Assessment of cerebral drug occupancy in humans using a single PET-scan: A [11C]UCB-J PET study.

PURPOSE: Here, we evaluate a PET displacement model with a Single-step and Numerical solution in healthy individuals using the synaptic vesicle glycoprotein (SV2A) PET-tracer [11C]UCB-J and the anti-seizure medication levetiracetam (LEV). We aimed to (1) validate the displacement model by comparing the brain LEV-SV2A occupancy from a single PET scan with the occupancy derived from two PET scans and the Lassen plot and (2) determine the plasma LEV concentration-SV2A occupancy curve in healthy individuals. METHODS: Eleven healthy individuals (five females, mean age 35.5 [range: 25-47] years) underwent two 120-min [11C]UCB-J PET scans where an LEV dose (5-30 mg/kg) was administered intravenously halfway through the first PET scan to partially displace radioligand binding to SV2A. Five individuals were scanned twice on the same day; the remaining six were scanned once on two separate days, receiving two identical LEV doses. Arterial blood samples were acquired to determine the arterial input function and plasma LEV concentrations. Using the displacement model, the SV2A-LEV target engagement was calculated and compared with the Lassen plot method. The resulting data were fitted with a single-site binding model. RESULTS: SV2A occupancies and VND estimates derived from the displacement model were not significantly different from the Lassen plot (p = 0.55 and 0.13, respectively). The coefficient of variation was 14.6% vs. 17.3% for the Numerical and the Single-step solution in Bland-Altman comparisons with the Lassen plot. The average half maximal inhibitory concentration (IC50), as estimated from the area under the curve of the plasma LEV concentration, was 12.5 µg/mL (95% CI: 5-25) for the Single-Step solution, 11.8 µg/mL (95% CI: 4-25) for the Numerical solution, and 6.3 µg/mL (95% CI: 0.08-21) for the Lassen plot. Constraining Emax to 100% did not significantly improve model fits. CONCLUSION: Plasma LEV concentration vs. SV2A occupancy can be determined in humans using a single PET scan displacement model. The average concentration of the three computed IC50 values ranges between 6.3 and 12.5 µg/mL. The next step is to use the displacement model to evaluate LEV occupancy and corresponding plasma concentrations in relation to treatment efficacy. CLINICAL TRIAL REGISTRATION: NCT05450822. Retrospectively registered 5 July 2022 https://clinicaltrials.gov/ct2/results? term=NCT05450822&Search=Search.

Supporting Patient-Clinician Interaction in Chronic HIV Care: Design and Development of a Patient-Reported Outcomes Software Application.

BACKGROUND: The consideration of health-related quality of life (HRQL) is a hallmark of best practice in HIV care. Information technology offers an opportunity to more closely engage patients with chronic HIV infection in their long-term management and support a focus on HRQL. However, the implementation of patient-reported outcome (PRO) measures, such as HRQL in routine care, is challenged by the need to synthesize data generated by questionnaires, the complexity of collecting data between patient visits, and the integration of results into clinical decision-making processes. OBJECTIVE: Our aim is to design and pilot-test a multimedia software platform to overcome these challenges and provide a vehicle to increase focus on HRQL issues in HIV management. METHODS: A multidisciplinary team in France and Australia conducted the study with 120 patients and 16 doctors contributing to the design and development of the software. We used agile development principles, user-centered design, and qualitative research methods to develop and pilot the software platform. We developed a prototype application to determine the acceptability of the software and piloted the final version with 41 Australian and 19 French residents using 2 validated electronic questionnaires, the Depression, Anxiety and Stress Scale-21 Items, and the Patient Reported Outcomes Quality of Life-HIV. RESULTS: Testing of the prototype demonstrated that patients wanted an application that was intuitive and without excessive instruction, so it felt effortless to use, as well as secure and discreet. Clinicians wanted the PRO data synthesized, presented clearly and succinctly, and clinically actionable. Safety concerns for patients and clinicians included confidentiality, and the potential for breakdown in communication if insufficient user training was not provided. The final product, piloted with patients from both countries, showed that most respondents found the application easy to use and comprehend. The usability testing survey administered found that older Australians had reduced scores for understanding the visual interface (P=.004) and finding the buttons organized (P=.02). Three-fourths of the respondents were concerned with confidentiality (P=.007), and this result was more prevalent in participants with higher anxiety and stress scores (P=.01), as measured by the Depression, Anxiety and Stress Scale-21 Items. These statistical associations were not observed in 15 French patients who completed the same questionnaire. CONCLUSIONS: Digital applications in health care should be safe and fit for purpose. Our software was acceptable to patients and shows potential to overcome some barriers to the implementation of PROs in routine care. The design of the clinicians' interface presents a solution to the problem of voluminous data, both synthesizing and providing a snapshot of longitudinal data. The next stage is to conduct a randomized controlled trial to determine whether patients experience increased satisfaction with care and whether doctors perceive that they deliver better clinical care without compromising efficiency.

