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Supplementary Figure 1 from Three-Dimensional Culture of Mouse Renal Carcinoma Cells in Agarose Macrobeads Selects for a Subpopulation of Cells with Cancer Stem Cell or Cancer Progenitor Properties

Supplementary Table 4 from Three-Dimensional Culture of Mouse Renal Carcinoma Cells in Agarose Macrobeads Selects for a Subpopulation of Cells with Cancer Stem Cell or Cancer Progenitor Properties

Supplementary Movie 1 from Hydrophilic Agarose Macrobead Cultures Select for Outgrowth of Carcinoma Cell Populations That Can Restrict Tumor Growth

Supplementary Table 2 from Three-Dimensional Culture of Mouse Renal Carcinoma Cells in Agarose Macrobeads Selects for a Subpopulation of Cells with Cancer Stem Cell or Cancer Progenitor Properties

Supplementary Table 3 from Three-Dimensional Culture of Mouse Renal Carcinoma Cells in Agarose Macrobeads Selects for a Subpopulation of Cells with Cancer Stem Cell or Cancer Progenitor Properties

Cancer cells and their associated tumors have long been considered to exhibit unregulated proliferation or growth. However, a substantial body of evidence indicates that tumor growth is subject to both positive and negative regulatory controls. Here, we describe a novel property of tumor growth regulation that is neither species nor tumor-type specific. This property, functionally a type of feedback control, is triggered by the encapsulation of neoplastic cells in a growth-restricting hydrogel composed of an agarose matrix with a second coating of agarose to form 6- to 8-mm diameter macrobeads. In a mouse cell model of renal adenocarcinoma (RENCA cells), this process resulted in selection for a stem cell–like subpopulation which together with at least one other cell subpopulation drove colony formation in the macrobeads. Cells in these colonies produced diffusible substances that markedly inhibited in vitro and in vivo proliferation of epithelial-derived tumor cells outside the macrobeads. RENCA cells in monolayer culture that were exposed to RENCA macrobead-conditioned media exhibited cell-cycle accumulation in S phase due to activation of a G2/M checkpoint. At least 10 proteins with known tumor suppression functions were identified by analysis of RENCA macrobead-conditioned media, the properties of which offer opportunities to further dissect the molecular basis for tumor growth control. More generally, macrobead culture may permit the isolation of cancer stem cells and other cells of the stem cell niche, perhaps providing strategies to define more effective biologically based clinical approaches to treat neoplastic disease. Cancer Res; 71(3); 725–35. ©2011 AACR.

Supplementary Table 1 from Three-Dimensional Culture of Mouse Renal Carcinoma Cells in Agarose Macrobeads Selects for a Subpopulation of Cells with Cancer Stem Cell or Cancer Progenitor Properties

Nutritional status has a profound effect on immune function, and vitamin D, omega-3 fats, and other nutrients have long been suspected of influencing the development of the allergic march, and the incidence and severity of allergic conditions. This review provides an overview of research and opinion published during the past decade and summarizes clinically actionable information about dietary and nutritional influences on allergy prevention. There is now enough evidence of effects of vitamin D, omega-3 fats, the Mediterranean diet, and antioxidants to recommend their use in routine clinical practice for prevention and treatment of allergic conditions. Uncertain areas remain, especially in methods to best identify persons with specific nutritional deficiencies, and in determining optimal target levels for all essential nutrients. The potential gain in individual quality of life, especially by asthma reduction, makes achieving good nutrition a key goal for every clinician.

Background PCSK9 (Proprotein Convertase Subtilisin/Kexin Type 9) increases LDL cholesterol (LDL-C) levels by stimulating the degradation of Low Density Lipoprotein receptors (LDL-r). This protein is now of high interest because antibodies which inhibit its effect on LDL-r are being developed. A severe hypercholesterolemia and / or an elevation of lipoprotein(a) can be treated with lipoprotein apheresis (LA) in high-risk patients. Methods We measured serum PCSK9 levels in patients eligible for the extracorporeal treatment: in 40 patients (Cohort I) who were treated with different systems before and after apheresis sessions and in the intervals between sessions. 10 patients (Cohort II) who were eligible but did not start LA yet served as controls. Results Patients' baseline serum PCSK9 levels were elevated relative to healthy volunteers and LA sessions acutely reduced the mean PCSK9 concentrations by 51%. Comparison of the effectiveness of the different LA methods demonstrated the DSA and HELP were more effective than the DALI system. After 24 h PCSK9 levels had returned to baseline compared to 8 days for the LDL-C concentrations to return to its pre-apheresis levels. In Cohort II baseline PCSK9 levels were similar to those in Cohort I. Conclusion The acute reductions of PCSK9 by apheresis may be beneficial with respect to increasing the effectiveness of lipid-lowering drugs and with respect to an anti-atherosclerotic effect. In the future, antagonists to PCSK9 will probably be combined with or possibly replace LA in patients with a very high cardiovascular risk.

Chronic rhinosinusitis (CRS) is a complex inflammatory disease with variable disease manifestation. Though external risk factors are associated with development and/or persistence of CRS, the host mucosal response is also important, as nasal epithelium acts as a physical and immune barrier. Under inflammatory stress, the nasal epithelium can undergo injury, followed by a rapid remodeling response ranging from epithelial hyperplasia, to goblet-cell metaplasia, to denudation, loss of cilia, fibrosis, and basement membrane thickening. Identification of gene expression signatures and molecular pathways in CRS pathogenesis have now begun to contribute significantly to a better understanding of the genetic and molecular alterations underlying CRS development and progression. Genetic studies are especially illuminating when multiple gene variants synergize within a permissive environmental context, and are expected to guide development of more effective therapeutic targets for CRS treatment.

