1S, however, showing the highest absolute values in both fluids. In the fractionated ejaculate an association of free oestrogens, particularly E2beta, with spermatozoa was observed.
Androgens↗
The aroC gene of Aspergillus nidulans codes for a monofunctional, allosterically regulated chorismate mutase.
The cDNA and the chromosomal locus of the aroC gene of Aspergillus nidulans were cloned and is the first representative of a filamentous fungal gene encoding chorismate mutase (EC 5.4.99.5), the enzyme at the first branch point of aromatic amino acid biosynthesis. The aroC gene complements the Saccharomyces cerevisiae aro7Delta as well as the A. nidulans aroC mutation. The gene consists of three exons interrupted by two short intron sequences. The expressed mRNA is 0.96 kilobases in length and aroC expression is not regulated on the transcriptional level under amino acid starvation conditions. aroC encodes a monofunctional polypeptide of 268 amino acids. Purification of this 30-kDa enzyme allowed determination of its kinetic parameters (k(cat) = 82 s(-1), n(H) = 1. 56, [S](0.5) = 2.3 mM), varying pH dependence of catalytic activity in different regulatory states, and an acidic pI value of 4.7. Tryptophan acts as heterotropic activator and tyrosine as negative acting, heterotropic feedback-inhibitor with a K(i) of 2.8 microM. Immunological data, homology modeling, as well as electron microscopy studies, indicate that this chorismate mutase has a dimeric structure like the S. cerevisiae enzyme. Site-directed mutagenesis of a crucial residue in loop220s (Asp(233)) revealed differences concerning the intramolecular signal transduction for allosteric regulation of enzymatic activity.
Allosteric Regulation↗
Soft docking an L and a D peptide to an anticholera toxin antibody using internal coordinate mechanics.
BACKGROUND: The tremendous increase in sequential and structural information is a challenge for computer-assisted modelling to predict the binding modes of interacting biomolecules. One important area is the structural understanding of protein-peptide interactions, information that is increasingly important for the design of biologically active compounds. RESULTS: We predicted the three-dimensional structure of a complex between the monoclonal antibody TE33 and its cholera-toxin-derived peptide epitope VPGSQHID. Using the internal coordinate mechanics (ICM) method of flexible docking, the bound conformation of the initially extended peptide epitope to the antibody crystal or modelled structure reproduced the known binding conformation to a root mean square deviation of between 1.9 A and 3.1 A. The predicted complexes are in good agreement with binding data obtained from substitutional analyses in which each epitope residue is replaced by all other amino acids. Furthermore, a de novo prediction of the recently discovered TE33-binding D peptide dwGsqhydp (single-letter amino acid code where D amino acids are represented by lower-case letters) explains results obtained from binding studies with 172 peptide analogues. CONCLUSIONS: Despite the difficulties arising from the huge conformational space of a peptide, this approach allowed the prediction of the correct binding orientation and the majority of essential binding features of a peptide-antibody complex.
Antibodies, Monoclonal↗
Identification of genes regulated by muscarinic acetylcholine receptors: application of an improved and statistically comprehensive mRNA differential display technique.
In order to identify genes that are regulated by muscarinic acetylcholine receptors, we developed an mRNA differential display technique (DD) approach. By increasing redundancy and by evaluating optimised reagents and conditions for reverse transcription of total RNA, PCR and separation of PCR products, we generated a DD protocol that yields highly consistent results. A set of 64 distinct random primers was specifically designed in order to approach a statistically comprehensive analysis of all mRNA species in a defined cell population. This modified DD protocol was applied to total RNA of HEK293 cells stably expressing muscarinic m1 acetylcholine receptors and cells stimulated with the receptor agonist carbachol were compared to identical but non-stimulated cells. In 81 of 192 possible PCR experiments, 38 differential bands were identified. Sequence analysis followed by northern blot analyses confirmed differentially expressed genes in 19 of 23 bands analysed. These represented 10 distinct immediate-early genes that were up-regulated by m1AChR activation: Egr-1, Egr-2, Egr-3, NGFi-B, ETR101, c- jun, jun -D, Gos-3 and hcyr61, as well as the unknown gene Gig-2. These data show that this improved DD protocol can be readily applied to reliably identify differentially expressed genes.
Base Sequence↗
The use of noninvasive pressure support ventilation for severe respiratory insufficiency due to pulmonary oedema.
