Detection of type III secretion genes as a general indicator of bacterial virulence

Type III secretion systems of Gram-negative bacteria are specific export machineries for virulence factors which allow their translocation to eukaryotic cells. Since they correlate with bacterial pathogenicity, their presence is used as a general indicator of bacterial virulence. By comparing the genetic relationship of the major type III secretion systems we found the family of genes encoding the inner-membrane channel proteins represented by the Yersinia enterocolitica lcrD (synonym yscV) and its homologous genes from other species an ideal component for establishing a general detection approach for type III secretion systems. Based on the genes of the lcrD family we developed gene probes for Gram-negative human, animal and plant pathogens. The probes comprise lcrD from Y. enterocolitica, sepA from enteropathogenic Escherichia coli, invA from Salmonella typhimurium, mxiA from Shigella sonnei, as well as hrcV from Erwinia amylovora. In addition we included as a control probe the flhA gene from E. coli K-12 to validate our approach. FlhA is part of the flagellar export apparatus which shows a high degree of similarity with type III secretions systems, but is not involved in pathogenicity. The probes were evaluated by screening a series of pathogenic as well as non-pathogenic bacteria. The probes detected type III secretion in pathogens where such systems were either known or were expected to be present, whereas no positive hybridization signals could be found in non-pathogenic Gram-negative bacteria. Gram-positive bacteria were devoid of known type III secretion systems. No interference due to the genetic similarity between the type III secretion system and the flagellar export apparatus was observed. However, potential type III secretion systems could be detected in bacteria where no such systems have been described yet. The presented approach provides therefore a useful tool for the assessment of the virulence potential of bacterial isolates of human, animal and plant origin. Moreover, it is a powerful means for a first safety assessment of poorly characterized strains intended to be used in biotechnological applications.     

Pharmacokinetics of 10 mg of mifepristone

The results of several randomized studies have verified the efficacy of 10 mg mifepristone in emergency contraception. In the present study we characterized the pharmacokinetics of 10 mg mifepristone. Eight healthy female volunteers received a single oral dose of mifepristone on the day 10 or 11 of their menstrual cycle. Blood samples were collected at 0, 1, 2, 4 and 8 h, daily for the next 6 days and on day 10 after mifepristone. Mifepristone concentrations were determined by radioimmunoassay preceded by column chromatography. A peak level of 1.41 ± 0.31 μmol/L (mean ± SD) was measured at 1 h. Individual elimination phase half-lives varied from 15.3 to 26.8 h, the mean (± SD) value being 19.6 ± 4.50 h. Serum mifepristone concentrations exceeded 10 nmol/L in all volunteers for an average of 4.9 days. The pharmacokinetic data on 10 mg mifepristone are in line with previous pharmacokinetic and clinical data, and encourage further development of the 10-mg dose in emergency contraception.     

Contrast adaptation in human retina and cortex

PURPOSE: Although cortical contrast adaptation has been extensively studied with both psychophysical and electrophysiological techniques, little is known about retinal contrast adaptation in humans. METHODS: Retinal and cortical long-term contrast adaptation was assessed with simultaneous measurement of pattern electroretinogram (PERG) and cortical visual evoked potentials (VEPs). This study involved three approaches: sampling of the contrast transfer function from 2.7% to 98% with adaptation to high (98%) and low (7.3%) contrasts, linearity of adaptation effects, and transfer of contrast adaptation between parallel and orthogonal grating orientations. RESULTS: Contrast adaptation affected retinal and cortical recordings quite differently. The VEP showed a sigmoid contrast transfer function, which was shifted toward higher contrasts (by a factor of 1.9), whereas amplitudes at higher test contrasts were enhanced to 127%. The PERG decreased in amplitude to approximately 90%, and the latency was significantly reduced by 4 to 6 msec (P < 0.05). All measured effects were linear with adaptation contrast. Orientation played no role in the PERG results, whereas the VEP was enhanced to 125% when tested parallel and to 150% when tested orthogonal to adaptation. CONCLUSIONS: VEP results confirm and extend previous findings and fit well with single-cell recordings. The PERG findings suggest that retinal contrast adaptation occurs and mainly operates in the temporal domain, comparable to rapid gain-control findings in cats and primates.     

