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BACKGROUND: Chronic fatigue syndrome (CFS) is an illness of unknown origin that may have familial risks. Low natural killer (NK) lymphocyte activity was proposed as a risk for familial CFS in 1998. Since then, there have been many studies of NK lymphocytes in CFS in general populations but few in familial CFS. Antibody-dependent cell-mediated cytotoxicity (ADCC) by NK lymphocytes helps control viral infections. ADCC is affected by variant CD16A receptors for antibody that are genetically encoded by FCGR3A. METHODS: This report characterizes ADCC effector NK cell numbers, ADCC activities, and FCGR3A variants of five families each with 2–5 CFS patients, their family members without CFS and unrelated controls. The patients met the Fukuda diagnostic criteria. We determined: CD16Apositive blood NK cell counts; EC50s for NK cell recognition of antibody; ADCC lytic capacity; FCGR3A alleles encoding CD16A variants, ROC tests for biomarkers, and synergistic risks. RESULTS: CFS patients and their family members had fewer CD16Apositive NK cells, required more antibody, and had ADCC that was lower than the unrelated controls. CFS family members were predominantly genetically CD16A F/F s for the variant with low affinity for antibodies. ROC tests indicated unsuitability of ADCC as a biomarker for CFS because of the low ADCC of family members without CFS. Familial synergistic risk vs. controls was evident for the combination of CD16Apositive NK cell counts with ADCC capacity. CONCLUSIONS: low ADCC may be a risk factor for familial CFS. Furthermore, characterization of familial CFS represents an opportunity to identify pathogenic mechanisms of CFS.

Background The search for a biomarker specific for Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS) has been long, arduous and, to date, unsuccessful. Researchers need to consider their expenditures on each new candidate biomarker. In a previous study of antibody-dependent cell-mediated cytotoxicity (ADCC) by natural killer lymphocytes, we found lower ADCC for ME/CFS patients vs. unrelated donors but ruled against low ADCC as a biomarker because of similar ADCC for patients vs. their family members without ME/CFS. Objective We applied inclusion of family members without ME/CFS, from families with multiple CFS patients, as a second non-ME/CFS control group in order to re-examine inflammation in ME/CFS. Method Total and CD16A-positive 'non-classical' anti-inflammatory monocytes were monitored. Results Non-classical monocytes were elevated for patients vs. unrelated healthy donors but these differences were insignificant between patients vs. unaffected family members. Conclusions Inclusion of family members ruled against biomarker considerations for the monocytes characterized. These pilot findings for the non-classical monocytes are novel in the field of ME/CFS. We recommend that occupational therapists advocate and explain to family members without ME/CFS the need for the family members' participation as a second set of controls in pilot studies to rapidly eliminate false biomarkers, optimize patient participation, and save researchers' labor.

ADCC (antibody-dependent cell-mediated cytotoxicity) is dependent on the varying capacity of NK cells to kill, the affinities ofFCGR3A-encoded CD16A receptors for antibody, and the presence of antigen-specific antibodies.In vivoADCC depends on the number of CD16A receptor-positive NK cells in blood. We hypothesized that low ADCC cell function or low effector cell numbers could be biomarkers or risk factors for myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS). We measured NK cell ADCC lytic capacity and antibody recognition, CD16Apositive NK cells/µl blood, andFCGR3Ahomozygosity for the F allele that encodes low affinity CD16A antibody receptors. ME/CFS patients met the Fukuda 1994 diagnostic criteria. In this pilot report, we examined 5 families, each with 2 to 5 ME/CFS patients, and compared 11 patients, 22 family members without ME/CFS, and 16 unrelated healthy controls. ADCC was measured as CX1:1 cytotoxic capacity (the percentage of51Cr-Daudi tumors with obinutuzumab anti-CD20 antibody that were killed at a 1:1 ratio of CD16Apos NKs to Daudis) and CX-slope. Individual CX1:1 capacities varied from 16.2% to 81.8% and were comparable between patients and unaffected family members, while the ADCC of both family groups was lower than the unrelated healthy controls. The lack of difference between patients and their unaffected family members indicates that low ADCC is unsuitable as a diagnostic biomarker for ME/CFS. Familial CD16Apos NK blood cell counts were lower than unrelated healthy controls. The potential for synergistic effects of combined low CX1:1 and low effector cell counts occurring in the same individual was 24-fold greater for CFS family members than for unrelated controls.FCGR3Aof the families was predominantly F/F homozygous, correlating with the observed low EC50 for NK recognition of target cell-bound antibody. In summary, low ADCC is unsuitable as a biomarker, but could be a familial risk factor, for ME/CFS.