Epithelial remodeling is a part of our natural defense mechanisms, and includes migration, proliferation, and differentiation of epithelial cells, as well as the interactions between epithelial and stromal cells. It is not yet possible to distinguish between cause and effect during epithelium remodeling, and are there no clear roles for the many factors involved in respiratory infectious and inflammatory diseases due to a lack of critical information about epithelial cell responses. Most reported data are from lower airway studies or animal models. Therefore, research based on human nasal epithelial stem/progenitor cells can illuminate the pathophysiology of nasal airway disease from a different, more specific perspective. In this review, we discuss epithelial stem/progenitor cell research throughout the airway, with special attention to phenotypes and characterization of these cells from the nasal airway. Recently, we have isolated and cultured P63-positive human epithelial stem/progenitor cells from turbinate biopsies of healthy volunteers and from inflamed mucosa of patients with chronic rhinosinusitis with and without nasal polyposis. These cells propagate in serum-free, growth factor-supplemented, Dulbecco's modified Eagle's medium/F12 media, on either human fibroblast or 3T3 feeder layers. Self-renewal, proliferation, and differentiation potential at an air-liquid interface are being investigated to understand the molecular pathways underlying nasal inflammation. This in vitro culture system for nasal epithelial regeneration will allow molecular studies of human nasal epithelial cell interactions, differentiation, and repair, as well as responses to both environmental agents and to potential anti-inflammatory treatments.

To test the hypothesis that uvulopalatopharyngoplasty (UPPP) improves sleep apnea-related quality of life (measured on the Functional Outcomes of Sleep Questionnaire [FOSQ]) at 3-month follow-up. Secondary objectives were to test (1) the stability of the outcomes at 6 months, (2) the effect on global sleep apnea quality-of-life change, and (3) the effect on sleep apnea symptoms.Multicenter, prospective, longitudinal case series.Diverse university- and community-based otolaryngology practices.The cohort included 68 patients from 17 practices, with a mean ± standard deviation age of 44 ± 12 years and mean apnea-hypopnea index of 35 ± 32 events/hour. All patients underwent UPPP, defined as an open procedure modifying the shape and size of the palate, pharynx, and uvula, with or without tonsillectomy. Baseline data were collected on site before surgery, and outcome data were collected by mail 3 and 6 months after surgery, with follow-up rates of 51% and 50%, respectively.FOSQ scores improved from 14.3 ± 3.4 (scale 5-20, normal ≥17.9) at baseline to 17.2 ± 2.7 at 3 months (mean improvement 2.9; 95% confidence interval, 1.8-4.0; P.001) and 17.5 ± 2.5 at 6 months (mean improvement 3.1; 95% confidence interval, 2.0-4.2; P.001). All quality-of-life and symptom measures improved significantly at 3 and 6 months (all P.05).This prospective, multicenter, university- and community-based study provides evidence that UPPP significantly improves disease-specific quality of life and sleep apnea symptoms in patients with sleep apnea. Validity may be limited by significant loss to follow-up and absence of an unoperated control group.

The culture of tumor cell lines in three-dimensional scaffolds is considered to more closely replicate the in vivo tumor microenvironment than the standard method of two-dimensional cell culture. We hypothesized that our method of encapsulating and maintaining viable and functional pancreatic islets in agarose–agarose macrobeads (diameter 6–8 mm) might provide a novel method for the culture of tumor cell lines. In this report we describe and characterize tumor colonies that form within macrobeads seeded with mouse renal adenocarcinoma cells. Approximately 1% of seeded tumor cells survive in the macrobead and over several months form discrete elliptical colonies appearing as tumor cell niches with increasing metabolic activity in parallel to colony size. The tumor colonies demonstrate ongoing cell turnover as shown by BrdU incorporation and activated caspase-3 and TUNEL staining. Genes upregulated in the tumor colonies of the macrobead are likely adaptations to this novel environment, as well as an amplification of G1/S cell-cycle checkpoints. The data presented, including SCA-1 and Oct4 positivity and the upregulation of stem cell–like genes such as those associated with the Wnt pathway, support the notion that the macrobead selects for a subpopulation of cells with cancer stem cell or cancer progenitor properties. Cancer Res; 71(3); 716–24. ©2011 AACR.

Cancer cells and their associated tumors have long been considered to exhibit unregulated proliferation or growth. However, a substantial body of evidence indicates that tumor growth is subject to both positive and negative regulatory controls. Here, we describe a novel property of tumor growth regulation that is neither species nor tumor-type specific. This property, functionally a type of feedback control, is triggered by the encapsulation of neoplastic cells in a growth-restricting hydrogel composed of an agarose matrix with a second coating of agarose to form 6- to 8-mm diameter macrobeads. In a mouse cell model of renal adenocarcinoma (RENCA cells), this process resulted in selection for a stem cell–like subpopulation which together with at least one other cell subpopulation drove colony formation in the macrobeads. Cells in these colonies produced diffusible substances that markedly inhibited in vitro and in vivo proliferation of epithelial-derived tumor cells outside the macrobeads. RENCA cells in monolayer culture that were exposed to RENCA macrobead-conditioned media exhibited cell-cycle accumulation in S phase due to activation of a G2/M checkpoint. At least 10 proteins with known tumor suppression functions were identified by analysis of RENCA macrobead-conditioned media, the properties of which offer opportunities to further dissect the molecular basis for tumor growth control. More generally, macrobead culture may permit the isolation of cancer stem cells and other cells of the stem cell niche, perhaps providing strategies to define more effective biologically based clinical approaches to treat neoplastic disease. Cancer Res; 71(3); 725–35. ©2011 AACR.