The International development of PROQOL-HCV: An instrument to assess the health-related quality of life of patients treated for Hepatitis C virus.

BACKGROUND: Hepatitis C virus (HCV) compromises Health-related Quality of Life (HRQL) with detriments to Physical, Mental and Social health domains. Treatment with interferon and ribavirin is associated with side effects which further impair HRQL. New treatments appear potent, effective and tolerable. However, Patient Reported Outcomes instruments that capture the impact on HRQL for people with hepatitis C are largely non-specific and will be needed in the new treatment era. Therefore, we developed a conceptually valid multidimensional model of HCV-specific quality of life and pilot survey instrument, the Patient Reported Outcome Quality of Life survey for HCV (PROQOL-HCV). METHODS: HCV patients from France (n = 30), Brazil (n = 20) and Australia (n = 20) were interviewed to investigate HCV-HRQL issues raised in the scientific literature and by treatment specialists. Interviews were recorded, transcribed and translated into English and French. RESULTS: Fifteen content dimensions were derived from the qualitative analysis, refined and fitted to four domains: (1) Physical Health included: fatigue, pain, sleep, sexual impairment and physical activity; (2) Mental Health: psychological distress, psychosocial impact, and cognition; (3) Social Health: support, stigma, social activity, substance use; (4) TREATMENT: management, side effects, and fear of treatment failure. The impact of some dimensions extended beyond their primary domain including: physical activity, cognition, sleep, sexual impairment, and the three treatment dimensions. A bank of 300 items was constructed to reflect patient reports and, following expert review, reduced to a 72-item pilot questionnaire. CONCLUSION: We present a conceptually valid multidimensional model of HCV-specific quality of life and the pilot survey instrument, PROQOL-HCV. The model is widely inclusive of the experience of hepatitis C and the first to include the treatment dimension.

Psychometric assessment of health-related quality of life and symptom experience in HIV patients treated with antiretroviral therapy.

PURPOSE: Symptoms which are found to cluster consistently can have synergistic effects on patient outcomes and therefore may serve to predict morbidity or disentangle disease progression from comorbid conditions. Self-report HIV-specific symptom and HRQL measures were jointly analyzed in HIV-positive patients under different antiretroviral treatment regimens. METHODS: The responses of N = 365 patients from four countries to the 9-item Physical Health and Symptom dimension of the PROOQL-HIV questionnaire and an HIV Symptom checklist were analyzed. Item response modeling and multidimensional scaling were used to derive HRQL scores free of any differential item functioning related to gender and target language and to summarize symptom co-expression in patients under protease inhibitor treatment(PI, N = 164, 45%) versus other medication (Non-PI). RESULTS: Women reported poorer HRQL (p = 0:037), and HRQL did not differ between the target languages of French, English, and Thai. Fatigue, muscular pain, or difficulties falling asleep was the most frequently reported symptoms [35%). PI versus Non-PI patients exhibited different pattern of symptoms with lipodystrophy-related and gastrointestinal symptoms forming well-separated clusters in the PI group. A higher number of symptoms were associated with lower HRQL (p < 0:001), and patients taking PIs reported lower HRQL (p = 0:003). Patients in both groups who reported fatigue, sexual dysfunction, or several lipodystrophy-related symptoms had poorer quality of life.

