Prediction of cortical theta oscillations in humans for phase-locked visual stimulation.

BACKGROUND: The timing of an event within an oscillatory phase is considered to be one of the key strategies used by the brain to code and process neural information. Whereas existing methods of studying this phenomenon are chiefly based on retrospective analysis of electroencephalography (EEG) data, we now present a method to study it prospectively. New method: We present a system that allows for the delivery of visual stimuli at a specific phase of the cortical theta oscillation by fitting a sine to raw surface EEG data to estimate and predict the phase. One noteworthy feature of the method is that it can minimize potentially confounding effects of previous trials by using only a short sequence of past data. RESULTS: In a trial with 10 human participants we achieved a significant phase locking with an inter-trial phase coherence of 0.39. We demonstrated successful phase locking on synthetic signals with a signal-to-noise ratio of less than - 20 dB. Comparison with existing method(s): We compared the new method to an autoregressive method published in the literature and found the new method was superior in mean phase offset, circular standard deviation, and prediction latency. CONCLUSIONS: By fitting sine waves to raw EEG traces, we locked visual stimuli to arbitrary phases within the theta oscillatory cycle of healthy humans.

Real-Time RT-PCR for the Detection of Lyssavirus Species.

The causative agents of rabies are single-stranded, negative-sense RNA viruses in the genus Lyssavirus of Rhabdoviridae, consisting of twelve classified and three as yet unclassified species including classical rabies virus (RABV). Highly neurotropic RABV causes rapidly progressive encephalomyelitis with nearly invariable fatal outcome. Rapid and reliable diagnosis of rabies is highly relevant for public and veterinary health. Due to growing variety of the genus Lyssavirus observed, the development of suitable molecular assays for diagnosis and differentiation is challenging. This work focused on the establishment of a suitable real-time RT-PCR technique for rabies diagnosis as a complement to fluorescent antibody test and rabies tissue culture infection test as gold standard for diagnosis and confirmation. The real-time RT-PCR was adapted with the goal to detect the whole spectrum of lyssavirus species, for nine of which synthesized DNA fragments were used. For the detection of species, seven probes were developed. Serial dilutions of the rabies virus strain CVS-11 showed a 100-fold higher sensitivity of real-time PCR compared to heminested RT-PCR. Using a panel of thirty-one lyssaviruses representing four species, the suitability of the protocol could be shown. Phylogenetic analysis of the sequences obtained by heminested PCR allowed correct classification of all viruses used.

[Microcirculatory monitoring of sepsis]. / Mikrozirkulatorisches Monitoring der Sepsis.

Microcirculatory dysfunctions play a central role in the pathophysiology of sepsis and shock. Modern methods enable microvascular monitoring in man and offer the possibility to test the effect of novel therapeutical strategies for sepsis. Furthermore, these techniques may be future tools for the monitoring of critically ill patients. In this review, we will describe four microvascular monitoring devices and give an overview of the microcirculatory changes observed during the course of sepsis. Laser Doppler fluxmetry is an easy to use noninvasive technique to measure tissue perfusion enabling monitoring of the effect of different catecholamines on the gastric perfusion during sepsis. Increased microvascular permeability and altered blood flow in septic patients can be quantified by venous congestion plethysmography. Alterations in sublingual microvascular blood flow are detected by intravital microscopy in septic patients and were identified as an outcome predictor. Furthermore, the role of gastrointestinal pCO2-tonometry for microcirculatory monitoring of the perfusion of splanchnic organs during sepsis is discussed. The true clinical value of these techniques has yet to be established and will depend on larger clinical trials showing an impact on diagnostics and patient management.

Influence of different cuff inflation protocols on capillary filtration capacity in human calves -- a congestion plethysmography study.

It has been suggested that venous congestion plethysmography (VCP) substantially underestimates microvascular permeability by activation of a veni-arteriolar constrictor mechanism, even when using small (< 25 mmHg) congestion pressure steps. We studied human lower limbs of 18 young healthy volunteers to test whether the congestion pressure step size of the VCP protocol has an influence on the values of the capillary filtration capacity (CFC) and isovolumetric venous pressure (P(vi)). Two different dual stage VCP pressure step protocols, with 3 and 10 mmHg steps, were used in randomised order and separated by a transient reduction in congestion pressure. Since lymph flow is known to increase after venous congestion, we also looked to see if changes in the estimated lymph flow (J(v)L) occur as a result of these VCP protocols. The measured CFC (median [25th; 75th percentile]) was 2.6 [2.5; 3.2] x 10(-3) ml (100 ml)(-1) min(-1) mmHg(-1) with the 3 mmHg pressure step protocol, which was not different from the value of 2.9 [2.7; 3.4] x 10(-3) ml (100 ml)(-1) min(-1) mmHg(-1) obtained with 10 mmHg pressure steps. However, when either of these step sizes was applied after a transient venous decongestion, significantly higher values of CFC, 4.0 [3.4; 4.1] x 10(-3) and 3.5 [3.1; 4.5] x 10(-3) ml (100 ml)(-1) min(-1) mmHg(-1), respectively, were obtained (P < 0.05). The assessment of P(vi) was also independent of the pressure protocol (10 mmHg: 8.0 [5.7; 13.2] mmHg and 3 mmHg: 15.7 [12.5; 18.5] mmHg), but when P(vi) was measured after the transient deflation, significantly higher values were found with both 10 and 3 mmHg steps (24.1 [20.9; 27.3] and 30.4 [28.9; 30.9] mmHg, respectively; P < 0.01). The transient pressure reduction was associated with a rise in estimated J(v)L from 0.04 [0.03; 0.05] to 0.12 [0.08; 0.18] and 0.04 [0.04; 0.05] to 0.09 [0.07; 0.10] ml (100 ml)(-1) min(-1), respectively (P < 0.01). The first stage data from these protocols shows that the value of CFC is not influenced by the size of the cumulative venous pressure steps, providing they are of 10 mmHg or less. The data also show that J(v)L can be estimated with small step VCP protocols. We hypothesise that the sudden reduction in cuff pressure after venous congestion is associated with a temporary upregulation of lymph flow. As the congestion pressure is raised again, there is a modulation of the enhanced lymph flow, such that the resulting CFC slope appears greater than that obtained in the first stage of the protocol.

