A class C CpG toll-like receptor 9 agonist successfully induces robust interferon-alpha production by plasmacytoid dendritic cells from patients chronically infected with hepatitis C.

Immunomodulators that induce local endogenous interferon-alpha (IFN-alpha) production by plasmacytoid dendritic cells (pDCs) may offer new strategies for the treatment of patients chronically infected with the hepatitis C virus (HCV). However, such an approach may be compromised if reports are true that IFN-alpha production by pDCs from patients with chronic HCV (cHCV) is profoundly impaired. To address the question of pDC dysfunction in cHCV more definitively, in the present study a panel of four prototypic synthetic agonists of toll-like receptor 7 (TLR7) or TLR9 were administered in vitro to pDCs purified from cHCV patients and from normal uninfected donors and their responses compared in terms of not only IFN-alpha production but also the global expression of other cytokines and phenotypic maturation. Plasmacytoid DCs from uninfected donors produced substantial levels of IFN-alpha in response to three of the four agonists and yet only one TLR9 agonist, a class C CpG oligodeoxynucleotide (ODN), induced robust IFN-alpha production by pDCs from cHCV patients. Proinflammatory cytokine production and phenotypic maturation in response to all four agonists was equivalent in infected and uninfected pDCs. These data point to a profound but selective defect in IFN-alpha production by pDCs from cHCV donors. Nonetheless, a class C CpG ODN successfully induced robust IFN-alpha production, suggesting that this class of TLR9 agonist may have utility as a future immunotherapeutic for the treatment of chronic HCV infection.

A novel, helminth-derived immunostimulant enhances human recall responses to hepatitis C virus and tetanus toxoid and is dependent on CD56+ cells for its action.

We have described previously an immunostimulant derived from Onchocerca volvulus, the helminth parasite that causes onchocerciasis. Recombinant O. volvulus activation-associated secreted protein-1 (rOv-ASP-1) was a potent adjuvant for antibody and cellular responses to protein, polypeptide and small peptide antigens. Our aims were to determine whether rOv-ASP-1 is immunostimulatory for human peripheral blood mononuclear cells (PBMC) and, if so, whether it could augment cellular responses against human pathogen antigens in vitro. Cytokines from rOv-ASP-1-stimulated human PBMC were measured by a fluorescence activated cell sorter-based multiplex assay. Recall responses of normal healthy donor (NHD) and chronic hepatitis C virus (c-HCV)-infected patient PBMC to tetanus toxoid (TT) or HCV core (HCVco) antigen, respectively, were measured by interferon-gamma enzyme-linked immunospot assays. Interferon-gamma was the predominant cytokine induced by rOv-ASP-1. 77.3% of NHD anti-TT and 88.9% of c-HCV anti-HCVco responses were enhanced by rOv-ASP-1. The immunostimulant effect was dependent upon contact between CD56+ and CD56- fractions of PBMC. We have described a helminth-derived protein that can act as an immunostimulant for human recall responses in vitro to TT and, perhaps more importantly, HCV antigens in patients with chronic HCV infection. Our longer-term goal would be to boost anti-viral responses in chronic infections such as HCV.

A mutant in Lycopersicon esculentum Mill. with highly reduced VA mycorrhizal colonization: isolation and preliminary characterisation.

This paper reports the successful isolation and preliminary characterisation of a mutant of Lycopersicon esculentum Mill. with highly reduced vesicular-arbuscular (VA) mycorrhizal colonization. The mutation is recessive and has been designated rmc . Colonization by G. mosseae is characterised by poor development of external mycelium and a few abnormal appressoria. Vesicles were never formed by this fungus in association with the mutant. Gi. margarita formed large amounts of external mycelium, complex branched structures and occasional auxiliary cells. Small amounts of internal colonization also occurred. Laser scanning confocal microscopy (LSCM) gave a clear picture of the differences in development of G. intraradices and Gi. margarita in mutant and wild-type roots and confirmed that the fungus is restricted to the root surface of the mutants. The amenability of tomato for molecular genetic characterisation should enable us to map and clone the mutated gene, and thus identify one of the biochemical bases for inability to establish a normal mycorrhizal symbiosis. The mutant represents a key advance in molecular research on VA mycorrhizal symbiosis.

Automated system for capture and detection of nucleic acids.

A fully automated nucleic acid analysis system is described, which offers positive sample identification, improved sensitivity and reduced user interaction compared to conventional techniques. The system relies on the sequence-specific capture of DNA onto solid-phase particles, confirming product identity without the problems of interpretation and lack of sequence information inherent in gel-based analyses. The system can be used for sequence confirmation, mutation analysis and semiquantitative detection of PCR products.