Safety of poly-L-lactic acid (New-Fill®) in the treatment of facial lipoatrophy: a large observational study among HIV-positive patients.

BACKGROUND: Facial lipoatrophy is a frequently reported condition associated with use of antiretroviral (ARV) drugs. Poly-L-lactic acid (PLLA) acid has been used to correct facial lipoatrophy in people with HIV since 2004 both in Europe and the United States. The objective of this study was to establish, in real life conditions and in a large sample, the safety of PLLA (New Fill®, Valeant US, Sinclair Pharma Paris, France) to correct facial lipoatrophy among HIV-positive patients. METHODS: A longitudinal study was conducted between 2005 and 2008 in France. Data from 4,112 treatment courses (n = 4,112 patients) and 15,665 injections sessions (1 to 5 injection sessions per treatment course) were gathered by 200 physicians trained in the use of PLLA. RESULTS: The average age of patients (88.3% males) treated for lipoatrophy was 47.1 ± 8.1 years (Mean ± SD); 91.2% of patients had been receiving ARV treatment for 10.9 (±4.2) years; CD4 T-cell count was 535 ± 266 cells/mm3. The duration of facial lipoatrophy was 5 ± 2.8 years and the severity was such that 47.3% of patients required five injection sessions of PLLA and 81.9% of the sessions required two vials of the preparation. The final visit, scheduled two months after the last injection session, was attended by 66.0% of patients (n = 2,713). 48 treatment courses (2.8%) were discontinued due to adverse events (AEs). The overall incidence of AEs per course was 18.8%. Immediate AEs, bleeding (3.4%), bruising (2.3%), pain (2.0%), redness at injection site (1.6%), and swelling of the face (0.7%), occurred in 15.4% of courses and 7.0% of sessions (usually during the first session). Non-immediate AEs, mainly nodules (5.7%), inflammation (0.7%), granuloma (0.3%), discolouration (0.2%), and skin hypertrophy (0.1%), occurred in 6.7% of courses. Non-immediate AEs occurred within a time ranging from 21 days (inflammation) to 101 days (granuloma) and all but three of the 13 cases of granuloma resolved. Product efficacy was rated satisfactory by 95% of the patients and physicians. CONCLUSIONS: This study demonstrated, in real-life conditions and on a large sample, that PLLA injections were feasible, efficient, and safe when performed by trained physicians.

Electronic versus paper-based assessment of health-related quality of life specific to HIV disease: reliability study of the PROQOL-HIV questionnaire.

BACKGROUND: Electronic patient-reported outcomes (PRO) provide quick and usually reliable assessments of patients' health-related quality of life (HRQL). OBJECTIVE: An electronic version of the Patient-Reported Outcomes Quality of Life-human immunodeficiency virus (PROQOL-HIV) questionnaire was developed, and its face validity and reliability were assessed using standard psychometric methods. METHODS: A sample of 80 French outpatients (66% male, 52/79; mean age 46.7 years, SD 10.9) were recruited. Paper-based and electronic questionnaires were completed in a randomized crossover design (2-7 day interval). Biomedical data were collected. Questionnaire version and order effects were tested on full-scale scores in a 2-way ANOVA with patients as random effects. Test-retest reliability was evaluated using Pearson and intraclass correlation coefficients (ICC, with 95% confidence interval) for each dimension. Usability testing was carried out from patients' survey reports, specifically, general satisfaction, ease of completion, quality and clarity of user interface, and motivation to participate in follow-up PROQOL-HIV electronic assessments. RESULTS: Questionnaire version and administration order effects (N=59 complete cases) were not significant at the 5% level, and no interaction was found between these 2 factors (P=.94). Reliability indexes were acceptable, with Pearson correlations greater than .7 and ICCs ranging from .708 to .939; scores were not statistically different between the two versions. A total of 63 (79%) complete patients' survey reports were available, and 55% of patients (30/55) reported being satisfied and interested in electronic assessment of their HRQL in clinical follow-up. Individual ratings of PROQOL-HIV user interface (85%-100% of positive responses) confirmed user interface clarity and usability. CONCLUSIONS: The electronic PROQOL-HIV introduces minor modifications to the original paper-based version, following International Society for Pharmacoeconomics and Outcomes Research (ISPOR) ePRO Task Force guidelines, and shows good reliability and face validity. Patients can complete the computerized PROQOL-HIV questionnaire and the scores from the paper or electronic versions share comparable accuracy and interpretation.

