A systematic review and meta-analysis of patient data from the West Africa (2013-16) Ebola virus disease epidemic.

BACKGROUND: Over 28 000 individuals were infected with Ebola virus during the West Africa (2013-2016) epidemic, yet there has been criticism of the lack of robust clinical descriptions of Ebola virus disease (EVD) illness from that outbreak. OBJECTIVES: To perform a meta-analysis of published data from the epidemic to describe the clinical presentation, evolution of disease, and predictors of mortality in individuals with EVD. To assess the quality and utility of published data for clinical and public health decision-making. DATA SOURCES: Primary articles available in PubMed and published between January 2014 and May 2017. ELIGIBILITY: Studies that sequentially enrolled individuals hospitalized for EVD and that reported acute clinical outcomes. METHODS: We performed meta-analyses using random-effect models and assessed heterogeneity using the I2 method. We assessed data representativeness by comparing meta-analysis estimates with WHO aggregate data. We examined data utility by examining the availability and compatibility of data sets. RESULTS: In all, 3653 articles were screened and 34 articles were included, representing 16 independent cohorts of patients (18 overlapping cohorts) and at least 6168 individuals. The pooled estimate for case fatality rate was 51% (95% CI 46%-56%). However, pooling of estimates for clinical presentation, progression, and predictors of mortality in individuals with EVD were hampered by significant heterogeneity, and inadequate data on clinical progression. Our assessment of data quality found that heterogeneity was largely unexplained, and data availability and compatibility were poor. CONCLUSIONS: We have quantified a missed opportunity to generate reliable estimates of the clinical manifestations of EVD during the West Africa epidemic. Clinical data standards and data capture platforms are urgently needed.

The BIOSAFEPAPER project for in vitro toxicity assessments: preparation, detailed chemical characterisation and testing of extracts from paper and board samples.

Nineteen food contact papers and boards and one non-food contact board were extracted following test protocols developed within European Union funded project BIOSAFEPAPER. The extraction media were either hot or cold water, 95% ethanol or Tenax, according to the end use of the sample. The extractable dry matter content of the samples varied from 1200 to 11,800 mg/kg (0.8-35.5 mg/dm2). According to GC-MS the main substances extracted into water were pulp-derived natural products such as fatty acids, resin acids, natural wood sterols and alkanols. Substances extracted into ethanol particularly, were diisopropylnaphthalenes, alkanes and phthalic acid esters. The non-food contact board showed the greatest number and highest concentrations of GC-MS detectable compounds. The extracts were subjected to a battery of in vitro toxicity tests measuring both acute and sublethal cytotoxicity and genotoxic effects. None of the water or Tenax extracts was positive in cytotoxicity or genotoxicity assays. The ethanol extract of the non-food contact board gave a positive response in the genotoxicity assays, and all four ethanol extracts gave positive response(s) in the cytotoxicity assays to some extent. These responses could not be pinpointed to any specific compound, although there appeared a correlation between the total amount of extractables and toxicity.

Embryonic Lethals and T-DNA Insertional Mutagenesis in Arabidopsis.

T-DNA insertional mutagenesis represents a promising approach to the molecular isolation of genes with essential functions during plant embryo development. We describe in this report the isolation and characterization of 18 mutants of Arabidopsis thaliana defective in embryo development following seed transformation with Agrobacterium tumefaciens. Random T-DNA insertion was expected to result in a high frequency of recessive embryonic lethals because many target genes are required for embryogenesis. The cointegrate Ti plasmid used in these experiments contained the nopaline synthase and neomycin phosphotransferase gene markers. Nopaline assays and resistance to kanamycin were used to estimate the number of functional inserts present in segregating families. Nine families appeared to contain a T-DNA insert either within or adjacent to the mutant gene. Eight families were clearly not tagged with a functional insert and appeared instead to contain mutations induced during the transformation process. DNA gel blot hybridization with internal and right border probes revealed a variety of rearrangements associated with T-DNA insertion. A general strategy is presented to simplify the identification of tagged embryonic mutants and facilitate the molecular isolation of genes required for plant embryogenesis.

Automatic implantable cardioverter-defibrillator implantation without thoracotomy using an endocardial and submuscular patch system.

