Directing Biosynthesis: Practical Supply of Natural and Unnatural Cyanobactins.

The increasingly rapid accumulation of genomic information is revolutionizing natural products discovery. However, the translation of sequence data to chemical products remains a challenge. Here, we detail methods used to circumvent the supply problem of cyanobactin natural products, both by engineered synthesis in Escherichia coli and by using purified enzymes in vitro. Such methodologies exploit nature's strategies of combinatorial chemistry in the cyanobactin class of RiPP natural products. As a result, it is possible to synthesize a wide variety of natural and unnatural compounds.

Synthesis and bioactivity of nobilamide B.

An alternative and facile solution/solid-phase approach is reported for the total synthesis of neuroactive peptide, nobilamide B. Z-Dhb was formed in solution via EDC/CuCl induced elimination. The solid-phase synthesis employed HBTU/Oxyma Pure™ coupling using Barlos resin. Synthetic nobilamide B was also found to be neuroactive in primary cultures of dorsal root ganglion (DRG) neurons.

The differences in sleep profile changes under continuous positive airway pressure (CPAP) therapy between non-obese, obese and severely obese sleep apnea patients.

Sleep disturbances in obstructive sleep apnea are caused mainly by repetitive apneas and hypopneas. An alternative factor contributing to disordered sleep may be the obesity, which is frequently associated with sleep apnea. The sleep disturbing effect of obesity was found previously in obese nonapneic subjects. The aim of this study was to evaluate the effect of obesity on sleep quality in sleep apnea patients in particular in patients under continuous positive airway pressure (CPAP) with successfully normalized respiration. We reviewed the archive data of 18 non-obese, 18 obese and 17 severely obese age and gender matched sleep apnea patients treated with CPAP. The polysomnographic parameters from the diagnostic night, from the second night under CPAP and from the follow up night (after three months of CPAP use) were compared. Before CPAP the apnea hypopnea index was worse in obese and in severely obese group and it normalised under CPAP in all groups. The severely obese group showed more light sleep and less REM sleep before CPAP and inversely - less light and more REM sleep in the second night under CPAP than the non-obese group. In the follow up, there was no differences in sleep profile between groups. This study indicates obesity does not affect the sleep independently of respiratory disorders. Before therapy it is associated with more severe sleep apnea and indirectly with worse sleep quality.

Isotope shift in the dielectronic recombination of three-electron ANd57+.

Isotope shifts in dielectronic recombination spectra were studied for Li-like (A)Nd(57+) ions with A=142 and A=150. From the displacement of resonance positions energy shifts deltaE(142 150)(2s-2p(1/2))=40.2(3)(6) meV [(stat)(sys)] and deltaE(142 150)(2s-2p(3/2))=42.3(12)(20) meV of 2s-2p(j) transitions were deduced. An evaluation of these values within a full QED treatment yields a change in the mean-square charge radius of (142 150)deltar(2)=-1.36(1)(3) fm(2). The approach is conceptually new and combines the advantage of a simple atomic structure with high sensitivity to nuclear size.

Screened radiative corrections from hyperfine-split dielectronic resonances in lithiumlike scandium.

Term energies for dielectronic-recombination Rydberg resonances below 0.07 eV are determined for Sc18+ with absolute accuracies below 0.0002 eV by electron collision spectroscopy in an ion storage ring, using the twin-electron-beam technique and a cryogenic photocathode. The lithiumlike 2s_{1/2}-2p_{3/2} transition energy for Z=21 is determined to 4.6 ppm, less than 1% of the few-body effects on radiative corrections. Features from the hyperfine structure of the 2s state could be resolved in the dielectronic-recombination spectrum.

The influence of obesity on sleep quality in male sleep apnea patients before and during therapy.

Evidence exists that obesity, even in the absence of sleep related respiratory disorders, affects sleep negatively. In this study we examined the influence of obesity on sleep quality of male sleep apnea patients before and after breathing normalization with continuous positive airway pressure (CPAP). We compared the polysomnography from the diagnostic night, second night with CPAP, and a control night (three months later) in 13 non-obese, 13 obese, and 12 severely obese male obstructive sleep apnea patients. In the diagnostic polysomnography, obese and severely obese subjects showed increases in apnea-hypopnea index (AHI) and NREM-1 sleep, and decreases in min SaO(2), REM sleep, and partially slow wave sleep (SWS), when compared with the non-obese group. In the second night under CPAP, normalization of the AHI and a rebound of REM and SWS occurred, which was more pronounced in severely obese than in the non-obese and obese group. The polysomnography recorded three months thereafter revealed no differences in sleep stages between the groups. We conclude that after the long-term CPAP therapy, no effect of obesity on sleep quality is apparent.