Natural killer (NK) lymphocyte ADCC supports anti-viral protection and monoclonal antibody (mAb) anti-tumor therapies. To predict in vivo ADCC therapeutic responses of different individuals, measurement of both ADCC cellular lytic capacity and their NK cellular receptor recognition of antibodies on 'target' cells are needed, using clinically available amounts of blood. Twenty ml of blood provides sufficient peripheral blood mononuclear cells (PBMCs) for the new assay for lytic capacity described here and for an antibody EC50 assay for Fc-receptor recognition. For the lytic capacity assay, we employed flow cytometry to quantify the CD16A IgG Fc-receptor positive NK effector cells from PBMCs to avoid loss of NKs during isolation. Targets were 51Cr-labeled Daudi B cells pretreated with excess obinutuzumab type 2 anti-CD20 mAb and washed; remaining free mAb was insufficient to convert B cells in the PBMCs into 'targets'. We calculated: the percentage Daudis killed at a 1:1 ratio of CD16A-positive NK cells to Daudis (CX1:1); lytic slopes; and ADCC50 lytic units. Among 27 donors, we detected wide ranges in CX1:1 (16-73% targets killed) and in lytic slopes. Slope variations prevented application of lytic units. We recommend CX1:1 to compare individuals' ADCC capacity. CX1:1 was similar for purified NK cells vs. PBMCs and independent of CD16A V & F genotypes and antibody EC50s. With high mAb bound onto targets and the high affinity of obinutuzumab Fc for CD16A, CX1:1 measurements discern ADCC lytic capacity rather than antibody recognition. This assay allows ADCC to be quantified without NK cell isolation and avoids distortion associated with lytic units.

Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is characterized by fatigue that does not improve with rest and worsens with exertion. ME/CFS may involve inflammation in the brain and/or be associated with viral infections. Increased numbers of pro-inflammatory monocytes in blood are a hallmark of infection. Biomarkers for, and information about, the CFS disease process are greatly needed. Here we queried circulating monocytes. In a pilot study, CFS patients and family members without CFS from three families were compared in order to reduce genetic and environmental variables. The patients conformed to the Fukuda 1994 standards. Unrelated healthy control subjects matched to the patients were included in the study. Blood monocytes were stained and analyzed by flow cytometry: counted with TruCountR beads, or stained after isolation and overnight culture of isolated peripheral blood monocytes. M1 pro-inflammatory monocytes are CD16A negative, while anti-inflammatory monocytes are CD16A positive; CD2 is a marker for dendritic monocytes, and changes in forward scatter could indicate in vitro activation. Notably, there were statistically significant differences between the lower percentages of M1 monocytes of the CFS patients compared to the controls; however, these differences were not significant between the patients and their family members without CFS. Also, the CD16A MFIs of the patients' monocytes were lower compared to controls but not to family members. One conclusion is that it is important to include family members without CFS as a control group in the search for biomarkers to diagnose CFS.

The DØ experiment enjoyed a very successful data-collection run at the Fermilab Tevatron collider between 1992 and 1996. Since then, the detector has been upgraded to take advantage of improvements to the Tevatron and to enhance its physics capabilities. We describe the new elements of the detector, including the silicon microstrip tracker, central fiber tracker, solenoidal magnet, preshower detectors, forward muon detector, and forward proton detector. The uranium/liquid-argon calorimeters and central muon detector, remaining from Run I, are discussed briefly. We also present the associated electronics, triggering, and data acquisition systems, along with the design and implementation of software specific to DØ. © 2006 Elsevier B.V. All rights reserved.