The automatic cardioverter-defibrillator lead system is implanted by a thoracotomy procedure that may result in atelectasis, pleural effusion, cardiac tamponade and lengthy convalescence. A new defibrillator lead system that allows selection of different defibrillating current pathways is implanted without a thoracotomy. Ten patients requiring a cardioverter-defibrillator for recurrent sustained ventricular tachycardia (five patients) or aborted sudden cardiac death (five patients) were evaluated for implantation of this lead system. A lead configuration with a bidirectional defibrillating current pathway was implanted in nine patients. The defibrillation threshold with this lead configuration was 15 J in five patients, 20 J in three and 30 to 35 J in one patient. In the remaining patient the lead system had a 40 J defibrillation threshold and was not implanted. No perioperative complications occurred. Induced ventricular fibrillation was successfully terminated at the predischarge and intermediate follow-up (8 to 12 weeks) electrophysiologic studies. During the follow-up period, there were three deaths (one sudden, two due to heart failure) and two lead system failures (oversensing with inappropriate shocks in one patient and patch lead fracture in another). Implantation of the cardioverter-defibrillator lead system by a nonthoracotomy approach is feasible, has no significant perioperative complications and is well tolerated by patients. Effective defibrillation was demonstrated immediately as well as at intermediate follow-up study. The occurrence of patch lead fracture and oversensing requires improvement in the present (nonthoracotomy) lead system technology.

Long-term multicenter experience with a second-generation implantable pacemaker-defibrillator in patients with malignant ventricular tachyarrhythmias. The Guardian Multicenter Investigators Group.

A second-generation implantable pacemaker-cardioverter-defibrillator was evaluated in 200 patients with sustained ventricular tachycardia, ventricular fibrillation or prior cardiac arrest. The device permits demand ventricular pacing for bradyarrhythmias and for long QT interval or tachycardia suppression, uses programmable (3 to 30 J) energy shocks for conversion of ventricular tachycardia and ventricular fibrillation and is used with conventional pacing and defibrillation leads. Ventricular tachycardia/fibrillation recognition is based on the ventricular electrogram rate and requires reconfirmation before shock delivery. Two hundred patients (mean age 62 years, mean left ventricular ejection fraction 36%) were enrolled and followed up for 0 to 23 months (mean 12). Epicardial lead system implantation was performed with use of an anterolateral thoracotomy (38%), median sternotomy (26%) and subxiphoid (20%) or subcostal (16%) approach. Perioperative mortality rate was 5.5% (all nonarrhythmic deaths). Implant defibrillation threshold ranged from 3 to 30 J (mean 15), with initial programmed shock energy ranging from 3 to 30 J (mean 22). Ventricular tachycardia/fibrillation sensing threshold ranged from 0.7 to 1.8 mV (median 1) and the tachycardia detection interval from 288 to 416 ms (median 320). Reprogramming of implant variables was necessary for reliable electrographic sensing (54 patients), programmed shock therapy (61 patients) and tachycardia detection rate (63 patients). Device activation for potential shock delivery occurred in 111 patients (55.5%) with actual shock delivery after ventricular tachycardia/fibrillation reconfirmation in 66 patients (33%). During follow-up study, there was a 1% arrhythmia mortality rate, 6.5% cardiac mortality rate and 10.5% total mortality rate. This study demonstrates that the programmable implantable pacemaker-cardioverter-defibrillator is effective in preventing arrhythmic death, yet reduces patient exposure to repeated shock therapy. Reprogramming is usually necessary during follow-up for optimal function.

Agouti related protein (AgRP) is upregulated in Cushing's syndrome.

Alpha melanocyte-stimulating hormone (alpha-MSH) is an agonist at the melanocortin 3 (MC3-R) and melanocortin 4 (MC4-R) receptors. Alpha-MSH stimulates corticosterone release from rat and human adrenal cells. Patients with Cushing's syndrome have elevated levels of serum alpha-MSH. Agouti related protein (AgRP) is an endogenous antagonist at the MC3-R and MC4-R and is expressed in the rat adrenal cortex. AgRP antagonises alpha-MSH-induced corticosterone release from rat and bovine adrenal cells. This suggests that AgRP may have an inhibitory paracrine role in the adrenal gland. We measured adrenal AgRP mRNA expression and circulating AgRP in 2 patients with Cushing's syndrome and controls. Adrenal AgRP mRNA expression and plasma AgRP were higher in the patients with Cushing's syndrome compared to controls. Plasma AgRP in the patients with Cushing's syndrome following bilateral adrenalectomy and hydrocortisone replacement were similar to the levels seen in controls. Our results suggest that AgRP may have a novel inhibitory paracrine role in the human adrenal gland.