Storage-ring measurement of the hyperfine induced 47Ti18+(2s2p 3P0 --> 2s2 1S0) transition rate.

The hyperfine induced 2s2p (3)P(0) --> 2s(2) (1)S(0) transition rate A(HFI) in berylliumlike (47)Ti(18+) is measured. Resonant electron-ion recombination in a heavy-ion storage ring is employed to monitor the time dependent population of the (3)P(0) state. The experimental value A(HFI)=0.56(3) s(-1) is almost 60% larger than theoretically predicted.

Early tandem high-dose ifosfamide, carboplatin, etoposide therapy with stem cell rescue for small-cell lung cancer: brief report on the results of a phase-I/II trial.

OBJECTIVE: High-dose therapy (HDT) for small-cell lung cancer is experimental. Late intensification HDT for chemosensitive disease can increase the number and quality of remissions and prolong relapse-free survival, but has not yet shown impact on overall survival. This is possibly due to resistant residual disease. To overcome the development of resistance, we have tested early intensification tandem HDT. METHODS: We performed a phase-I/II trial using 1 conventional cycle of ifosfamide, carboplatin, etoposide (ICE) plus granulocyte colony-stimulating factor for stem cell recruitment followed by 2 cycles of high-dose ICE with rescue by CD34+ cell-enriched peripheral blood mononuclear cells. Dose escalation was performed for the 2 high-dose ICE cycles. Radiotherapy for limited disease was according to standard protocols. RESULTS: 17 patients were entered: 2 female patients; 15 male patients; median age 53 (range 36-65) years; 2 patients with limited disease, and 15 patients with extensive disease. We treated 4 patients at dose level 1, 11 patients at level 2, and 2 patients at level 3. The maximum tolerable dose was at level 2 with neuropathy being dose-limiting. Overall, toxicity was < or = grade 2 for all patients up to dose level 2 with hematotoxicity being grade 4 for all patients. There were 15 partial remissions (88%), 1 no change (6%), and 1 progressive disease (6%). Median time to progression was 7.9 months. Overall survival was 12.9 months (median). CONCLUSIONS: Early intensification with this protocol is feasible. Although a comparatively good response rate and median time to progression have been observed in this group dominated by patients with extensive disease, overall survival is short and no substantial long-term survival was found.

[Onset of action of formoterol in patients with moderate to severe, partially reversible airflow obstruction assessed by bodyplethysmography]. / Bodyplethysmographischer Wirkeintritt von Formoterol bei Patienten mit mittel- bis schwergradiger partiell reversibler Atemwegsobstruktion.

BACKGROUND: Formoterol is a long acting beta2-agonist used for the treatment of obstructive airway diseases. Compared with Salmeterol, Formoterol has a rapid onset of bronchodilation. There are only scant data regarding the comparative onset of action using bodyplethysmography in moderate to severely obstructive patients. METHODS: In a mono-center, single-blinded parallel group study 60 patients (age: 61.9 +/- 12.8 years, 65 % male) with moderate to severe (mean FEV(1) 40.6 +/- 15.3 % of predicted), partially reversible (FEV(1) post bronchodilator > 15 % from baseline) airway obstruction were treated with either formoterol 12 microg bid or salmeterol 50 microg bid over a period of two weeks. Onset of action was measured by airway resistance (sRaw) before and after two weeks of treatment. RESULTS: Compared with Salmeterol, Formoterol had a significantly faster onset of action (10 % decrease of Raw) at baseline (F: 1.4 +/- 0.9 vs. S: 15.1 +/- 34.5 min., p < 0.0001) and after two weeks of treatment (F: 6.2 +/- 21.6 vs. S: 51 +/- 135 min., p < 0.0001). Morning FEV(1) improved similarily during treatment in both study groups, when compared with baseline lung function (F: 1.38 +/- 0.64 vs. 1 +/- 0.41 l; S: 1.43 +/- 0.67 vs. 1 +/- 0.4 l, p < 0.05, both comparisons). Both treatments were well tolerated. CONCLUSION: Formoterol produces a rapid improvement of airway resistance in patients with moderate to severe, partially reversible airway obstruction. The onset of bronchodilation was significantly faster for Formoterol compared with Salmeterol. Both drugs improved lung function similarily after two weeks of treatment. It is important to distinguish beta2-agonists not only into short- and long-acting but also into fast- and slow-acting.

Breast cancer screening in a rural Wisconsin community.