Purpose of review Patch testing has rapidly become an important clinical allergy tool, but is underutilized in the evaluation of complex patients, especially when these patients have skin disorders and respiratory allergies, food allergies, or eosinophilic enteritis. Learning when and how to use patch tests thus adds to any practitioner's diagnostic abilities. Recent findings This review discusses selected studies from the past year, grouped into immunology, pediatric testing, contact allergy, food allergy, and drug allergy. Summary Patch tests can detect a wide range of sensitivities to inorganic and organic chemicals, drugs, biologic molecules, inhalants acting as contactants, food allergens, allergens that have not been commercially extracted, and solid allergens. Because patch tests detect the full range of immunologic reactions, Gell and Coombs type I to IV, they may be uniquely reactive when other allergy tests are negative. Because of the large number of published studies that utilize patch testing, clinicians often can use a literature search of prior studies of similar problems, or for particular allergens, to choose technical details that make successful patch testing more likely.

Patients undergoing dialysis treatment have a high incidence of dyslipidemia. Rosuvastatin is a potent statin drug that improves overall lipid profiles in dyslipidemic patients. However, the pharmacokinetics of rosuvastatin has not been studied in patients with end-stage kidney disease undergoing chronic peritoneal dialysis (PD). The goals of this study are to determine the pharmacokinetics and tolerability of a single oral dose of rosuvastatin in patients undergoing continuous ambulatory PD (CAPD). This was a nonrandomized, open-label, 1-week trial. Ten stable PD patients were given a single oral dose of rosuvastatin (10 mg). Serial blood samples were obtained over the next 48 hours, and the patients were followed for 1 week while they underwent CAPD. Rosuvastatin plasma concentration peaked (Cmax) at 3.68 +/- 2.3 ng/ml (geometric mean), 4.5 hours (median; range 2 - 6 hours) after oral dosing. The plasma concentration of rosuvastatin was 0.44 +/- 0.23 ng/ml at 24 hours (C24) and 0.14 +/- 0.07 ng/ml, with levels below the detectable range in 5 of 10 subjects, at 48 hours (C48). The area under the plasma concentration-time from 0 to 48 hours (AUC0-48) was 32.6 +/- 1.6 ng/ml/h. These pharmacokinetic profiles of rosuvastatin in CAPD patients are very similar to those observed in healthy volunteers, but different from patients with Stages 4 - 5 chronic kidney disease. A single oral dose of rosuvastatin was well tolerated in this small number of patients. We conclude that pharmacokinetic profiles of rosuvastatin in patients undergoing CAPD are similar to those observed in healthy volunteers. These findings suggest that a lower dose of rosuvastatin (<

Allergic rhinitis (AR), a chronic inflammatory disease of the upper airway, is one of the most common chronic diseases in the United States and is estimated to affect up to 60 million people. Pediatric Allergies in America is the largest and most comprehensive survey to date of pediatric patients and parents of patients with allergy, as well as health care providers (HCPs), regarding AR in children and its treatment. The goals of the survey were to determine the prevalence of AR in the US pediatric population and to collect information on what effect the condition has on patients in terms of symptom burden, quality of life, productivity, disease management, and pharmacologic treatment. This national survey screened 35,757 households to identify 500 children with HCP-diagnosed nasal allergies and 504 children without nasal allergies who were between the ages of 4 and 17 years. Parents of young children, as well as children 10 to 17 years of age, were questioned about the condition and its treatment. In parallel, 501 HCPs were interviewed. This survey has captured previously unavailable data on the prevalence of nasal allergies and their most common and most bothersome symptoms, on the effect of nasal allergies on the quality of life of children, and on medication use, including both over-the-counter and prescription medications, and has identified factors affecting satisfaction with treatment. The Pediatric Allergies in America survey also identifies distinct areas for improvement in the management of AR in children. In fact, based on the results of this survey, it appears that HCPs overestimate patients' and parents' satisfaction with disease management and the benefit of medications used for the treatment of nasal allergies in children. Findings from this national survey have identified important challenges to the management of AR, suggesting that its burden on children in the United States has been significantly underestimated.

Untreated pediatric patients with homozygous familial hypercholesterolemia usually have myocardial infarctions, heart failure, or death by the teenage years. Low-density lipoprotein (LDL) apheresis effectively lowers LDL cholesterol in the short term, but there is little published information on the long-term safety and efficacy of this treatment in children. An analysis was performed of a registry of all 29 patients who began LDL apheresis before 18 years of age at 15 sites during the 11 years since approval by the United States Food and Drug Administration. A chart review of 9 patients treated at The Rogosin Institute was also performed to obtain additional details about lipid lowering, adverse events, and cardiovascular status. Of the 29 patients, 20 are currently treated, with a mean age at the start of treatment of 9 +/- 4 years (range 3 to 15) and a mean treatment duration of 6 +/- 4 years (range 2 to 21). The baseline LDL cholesterol (521 +/- 126 mg/dl) is acutely lowered by 75% and chronically lowered by 48% with biweekly sessions. Systemic adverse events have been uncommon. Atherosclerotic disease of the coronary arteries and/or aorta or aortic valve was evident by angiography and/or echocardiography in 12 patients (60%) at baseline and progressed to more severe, symptomatic disease in 6 (30%). In conclusion, LDL apheresis is well tolerated for decades by even very young pediatric patients with homozygous familial hypercholesterolemia. It effectively lowers LDL cholesterol, but target LDL levels are not achieved, and some patients will show progression of cardiovascular disease.