Cloning and characterization of a novel cDNA that is IFN-gamma-induced in mouse peritoneal macrophages and encodes a putative GTP-binding protein.

Macrophage activation by IFN-gamma results in a cascade of gene expression. To identify genes activated in mouse peritoneal macrophages by IFN-gamma, we created a cDNA subtraction library of IFN-gamma-induced genes. We have isolated from this subtraction library a novel cDNA clone, called Mg21, whose mRNA is absent in unstimulated mouse peritoneal macrophages and is induced to high levels within 4 h following the addition of IFN-gamma. Induction of Mg21 mRNA by IFN-gamma occurred in the presence of cycloheximide, indicating that expression of Mg21 mRNA does not require protein synthesis. A small amount of Mg21 mRNA was also induced by LPS, but not by IL-2, IL-4, IL-10, or TNF-alpha. The DNA sequence of Mg21 is 1617 nucleotides and contains an open reading frame that codes for a protein of 415 amino acids with a predicted molecular weight of 47,106 Da. The predicted amino acid sequence lacks a signal sequence or transmembrane segments, indicating that the protein is an intracellular protein. Computer search of the GenBank and EMBL databases indicates that this cDNA clone is unique but has 57% sequence identity with IRG-47, which is a mouse gene induced by IFN-gamma in pre-B and B lymphocyte cell lines. IRG-47 encodes an intracellular protein that contains three conserved protein motifs present in GTP-binding proteins. Analysis of the protein sequence of Mg21 showed that these three conserved protein motifs are also present in Mg21.

IFN-gamma increases cathepsin H mRNA levels in mouse macrophages.

Expression of major histocompatibility complex (MHC) class II molecules and ability to present antigen to T lymphocytes is acquired upon activation of the macrophage by interferon-gamma (IFN-gamma). Little information is available concerning immune regulation of protease gene expression in mouse macrophages. We have isolated a cDNA clone for cathepsin H, a lysosomal cysteine proteinase from a cDNA subtraction library of mouse macrophage genes induced by IFN-gamma, and have characterized its expression. The level of cathepsin H mRNA increased in mouse peritoneal macrophages following addition of IFN-gamma. Cathepsin H mRNA levels began to increase 8 h after the addition of IFN-gamma and was maximal at 24-48 h. This increase was concordant in time with appearance of MHC class II E beta mRNA and Ia invariant chain mRNA. The increase in cathepsin H mRNA levels by IFN-gamma was dose dependent. Cycloheximide treatment of peritoneal macrophages inhibited the increase in cathepsin H mRNA levels induced by IFN-gamma, suggesting that the increase in cathepsin mRNA levels requires de novo protein synthesis. Lipopolysaccharide and cytokines interleukin-2 (IL-2), IL-4, IL-10, and tumor necrosis factor alpha were found to have no effect on cathepsin H mRNA levels in mouse peritoneal macrophages.

Inducible expression of cytokinin biosynthesis in Agrobacterium tumefaciens by plant phenolics.

Nopaline strains of Agrobacterium tumefaciens contain a gene, tzs, that encodes a cytokinin biosynthetic prenyl transferase. The gene is located adjacent to the Ti plasmid virulence region and is constitutively expressed at low levels. As a result, bacteria containing tzs secrete low levels of zeatin into the medium. We find zeatin secretion to be induced more than 100-fold by acetosyringone, one of a number of naturally occurring phenolics produced by plants in response to wounding. Induction was very sensitive to the pH of the medium (optimum pH 5.5) and was due to massive overexpression of tzs-encoded cytokinin prenyl transferase activity. The relative ability of members of a set of phenols to induce tzs expression was examined and found to be parallel to that reported for activation of other virulence genes. A series of molecular cloning experiments established that virA and virG, two genes known to be essential to the virulence induction process, were necessary and sufficient for phenolic-induced tzs expression. Sequences present in the promoter region of tzs were found to be similar to those present in genes regulated by bacterial two-component positive regulatory systems.

Therapy of ventricular tachycardia.