In vivo tumor oxygen tension measurements for the evaluation of the efficiency of photodynamic therapy.

Among the sequence of events which occur during photodynamic therapy (PDT) are depletion of oxygen and disruption of tumor blood flow. In order to more clearly understand these phenomena we have utilized transcutaneous oxygen electrodes to monitor tissue oxygen disappearance. These results provide, for the first time, non-invasive real-time information regarding the influence of light dose on tissue oxygenation during irradiation. Measurements were conducted on transplanted VX-2 skin carcinomas grown in the ears of New Zealand white rabbits. Rabbits were treated with Photofrin II and tumors were irradiated with up to 200 kJ/m2 (500 W/m2) of 630-nm light. Substantial reductions in tumor oxygen tension were observed upon administration of as little as 20 kJ/m2. For a series of brief irradiations, oxygen tension was modulated by the appearance of laser light. Tissue oxygen reversibility appeared to be dependent upon PDT dose. Long-term, irreversible tissue hypoxia was recorded in tumors for large (200 kJ/m2) fluences. These results suggest that transcutaneous oxygen tension may be useful as a general indicator of the effectiveness of PDT and as an in situ predictor of the energy required to elicit tumor damage.

A comparison of bag mask and mouth mask ventilation in anaesthetised patients.

Anaesthetic residents used bag valve mask (BVM) or mouth mask (MM) ventilation, both with an O2 flow of 15 l min-1 to ventilate 30 ASA I or II anaesthetised patients for 4 min prior to endotracheal intubation. Mean nasopharyngeal O2 was higher with BVM (BVM 95% (S.D. 3%) MM 54% (S.D. 12%)). End tidal CO2 (ETCO2) was similar in both groups (ETCO2% at 4 min: BVM 4.65 (S.D. 0.84) MM 4.53 (S.D. 0.54)) but respiratory rate was faster with BVM (BVM 17 min-1 (S.D. 5) MM 12 min-1 (S.D. 4)). Peak (Paw) and mean (Paw) airway pressures were higher with MM and MM produced significant expiratory pressure [cmH2O: BVM Paw 16.7 (S.D. 5.3) Paw 4.2 (S.D. 2.1) MM: Paw 20.9 (S.D. 5.2) Paw 7.8 (S.D. 2.1)) minimum expiratory pressure: MM 2.4 (S.D. 1.1) BVM 0.2 (S.D. 0.4). Gastric insufflation was detected in two MM and two BVM patients. This tended to be more severe with MM ventilation. Although MM ventilation has some important disadvantages it can be used effectively by resuscitators with little or no experience in its use.

Cardiovascular effects of anesthesia and operation.

Anesthesia and surgery have a wide range of effects on the cardiovascular system. Even in healthy patients having minor operations, anesthetic agents can cause significant cardiac depression and hemodynamic instability. Virtually all anesthetic agents have intrinsic myocardial depressant properties, although some may mask this with sympathetic stimulation. The vasodilatory effects of the volatile agents can result in serious hypotension when combined with this negative inotropy. In the patient with pre-existing cardiac disease, these cardiovascular anesthetic effects become much more serious. These patients will not tolerate wide swings of hemodynamic variables, and the cardiodepressant effects of anesthetics are more pronounced in them. The stress of anesthesia and surgery frequently unmasks previously undiagnosed heart disease. Surgery itself provides many insults to the cardiovascular system, and these may be additive with the effects of anesthesia. These include loss of blood and other volume shifts, release of various substances into the circulation, hypothermia, sudden changes in cardiac preload and afterload, myocardial ischemia, and effects of drugs or blood products given for surgical reasons. The signs and symptoms of these surgical stresses to the cardiovascular system are often masked by anesthesia.

Intravenous beta2-agonists for acute asthma in the emergency department.