ADCC (antibody-dependent cell-mediated cytotoxicity) requires the cytotoxic subset of NK cells that can also recognize antibody. In vivo function may depend on the number of these ADCC NK cells. We report on ADCC activity and the number of IgG FcR-positive NK cells/ul blood as risk factors for chronic fatigue syndrome (CFS). ADCC capacity was measured as CX1:1 (PMID 29113954), the % Daudi B tumor cells killed at a 1:1 ratio of CD16A FcR-pos CD56pos NK effector cells to Daudi targets with obinutuzumab anti-CD20. CFS patients met the 1994 Fukuda diagnostic guidelines. We examined 5 families with 2 or more patients per family, comparing patients with their non-CFS family members and with unrelated healthy controls. ADCC activity and effector cell counts were similar between CFS patients and their family members. Surprisingly, the CX1:1 of both the CFS patients and their non-CFS family members was low, 77% of the controls (P=0.03). The ADCC effector cell counts/ul blood were also lower for both (two-tailed P=0.09, insignificant). The potential for synergistic interaction of low CX1:1 and low effector cell counts was 24-fold greater for CFS family members than for unrelated healthy controls (P

We would like to discuss two topics, one speci “c, the other more general.We “rst track an emerging trend in current research on the relationbetween morality and judgments of intentionality. Then we take up thegeneral question of how we can study such folk concepts as intentionality.From badness to broader hypothesesThe multitude of research examining folk judgments of intentionalityand morality following Joshua KnobeOs (2003a) intriguing “ndings haslargely sought to explain the disparity in intentionality judgments forimmoral vs. nonmoral (neutral) actions. Knobe discovered that peopledo not regard the presence of skill (or even intention) as necessary forjudging an action intentional when the action is morally bad. The essenceof the question is why , when considering a morally charged action, peo-ple seem to abandon their basic concept of intentionality (Malle & Knobe,1997) Ð in which, it had appeared, desire, belief, intention, awareness,and skill must each be present to consider an action intentional. Toanswer this question, many researchers have focused on the role of blame.

PURPOSE: Patterns of use and compliance with guidelines for use of lipid-based amphotericin B (LBA) and caspofungin at two medical institutions were studied. METHODS: Charts of all adult patients receiving LBA or caspofungin or both in combination at the University of California at San Francisco Medical Center (UCSF) and the University of California at San Diego Medical Center (UCSD) between January 1 and October 22, 2001, were reviewed. RESULTS: A total of 140 episodes of use of LBA, caspofungin, or the combination were identified at the two institutions. The primary indication for therapy with LBA was candidiasis (34% of episodes), whereas caspofungin and the drug combination were most commonly used for aspergillosis (43% and 71% of episodes, respectively). UCSF used the antifungal agents in accordance with its guidelines in 25% of the episodes of use and UCSD in 56%. Despite a higher qualifying serum creatinine (SCr) concentration for use of LBA at UCSF than at UCSD, the mean SCr concentration upon use was lower at UCSF. Among UCSF patients who did not meet the strict SCr criteria for use of either agent, most had concomitant renal failure as defined by a creatinine clearance of

The association of amphotericin B with nephrotoxicity is well known, but risk factors for this complication are not well characterized. One hundred and seventy-eight patients who received3 days of intravenous amphotericin B and a minimal total cumulative dose100 mg were reviewed retrospectively. The mean age, average cumulative dose of amphotericin B and duration of therapy were 46 +/- 22 years, 536 +/- 547 mg and 16.6 +/- 8.2 days, respectively. Eighty-six percent of patients received amphotericin B for empirical therapy of febrile neutropenia. Various definitions of nephrotoxicity were used; these were as follows (the incidence of nephrotoxicity as determined by the given definition is given in parentheses): definition 1, a change in creatinine of46 mumol/L over baseline (50%); definition 2, a doubling of creatinine over baseline (49%); definition 3, a change in creatinine of92 mumol/L (29%); definition 4, a doubling and/or a change in creatinine of92 mumol/L (49%); definition 5, an increase in creatinine to230 mumol/L (8%). Multivariate analysis showed that nephrotoxicity was associated with a greater cumulative dose of amphotericin B and receipt of concomitant nephrotoxic drugs for all definitions (P0.05). In those patients who experienced severe nephrotoxicity (creatinine increased to230 mumol/L), cyclosporin therapy was the most significant risk factor (odds ratio 18.8, P = 0.022). Haemodialysis was necessary in one patient, but multiple concomitant risk factors for renal dysfunction were present. No patient experienced irreversible nephrotoxicity. These findings allow for stratification of patients at risk for amphotericin B-induced nephrotoxicity and rational use of alternative agents.