Homozygous familial hypercholesterolemia (HoFH) is a rare, inherited, life-threatening, metabolic disorder of low-density lipoprotein (LDL) receptor function characterized by elevated serum LDL cholesterol (LDL-C) and rapidly progressive atherosclerotic cardiovascular disease (ACVD). Since LDL receptors are predominantly found on hepatocytes, orthotopic liver transplantation (OLT) has emerged as a viable intervention for HoFH because LDL receptor activity is restored. This study assessed the effects of OLT on ACVD and ACVD risk factors in pediatric patients with HoFH. We analyzed lipids, lipoproteins, body mass index, glucose, blood pressure, and cardiovascular imaging in 8 pediatric patients who underwent OLT for HoFH. Total serum cholesterol, LDL-C, lipoprotein (a), and apolipoprotein B/apolipoprotein A1 ratio decreased to normal values in all subjects (p values0.001) at 1 month after OLT and were maintained for the length of follow-up (2 to 6 years). There were few complications related to surgery or immunosuppressive therapy. Two patients developed mild hypertension. In the first 4 subjects monitored for 4 to 6 years after OLT, coronary artery disease did not develop or progress except in 1 minor artery in 1 subject and actually regressed in 2 subjects with50% stenosis. However, aortic valve stenosis progressed in 2 of 4 subjects. In conclusion, OLT is an effective therapeutic option for patients with HoFH with coronary artery disease and persistently elevated serum LDL-C despite maximum medical therapy. Aortic valvular disease may progress. Long-term data are needed to evaluate the true risk-benefit ratio of this surgical approach.

Monocyte chemoattractant protein-1 (MCP-1) is essential in atherogenesis. Oxidized lipids regulate MCP-1 expression and release from mononuclear cells. In this study we investigated (1) whether statin therapy reduces lipopolysaccharide (LPS)–stimulated MCP-1 production in human whole-blood samples and (2) the relationships between in vitro low-density lipoprotein (LDL) oxidation and MCP-1 production. Fasting blood samples were obtained from 55 healthy nonsmoking adults with moderate hypercholesterolemia who were participating in a randomized double-blind 8-week trial comparing the effects of statin therapy with those of placebo on cytokine production. Samples were analyzed for resistance to copper-mediated LDL oxidation (lag time in minutes), as well as MCP-1– and interleukin-8 (IL-8)–stimulated production. Statin therapy reduced MCP-1 production (mean ± SD) −161 ± 399 pg/mL/mm 3 white cells) compared with 267 ± 985 pg/mL/mm 3 in the placebo group, but changes were not different between active and placebo groups ( P = .13). Statin therapy also increased lag times (median [interquartile range]; 20.5 [7.0–51.2] minutes vs −17.0 [−5.3–16.5] minutes; P = .067 for group difference). Inhibition of MCP-1 production correlated with prolongation of lag time ( r = .46, P = .0056) in statin-treated subjects. Statin therapy reduced MCP-1 production in the whole blood of human subjects and these changes were correlated with improvement in LDL oxidative resistance.

Objective This study was undertaken to demonstrate the safety and efficacy of in vitro, radioallergosorbent test (RAST)-based inhalant allergen immunotherapy. Study design and setting Prospective 22 year single site clinical study, with outcome evaluations of 480 perennial allergic rhinitis patients, including 96 with concomitant asthma. Results Rhinitis symptom control after 2 years of immunotherapy was excellent in 32.5% of patients, good in 45.6%, and fair in 14.2%. There was no improvement in 7.7%. For patients with asthma, 81% had good or excellent pulmonary symptom improvement, and no patient failed to improve. No severe reactions occurred, but there were 5 limited systemic reactions, or 0.008% of injections, during a 2.5-year mean immunotherapy treatment course. Conclusion RAST-based immunotherapy is safe and effective for patients with perennial allergic rhinitis, with or without concomitant asthma. Significance This is the first large, multiyear study of safety and efficacy of RAST-based immunotherapy for treatment of perennial allergic rhinitis and asthma. EBM rating: C.

PURPOSE OF REVIEW Nasal obstruction, the cardinal symptom of persistent (perennial) allergic rhinitis, is one of the most common symptoms encountered in primary care and in specialist clinics. It is difficult to quantify by clinical examination, and, hence, objective assessment of the nasal airway is critical to rhinologic research. Nasal obstruction in persistent allergic rhinitis must be treated the year round, and therefore treatment choices, costs, and compliance all become important public health issues. RECENT FINDINGS Many inflammatory and neurogenic mediators released during allergic reactions are able to cause plasma exudation and vasodilatation, with resultant edema and swelling of the nasal mucosa. Recently, technological advancements have made it possible to qualitatively and quantitatively study the nasal airway, providing greater insights into the understanding of physiological fluctuation and pathophysiological manifestations of nasal patency. From recent international guidelines, the management of allergic rhinitis includes combining treatments of the upper and lower airways, by using patient education, allergen avoidance, pharmacological treatment, and specific immunotherapy. Surgery may be needed as an adjunctive intervention. Multiple methods have been introduced to treat turbinate hypertrophy. However, preservation of adequate nasal mucosal function is important, together with long-term results. SUMMARY It is important that consensus recommendations for the management of allergic rhinitis be designed and implemented by all levels of medical specialists in order to improve treatment outcomes.