BACKGROUND: Inhaled beta-agonist therapy is central to the management of acute asthma. The use of intravenous beta-agonist agents may also be beneficial in this setting. OBJECTIVES: To determine the benefit of intravenous (IV) beta2-agonists for severe acute asthma treated in the emergency department. SEARCH STRATEGY: Randomised controlled trials (RCT) were identified using the Cochrane Airways Group Register which is a compilation of systematic searches of MEDLINE, EMBASE, CINAHL, and CENTRAL as well as hand searching of 20 respiratory journals. Bibliographies from included studies and known reviews were also searched. Primary authors and content experts were contacted to identify eligible studies. SELECTION CRITERIA: Only RCTs were considered for inclusion. Studies were included if patients presented to the emergency department with acute asthma and were treated with IV selective or nonselective beta2-agonists versus placebo, inhaled beta2-agonists, or other standard of care. Pulmonary function, vital signs, arterial gasses, adverse effects, and/or clinical success could be reported as outcome measures. Two reviewers independently selected potentially relevant articles and selected articles for inclusion. Methodological quality was independently assessed using two scoring systems and two reviewers. DATA COLLECTION AND ANALYSIS: Data were extracted independently by two reviewers, and confirmed with corresponding authors. Missing data were obtained from authors or calculated from data present in the papers. Trials were combined using a random effects model for odds ratios (OR) or weighted mean differences (WMD) and reported with 95% confidence intervals (95% CI). MAIN RESULTS: From 746 identified references, 55 potentially relevant articles were identified and 15 were included. The trials included 584 patients. Overall, selective IV beta2-agonist use conferred no advantage over the comparator regimes. For example, it was associated with a lower PEFR after 60 mins compared to inhaled beta2-agonist, although the difference was not statistically significant (-24.7 l/min; 95%CI 2.9, -52.3). There was no difference in heart rate (4.5 bpm; 95% CI -4.9, 14.0). In the well performed blinded studies there was no difference in autonomic side effects between treatments (Odds Ratio 2.2 (95%CI 0.9, 5.7). REVIEWER'S CONCLUSIONS: There is no evidence to support the use of IV beta2-agonists in patients with severe acute asthma. These drugs should be given by inhalation. No subgroups were identified in which the IV route should be considered.

Effects of motion, ambient light, and hypoperfusion on pulse oximeter function.

STUDY OBJECTIVE: To compare the performance of five pulse oximeters during hypoperfusion, probe motion, and exposure to ambient light interference. DESIGN: Prospective study. SETTING: Laboratory facility at a university medical center. PATIENTS: 8 unanesthetized, ASA physical status I volunteers. INTERVENTIONS: We evaluated five common pulse oximeters with respect to three scenarios: (1) an operating room light was shone on oximeter probes, (2) a motion generator was used to generate 2 Hz and 4 Hz hand motion, and (3) a pneumatic compression device overlying the brachial artery was used to simulate hypoperfusion. Electrocardiographic (ECG) and arterial blood gas values were considered gold standards for heart rate (HR) and oxygen saturation (SpO2) respectively. SpO2 nondisplay and values greater than 4% from simultaneous arterial SaO2-oximeter values were defined as errors. Nondisplay of HR, or HR greater than 5% from ECG values, were also considered errors. MEASUREMENTS AND MAIN RESULTS: The Ohmeda and Nellcor N200 with finger probe had the highest total failure rates with respect to both SpO2 and HR due to ambient light interference (p < 0.05). The Nellcor N200 with finger probe and N200 with C lock were the most accurate with regard to SpO2 during 2 Hz and 4 Hz motion (p < 0.05). However, all oximeters failed dramatically during 4 Hz motion when measuring HR. In the hypoperfusion model, the Nellcor N200 with finger probe and the Nellcor C Lock oximeters performed significantly better than all others in terms of both HR and SpO2 (P < 0.05), while the Criticare oximeter failed 100% of the time. CONCLUSION: There are significant differences in the accuracy of commercially available pulse oximeters during nonideal circumstances, with failure rates varying from approximately 5% to 50% depending on the oximeter and source of interference. Furthermore, no single oximeter performed the best under all conditions.

Pulse oximeter performance during desaturation and resaturation: a comparison of seven models.

STUDY OBJECTIVE: To compare pulse oximeter performance during induced hypoxemia. DESIGN: Prospective investigation in human volunteers. SETTING: Laboratory facility at a university medical center. PATIENTS: 8 unanesthetized, healthy ASA physical status I volunteers. INTERVENTIONS: We evaluated the accuracy and response times of seven popular pulse oximeters during induced hypoxemia. Arterial blood fractional oxygen saturation (SaO2) measurements were performed simultaneously and considered a gold standard. MEASUREMENTS AND MAIN RESULTS: All oximeters were accurate (+/-2%) while subjects were breathing room air. During maximal hypoxemia (induced by breathing a FIO2 = 10% in nitrogen), large differences were noted between oxygen saturation as measured by pulse oximetry (SpO2) and SaO2 values, with pulse oximeters consistently underreporting SpO2 when actual SaO2 values were 75% or less. The Ohmeda 3740 (Ohmeda, Boulder, CO) using an ear probe was the first to detect desaturation (change in SpO2 > 3%) in 4 of 8 subjects (p < 0.05), and the Nellcor N200 reflectance oximeter (Nellcor, Inc., Pleasanton, CA) was first in 3 of 8 subjects (p < 0.05). During resaturation (after administering 100% oxygen), the Novametrix Oxypleth (Novametrix, Wallingford, CT) was significantly faster than other oximeters (p < 0.05) to return to baseline (SpO2 = 98%). CONCLUSION: Most models of oximeters tested performed well when hemoglobin oxygen saturation was high, but all were inaccurate when SaO2 was approximately 75%. During induced hypoxemia, there were significant differences in the response times of oximeters tested, with no model demonstrably superior to others in all measures of performance.