Plastics in human diets: development and evaluation of the 24-h Dietary Recall - Plastic Exposure and the Dietary Plastics Score.

Background: Humans are commonly exposed to plastic through their dietary intake and food consumption patterns. Plastic-associated chemicals (PAC), such as bisphenols and phthalates, are recognized as endocrine-disrupting and are associated with increased risk of cardiovascular disease and metabolic syndrome. However, accurate methods to assess dietary exposure to plastic products and PAC are inadequate, limiting interrogation of health impacts. Aim: To develop a tool that captures complete dietary exposure to plastics and establish a diet quality score to measure adherence to a low plastic dietary pattern. Methods: We developed the 24-h Dietary Recall - Plastic Exposure (24DR-PE) and administered it to healthy adults (n = 422). This computer-assisted, interviewer-administered tool systematically collects data on food volumes and types, packaging materials, storage, processing, cooking, and consumption methods to assess a food's exposure to plastic. Specifically, the 24DR-PE incorporates predefined criteria for identifying high-risk practices and food characteristics, such as individually packaged items or those microwaved in plastic, enabling the assignment of scores based on a theoretically derived Dietary Plastics Scoring Matrix. Conclusion: The 24DR-PE is the first tool specifically designed to capture detailed data on dietary exposures to plastic products. The next step is to validate the score using laboratory results of urine samples we collected contemporaneous to the dietary information. Once validated, the tool has potential for widespread distribution making it valuable for population monitoring, intervention guidance, and future research investigating the interplay between plastics, diet, and human health.

Patient-related factors impact the implementation of inpatient antibiotic allergy delabeling.

Background: The clinical consequences of an antibiotic allergy label are detrimental, impacting health care delivery and patient outcomes. We assessed hospital inpatients with intent to offer free antibiotic allergy labeling (AAL) assessment within a randomized controlled trial. Objective: We sought to determine the feasibility of establishing an adult antibiotic allergy delabeling service in a Western Australian tertiary public hospital. Methods: Inpatients (N = 1503) with AAL were identified through medical records and screened for eligibility to participate in a randomized controlled trial. Those recruited were randomized to undergo assessment by skin testing ± oral challenge, or direct oral challenge. A control group received usual care. Results: Of the 1503 inpatients with an AAL, 429 (28.5%) were eligible for AAL assessment. The primary excluding factor (1074 [71.5%]) was contraindicated medication use (387 [36.0%]), followed by cognitive impairment (298 [27.9%]). Thirty-nine patients were randomized, of which 20 received allergy testing and 19 usual care; all patients were followed up for 5 years. Older patients were less likely to be eligible (10-year increase: odds ratio, 0.82; 95% CI, 0.77-0.88; P < .0001), whereas surgical patients were more likely to be eligible than medical patients (odds ratio, 2.49; 95% CI, 1.97-3.16; P < .0001). Conclusions: Antibiotic allergy delabeling in the acute care context is not straightforward. Competing clinical concerns and patient acceptance are some barriers to an inpatient service. Nor is it apparent that inpatient versus outpatient testing is cost saving although select patient groups may benefit. Testing younger people and those with predicted high antibiotic usage will derive maximal individual and health system benefits.

Comparing Skin and Serum Testing to Direct Challenge Outcomes in Children With ß-Lactam Allergies.