Decreased concentrations of total cholesterol, lipoproteins, and lipoprotein cholesterols occur early in the course of critical illness. Low cholesterol concentrations correlate with high concentrations of cytokines such as interleukin (IL)-6 and IL-10, and may be due to decreased synthesis or increased catabolism of cholesterol. Low cholesterol concentrations have been associated clinically with several adverse outcomes, including the development of nosocomial infections. The study was performed to test the hypothesis that a low cholesterol concentration predicts mortality and secondarily predicts the development of organ dysfunction in critical surgical illness.A prospective study was undertaken of 215 patients admitted to a university surgical ICU with systemic inflammatory response syndrome (SIRS). Serial blood samples were collected within 24 h of admission, as well as on the morning of days 2, 4, and 7 of the ICU stay for as long as the patients were in the ICU. Demographic data and predetermined outcomes were noted.One hundred nine patients had at least two samples drawn and form the population for analysis. Sixty-two of the patients had three samples obtained, whereas 42 patients had four samples obtained. By univariate analysis, non-survivors were more severely ill on admission (APACHE III), more likely to have been admitted to the ICU as an emergency, more likely to develop a nosocomial infection, and more likely to develop severe organ dysfunction (MODS) (all, p0.05). Death was associated on day 1 with increased concentrations of sIL2R, IL-6, IL-10, and sTNFR-p75 (all, p0.01), but there were initially no differences in serum lipid concentrations. However, by day 2, concentrations of IL-6, IL-10, and cholesterol had decreased significantly (all, p0.05) from day 1 in non-survivors but not in survivors; the difference in serum cholesterol concentration persisted to day 7 (p0.05). Persistently elevated concentrations of IL-6 and IL-10 were observed in patients who developed severe MODS. By logistic regression, increased APACHE III score, development of a nosocomial infection, and decreased cholesterol concentration were independently associated with mortality.Decreased serum cholesterol concentration is an independent predictor of mortality in critically ill surgical patients. Repletion of serum lipids is a feasible therapeutic approach for the management of critical illness.

Testing for food and chemical sensitivities usually becomes necessary as part of the evaluation of otolaryngology patients who have chronic illness. The more complex the patient, and the more recalcitrant the problem is to treatment, the more likely it is that allergies, and especially food or chemical sensitivities, are involved in the pathogenesis of the illness. Failure to consider all major allergen contacts, including foods and chemicals, can lead to inadequate therapy. Similarly, failure to understand total allergic and oxidant load and the effects of chemical toxicity can lead to inappropriate or ineffective treatment. Clinically, food allergies occur in two different types: immediate, anaphylactic, fixed reactions and delayed, chronic, cyclic reactions. Different test methods have been developed for the two types. Fixed food allergies can be safely and efficiently detected by in vitro specific IgE or histamine release tests. Cyclic food allergies are best detected by either oral food challenges or by the IPDFT test. Choosing the best test for a particular patient requires a clear understanding of the two food allergy types and how their clinical presentations differ. Other tests for food allergies are compared and contrasted with these primary tests. Chemical sensitivity also occurs in two different clinical types: allergic, and toxic. True allergy to chemical haptens, either type I, IgE-mediated, or type IV, delayed hypersensitivity, occurs with significant frequency but is often unsuspected. Chemical toxicity can be caused by the aftereffects of an acute exposure or as a result of chronic, low-level exposure, but is even more frequently unsuspected and will not be diagnosed without a high index of suspicion. Both types of chemical sensitivity need to be addressed in any patients who have either a high allergen or chemical exposure load [105]. Either in vitro or in vivo tests can be used for chemical allergy detection; the advantages of each are outlined. Chemical toxicity screening tests are available and useful but do not detect all possible toxicants. Definitive toxic chemical tests usually require specialized laboratory facilities and expert consultation, for which possible sources are specified. The most important point in testing for food or chemical sensitivity is to be aware that food or chemical sensitivity can be contributing to a specific patient's clinical problems. Only then can appropriate investigations be undertaken to understand and then, perhaps, to intervene successfully in that illness.

Endotoxemia is associated with rapid and marked declines in serum levels of LDL and HDL by unknown mechanisms. Six normal volunteers received a single, small intravenous (iv) dose of endotoxin (Escherichia coli 0113, 2 ng/kg) or saline in a random order, cross-over design. After endotoxin treatment, volunteers had mild, transient flu-like symptoms and markedly increased serum levels of tumor necrosis factor and its soluble receptors, interleukin-6, cortisol, serum amyloid A, and C-reactive protein. Triglyceride (TG), VLDL-TG, and nonesterified fatty acid increased (peak at 3–4 h), then TG declined (nadir at 9 h), and then cholesterol, LDL cholesterol, apolipoprotein B (apoB), and phospholipid declined (nadirs at 12–24 h). HDL cholesterol and apoA-I levels were not affected, but half of the decrease in phospholipid was HDL phospholipid. Lipopolysaccharide binding protein (LBP) rose 3-fold (peak at 12 h), with smaller and later decreases in the activities of phospholipid transfer protein and cholesteryl ester transfer protein. In conclusion, a decline in LDL was rapidly induced in normal volunteers with a single iv dose of endotoxin. The selective loss of phospholipid from HDL may have been mediated by LBP and, after more intense or prolonged inflammation, could result in increased HDL clearance and reduced HDL levels.