Therapeutic modalities for ventricular tachycardia include antiarrhythmic drugs, direct current cardioversion, electrical pacing and surgical intervention. Lidocaine, procainamide and bretylium are all capable of controlling recurrent ventricular tachycardia; bretylium has the advantage of also being antifibrillatory and of raising the threshold for ventricular fibrillation. Lidocaine and bretylium are available only in i.v. form. Procainamide is available in i.v. as well as oral form. Other oral antiarrhythmic agents include quinidine, disopyramide, beta-blockers such as propranolol and verapamil. The latter may be useful in ventricular arrhythmias induced by ischemia; of these, only beta-blockers appear to significantly raise the threshold for ventricular fibrillation. Control of ventricular ectopy does not always preclude ventricular tachycardia and ventricular fibrillation. In treating ventricular tachycardia, bretylium tosylate is generally given 5 to 10 mg/kg i.v. over 10 to 20 minutes. Given too rapidly, it may cause nausea and vomiting. Orthostatic hypotension, a common side effect, generally abates with continued use and may be ameliorated with tricyclic antidepressants such as protriptyline. Significant supine hypotension may be encountered in patients with acute myocardial infarction and may be managed with pressor agents or fluids, or both. The antiarrhythmic efficacy of bretylium was analyzed in 40 patients. Five etiologic groups were defined by cardiac catheterization: 19 patients had atherosclerotic heart disease, 6 had primary myocardial disease, 4 had mitral valve prolapse, 4 had rheumatic heart disease and 7 had miscellaneous or no heart disease. All patients had recurrent ventricular tachycardia (VT); 23 had ventricular fibrillation (VF) as well. Other antiarrhythmic agents had failed in 38 patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Single-chamber ventricular pacing is not associated with worsening heart failure in sick sinus syndrome.

Ventricular pacing (with loss of normal atrioventricular synchrony) has been considered to have a role in the development or progression of congestive heart failure (CHF) in patients with sick sinus syndrome (SSS). No rigorous study has tested this hypothesis. Five hundred seven consecutive patients with SSS who received an initial pacemaker from January 1980 to December 1989 were studied. Atrial or dual-chamber pacemakers were implanted in 395 patients and ventricular pacemakers in 112. After a mean follow-up of 65 +/- 37 months, 97 patients (19%) developed new CHF or increased their New York Heart Association functional class. By univariate analysis, preimplant predictors for these events were left ventricular dysfunction (p < 0.001), valvular heart disease (p = 0.004), peripheral vascular disease (p = 0.005), diabetes (p = 0.02), coronary artery disease (p = 0.02), high New York Heart Association functional class (p = 0.03) and complex ventricular arrhythmia (p = 0.03). By multivariate analysis (logistic regression), the only predictors for CHF were valvular heart disease (p = 0.002; odds ratio [OR] 2.51), peripheral vascular disease (p = 0.003; OR 1.7) and complex ventricular arrhythmia (p = 0.027; OR 2.74). When the analysis was restricted to patients who had preimplant assessment of left ventricular function, independent predictors for CHF were left ventricular dysfunction (p < 0.001; OR 1.66), and complex ventricular arrhythmia (p < 0.001; OR 1.75). In conclusion, progressive or new-onset CHF is a consequence of the underlying cardiovascular disease. In the present population of patients with SSS, ventricular pacing mode was not associated with an increased incidence of CHF.

Three cases of hypotension and syncope with ventricular pacing: possible role of atrial reflexes.

Hypotension with lightheadedness and near syncope occurred in three patients during effective ventricular pacing. Hemodynamic studies demonstrated a decrease in cardiac output ranging from almost no decrease to 15 percent, presumably related to the loss of effective atrial contraction. The decrease in output was too small to explain by itself the reduced blood pressure, which resulted from paradoxic reduction of total peripheral resistance in one patient and from failure of resistance to increase in two. Baroceptor reflexes (Valsalva response) were normal in all three patients; hence it is suggested that the failure of compensatory increases in total peripheral resistance may be due to a reflex from the sudden atrial distension that occurs during atrioventricular (A-V) dissociation. The fluctuations in arterial pressure during ventricular pacing were synchronous with the appearance of cannon waves in the right atrial pressure tracing. Arterial pressure during A-V dissociation thus appears to be balanced by two opposite reflexes: the baroceptor reflex, which attempts to compensate for reduction in output, and atrial distension, which reduces peripheral resistance.

Chronic transvenous pacemaker lead removal using a unique, sequential transvenous system.