Antimicrobial agents are widely utilized in the prevention and treatment of infection in patients with cardiovascular diseases. Knowledge of spectrum of activity, pharmacology, adverse effect and toxicity profile, and emergence of resistant pathogens is critical to the appropriate use of these agents. The following review will emphasize the safe, effective use of antimicrobial drugs in the cardiovascular patient.

Commercially available lipid formulations of amphotericin B (Abelcet, Amphotec, and AmBisome) represent a significant advance in drug delivery technology. Differences in biochemical, pharmacokinetic, and pharmacodynamic properties among the lipid products have been shown in in vitro and in vivo models. Clinical experience with these products has been primarily in patients either refractory to or intolerant of conventional amphotericin B deoxycholate (AmBd). None of the lipid-based products demonstrates superior efficacy when prospectively compared with AmBd in the treatment of documented infections. When used for the empirical treatment of febrile neutropenia, AmBisome significantly reduced the incidence of proven emergent fungal infections but did not improve short-term survival rates, in comparison with AmBd. Acute infusion-related adverse events vary, whereas nephrotoxicity is reduced with all three lipid formulations. Until superior efficacy is clearly shown (for documented infections) or pharmacoeconomic analyses document the value of these drugs, use of such expensive agents should be highly restricted to those who are intolerant of or refractory to AmBd.

A new high-lift airfoil design philosophy has been developed and experimentally validated through wind-tunnel tests. A key element of the high-lift design philosophy was to make use of a concave pressure recovery with aft loading. Three codes for airfoil design and analysis (PROFOIL, the Eppler code, and ISES) were used to design the example S1223 high-lift airfoil for a Reynolds number of 2 3 10 5 . In windtunnel tests, the new airfoil yielded a maximum lift coefe cient of 2.2. With vortex generators and a 1% chord Gurney eap (used separately), the Cl,max increased to 2.3. The airfoil demonstrates the rather dramatic gains in Cl,max over those airfoils previously used for high-lift low Reynolds number applications.

In a nominally two-dimensional flow, extensive wake surveys were performed on two airfoils at low Reynolds numbers to quantify the profile drag variation along the airfoil model span. Wake profile measurements were made at 57 spanwise stations spaced 2% chord apart and 1.25 chord lengths downstream of the trailing edge. Results at a Reynolds number (Re) of 2 X 10' show that variations on the order of 5-40% are typical. In an extreme case, however, over a spanwise distance of less than 12% chord, the profile drag coefficient changed from approximately 0.006 to 0.016, which illustrates the sometimes rather dramatic three-dimensional nature of the flow. Measurements taken at higher Reynolds numbers and closer to the trailing edge showed significant reductions in the spanwise drag variation, which suggests that the laminar separation bubble and the developing wake play an important role. A zigzag boundary-layer trip and an isolated roughness element were also investigated to examine their effects as compared with their respective undisturbed cases. Finally, the results suggest that the lack of good agreement in profile drag measurements between different wind-tunnel facilities can partly be traced to the associated measurement techniques used as weD as the flowfield three dimensionality. Differences in agreement between facilities can be expected when only one spanwise wake profile is taken per angle of attack, as has often been done in the past.

Objective To examine the impact of a change in the empiric gentamicin dose from 5 mg/kg every 24h to 5 mg/kg every 36h on target drug concentration achievement in neonates with hypoxic ischemic encephalopathy (HIE) receiving therapeutic hypothermia. Study Design Gentamicin drug concentrations in neonates with HIE receiving therapeutic hypothermia were examined during two time periods in a retrospective chart review. During the initial treatment period (November 2007 to March 2010; n=29), neonates received gentamicin 5 mg/kg every 24h (Q24h period). During the second treatment period (January 2011 to May 2012; n=23), the dose was changed to 5 mg/kg every 36h (Q36h period). Cooling criteria and protocol remained the same between treatment periods. Gentamicin drug concentrations including achievement of target trough concentrations (2 mg/L decreased from 38% to 4% with implementation of a Q36h dosing interval (P