BACKGROUND: There is a scarcity of prospective studies investigating the relative roles of skin prick and intradermal testing, serum specific IgE, and extended oral challenges in diagnosing children with reported ß-lactam allergies. OBJECTIVE: To determine the sensitivity and specificity of skin testing and serum specific IgE in children with ß-lactam allergies, with immediate and nonimmediate historic reactions. METHODS: Four hundred children with parent-reported ß-lactam allergies were recruited into an open-label prospective study. Detailed allergy histories were collected. Those with medically observed and documented histories of anaphylaxis, requiring epinephrine, or severe cutaneous adverse reactions were excluded. In total, 380 children underwent all testing modalities and a direct provocation test. Each child was followed up for a minimum of 3 years. RESULTS: True allergy in children was uncommon; 8.3% reacted to the direct provocation challenge or the 5-day extended oral provocation challenge. Children reporting cephalosporin allergy or a reaction within 1 year were more likely to react to direct provocation testing. The sensitivity, specificity, and positive predictive value of skin testing were 12.5%, 98.8%, and 20.0% for direct challenge outcomes, 4.76%, 99.0%, and 25.0% for extended challenge outcomes, and 6.9%, 99.0%, and 40.0% for both challenges combined, respectively. Follow-up investigations revealed that 5.7% of children had a mild repeat reaction and 2.7% continued to avoid the culprit despite successful delabeling. The relabeling rate for children readmitted to hospital was 15%, with the relabeing being unfounded. CONCLUSIONS: Genuine ß-lactam allergies were rare, with over 90% of children effectively delabeled. Skin and serum specific IgE testing did not aid the diagnosis of ß-lactam antibiotic allergy in children, regardless of medical history. Extended oral challenges proved valuable in confirming allergies and boosted parental confidence.

HIV-related stigma and physical symptoms have a persistent influence on health-related quality of life in Australians with HIV infection.

BACKGROUND: The health-related quality of life (HRQL) of people living with HIV infection is an important consideration in HIV management. The PROQOL-HIV psychometric instrument was recently developed internationally as a contemporary, discriminating HIV-HRQL measure incorporating influential emotional dimensions such as stigma. Here we present the first within-country results of PROQOL-HIV using qualitative and quantitative data collected from a West Australian cohort who participated in the development and validation of PROQOL-HIV, and provide a comprehensive picture of HRQL in our setting. METHODS: We carried out a secondary analysis of data from Australian patients who participated in the international study: 15 in-depth interviews were conducted and 102 HRQL surveys using the PROQOL-HIV instrument and a symptom questionnaire were administered. We employed qualitative methods to extract description from the interview data and linear regression for exploration of the composite and sub-scale scores derived from the survey. RESULTS: Interviews revealed the long-standing difficulties of living with HIV, particularly in the domains of intimate relationships, perceived stigma, and chronic ill health. The novel PROQOL-HIV instrument discriminated impact of treatment via symptomatology, pill burden and treatment duration. Patients demonstrated lower HRQL if they were: newly diagnosed (p=0.001); naive to anti-retroviral treatment (p=0.009); reporting depression, unemployment or a high frequency of adverse symptoms, (all p<0.001). Total HRQL was notably reduced by perceived stigma with a third of surveyed patients reporting persistent fears of both disclosing their HIV status and infecting others. CONCLUSIONS: The analysis showed that psychological distress was a major influence on HRQL in our cohort. This was compounded in people with poor physical health which in turn was associated with unemployment and depression. People with HIV infection are living longer and residual side effects of the earlier regimens complicate current clinical management and affect their quality of life. However, even for the newly diagnosed exposed to less toxic regimens, HIV-related stigma exerts negative social and psychological effects. It is evident that context-specific interventions are required to address persistent distress related to stigma, reframe personal and public perceptions of HIV infection and ameliorate its disabling social and psychological effects.

Levels of nitrate, nitrite and nitrosamines in model sausages during heat treatment and in vitro digestion - The impact of adding nitrite and spinach (Spinacia oleracea L.).