BACKGROUND: Screening tests have been shown to play an important role in earlier disease stage detection of breast cancer. AIM: The aim of this study was to assess the acceptance of screening mammography in a rural community in Wisconsin. METHODS: A cohort of 60 patients with breast cancer was analyzed retrospectively for disease stage, screening mammography, and patient compliance. RESULTS: Of 60 patients with breast cancer, only 25 (42%) underwent screening mammography. Compliance was suboptimal among patients; especially poor in the elderly population. CONCLUSION: Promotional steps to increase acceptance of screening tests need to be emphasized especially in patients > or = 60 years in age to enhance early detection of breast cancer and prolong disease-free interval.

Functional variables associated with the clinical grade of dyspnoea in coal miners with pneumoconiosis and mild bronchial obstruction.

OBJECTIVES: Dyspnoea is a common symptom in coal miners with pneumoconiosis. Among others, gas exchange disturbances due to airway obstruction or mismatch between ventilation and perfusion may be underlying mechanisms. The validation of dyspnoea by the degree of airway obstruction is controversial, because the extent of airway obstruction often does not correlate with the clinical grade of breathlessness. METHODS: The association was investigated between breathlessness (self reported, on a six point scale) and indices of submaximal spiroergometry in 66 coal workers with radiographically confirmed pneumoconiosis (International Labour Organisation (ILO) grade of profusion > or =1/0, mean (SD) age 64 (5.5) years, mean (SD) forced expired volume in 1 second (FEV(1)) 77.5 (22.9) % predicted). RESULTS: The clinical degree of breathlessness was independently associated with minute ventilation/oxygen consumption (VE/VO(2)) ratio (beta 0.423, 95% confidence interval (95% CI) 0.18 to 0.67, p=0.001) and smoking (beta 0.318, 95% CI 0.21 to 1.79, p=0.014) in a multiple linear regression analysis. The VE/VO(2) ratio (beta 0.556, 95% CI 0.20 to 0.90, p=0.003) was also the best predictor of breathlessness when only coal miners with airway obstruction (FEV(1) < 80% predicted) were analyzed. CONCLUSION: The VE/VO(2) ratio as a measurement of mismatch between ventilation and perfusion predicted the clinical grade of breathlessness better than measurements of bronchial obstruction at rest in coal workers with pneumoconiosis.

Hexanoate synthase, a specialized type I fatty acid synthase in aflatoxin B1 biosynthesis.

In fungi, fatty acids are biosynthesized by large multifunctional enzyme complexes, the fatty acid synthases (FASs), which catalyze chain assembly in an iterative manner. Many fungal secondary metabolites contain fatty acid moieties, and it is often unclear whether they are recruited from primary metabolism or are biosynthesized de novo by secondary metabolic FASs. The most convincing evidence of such a dedicated FAS comes from the biosyntheses of aflatoxin (AF) and sterigmatocystin (ST) in certain species of the filamentous fungus Aspergillus. Incorporation studies in AF and genetic analyses of ST and AF biosynthesis strongly suggest that their biosyntheses begin with the production of a C6 fatty acid by a specialized FAS. The genes encoding the alpha (hexA) and beta (hexB) subunits of this hexanoate synthase (HexS) from the AF pathway in Aspergillus parsiticus SU-1 were cloned and both their gDNAs and cDNAs were sequenced and their transcriptional ends analyzed. Translated amino acid sequences are predicted to result in proteins of 181.3 and 210.5 kDa, for HexA and HexB, respectively. Comparison of the HexA and HexB sequences with those of the ST FAS subunits and primary metabolic FASs indicated that the secondary metabolic enzymes are members of a well-defined subclass of the FAS family. Phylogenetic predictions and an analysis of GC-bias in AF and ST pathway genes compared with primary metabolic Aspergillus genes were used as a basis to propose a route for the evolution of the AF and ST clusters.

Microbial symbionts of marine invertebrates: opportunities for microbial biotechnology.

Marine invertebrates are sources of a diverse array of bioactive metabolites with great potential for development as drugs and research tools. In many cases, microorganisms are known or suspected to be the biosynthetic source of marine invertebrate natural products. The application of molecular microbiology to the study of these relationships will contribute to basic biological knowledge and facilitate biotechnological development of these valuable resources. The bryostatin-producing bryozoan B. neritina and its specific symbiont "Candidatus Endobugula sertula" constitute one promising model system. Another fertile subject for investigation is the listhistid sponges that contain numerous bioactive metabolites, some of which originate from bacterial symbionts.

[Dyspnea and lung function. Results of a multicenter study]. / Dyspnoe und Lungenfunktion. Ergebnisse einer Multicenterstudie.