Incidence of perioperative myocardial ischemia in TURP patients.

STUDY OBJECTIVE: To determine the incidence of new episodes of myocardial ischemia in patients undergoing transurethral resection of the prostate (TURP). DESIGN: Prospective, nonrandomized study. SETTING: Veterans Administration medical center. PATIENTS: 39 patients undergoing elective TURP. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Myocardial ischemia was detected with a 3-channel ambulatory ECG recorded. The ambulatory ECG recorder was applied preoperatively and removed when the patient left the recovery room. New myocardial ischemia was defined as a 1 mm or greater ST depression or a 2 mm or greater ST elevation from baseline, lasting for 1 minute or longer in at least one lead at the J point plus 60 msec unless this point fell within the T wave, in which case the J point 40 msec or greater was used. ST changes consistent with myocardial ischemia were confirmed by a cardiologist blinded to the patient's clinical course. Seven of 39 TURP patients (18%) had ST segment changes indicative of new myocardial ischemia. These seven patients had more prostate tissue resected and more blood loss than the 32 patients who did not have any myocardial ischemia (p < 0.05). CONCLUSIONS: Patients undergoing TURP have an 18% incidence of myocardial ischemia. Patients undergoing TURP with more prostate tissue resected and greater blood loss are at increased risk for perioperative myocardial ischemia.

Lord or vassal? Academic anesthesiology finances in 2000.

UNLABELLED: This article examines recent trends in the management of academic physician practice groups, and in particular the allocation of revenues and expenses to anesthesiology departments. The history of academic group practice is traced, beginning with the "corporate model," in which each department functioned in financial independence from the others. This evolved gradually into the "feudal system," in which departments were ostensibly independent, but paid variable and often large "assessments" to the central group. The final stage in this evolution is the "big bag," in which all clinical revenue is pooled by the central practice group, and then distributed by the group to departments or individuals according to some compensation plan formula. The advantages and disadvantages of each of these systems are discussed as they apply to anesthesiology departments. A productivity-based compensation plan formula under the big bag system is calculated for a typical anesthesiology department. This calculation shows that if the compensation formula is truly based on measured clinical productivity, anesthesiology departments may actually fare better under the big bag than under the feudal system. Finally, options for survival in the academic practice groups of the future are discussed. IMPLICATIONS: The history, current status, and trends of finances in academic anesthesiology departments are reviewed. Knowledge of these issues will help departments develop funds allocation methods to ensure that they receive an appropriate share of their faculty practice group's clinical income.

Continuous measurement of intraarterial pHa, PaCO2, and PaO2 in the operating room.

Miniaturized sensors based upon the principles of optical fluorescence can measure the pH, PCO2, and PO2 of liquid or gas media. A prototype of a three-component fiberoptic sensor has been developed for intraarterial application by CDI, 3M Health Care, Irvine, California. We report the first study of this continuous intraarterial monitor in patients undergoing surgical procedures under general anesthesia. Fourteen patients participated in the study. The fiberoptic sensor was calibrated before insertion and then passed through an existing 18-gauge radial artery cannula. Blood samples were drawn at frequent intervals through the same cannula for in vitro blood gas analysis. For each of the 87 arterial blood gas samples obtained, the in vitro values of pHa, PaCO2, and PaO2 were compared with simultaneous readings from the fiberoptic sensor. For pHa, the mean error (error = fiberoptic value minus in vitro value) or "bias" of the fiberoptic data was -0.032 and the standard deviation of error or "precision" was 0.042. For PaCO2, the bias was -3.8 mm Hg and the precision was 4.7 mm Hg. For PaO2, the bias was -9.0 mm Hg and the precision was 23.3 mm Hg. For PaO2 values less than 175 mm Hg, the bias was -8.5 mm Hg and the precision was 8.3 mm Hg. Expressed in terms of percentage errors, the bias +/- precision values were -11.5% +/- 13.3% for PaCO2, and -6.2% +/- 10.0% for PaO2. The duration of the surgical procedures ranged from 1.6 to 8 h with an average of 4.2 h.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of carbon monoxide inhalation on pulse oximetry and transcutaneous PO2.