Transvenous removal of 43 consecutive chronic pacemaker leads was successful in 28 patients. For leads not removed by traction at the pacemaker connection terminal, a unique locking stylet was advanced through the inner coil lumen and engaged at the tip to allow traction without lead elongation. Leads not extracted with the locking stylet alone had traction maintained on the stylet as sheaths were advanced over the lead to dilate and detach any fibrous tissue adherent to the lead. By applying traction at the pacemaker connection terminal, 2 leads were removed. The locking stylet alone extracted 9 leads. Both the locking stylet and sheaths were necessary to explant 32 leads. There were 15 right atrial and 22 right ventricular leads completely removed. Additionally, 6 right ventricular leads were subtotally removed leaving only the tip in the right ventricular apex. Avulsed myocardium was removed with the lead in 1 patient without sequelae. A subacute hemothorax developed in 1 patient 18 days after discharge requiring drainage, and subclavian vein thrombosis developed in another, which was successfully treated with anticoagulation. Hypotension occurred in 1 patient during final positioning of new leads, which responded to conservative treatment. Chronic pacemaker leads can be reliably removed without thoracotomy. Both a unique locking stylet to allow traction without lead elongation and a sheath to dilate and detach adherent fibrous tissue are needed for consistent success. Recognized complications included myocardial avulsion without sequelae, subacute hemothorax, subclavian vein thrombosis and transient hypotension.

Long-term control of congestive heart failure with captopril.

The long-term effects of captopril therapy were assessed by sequential hemodynamic studies over a 6 month period in 19 patients with resistant congestive heart failure. Initial improvement during the first week of therapy was noted only in 11 and was marked by significant (p less than 0.005) increases in cardiac output and stroke volume, slowing of heart rate, and reduction of total peripheral resistance. Of the remaining eight patients, seven improved subsequently with maintained therapy so that by the end of 3 months of treatment only one patient failed to respond significantly. The hemodynamic index that reflected response most consistently was the shortening in pulmonary mean transit time. Simultaneously with clinical improvement there was a reduction in cardiopulmonary volume that reflected a reduction in pulmonary congestion and probably systemic vasodilation. Associated with these hemodynamic changes there was an increase in plasma renin activity and a significant reduction in plasma aldosterone, but these changes did not differ significantly between patients who responded markedly and those who responded moderately to converting enzyme inhibition. These results suggest that the response of congestive heart failure to captopril can occur gradually. Improvement was related to peripheral hemodynamic changes which led to a reduction in both total peripheral resistance and cardiopulmonary volume. The reduction in the plasma aldosterone/renin activity ratio was an effective marker of compliance.

Initial experience with the maze procedure for atrial fibrillation.

From January 1991 until May 1992, a total of 14 patients (mean age 48 years) underwent the maze procedure for refractory atrial fibrillation (mean duration, 7 years; mean number of antiarrhythmic medications, six). Three patients had had embolic events, one patient had had a cardiac arrest from flecainide, one had pulmonary fibrosis from amiodarone, and six of ten who were employed were temporarily disabled. Two patients underwent successful mitral valve repair in which the maze procedure was added as a secondary goal of the operation. Postoperative fluid retention was a problem in five patients (36%). Six patients (43%) were temporarily treated with an antiarrhythmic medication. Two patients (14%) with preoperative sick sinus syndrome required pacemakers. One patient was discharged from the hospital but died suddenly less than 1 month after the operation (7% operative mortality) of hyperkalemia caused by acute renal failure. All patients beyond 3 postoperative months (100% "cure") are receiving no antiarrhythmic medications, have sinus rhythm, or have p-wave tracking with ventricular pacing. Atrial contraction has been documented by cinegraphic magnetic resonance imaging studies and by Doppler echocardiography performed when sinus rhythm had resumed. The maze procedure is an extensive operation but is indicated for selected patients who have the severe sequelae of atrial fibrillation.

Sedation and pain management during pacemaker implantation, electrophysiologic testing, and related procedures.

A technique for the management of pain and anxiety during pacemaker implantation, electrophysiologic studies, and related procedures is described. The technique involves the intravenous administration of small amounts of the narcotic agonist-antagonist nalbuphine and the benzodiazepine diazepam. The small amounts of medication used induce relaxation while allowing the patient to interact and cooperate during the procedure. The absence of side effects facilitates outpatient treatment. This method of pain management not only improves the quality of patient care but also encourages earlier hospital discharge.

Determination of bisphenol-type contaminants from food packaging materials in aqueous foods by solid-phase microextraction-high-performance liquid chromatography.