OBJECTIVE: Experimental use of noninvasive pressure support ventilation (NIPSV) in patients with severe pulmonary oedema who would have been intubated if noninvasive ventilation were not available. DESIGN: Open, prospective, within patients non comparative study. SETTING: Internal intensive care unit (11 beds) at a university hospital. PATIENTS: 29 patients with severe respiratory distress and confirmed pulmonary oedema. INTERVENTIONS: NIPSV was applied via a tight fitting face mask delivering between 13 and 24 cm H2O inspiratory airway pressure and 2 to 8 cm H2O expiratory airway pressure. MEASUREMENTS AND RESULTS: One patient required endotracheal intubation. Mean plethysmographic oxygen saturation rose significantly within 30 min from 73.8+/-11 to 90.3+/-5%, while the oxygen supply was reduced from 7.3+/-3.7 to 5.1+/-3 l/min. Mean pH increased significantly (p<0.01) from 7.22+/-0.1 before NIPSV to 7.31+/-0.07 after 60 min of NIPSV. Partial pressure of carbon dioxide was 62+/-18.5 mmHg but decreased significantly within 60 min to 48.4+/-11.5 mm Hg. Heart rate and-blood pressure stabilised continuously during the observation time. Mean duration of NIPSV was 6 h 9 min (range 60 min to 24 h). There were no serious side effects. Four patients died from underlying diseases between 1 and 28 days after NIPSV. CONCLUSION: NIPSV is a highly effective technique with which to treat patients with severe cardiogenic pulmonary oedema.
Aged↗
Sexual development of Aspergillus nidulans in tryptophan auxotrophic strains.
The interplay between sexual development and amino acid biosynthesis in the ascomycete Aspergillus nidulans was studied. The growth and differentiation of four tryptophan auxotrophic strains that are unable to regulate their tryptophan biosynthesis, were examined. These strains are sterile on medium containing low tryptophan concentrations. Fruitbody formation was restored by supplementation with high concentrations of tryptophan and was promoted by supplementation with indole or auxin. Tryptophan supplementation resulted in auxin production of A. nidulans. Fertility of ascospores of the tryptophan auxotrophic strains could only be partially re-established. trpC transcript levels and enzyme activities remained stable in cleistothecia and conidiophore extracts as compared to those of mycelium, while levels of gene transcripts involved in glycolysis were lower in fruitbodies and conidiospores. Auxotrophic strains unable to form fruitbodies at intermediate amino acid supplementation levels turned on the cross-pathway regulatory system that is induced by amino acid starvation. We conclude that there is a connection between the genetic network of cross-pathway control and sexual development in A. nidulans.
Aspergillus nidulans↗
Tripolar acytokinetic mitosis and formation of feto-maternal syncytia in the bovine placentome: different modes of the generation of multinuclear cells.
The vast majority of trophoblast giant cells in the ruminant placenta are binuclear and are believed to derive from mononuclear trophoblastic cells by a single acytokinetic mitosis. There is no satisfactory explanation for the generation of the small proportion of trophoblast giant cells with one, three, or more nuclei. In this light-and electronmicroscopic study of bovine placentomal tissue from the second half of gestation, developmental stages of the trophoblast giant cells are investigated. Large mitotic figures indicate mitotic polyploidization, which is proposed to be due to two subsequent acytokinetic mitoses. Tripolar mitoses offer an explanation for the development of trinucleate trophoblast giant cells. Measurements of nuclear volumes in a series of semithin sections revealed that three size classes of trophoblast giant cells occur. The approximately doubling of nuclear volume between each class is thought to reflect different levels of DNA content that result from polyploidization in this cell type. Although trinuclear feto-maternal hybrid cells are the standard outcome of the fusion of binuclear trophoblast giant cells with uterine epithelial cells, some syncytia with at least five nuclei were observed in the uterine epithelium.
Animals↗
Spot synthesis: observations and optimizations.