UNLABELLED: In seven pneumological centres 266 patients with different pneumological diseases were investigated. After having clarified several questions regarding the severity of the dyspnoea, cough intensity and the volume of sputum, as well as basic clinical investigation and after an x-ray of the thorax, the diagnosis was arrived at. Subsequently the lung function investigation with the flow-volume curve (including IVC, FVC, PEF, FEV1, MEF50%) and the body plethysmographic Rt and IGV were carried out. Different quality control procedures at and between the different centres ensured comparable results. All centres agreed to using methods well compatible with each other. The question as to which kind of parameters of lung function would agree best with the amount of the dyspnoea, was resolved. The causes for the large scatter of the results are described. Cough and sputum exercise an influence even on the degree of dyspnoea, but not by deteriorating the lung function. The results are shown for the entire collective (Part I) in respect of the different diagnoses (Part II). With different diagnosis the same significant correlations exist but the curves are positioned at different levels of the coordinate system. CONCLUSION: Significant correlations exist between the dyspnoea scale and function parameters. There are individual differences between the dyspnoea scale and disturbances of the function parameters. Carefully performed lung function analyse definitely important in any case.

Pharmacokinetics of beta2-sympathomimetics at the example of fenoterol and conclusion for the administration.

The pharmacokinetics of fenoterol (salbutamol, terbutaline) after systemic administration (i.v., infusion, nasal administration) can be best described by a three-compartmental model. Nasal administration causes an effect-time profile between that of infusion and inhalation. The effects of nasal administration on the lung function and the heart rate depend on the plasma levels of the beta2-agonist. The absorption rate after inhalation differs individually in a large variability (1-27% of the dosage). After inhalation the effect on the lung function does not depend on the absorption rate nor on the plasma levels of the beta2-agonist. After inhalation the effect on the lung function is three- to fivefold more expressed than predictable from the plasma level. From the pk/pd data it can be assumed, that there are 10-20 fold higher concentrations in the airways than in the plasma. It is assumed that there are structures nearby the beta2-receptor responsible for the long-lasting effect observed after inhalation. These depot structures cannot be reached from the plasma in concentrations needed for bronchodilation under in vivo conditions. In respect to the effect/side effect relations, there is no doubt that inhalation is the optimal solution for administering beta2-agonists. In respect to the effect/side effect relations more frequent administration of small doses seems to be more favourable than high doses inhaled in long periods.

[Submaximal spiroergometric stress study in patients with mixed dust pneumoconiosis]. / Submaximale spiroergometrische Belastungsuntersuchung bei Patienten mit Mischstaub-Pneumokoniosen.

Disabilities of patients with coal-workers' pneumoconiosis (CWP) are currently estimated by changes in lung function at rest and the degree of dust equivalents in chest x-ray (ILO-classification 1980). Functional disturbances during exercise are not taken into consideration on a regular basis. We assumed that standardised sub-maximal exercise tests might be useful even in disabled patients to give additional information on functional disabilities of patients with CWP. The impact of low-grade anthracosilicosis on ventilation and gas exchange during exercise was assessed in 20 patients (all male, age 64.55 +/- 3.78 years) and 24 healthy volunteers (all male, age 58.13 +/- 4.68 years, never dust exposed). Data were also analysed according to ILO classification subgroups (group 1: ILO-classification 1/0 to 1/2, n = 11; group 2: ILO-classification 2/1 to 2/2, n = 9). Heart rate (HR), minute ventilation (VE), ventilatory equivalent for O2 (VE/VO2), ventilatory equivalent for CO2 (VE/VCO2), O2 uptake (VO2)CO2 output (VCO2), end-tidal oxygen partial pressure (PetO2), end-tidal carbon dioxide partial pressure (petCO2) and estimated dead space to tidal volume ratio (VD/VT) were determined breath-by-breath during a 50 watts constant work load protocol on an exercise bicycle. The VE/VO2 (patients: 32.9 +/- 4.2; controls: 25.7 +/- 2.9; p < 0.001), the VE/CO2 (patients: 39.4 +/- 4.6; controls; 31.0 +/- 3.9), the VE (patients: 30.1 +/- 5.6, controls: 23.5 +/- 3.0; p < 0.001) and the PetO2 (patients 115.6 +/- 4.8; controls: 99.1 +/- 27.4; p < 0.05) at an exercise of 50 watts were significantly higher in the patients' group, the PetCO2 (patients: 38.4 +/- 4.5; controls: 44.0 +/- 4.1) were significantly lower in the patients' group. The comparison of patients subgroups did not reveal any significant differences between group 1 and group 2. A higher VE/VO2 during 50 watts of exercise is attributable to an increased ventilation/perfusion mismatch. There was no correlation between the radiological grade of the CWP (ILO classification) and the ventilation and gas exchange during exercise. We conclude that a sub-maximal spiroergometry with a 50 watts constant work load might serve as a sensitive and easy to apply procedure add information about the functional impairment in CWP. The exercise test should be included in the evaluation for disability benefits.