Nitrite derivatives react with endogenous precursors forming N-nitrosamines associated with development of colorectal cancer. The present study aims to investigate the formation of N-nitrosamines in sausage during processing and in vitro gastrointestinal digestion after adding sodium nitrite and/or spinach emulsion. The INFOGEST digestion protocol was used to simulate the oral, gastric, and small intestinal phases of digestion, and sodium nitrite was added in the oral phase to mimic the input of nitrite from saliva as it has shown to affect the endogenous formation of N-nitrosamines. The results show that the addition of spinach emulsion, in spite of it being a source of nitrate, did not affect the nitrite content in either batter, sausage, or roasted sausage. The levels of N-nitrosamines increased with the added amount of sodium nitrite, and further formation of some volatile N-nitrosamines was observed during roasting and in vitro digestion. In general, N-nitrosamine levels in the intestinal phase followed the same trend as in the undigested products. The results further indicate that nitrite present in saliva may cause a significant increase in N-nitrosamine levels in the gastrointestinal tract and that bioactive components in spinach may protect against the formation of volatile N-nitrosamines both during roasting and digestion.

The role of endocrine-disrupting phthalates and bisphenols in cardiometabolic disease: the evidence is mounting.

PURPOSE OF REVIEW: There is substantive and accumulating evidence that endemic exposure to plastic-associated chemicals (PACs) contribute to the pathophysiology of metabolic conditions, like obesity, diabetes, and heart disease. The consequences of this endemic exposure in inducing a pro-inflammatory state in adipose tissues as a critical link between exposure and disease is reviewed. RECENT FINDINGS: In general, PACs are classified as nonpersistent in vivo because of their rapid metabolism to easily excreted forms. The parental chemicals, however, are typically lipophilic, with the potential to bioaccumulate. Recent data from selected association studies suggest exposure to PACs drive predisease states like obesity and inflammation of the adipose tissues. A range of experimental studies are discussed with a focus on biological mechanisms that are susceptible to the influence of PACs and which may promote metabolic disease, the detection of PACs within susceptible tissues and biological effects that are detectable at doses that correspond to real-life exposures to these chemicals. SUMMARY: If we hypothesize the toxic pressure from chronic exposure to PACs will progress disease processes, then individuals with comprehensively characterized indicators of premetabolic disease could undergo trials of quantifiable interventions to reduce exposure to PACs to test if the trajectory of disease-associated analytes, is altered.

Chlorate and perchlorate residues in food products on the Danish market.

Upon the European commission's request in 2013, Denmark initiated a survey of the levels of chlorate and perchlorate in agriculture products from the Danish market over a period of 4 years from 2014 to 2017. The results of the survey were used to guide decision making with a view to setting maximum levels for intra-Union trade in 2013 and reviewing and updating the levels from 2020. The results of the Danish survey and the employed analytical method are documented in this paper. In addition to the 89 domestic samples, 30 samples of foreign origin were analysed for chlorate and perchlorate, together with ten samples of baby food and seven samples of infant formula collected from the Danish market. Samples were extracted using the Quick Polar Pesticides extraction method (QuPPe) and analysed on a triple quadrupole LC-MS/MS system. The levels of both chlorate and perchlorate detected in samples originating from Denmark were lower than the current MRLs except for a single sample of lettuce. Moreover, the percentage of foreign samples that were positive for chlorate and perchlorate residues was higher than the percentage of domestic samples. The reason for the generally low levels and low frequency of chlorate and perchlorate in Danish products is most likely linked to the fact that the water supply in Denmark is 100% unchlorinated ground water. The present study shows that residues of perchlorate and chlorate are found in Danish food products at a relatively low frequency and that the levels found are below the EU's MRLs implemented in 2020.

Development of a new generic extraction method for the analysis of pesticides, mycotoxins, and polycyclic aromatic hydrocarbons in representative animal feed and food samples.

Various generic extraction methods have been used to determine pesticide residues, mycotoxins, and polycyclic aromatic hydrocarbons (PAHs) in food and animal feed to ensure consumer safety. However, these methods cannot extract all relevant compounds at an acceptable rate of recovery. This study presents a new extraction method. This new method facilitated the identification of 231 compounds, including 196 pesticides, 11 mycotoxins, and 24 PAHs over a broad range of polarities. These compounds were identified in various sample matrices, including those that are lipid-rich. The processed sample is first extracted with water, acetonitrile, formic acid, and heptane. The addition of ammonium formate results in separation into three phases and enables analysis of the aqueous phase. Solid-phase extraction clean-up procedures were performed as necessary followed by analysis by liquid or gas chromatography and mass spectrometry. Analyte recoveries were typically in the range of 70 - 120% with relative standard deviations below 20%.