BACKGROUND:Lipids and lipoproteins have been shown to bind and neutralize endotoxin and to improve outcomes in animal models of sepsis.OBJECTIVE:To provide safety and pharmacokinetic data for a protein-free, phospholipid-rich emulsion developed as an agent to neutralize endotoxin, and to study the changes in lipids and lipoproteins following emulsion administration.METHODS:Thirty healthy male volunteers (aged 18–45 y) were given an emulsion containing 92.5% soy phospholipid, 7.5% soy triglyceride, and 18 mM sodium cholate using a double-blind, placebo-controlled crossover protocol. Emulsion at 3 escalating doses (75, 150, 300 mg/kg) based on phospholipid content was administered by intravenous infusion over 2 hours in the low- and mid-dose groups and 6 hours in the high-dose group.RESULTS:All subjects completed the protocol without significant toxicities. A slight dose-dependent increase in indirect bilirubin at the 24-hour time point was observed in the emulsion treatment period, with a maximum difference between placebo and emulsion of 0.9 mg/dL. Mean ± SD peak phospholipid levels were 316 ± 30, 533 ± 53, and 709 ± 86 mg/dL, and phospholipid half-lives were 5.4 ± 0.6, 5.4 ± 0.5, and 8.0 ± 0.8 hours for the low, mid, and high doses, respectively. Increases in total cholesterol, low-density lipoprotein cholesterol and apolipoprotein A-I and B levels were observed. High-density lipoprotein cholesterol decreased immediately following emulsion infusion, but rebounded to above placebo levels by 24 hours.CONCLUSIONS:A unique phospholipid-rich emulsion was shown to have a favorable safety profile and to expand the blood lipid and lipoprotein pool without the use of human-derived blood products. Lipid levels expected to protect against the physiologic effects of bacterial endotoxin were achieved.

Lipoprotein phospholipid (PL) plays a major role in neutralization of endotoxin. This study tested the hypothesis that prophylactic administration of a PL-enriched emulsion (PRE), which augments PL content of serum lipoproteins and neutralizes endotoxin in vitro, would preserve cardiovascular function and improve survival in porcine septic peritonitis. A control group was compared with low-, mid-, and high-dose treatment groups that received PRE by primed continuous infusion for 48 h. A fibrin clot containing live Escherichia coli 0111.B4 was implanted intraperitoneally 30 min after the priming dose. Survival increased in a dose-dependent manner and was correlated with serum PL. Infused PL was associated with high-density lipoprotein in the low-dose group and all serum lipoproteins at higher doses. Treatment significantly lowered serum endotoxin and tumor necrosis factor (TNF)-α, preserved cardiac output and ejection fraction, and attenuated increases in systemic and pulmonary vascular resistances. This study demonstrated that augmentation of lipoprotein PL via administration of PRE improved survival and offered a novel therapeutic approach to sepsis.

To determine the relationship of hypolipidemia to cytokine concentrations and clinical outcomes in critically ill surgical patients.Consecutive, prospective case series.Surgical intensive care unit of an urban university hospital.Subjects were 111 patients with a variety of critical illnesses, for whom serum lipid, lipoprotein, and cytokine concentrations were determined within 24 hrs of admission to a surgical intensive care unit. Controls were 32 healthy men and women for whom serum lipid, lipoprotein, and cytokine concentrations were determined.Blood samples were drawn on admission to the intensive care unit. Predetermined clinical outcomes including death, infection subsequent to intensive care unit admission, length of intensive care unit stay, and magnitude of organ dysfunction were monitored prospectively.Measurements included total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, apolipoproteins A-I and B, phospholipid, triglyceride, interleukin-6, interleukin-10, soluble interleukin-2 receptor, tumor necrosis factor-alpha, and soluble tumor necrosis factor receptors p55 and p75. Mean serum lipid concentrations were extremely low: total cholesterol, 127 +/- 52 mg/dL; low-density lipoprotein cholesterol, 75 +/- 41 mg/dL; high-density lipoprotein cholesterol, 29 +/- 15 mg/dL. Total, low-density lipoprotein, and high-density lipoprotein cholesterol concentrations and apolipoprotein concentrations inversely correlated with interleukin-6, soluble interleukin-2 receptor, and interleukin-10 concentrations, whereas the triglyceride concentration correlated positively with tumor necrosis factor soluble receptors p55 and p75. Clinical outcomes were related to whether the admission cholesterol concentration was above (n = 56) or below (n = 55) the median concentration of 120 mg/dL. Each of the clinical end points occurred between 1.9- and 3.5-fold more frequently in the very low cholesterol (120 mg/dL) group. Nine patients (8%) died during the hospitalization. Seven of the nine patients who died had total cholesterol concentrations below the median concentration of 120 mg/dL.Low cholesterol and lipoprotein concentrations found in critically ill surgical patients correlate with interleukin-6, soluble interleukin-2 receptor, and interleukin-10 concentrations and predict clinical outcomes.

Treatment to low-density lipoprotein (LDL) cholesterol targets has become a focus in the management of patients with coronary heart disease (CHD). Many patients with familial hypercholesterolemia (FH) are unable to reach targets because of drug intolerance or extremely high baseline LDL cholesterol levels. Consequently, LDL apheresis has become a useful modality for the treatment of patients with severe hypercholesterolemia. Commonly used LDL apheresis systems utilize immunoadsorption columns, dextran sulfate cellulose columns, or heparin precipitation. A new and simpler treatment modality is emerging which uses whole blood compatible columns. All systems require systemic anticoagulation, extracorporeal processing of blood, and venous vascular access. Acute LDL lowering is 70% to 80% and time-averaged LDL lowering is 40% to 50%. Lipoprotein(a) is also substantially lowered. Clinical efficacy has been shown in several studies. Mechanisms for clinical improvement in addition to regression of atherosclerotic plaque have been suggested by recent research.