Positionally addressable syntheses of peptides on continuous cellulose membranes (spot synthesis) have often been reported in detail, but important questions dealing with synthesis quality, reproducibility and subsequent binding assays have largely been under-emphasized. In this report we have investigated some of these problems. The most important results were: (i) the signal intensity of ligate binding to cellulose-bound peptides and the affinity of the corresponding soluble peptides show good correlation, illustrated by three different ligate binding assays; (ii) reducing peptide density on the cellulose avoids the 'ring spot' effect, i.e. where less binding is observed in the spot-center compared to the rim. We recommend a peptide density of 10 nmol/cm2 as a reasonable starting point for further optimization; (iii) statistical analysis of binding assay reproducibility with more than 15000 peptides resulted in a mean standard signal deviation of 0.18; and (iv) optimization of side-chain deprotection revealed that a 30-min pretreatment of the cellulose with 90% trifluoroacetic acid followed by the standard deprotection protocol resulted in higher purity of the synthesized products.
Amino Acid Sequence↗
The WD protein Cpc2p is required for repression of Gcn4 protein activity in yeast in the absence of amino-acid starvation.
The CPC2 gene of the budding yeast Saccharomyces cerevisiae encodes a G beta-like WD protein which is involved in regulating the activity of the general control activator Gcn4p. The CPC2 gene encodes a premRNA which is spliced and constitutively expressed in the presence or absence of amino acids. Loss of CPC2 gene function suppresses a deletion of the GCN2 gene encoding the general control sensor kinase, but not a deletion in the GCN4 gene. The resulting phenotype has resistance against amino-acid analogues. The Neurospora crassa cpc-2 and the rat RACK1 genes are homologues of CPC2 that complement the yeast cpc2 deletion. The cpc2 delta mutation leads to increased transcription of Gcn4p-dependent genes under non-starvation conditions without increasing GCN4 expression or the DNA binding activity of Gcn4p. Cpc2p-mediated transcriptional repression requires the Gcn4p transcriptional activator and a Gcn4p recognition element in the target promoter. Frameshift mutations resulting in a shortened G beta-like protein cause a different phenotype that has sensitivity against amino-acid analogues similar to a gcn2 deletion. Cpc2p seems to be part of an additional control of Gcn4p activity, independent of its translational regulation.
Acetates↗
Structure and dynamics of the B12-binding subunit of glutamate mutase from Clostridium cochlearium.
Glutamate mutase (Glm) is an adenosylcobamide-dependent enzyme that catalyzes the reversible rearrangement of (2S)-glutamate to (2S, 3S)-3-methylaspartate. The active enzyme from Clostridium cochlearium consists of two subunits (of 53.6 and 14.8 kDa) as an alpha2beta2 tetramer, whose assembly is mediated by coenzyme B12. The smaller of the protein components, GlmS, has been suggested to be the B12-binding subunit. Here we report the solution structure of GlmS, determined from a heteronuclear NMR-study, and the analysis of important dynamical aspects of this apoenzyme subunit. The global fold and dynamic behavior of GlmS in solution are similar to those of the corresponding subunit MutS from C. tetanomorphum, which has previously been investigated using NMR-spectroscopy. Both solution structures of the two Glm B12-binding subunits share striking similarities with that determined by crystallography for the B12-binding domain of methylmalonyl CoA mutase (Mcm) from Propionibacterium shermanii, which is B12 bound. In the crystal structure a conserved histidine residue was found to be coordinated to cobalt, displacing the endogenous axial ligand of the cobamide. However, in GlmS and MutS the sequence motif, Asp-x-His-x-x-Gly, which includes the cobalt-coordinating histidine residue, and a predicted alpha-helical region following the motif, are present as an unstructured and highly mobile loop. In the absence of coenzyme, the B12-binding site apparently is only partially formed. By comparing the crystal structure of Mcm with the solution structures of B12-free GlmS and MutS, a consistent picture on the mechanism of B12 binding has been obtained. Important elements of the binding site only become structured upon binding B12; these include the cobalt-coordinating histidine residue, and an alpha helix that forms one side of the cleft accommodating the nucleotide 'tail' of the coenzyme.
Amino Acid Sequence↗
Increased release of transforming growth factor (TGF)-beta1, TGF-beta2, and chemoattractant mediators in pneumonia.