The flow-volume curve in patients with obstructive airway diseases partial analysis and functional importance.

Flow-volume curves in patients with obstructive airway disease differs from that observed in healthy subjects. Two types of pathological curves can be differentiated: these with clear sharp bend and intermediate forms characterised by the different grade of concavity of the descending segments plotted against X-axis. The aim of our present investigation is to elucidate the mechanisms which determines the forced expiratory airflow course in patients with obstructive airway diseases. Patients with sharp bend curves show changes of the several lung function data which are more advanced than in subjects with the intermediate forms of the flow-volume curves. In cases of bend curves the volume of the forced expiration can be differentiated on the two parts: circumferential and serial. Circumferential volume exhaled on the very beginning of the expiration (above the bend) amounts 0.118L in average. This volume depends on the expiratory narrowing of the bronchi from the 1-st to 9-th generation. The serial volume contained between the bend and the end of expiration amounts about 95% of the expired volume. Flow limitation occurs in 5-th to 9-th generations which is manifested by the strong increase of the flow resistance. The intermediate types of the flow-volume curves is caused by the inhomogenous emptying of the lung together with corresponding volume dependent narrowing of the bronchi. The same mechanisms can be detected even on the bodypletysmographic tidal breathing resistance curves. The concave and particularly bend flow-volume curves has been attributed to the pulmonary emphysema. This is not entirely truth. Other conditions leading to inhomogenic emptying of the lung due to airway and parenchymal changes (such as lung cicatrisation) can influence expiratory flow course resulting in concave or even bend flow-volume relationships.

[Correlation of radiologic and pathologic-anatomical findings in dust-induced pneumoconiosis in former coal miners]. / Korrelation radiologischer und pathologisch-anatomischer Befunde bei Mischstaub-Pneumokoniosen ehemaliger Kohlebergarbeiter.

The ILO classification of small and large opacities is the basis for the compensation of patients with pneumoconiosis. In order to validate the radiological findings, we compared the ILO classification with the gradings of pneumoconiosis in the post mortem investigation (n = 126). An ILO classification of 1/0 used as the threshold value for accepting the diagnosis of a pneumoconiosis was 100% sensitive with a specificity of 2%. With a value of 2/3 the sensitivity decreased to 60% and specificity increased to 74%. The grading of large opacities in the radiograph correlated well with the post mortem findings (rs = 0.71; p < 0.001). For accepting the diagnosis pneumoconiosis from the radiograph alone a threshold value of 1/0 is not specific. The ROC-plot revealed that specificity reached acceptable levels with an ILO classification of at least 2/3.

[Chronic bronchitis and its sequelae. Therapy--prognosis--insurance medicine aspects]. / Die chronische Bronchitis und ihre Folgen. Therapie--Prognose--Versicherungsmedizinische Aspekte.

Chronic bronchitis is of enormous epidemiological, socio-medical and economical importance. The main cause of chronic bronchitis is active but also passive cigarette smoking. Other etiologic factors are: viral or bacterial infections, chronic dust exposure in occupational settings and air pollution. Cease of cigarette smoking is considered the most valuable action in the treatment of the chronic bronchitis. In addition to drug therapy, physical therapy and other measures of rehabilitation may be a supportive benefit. ABout 10-20% of all patients with chronic bronchitis develop airway obstruction and/or lung emphysema. Obstructive bronchitis with or without lung emphysema should be treated with corticosteroids, beta 2-agonists and/or theophylline. Evidence for the socio-economical burden of chronic bronchitis and its complications are the enormous costs for the social economy (direct costs: in- and outpatient treatment; indirect costs: premature pensions, sick leave).

Makaluvamines H-M and damirone C from the pohnpeian sponge Zyzzya fuliginosa.

Seven new pyrroloiminoquinone alkaloids, makaluvamines H-M [19-24] and damirone C[25], together with the known compounds, makaluvamines C[13], D[14], and G[17], were isolated from the sponge Zyzzya fuliginosa collected at Nahpali Island, Pohnpei, Micronesia. The structures of the new compounds were elucidated by interpretation of spectral data. The chemotaxonomic relationships involving the makaluvamines and related pyrroloiminoquinone alkaloids are discussed.