The short-term effectiveness of low-density lipoprotein (LDL) apheresis using a dextran sulfate cellulose adsorption column technique was previously examined in a 9-center, 22-week controlled trial in 64 patients with familial hypercholesterolemia (FH) who did not adequately respond to diet and drug therapy. Forty-nine patients (40 treatment, 9 controls) subsequently received LDL apheresis procedures as part of an optional follow-up phase. This study reports on the long-term safety, lipid lowering, and clinical efficacy of LDL apheresis for the 5-year period that includes both the initial controlled study and follow-up phase. During this time, patients received a total of 3,902 treatments of which 3,314 treatments were given during the follow-up phase. Adverse events were infrequent, occurring in 142 procedures (3.6%). Immediate reduction in LDL cholesterol was 76% both in homozygotes and in heterozygotes. Patients with homozygous FH had a progressive decrease in pretreatment LDL cholesterol level along with an increase in high-density lipoprotein (HDL) cholesterol level. There was no appreciable change in pretreatment lipoprotein level over time in heterozygotes. The rate of cardiovascular events during therapy with LDL apheresis and lipid-lowering drugs was 3.5 events per 1,000 patient-months of treatment compared with 6.3 events per 1,000 patient-months for the 5 years before LDL apheresis therapy. These findings support the long-term safety and clinical efficacy of LDL apheresis in patients with heterozygous and homozygous FH who are inadequately controlled with drug therapy.

Many types of skin tests have evolved from Blackley's early scratch tests. This review highlights both the similarities between current skin tests and their differences. All current skin tests are capable of detecting allergic hypersensitivity, but the tests differ in their sensitivity, specificity, safety, reproducibility, and applications. Common factors in initial and final test doses, and in test dose increments, are identified. Test methods that quantitatively measure a range of allergen concentrations have diagnostic advantages in terms of safely detecting a wide range of allergic sensitivities. Failure to detect the full spectrum of allergic disease can lead to treatment failure; therefore, complete skin testing is desirable. This is especially important when dealing with exquisitely sensitive patients, such as many grass or ragweed sensitized patients, but is equally vital when evaluating low sensitivity patients, including many who are mold or food sensitive. Quantitation also improves test reproducibility, which is why it is used for antigen standardization. Finally, quantitation has advantages when used to initiate and escalate allergen immunotherapy. In vivo testing continues to evolve. New types of prick testing devices, and refinements of intradermal and patch test methods, continue to be reported. All allergists need to stay current with developments in vivo testing, so that they can offer their patients diagnosis that is appropriate to each individual situation.

Vitamin D (vitD3) deficiency occurs frequently and has profound effects on health, especially asthma. This article examines how current knowledge of vitD3 actions and the worldwide distribution of vitD3 deficiency influences everyday clinical allergy practice. Within the limits of current knowledge, the article concisely explains the molecular nature of vitD3 actions, reviews key vitD3 research as it applies to clinical care, answers questions about the potential clinical impact of low vitD3 levels, and discusses use and safety of vitD3 supplements.

The use of LDL-apheresis to treat patients with severe hypercholesterolemia has gained wider clinical acceptance during the past 2-3 years, particularly in patients with coronary artery disease. Systems utilizing immunoadsorption columns, dextran sulfate cellulose columns and heparin precipitation have been most commonly employed. New or improved technologies include whole-blood compatible columns, double-filtration plasmapheresis and lipoprotein (a)-apheresis. The mechanisms for clinical improvement extend beyond simple regression of atherosclerotic plaque.

Asthma is a chronic inflammatory disease of the lower respiratory tract which is triggered by exposure to allergens or other airway irritants, and is commonly encountered in otolaryngologic practice. This three-part review is designed to assist otolaryngologists in recognizing and managing asthmatic patients. Part one summarized current knowledge of the pathophysiology and increasing prevalence of asthma, and its assessment and diagnosis. Part two discussed asthma management by environmental controls, anti-inflammatory therapies, and patient education, and compared pharmacologic treatments which are not primarily anti-inflammatory. These include mucolytic, anticholinergic, antihistamine, theophylline and beta agonist drugs. In this final part, anti-inflammatory treatments for asthma control are critically reviewed, including a balanced discussion of cromolyn, nedocromil, glucocorticoids, allergy immunotherapy, and the appropriate indications, possible toxicities and reasonable precautions for their use.

Asthma is a chronic inflammatory disease of the lower respiratory tract which is triggered by exposure to allergens or other airway irritants, and is commonly encountered in otolaryngologic practice. This three-part review is designed to assist otolaryngologists in effectively managing their asthmatic patients. In Part I, current information on the pathophysiology and increasing prevalence of asthma, its clinical variability, the assessment of asthma severity and methods for diagnosis of asthma were summarized. Part 11 discusses a tripartite strategy for asthma management, based on environmental controls, the use of anti-inflammatory therapies and patient education. In addition, pharmacologic treatments which are not primarily anti-inflammatory are reviewed in detail. The uses, effectiveness, side effects and suitability for drug combination therapy for mucolytic, anticholinergic, antihistamine, theophylline and beta agonist drugs are compared, and the disadvantages of beta antagonists are summarized. Part III concludes with an overview of antiinflammatory therapies for asthma control.

Asthma is a chronic inflammatory disease of the lower respiratory tract which is triggered by exposure to allergens or other airway irritants. This inflammation results in airway hyperresponsiveness, bronchial muscle spasm, mucous gland hypersecretion and mucosal edema, which combine to create symptoms such as cough, wheezing and respiratory distress. Because the inflammatory process is highly variable, asthma is a disorder with many possible presentations. It may therefore proceed for years without clinical recognition, and may challenge the most astute diagnostician. It is important for otolaryngologists to be able to suspect, diagnose and treat asthma. This is so because asthma is a common disease in the otolaryngologic patient population, both as one of the options in the differential diagnosis of respiratory complaints, and as a comorbid condition which may complicate the treatment of other medical or surgical problems. Furthermore, both the understanding of asthma's pathophysiology, and its optimum treatment methods have undergone radical changes during the past decade. This three-part discussion reviews our modern understanding of asthma, and proposes diagnosis and treatment guidelines which can assist otolaryngologists in effectively managing their asthmatic patients. Part one summarizes current information on the pathophysiology and increasing prevalence of asthma, its clinical variability, the assessment of asthma sensitivity, and methods for diagnosis of asthma. Parts two and three cover the strategy for asthma management, and the use of both adjunctive and anti-inflammatory therapies for asthma control.