Microvascular changes during anesthesia: sevoflurane compared with propofol.

BACKGROUND: We have developed a non-invasive computer-assisted venous congestion plethysmograph to measure the microvascular parameters in the lower limbs. This enables the assessment of microvascular changes following the induction of standardized anesthesia with either sevoflurane or propofol. METHODS: In a prospective randomized trial we measured the capillary filtration coefficient (CFC), isovolumetric venous pressure (Pvi), an index of the balance of Starling forces, and limb blood flow 24 h preoperatively, immediately after induction of anesthesia and on the 1st and 2nd postoperative day. Anesthesia was maintained with either 1.0% sevoflurane and 5 microg/kg/h remifentanil or propofol (3 mg/kg/h), and 5 microg/kg/h remifentanil in 20 female patients undergoing breast surgery. RESULTS: Preoperatively we found no significant differences between the mean CFC values of the sevoflurane group (3.7+/-0.3 ml/min 100 ml tissue/mmHg x 10-3=CFCU) and the propofol group (3.5+/-0.3 CFCU). In the sevoflurane group CFC decreased significantly to 2.9+/-0.2 CFCU, whereas it was unchanged in the propofol group. Both groups revealed a significant reduction in Pvi during steady-state anesthesia. Limb blood flow remained unchanged. There was an overall significant positive correlation between the perioperative fluid substitution and the difference between the preoperative and intraoperative CFC values (r = 0.64, P<0.01). CONCLUSION: The decreased CFC in response to sevoflurane may result in less extravasation of fluids into the interstitial space, thereby reducing intraoperative fluid requirements. These data suggest that sevoflurane may be the preferred anesthetic agent in subjects susceptible to large intraoperative fluid shifts.

Characterization of human papillomavirus in airway papillomas by histologic and biochemical analysis.

The role of human papillomavirus (HPV) in airway papillomas has been well defined in recent literature. The chronicity and recurrence of papillomas has been postulated to be a result of residual viral genome in tissue treated with standard laser techniques. Thirteen patients with airway papillomas were selected for study with polymerase chain reaction (PCR) methods to detect viral DNA. Specimens taken prior to laser therapy and specimens taken at laser margins were consistently positive for HPV DNA by PCR. The HPV DNA is apparently present in tissues after macroscopic disease has been ablated by laser techniques. Histologic analysis of laser biopsies demonstrated fragments of squamous epithelium with cytologic features of HPV infection. Laser treatment is ineffective in eradicating HPV-infected tissues from airway papillomas, and this finding supports the notion that recurrence is a product of HPV incorporated into tissue not ablated by laser irradiation. Specific methods, results, and clinical correlation will be discussed.

Characterization of cyclooxygenase in laryngeal papilloma by molecular techniques.

OBJECTIVES: Demonstrate the induction of cyclooxygenase-2 (COX-2) in laryngeal papilloma Discuss the possible causal role of COX-2 in papilloma formation. Consider the potential for treatment of papilloma using selective COX-2 inhibitors. STUDY DESIGN: Molecular biological analysis of COX-1 and COX-2 in laryngeal papilloma. METHODS: Tissue samples from five patients with recurrent respiratory papillomatosis (RRP) were analyzed by in situ hybridization, immunohistochemical staining, and reverse transcription polymerase chain reaction (RT-PCR) techniques. RESULTS: In situ hybridization to COX-2 mRNA showed strong autoradiographic signal surrounding fibrovascular cores. COX-1 autoradiographic signal was low intensity or nondetectable. Normal buccal mucosa biopsies showed low-density or nondetectable autoradiographic signal for both COX-1 and COX-2 mRNAs. In situ hybridization results were corroborated by RT-PCR studies. Levels of COX-2 mRNA were 13-fold more than those in normal mucosa. Immunohistochemical staining for COX-1 and COX-2 showed a similar pattern to that seen with in situ hybridization in both normal and papilloma tissues. CONCLUSIONS: There is an elevation of COX-2 expression in papilloma tissues. This may represent a causal role of COX-2 in the formation and proliferation of laryngeal papilloma. There may also be a role for selective COX-2 inhibition for the treatment of

Third branchial anomalies. Avoiding recurrences.

Lateral cervical cysts, sinuses, and fistulas have been described as anomalies of the normal development of the branchial apparatus. Third branchial apparatus anomalies are rare and constitute less than 1% of all such cases. Three cases of third branchial cleft cysts and sinus tracts are presented. Two patients had previously undergone multiple attempts at extirpation. Complete removal of recurrent branchial anomalies is difficult because of scarring and fascial plane disruption. Recurrences were often the result of inadequate excision, possibly of the tract communicating with the piriform sinus. To avoid this we advocate endoscopy prior to initial resection of a suspected branchial cleft anomaly to identify any pharyngeal communication. A combined, simultaneous endoscopic identification of the piriform sinus tract with a lateral external cervical dissection facilitates complete resection. In recurrent cases, wide-field extirpation of the cyst, tract, and scar tissue is necessary to ensure complete removal of the branchial cleft anomaly. A review of the literature and of branchial apparatus embryology is also presented.