A fast screening method consisting of off-line solid-phase microextraction coupled to HPLC and fluorescence detection, suitable for the analysis of several bisphenol derivatives and their degradation products in aqueous solution, has been developed. Detection limits of 0.7 ng ml(-1) for 2,2-bis[4-(glycidyloxy)phenyl]propane, 0.9 ng ml(-1) for bisphenol A bis(3-chloro-2-hydroxypropyl)ether, 1.1 ng ml(-1) for 2,2-bis(4-hydroxyphenyl)propane and 2.4 ng ml(-1) for bisphenol F diglycidyl ether have been achieved working in the linear range 10-500 ng ml(-1). The good analytical features achieved make the proposed method an interesting option for the direct determination of these compounds in aqueous canned food such as peas, tuna, olives, maize, artichokes or palm hearts. Both the optimization process and the results, including the analysis of real samples, are given and discussed.

HIV/AIDS policies and sports: the National Football League.

With the assistance of highly respected experts in the field of HIV/AIDS, the NFL has developed a comprehensive policy that should decrease the spread of HIV and any other blood-borne pathogens among its players and medical staff. Transmission of HIV infection is likely to be rare in the NFL. This is supported by the fact that in over 10 yr of the AIDS epidemic, the CDC has not attributed one AIDS case to athletic competition. Whatever the rate of HIV infection, on-the-field transmission is certainly less frequent than hepatitis B, which is manyfold more transmissible than HIV. Based on these facts, a player with HIV infection poses virtually no threat to others or himself by further athletic participation in the NFL. In addition, the relatively short average playing career of an NFL player and the extended period between HIV contraction and development of AIDS symptoms decrease the prospect that a player's HIV-positive status would affect his athletic performance. Consequently, HIV testing should remain voluntary, and continued participation in the NFL of HIV-infected players should remain a private decision between the player and his physician.

The Cox-Maze procedure: the Cleveland Clinic experience.

The Cox-Maze procedure was designed to address the consequences of atrial fibrillation, tachycardia, hemodynamic impairment, and thromboembolism. From 1991 until June 1999, 100 patients underwent the Maze operation at the Cleveland Clinic Foundation. The group included 72 men with a mean age of 58 +/- 11 years (range, 23 to 78 years). Initially, the Maze-I procedure was performed primarily for patients with lone atrial fibrillation. However, since 1995, the Maze-III procedure has been performed exclusively, and it is typically combined with mitral valve repair. Twenty-three patients had only a Maze procedure, 60 patients had the Maze procedure/mitral valve repair, 10 patients had Maze procedure/coronary artery bypass, 6 had Maze procedure/mitral valve replacement, and 1 had Maze procedure/atrial septal defect repair. Chronic atrial fibrillation was present in 78% of patients for a mean of 8 +/- 9 years. There was a 1% perioperative mortality and 5% late mortality rate. Median hospital stay was 9 +/- 5 days. Six patients required new early permanent pacemaker insertion. With a mean follow-up of 3 years, 90.4% of patients are in sinus rhythm (or atrial pacing). Preoperative symptoms were reduced: 24% had preoperative syncope; none had syncope in follow-up; 14% of patients preoperatively had cerebral or systemic emboli; and there were no perioperative or late embolic events. The Maze procedure effectively addressed the major complications of atrial fibrillation and was associated with low perioperative and late morbidity rates.

Analysis of annatto (Bixa orellana) food coloring formulations. 2. Determination of aromatic hydrocarbon thermal degradation products by gas chromatography.

Twenty samples of commercial annatto formulations have been analyzed for m-xylene and toluene using ambient alkaline hydrolysis, followed by solvent extraction and capillary gas chromatography. Fifteen of the samples contained <5 mg/kg toluene, four samples contained between 5 and 10 mg/kg toluene, and one sample contained 12 mg/kg toluene. The amounts found of m-xylene were 200 mg/kg (one sample), 160 mg/kg (one sample), between 30 and 88 mg/kg (four samples), between 7 and 25 mg/kg (seven samples), and <5 mg/kg (seven samples). Bixin-in-oil formulations contained the highest m-xylene concentrations and also gave the largest increase in headspace m-xylene concentration when heated in closed systems. The results are evidence for the thermal degradation of annatto during source extraction and processing, resulting in contamination by internal generation of both bixin and norbixin types with aromatic hydrocarbons. Two samples of norbixin of known production history (i. e., thermal versus nonthermal processes) were analyzed specifically to identify possible differences in their degradation component profiles. They were found to differ significantly in m-xylene content, which is consistent with their respective production histories.