Transforming growth factor-beta (TGF-beta), interleukin-8 (IL-8), and leukotrienes are potent neutrophil chemoattractants that are released in several lung diseases. There is limited information about the release of TGF-beta in bronchoalveolar lavage fluid (BALF) of patients with pneumonia. Furthermore, it is not clear if TGF-beta is differentially expressed in different lung diseases. The aim of our study was to compare the concentrations of TGF-beta1 and TGF-beta2 in the BALF of patients with pneumonia and other lung diseases. Furthermore, correlation of the TGF-beta levels with the concentration of chemoattractant mediators as well as with indicators of macrophage and granulocyte activation should be investigated. Patients with pneumonia, interstitial lung disease (ILD), or chronic obstructive pulmonary diseases (COPD) were included. Patients with ischemic heart disease without pulmonary involvement served as controls. The concentrations of TGF-beta1 and TGF-beta2, of the chemoattractant cytokine IL-8, of leukotriene B4, and of the leukotrienes C4, D4, and E4 were measured. Neutrophil elastase and granulocyte content (PMN) were used as markers for granulocyte activation, and neopterin was used as a marker for the activation of macrophages. Significantly elevated levels of TGF-beta1 (mean = 0.216 ng/ml, p < 0.01) were found in patients with microbiologically positive pneumonia but not in patients with ILD or COPD. A significant (p < 0.001) correlation was found between the TGF-beta1 concentrations and the IL-8 levels and the percentage of granulocytes (r = 0.76, and r = 0.44, respectively). Elevated TGF-beta2 concentrations were measured in the BALF of patients with pneumonia (mean = 1.4 ng/ml, p < 0.01) and with ILD. Pneumonia was also associated with increased concentrations of leukotrienes C4, D4, and E4 (mean = 91.61 pg/ml, p < 0.05) and leukotriene B4 (mean = 203.9 pg/ml, p < 0.01), significantly elevated levels of PMN elastase (mean = 2958.26 ng/ml, p < 0.01), and neopterin (mean = 0.42 nmol/L). Our results strongly suggest that different lung diseases do differ with regard to the released cytokines. TGF-beta1 probably plays a key role in regulation of pulmonary inflammation, particularly in pneumonia.
Bronchoalveolar Lavage Fluid↗
Immunolocalization of progesterone receptors in bovine placentomes throughout mid and late gestation and at parturition.
The corpus luteum is the main source of progesterone (P(4)) responsible for maintenance of gestation in cattle. So far it has not been possible to assign any biological role to placental P(4), which contributes only marginally and temporarily to peripheral maternal blood levels. In order to identify possible P(4) target cells within the placenta, placentomes from 150-, 220-, 240-, and 270-day-pregnant cows and from parturient cows (3 animals per group) were screened immunohistochemically for expression of the progesterone receptor (PR). During gestation, PR-positive staining was found exclusively in the nuclei of caruncular stromal cells (CSC; maternal part of the placentome) and of caruncular vascular pericytes. In placentomes from parturient cows, occasional positive nuclear staining was also observed in the walls of small caruncular arteries. The percentage of PR-positive CSC increased slightly from 51.8 +/- 2.6% on Day 150 to 56.2 +/- 5.6% at Day 270 (p < 0.05) and was 58.9 +/- 1.8% at parturition. These results suggest that in pregnant cattle, CSC are under the control of P(4) of placental rather than luteal origin. Thus, whereas luteal P(4) may regulate "coarse" systemic progestational functions in the maternal compartment in the classical hormonal manner, placental P(4) may act as a paracrine factor involved in the local regulation of caruncular growth, differentiation, and functions.
Animals↗
Apoplastic pH and Fe(3+) reduction in intact sunflower leaves
It has been hypothesized that under NO(3)(-) nutrition a high apoplastic pH in leaves depresses Fe(3+) reductase activity and thus the subsequent Fe(2+) transport across the plasmalemma, inducing Fe chlorosis. The apoplastic pH in young green leaves of sunflower (Helianthus annuus L.) was measured by fluorescence ratio after xylem sap infiltration. It was shown that NO(3)(-) nutrition significantly increased apoplastic pH at distinct interveinal sites (pH >/= 6.3) and was confined to about 10% of the whole interveinal leaf apoplast. These apoplastic pH increases presumably derive from NO(3)(-)/proton cotransport and are supposed to be related to growing cells of a young leaf; they were not found in the case of sole NH(4)(+) or NH(4)NO(3) nutrition. Complementary to pH measurements, the formation of Fe(2+)-ferrozine from Fe(3+)-citrate was monitored in the xylem apoplast of intact leaves in the presence of buffers at different xylem apoplastic pH by means of image analysis. This analysis revealed that Fe(3+) reduction increased with decreasing apoplastic pH, with the highest rates at around pH 5. 0. In analogy to the monitoring of Fe(3+) reduction in the leaf xylem, we suggest that under alkaline nutritional conditions at interveinal microsites of increased apoplastic pH, Fe(3+) reduction is depressed, inducing leaf chlorosis. The apoplastic pH in the xylem vessels remained low in the still-green veins of leaves with intercostal chlorosis.