Five dogs were anesthetized, intubated, and ventilated with various mixtures of oxygen, nitrogen, and carbon monoxide. Each dog was monitored with arterial and pulmonary artery catheters, a transcutaneous PO2 analyzer, and two pulse oximeters. An IL-282 Co-oximeter was used to periodically measure arterial oxyhemoglobin (O2Hb) and carboxyhemoglobin (COHb) as percentages of the total hemoglobin. The PaO2, PaCO2, and pHa were measured in the same blood specimens using standard electrodes. When the inspired oxygen concentration was reduced in the absence of COHb, the pulse oximeter saturation (SpO2) estimated O2Hb with reasonable accuracy. COHb levels were then varied slowly from 0-75% in each dog. As the COHb level increased and oxyhemoglobin decreased, both pulse oximeters continued to read an oxygen saturation of greater than 90%, while the actual O2Hb fell below 30%. In the presence of COHb, the SpO2 is approximately the sum of COHb and O2Hb, and, thus, may seriously overestimate O2Hb. The pulse oximeter, as the sole indicator of blood oxygenation, should, therefore, be used with caution in patients with recent carbon monoxide exposure. On the other hand, transcutaneous PO2 falls linearly as COHb increases, and reaches about one-fifth of its initial value at the highest COHb levels despite the maintenance of constant arterial PO2.

Pulse oximetry: applications and limitations.

The pulse oximeter estimates arterial hemoglobin saturation by measuring the light absorbance of pulsating vascular tissue at two wavelengths. The relationship between measured light absorbances and saturation was developed empirically and is built into the oximeter software. Studies in human volunteers have shown good performance of the device in healthy adults for saturations in the range of 70 to 100%. Studies in the operating room and intensive care unit have established its clinical accuracy and usefulness. The pulse oximeter has already found a number of clinical applications outside of the operating room, such as monitoring during patient transport, respiratory monitoring during narcotic administration, and evaluation of home-oxygen therapy. To use this monitor to its full potential, we must be aware of its limitations as well as its advantages. Because of the nature of the HbO2 dissociation curve, saturation measurements will not be sensitive to changes in PaO2 when the PaO2 is greater than 100 torr. This also implies that the pulse oximeter may fail to detect an inadvertent endobronchial intubation in the operating room. It may take minutes to detect an esophageal intubation in a well-preoxygenated patient. When desaturation does occur, the pulse oximeter detects it quickly, accurately, and reliably. Since the pulse oximeter uses two wavelengths of light, it cannot distinguish more than two hemoglobin species. Thus, COHb and MetHb will cause errors in SpO2 if present in large amounts. Intravenously administered dyes can also cause errors because of their absorbance properties, particularly methylene blue and indocyanine green. The pulse oximeter may be unable to detect an adequate signal during abnormal hemodynamic conditions. The pulse oximeter is one of the most important advances in noninvasive monitoring because it provides a means of continuously and quickly assessing arterial blood oxygenation. It is easy to use and interpret, requires little setup time, and poses no additional risks to the patient. Pulse oximetry may soon be a standard of practice for routine monitoring in any clinical setting in which the patient is at risk of hypoxemia.

Transcutaneous oxygen measurement: experimental studies and adult applications.

Transcutaneous PO2 sensors have been developed over the past ten years from the same basic electrodes used in conventional blood-gas machines. The skin is heated to enable the skin surface sensors to respond quickly to the gas tensions beneath them. PtcO2 is a variable that reflects the PO2 in the peripheral tissue. PtcO2 has its own range of normal values, and it responds to cardiopulmonary changes that affect tissue oxygenation. In most patients, those without decreased cardiac output, PtcO2 follows the trend of PaO2 and decreases relative to PaO2 with increasing patient age (see Table 2). In the presence of severely reduced cardiac output and peripheral perfusion, the PtcO2 values will deviate from their relationship with the arterial tensions and become blood flow dependent, thus providing quantitative information regarding blood flow. The technique of PtcO2 monitoring likely will gain wider acceptance because it is a noninvasive and continuous monitor that provides useful information regarding tissue oxygenation.