Processing factors of pesticide residues in biscuits and their relation to the physicochemical properties of pesticides.

Agricultural commodities are generally consumed as processed food. Therefore, it is indispensable to assess pesticide residues in processed products rather than only in the raw agricultural commodity, in order to approach a more realistic scenario of dietary exposure. Processing factors are important tools for dietary exposure risk assessments. In this study, processing factors for the baking process were derived for 41 pesticides in cereal bran-based biscuits. The raw materials used consisted of wheat, rye, oat, and barley grains with incurred pesticides, which originally was produced for test material for European Union Proficiency Tests. Information on physicochemical properties of pesticides was collected for understanding the fate of pesticides during the baking process. Average processing factors varied between 0.67 and 1.6. Most pesticide residues exhibited a reduction of pesticide residues of less than 24%, which correspond to a processing factor (PF) range between 1 and 0.76, showing resistance to the baking process. However, for polar compounds such as carbendazim and volatile compounds (chlorpyrifos-methyl, malathion, and pirimiphos-methyl) larger reduction rates were observed, up to 33% (PF: 0.67). In general, a prolonged baking time did not significantly affect the PF, because the main degradation process takes place within the first 6 min. However, this was not the case for the highly volatile compounds, highly polar compounds, and compounds of low degradation temperature. These latter were significantly reduced with prolonged baking time, resulting in a reduction rate of up to 95%, which means an almost complete elimination.

Screening for HIV-Associated Neurocognitive Impairment: Relevance of Psychological Factors and Era of Commencement of Antiretroviral Therapy.

Neurocognitive impairment (NCI) is common in people aging with HIV and can adversely affect health-related quality of life. However, early NCI may be largely asymptomatic and neurocognitive function is rarely assessed in the context of routine clinical care. In this study, we considered the utility of two assessment tools as screens for NCI in patients attending a community-based clinic (N = 58; mean age = 57 years): the Montreal Cognitive Assessment (MoCA) and a 3-item cognitive concerns questionnaire derived from the HIV Dementia Scale. Health-related quality of life and depression/anxiety were also measured. Indication of NCI using the MoCA was more prevalent compared to the 3-item questionnaire and was associated with the patients' initial antiretroviral therapy commencing between the years of 1997 and 2001, independently of age. Findings of the MoCA were not confounded by existing mood disorders, unlike the 3-item questionnaire. Therefore, we suggest implementing the MoCA as an initial screen for NCI.

Role of Sample Processing Strategies at the European Union National Reference Laboratories (NRLs) Concerning the Analysis of Pesticide Residues.

The guidance document SANTE 11945/2015 recommends that cereal samples be milled to a particle size preferably smaller than 1.0 mm and that extensive heating of the samples should be avoided. The aim of the present study was therefore to investigate the differences in milling procedures, obtained particle size distributions, and the resulting pesticide residue recovery when cereal samples were milled at the European Union National Reference Laboratories (NRLs) with their routine milling procedures. A total of 23 NRLs participated in the study. The oat and rye samples milled by each NRL were sent to the European Union Reference Laboratory on Cereals and Feedingstuff (EURL) for the determination of the particle size distribution and pesticide residue recovery. The results showed that the NRLs used several different brands and types of mills. Large variations in the particle size distributions and pesticide extraction efficiencies were observed even between samples milled by the same type of mill.

Effects of milling on the extraction efficiency of incurred pesticides in cereals.