Journal Article↗
[Effect of a 10 day postpartum administration of a PGF2 alpha analog at different concentrations on the course of the puerperium and fertility in dairy cows].
In this double-blind Study 72 dairy cows were treated with injections of Luprostiol (Reprodin). 32 cows served as control. Gynaecological examinations in 10-day intervals up to day 40 post partum and determination of progesterone levels in milk samples taken in 3-day intervals up to day 73 p. p. were performed. The administration of PGF2 alpha did not improve the early onset of ovarian activity in cows with uncomplicated or complicated delivery. Ovaries without detectable function up to day 73 p. p. only occurred in the placebo group, representing a significant (p < 0.05) difference to the control group. In cows after uncomplicated as well as in animals after complicated delivery involution of the uterus up to day 20 p. p. progressed faster in the verum group compared to the control group. Cervical diameters were highest in the placebo group up to day 20 p. p. Until day 40 p. p. the cervical diameters of the probands equaled those of the control animals. In contrary to cows after uncomplicated delivery the injection of PGF2 alpha after complicated parturition lead to a significant (p < 0.05) reduction of puerperal endometritis. Neither in cows with uncomplicated nor complicated delivery the administration of PGF2 alpha resulted in an improved conception rate. There were no significant positive effects on fertility parameters to be detected in general. Partial results observed were not dose dependent.
Animals↗
[Non-invasive positive pressure ventilation in cardiogenic pulmonary edema].
PATIENTS AND METHOD: 30 patients being admitted to our intensive care unit with severe cardiogenic pulmonary edema received non-invasive positive pressure ventilation (NIPPV) via face mask. RESULT: 29 responded well, 1 patient had to be intubated. Within 30 minutes those who responded well showed a significant improvement of the following parameters: rise of peripheral saturation from 75.5 to 90.1% and of pH from 7.24 to 7.29, decrease of pCO2 from 60.7 to 48.8 mm Hg and of systolic blood pressure from 144 to 124 mm Hg. Mean duration of ventilation was 6 h 55 min. Mean stay in intensive care unit was 2 days. No patient required ventilator support within 24 hours after NIPPV. Four patients died during hospital stay as a result of their underlying disease but not due to pulmonary edema. Observed side effects were vomiting in 4 cases during NIPPV without aspiration and 3 cases of skin lesions which healed uneventfully. CONCLUSION: Key role for this highly effective method seems to be the rapid improvement of left ventricular function during NIPPV.
Aged↗
[Noninvasive pressure support ventilation (NIPSV) as therapy for severe respiratory insufficiency due to pulmonary edema].
OBJECTIVE: Experimental use of noninvasive pressure support ventilation (NIPSV) in patients with severe pulmonary oedema who would have been intubated if noninvasive ventilation were not available. DESIGN: Open, prospective, within patients non comparative study. SETTING: Internal intensive care unit (11 beds) at a university hospital. PATIENTS: 29 patients with severe respiratory distress and confirmed pulmonary oedema. INTERVENTIONS: NIPSV was applied via a tight fitting face mask delivering between 13 and 24 cm H2O inspiratory airway pressure and between 2 and 8 cm H2O expiratory airway pressure. MEASUREMENTS AND RESULTS: One patient required endotracheal intubation. Mean plethysmographic oxygen saturation rose significantly within 30 min from 73.8 +/- 11 to 90.3 +/- 5% while the oxygen supply was reduced from 7.3 +/- 3.7 to 5.1 +/- 3 l/min. Mean pH increased significantly (p < 0.01) from 7.22 +/- 0.1 before NIPSV to 7.31 +/- 0.01 after 60 min of NIPSV. Partial pressure of carbon dioxide was 62 +/- 18.5 mmHg, but decreased significantly within 60 min to 48.4 +/- 11.5 mm Hg. Heart rate and blood pressure established continuously during observation time. Mean duration of NIPSV was 6 h 9 min (range 120 min to 24 h). There were no serious side effects. Four patients died from underlying diseases between 1 and 28 days after NIPSV. CONCLUSION: NIPSV is a highly effective technique with which to treat patients with severe cardiogenic pulmonary oedema.
Aged↗