Fiberoptic Bronchoscopy of Intubated Patients with Life-threatening Hemoptysis

Bleeding in the tracheobronchial tree in intubated patients on an intensive care unit is a potentially life-threatening incident. The antecedent state of disease and frequent respiratory failure require immediate and effective therapeutic measures to avoid further respiratory and cardiocirculatory depression. We present our bronchoscopic management of endobronchial bleeding. Cardiorespiratory function must be maintained by modification of the mechanical ventilation and drug therapy owing to the patient's condition. Seven consecutive patients with acute endobronchial bleeding were treated with fiberoptic bronchoscopy and instillation of cold epinephrine-saline solution (1:10,000–100,000) during the period of July 1997 to December 1997. Control of bleeding was achieved after 1 to 20 (mean ± SEM: 5.86 ± 0.93) bronchoscopic interventions during a period of 0.5 hours to 10 days. One control bronchoscopy was performed additionally in every patient. Cardiocirculatory instability was observed in five patients. Six patients survived; one patient died of uncontrolled bleeding caused by severe pulmonary aspergillosis. Fiberoptic endobronchial epinephrine instillation is an effective therapy for life-threatening hemoptysis in critically ill patients. Widespread use of flexible bronchoscopy makes this procedure immediately applicable in critical situations. Intubated and mechanically ventilated patients with life-threatening hemoptysis especially benefit from this rapidly feasible procedure.     

The stress of birth enhances in vitro spontaneous and IL-8-induced neutrophil chemotaxis in the human newborn

The birth process induces fetal stress. Stress has profound effects on the immune system, also by acting on the trafficking of leukocytes, a process in which adhesion and chemotaxis are primordial and critical events for the development of effective antimicrobial defenses. The newborn is rapidly challenged by a microflora at the epithelia linings and therefore depending on early, innate immunity onset. The objective of the study was to investigate the immune response in cord blood from newborns in relation to different degrees of fetal stress, with focus on neutrophil chemotaxis. We analyzed in vitro transmigration ability of neutrophils and their CD11b expression, measured total white blood count (WBC) and the major leukocyte populations, interleukin (IL)-8, interferon (IFN)-gamma, and soluble E-Selectin, as well as relevant immuno-modulating hormones in infants born at term after Cesarean section prior to the start of labor (n = 55), normal vaginal delivery (n = 87), and assisted delivery (n = 26). Arterial pH and lactate were used as stress markers. We found that spontaneous and IL-8-induced transmigration ability of neutrophils from newborns after normal delivery was significantly higher compared with that of neutrophils from Cesarean section or from adults. With a progressive increase in fetal stress, there were significant elevations in total WBC, in particular neutrophils and monocytes, as well as an enhanced IL-8 and soluble E-Selectin level. Assisted delivery, associated with the highest degree of fetal stress in addition had an enhanced lymphocyte and monocytes count as well as an increased IFN-gamma level. There were significant direct correlations between neutrophils and monocytes, respectively, with cortisol, beta-endorphin, and prolactin. Interferon-gamma was directly related to dopamine, as well as to the lymphocyte and monocyte count. The setting of the HPA-axis at birth is a promoter of an alarm response and a surge of neuroendocrine immuno-modulating factors that enhances antimicrobial defenses of the newborn. We speculate that IL-8 induced by normal labor may be a priming factor for an increased neutrophil chemotaxis through the pre-activated endothelium of the fetus. Assisted delivery may trigger excessive recruitment of additional inflammatory cells and IFN-gamma release.     

Automatic evaluation of prosodic features of tracheoesophageal substitute voice

In comparison with laryngeal voice, substitute voice after laryngectomy is characterized by restricted aero-acoustic properties. Until now, an objective means of prosodic differences between substitute and normal voices does not exist. In a pilot study, we applied an automatic prosody analysis module to 18 speech samples of laryngectomees (age: 64.2 ± 8.3 years) and 18 recordings of normal speakers of the same age (65.4 ± 7.6 years). Ninety-five different features per word based upon the speech energy, fundamental frequency F₀ and duration measures on words, pauses and voiced/voiceless sections were measured. These reflect aspects of loudness, pitch and articulation rate. Subjective evaluation of the 18 patients’ voices was performed by a panel of five experts on the criteria “noise”, “speech effort”, “roughness”, “intelligibility”, “match of breath and sense units” and “overall quality”. These ratings were compared to the automatically computed features. Several of them could be identified being twice as high for the laryngectomees compared to the normal speakers, and vice versa. Comparing the evaluation data of the human experts and the automatic rating, correlation coefficients of up to 0.84 were measured. The automatic analysis serves as a good means to objectify and quantify the global speech outcome of laryngectomees. Even better results are expected when both the computation of the features and the comparison method to the human ratings will have been revised and adapted to the special properties of the substitute voices.     

Cardiotocography -- proposal of system solution (author's transl)]

A system solution with new technical details and new demands on space distribution is proposed for the use of antenatal cardiotocography and cardiotocographic monitoring of labour. Bedside equipment will be reduced and widely shifted to a centralised control room by introduction of the system. All installations are of high flexibility to leave room for future developments in medical engineering and for necessary services. -- Large-scale use of the new technique with capacity utilisation of the hardware involved will be ensured by appropriate upgrading of labour room personnel as well as by techno-organisational improvement of work and minimisation of expenses on diagnosis and monitoring. The measures proposed work according to expectation and lead to good perinatological results.