This study investigated the effects of particle size and milling temperature on the extraction efficiencies of pesticide residues from cereal flour. Samples of cereal grains (barley, oat, rye and wheat) were milled using a centrifugal mill with four different sieves (0.2, 1.0, 3.0 and 5.0 mm) or a knife mill both at room temperature and after freezing of the grain at -80°C overnight. The incurred pesticides in the test materials were extracted by the QuEChERS method and analysed by LC-MS/MS and GC-MS/MS. The particle size distribution for the milled samples was determined using a vibratory sieve shaker. Based on the pesticide levels recovered from each of the different millings and the corresponding particle size distributions, it was confirmed that smaller average particle sizes increase the extraction efficiency up to 31%, with all other factors equal. The cereals milled at room temperature produced lower pesticide extraction efficiencies compared with cereals milled when still frozen, especially for heat-sensitive pesticides. Furthermore, milling frozen grains was easier and resulted in more homogeneous samples with smaller relative particle sizes.

Clean-up of cereal extracts for gas chromatography-tandem quadrupole mass spectrometry pesticide residues analysis using primary secondary amine and C18.

The level of co-extracted matrix in wheat and oat extracts obtained by the QuEChERS method (EN 15662) is high and the occurrence of free fatty acids generates a major matrix peak in TIC chromatograms (rt. 13-22min). Matrix can compromise the analytical performance in pesticide analysis using GC-MS/MS. In order to reduce the amount and the effects of matrix we tested the effect of using six different amounts of primary secondary amine (PSA) (0, 25, 50, 100, 150 and 200mg/ml extract) with and without the addition of six different amounts of C18 (0, 25, 50, 100, 150 and 200mg/ml extract) in the dispersive solid phase extraction (dSPE) procedure. dSPE clean-up using 25mg/ml extract significantly reduced the major matrix peak observed for wheat extracts. Higher amounts of PSA reduced the analytical response for iprodione and malathion. For oat extract 50-150mg PSA/ml extract was needed to obtain equally low intensity of the matrix peak. For oat the analytical responses of the target pesticides generally increased with increasing amount of PSA. C18 had no significant effect on the intensity of the major matrix peaks and even resulted in lower analytical responses for several of the target pesticides. Based on the present study it is concluded that the optimal dSPE clean-up procedure employs 25mg PSA/ml extract for wheat and 150mg PSA/ml extract for oat.

Mitochondrial DNA depletion and morphologic changes in adipocytes associated with nucleoside reverse transcriptase inhibitor therapy.

BACKGROUND: Nucleoside analogue reverse transcriptase inhibitor (NRTI) therapy provides sufficient conditions for progressive subcutaneous fat wasting in HIV-infected patients. As NRTI-induced host toxicity is proposed to involve cellular mitochondrial DNA (mtDNA) depletion, determinants of cellular mtDNA copy number and mitochondrial mass in adipocyte samples from NRTI-treated HIV-infected patients and antiretroviral-naive controls were investigated. Adipose tissue morphology was also assessed. METHODS: Subcutaneous fat samples were obtained from NRTI-treated, HIV-infected patients (n = 21), antiretroviral therapy-naive HIV-infected controls (n = 11), and HIV-seronegative controls (n = 6). Non-adipocytes were removed by collagenase digestion. Adipocyte mtDNA copies/cell was measured using a real time PCR-based assay, and adipocyte mitochondrial protein content was also measured. Light and electron microscopy were performed on tissue samples. FINDINGS: Adipocyte mtDNA copies/cell values were similar (P = 0.56) in HIV seronegative and HIV-infected control groups. NRTI treatment was associated with reduced adipocyte mtDNA copies/cell, representing mean mtDNA depletion in NRTI-treated individuals of 77.7% compared with the mean value for the HIV-infected control group (P < 0001). Additionally, significant differences were found in adipocyte mtDNA copies/cell between patients receiving stavudine (n = 12, mean mtDNA depletion 87.1%) and zidovudine (n = 9, mean mtDNA depletion 52.1%) (P < 0.001). Adipocyte mitochondrial mass was increased in the stavudine group only (mean increase 289%, P < 0.01). INTERPRETATION: NRTI therapy is associated with mtDNA depletion and mitochondrial proliferation in adipocytes, consistent with the hypothesis that NRTI-induced mtDNA depletion contributes to the pathogenesis of subcutaneous fat wasting. Morphologic assessment also supports a role for NRTI therapy in inducing adipocyte metabolic dysfunction and cell death.