A subset of patients with familial hypercholesterolemia (FH) have an inadequate lipid-lowering response to diet and drug treatment and should be considered for low-density lipoprotein (LDL)-apheresis therapy. This procedure selectively removes apolipoprotein B-containing particles [LDL, very-low-density lipoprotein, lipoprotein(a)] from plasma independent of diet and drug therapy. Methods for performing LDL-apheresis include dextran sulfate cellulose adsorption, immunoadsorption, and heparin-induced extracorporeal precipitation. The Liposorber Study Group evaluated LDL removal using the Liposorber® LA-15 LDL-apheresis System in 64 patients with FH who had not responded adequately to diet and maximal drug therapy. Mean acute reductions in LDL cholesterol (LDL-C) were 76% in heterozygous FH (HtFH) patients and 81% in homozygous FH (HoFH) patients. Time-averaged levels of LDL-C were lowered 41% in HtFH and 53% in HoFH patients. Hypotension was the most frequent side effect, occurring in 3% of procedures. The Liposorber® LA-15 System has been approved by the Food and Drug Administration and is recommended for 1) patients with functional homozygous FH (LDL-C level >500 mg/dL; 2) patients with coronary artery disease (CAD) and LDL-C levels ≥200 mg/dL; 3) patients without CAD, but an LDL-C level ≥300 mg/dL. © 1996 Wiley-Liss, Inc.

Lipoprotein(a) [Lp(a)] is a genetically determined risk factor for vascular disease and a potential link between coagulation, lipoproteins, and the development of atherosclerosis. Its role in the vascular complications of patients with chronic renal disease is unclear. We review methodologic issues involved in measuring Lp(a), particularly as they relate to studies of patients with chronic renal disease. The accurate measurement of Lp(a) is difficult because all the commercially available assays are sensitive to apolipoprotein(a) isoform size, Lp(a) behaves like an acute phase reactant, and levels vary markedly among ethnic groups. The results of 12 studies that included data on median Lp(a) levels in controls and patients receiving renal replacement therapy were analyzed. Although there was variation among studies, most found elevated levels of Lp(a) in patients receiving hemodialysis (range of medians, 9.0 to 38.4 mg/dL) compared with controls (range of medians, 4.7 to 19.7 mg/dL). With the exception of one study, Lp(a) levels also were elevated in patients receiving continuous ambulatory peritoneal dialysis compared with controls and patients receiving hemodialysis. In one study, an elevated Lp(a) level in patients receiving hemodialysis correlated with subsequent development of vascular events. A separate study associated the occurrence of vascular access occlusion with Lp(a) level. Following renal transplantation, Lp(a) levels decreased in all four studies, which included data before and after transplantation. Although variability in results were seen, Lp(a) levels appear to be elevated in patients receiving renal replacement therapy. Renal transplantation at least partially reverses this effect. The variability in results is probably related to methodologic difficulties in measuring Lp(a) and failure to segregate ethnic groups in study design and analysis.(ABSTRACT TRUNCATED AT 250 WORDS)

Statins are highly effective drugs prescribed to millions of people to lower LDL-cholesterol and decrease cardiovascular risk. The benefits of statin therapy seen in randomized clinical trials will only be replicated in real-life if patients adhere to the prescribed treatment regimen. But, about half of patients discontinue statin therapy within the first year, and adherence decreases with time. Patient, physician and healthcare system-related factors play a role in this problem. Recent studies have focused more on the patients’ perspectives on non-adherence. Adverse events are cited as the most common cause of statin discontinuation; thus, the healthcare provider must be willing to ally and dialogue with patients to address concerns and assess the risks and benefits of continued statin therapy.

Objective: This evidence-based clinical practice guideline (CPG) aims to improve clinical decision making of general practitioners and specialists in the treatment of acute and chronic rhinosinusitis in adults. Support clinicians in clinical decision making for medical treatment with target in the pathophysiological process and evidence-based efficacy, safety, and tolerability.Method: The Pan-American Association of Otorhinolaryngology and Head and Neck Surgery, in partnership with the Ibero American Agency for Development and Assessment of Health Technologies, developed a clinical practice guideline on medical management of acute and chronic rhinosinusitis in adults. This document provides punctual evidence-based recommendations for primary care physicians and otolaryngologists on the medical management of these conditions and complies with evidence-based medicine fundamentals and with well-validated guidelines methodology as recommended by the National Institute for Clinical Excellence, the National Ins...

During the past decades, numerous computational fluid dynamics (CFD) studies, constructed from CT or MRI images, have simulated human nasal models. As compared to rhinomanometry and acoustic rhinometry, which provide quantitative information only of nasal airflow, resistance, and cross sectional areas, CFD enables additional measurements of airflow passing through the nasal cavity that help visualize the physiologic impact of alterations in intranasal structures. Therefore, it becomes possible to quantitatively measure, and visually appreciate, the airflow pattern (laminar or turbulent), velocity, pressure, wall shear stress, particle deposition, and temperature changes at different flow rates, in different parts of the nasal cavity. The effects of both existing anatomical factors, as well as post-operative changes, can be assessed. With recent improvements in CFD technology and computing power, there is a promising future for CFD to become a useful tool in planning, predicting, and evaluating outcomes of nasal surgery. This review discusses the possibilities and potential impacts, as well as technical limitations, of using CFD simulation to better understand nasal airflow physiology.