ADCC can be essential for anti-viral immunity and supports many tumor immunotherapies. It is a challenge to quantify and one wants a fast assay that uses few lymphocytes when patient blood is limited. Three considerations affected our new assay design: 1) normal B cells within PBMCs introduce ‘cold target’ competition of radiolabeled tumor B cells if anti-B cell antibodies are present in the ADCC assays; 2) type 1 antibodies to CD20 B cell epitopes are internalized and cleared from the cell surface while type 2 anti-CD20 antibodies remain external; and 3) target cells with MHC I initiate KIR-regulation of lysis that will vary among donors. To address these issues, we used Daudi B leukemia cells that lack MHC I and pretreated the Daudi cells with the humanized nonfucosylated/glycoengineered monoclonal type 2 anti-CD20 antibody obinutuzumab (GA101 G2). Target cell pretreatment prevented ‘cold target’ competition. We used 4 hr 51Cr-release for its sensitivity and varied the PBMC to target (E:T) cell ratios to obtain lytic units. Flow cytometric counts of CD16A Fc-receptor positive cells with TrueCountR beads determined the exact number of ADCC effectors within the unfractionated PBMCs. There was low NK activity to Daudi cells, which required subtraction of simultaneous controls without antibody. We plotted ADCC activity as a log function of the E:T ratios and observed that ADCC was dependent upon interactions of multiple CD16A-positive effectors per target cell, which was expressed as lytic units per 106 CD16A cells, that varied 50 fold among donors. In addition, the linear slopes of CD16A cell lytic cooperativity varied 8 fold. In summary, we have established a fast quantitative assay for comparison of human ADCC that uses as few as 8 million PBMCs.

In a prospective, randomized, blinded study, 826 patients undergoing clean neurosurgical procedures received single intravenous doses of ceftizoxime (2 g) (n = 422) or a combination of vancomycin (1 g) and gentamicin (80 mg) (n = 404) 1 hour before an incision was made. Patients with infected or contaminated wounds and those receiving shunts or other implants were excluded. Primary wound infections occurred within 30 days in five patients in each group and were most common after spinal surgery and procedures through previous incisions. Secondary infections (pneumonias, urinary tract infections, and intravenous line-related bacteremia) occurred in 24 patients in the ceftizoxime group and 25 in the vancomycin/gentamicin group. The infection rates after transsphenoidal procedures (n = 129) were remarkably low in both groups. Ceftizoxime caused no adverse drug reactions, but six patients in the vancomycin/gentamicin group had clinically significant infusion-related hypotension or flushing. Placement of a temporary external drain, use of an operating microscope, preoperative steroids, and diabetes were not associated with increased infection rates. Analysis of routinely encountered ventricular cerebrospinal fluid and simultaneously obtained peripheral blood showed low but detectable levels of all three antibiotics within 2 hours; only ceftizoxime, however, achieved cerebrospinal fluid levels sufficient to inhibit the staphylococcus and Gram-negative bacilli most often associated with postneurosurgical infections. We conclude that ceftizoxime is as effective as vancomycin and gentamicin in neurosurgical prophylaxis but is less toxic and penetrates cerebrospinal fluid better.

Descriptive natural-language captions can help organize multimedia data. We describe our MARIE system, which interprets English queries directing the retrieval of media objects. It is novel in the extent to which it exploits previously interpreted and indexed English captions for the media objects. Our routine filtering of queries through descriptively complex captions (as opposed to keyword lists) before retrieving data can actually improve retrieval speed, as media data are often bulky and time-consuming to retrieve and difficult upon which to perform content analysis, and even small improvements to query precision can often pay off. Handling the English of captions and queries about them is not as difficult as it might seem, as the matching does not require deep understanding, just a comprehensive type hierarchy for caption concepts. An important innovation of MARIE is “supercaptions” describing sets of captions, which can minimize caption redundancy.

Several controversies exist regarding the use of antibiotic prophylaxis in cesarean section. Studies have investigated effectiveness, choice of drug, timing, duration appropriate route of administration. Efficacy of antibiotic prophylaxis in preventing postoperative endometritis in certain high-risk patients has been well documented. Factors identifying patients at high risk for endometritis include: indigent socioeconomic status, labor prior to cesarean section, rupture of chorioamniotic membranes number of vaginal examinations. Although many different antibiotics have been tested, no particular regimen has been found to be superior. Administration of antibiotics after cord clamping reduces the risk of fetal exposure without compromising maternal risk of endometritis. A single dose of ampicillin or a first-generation cephalosporin (e.g., cefazolin or cephalothin sodium) given intravenously provides adequate prophylaxis. No additional benefit has been demonstrated with the use of multiple-dose regimens.