Benefit of inhalation aromatherapy as a complementary treatment for stress and anxiety in a clinical setting - A systematic review.

OBJECTIVE: The purpose of this systematic review is to ascertain the impact of inhalation aromatherapy on stress and anxiety in clinical settings. METHODS: A search strategy was developed using various databases. Randomised Controlled Trials (RCTs) as well as single and double-blind pilot clinical studies (non-RCT) using inhalation aromatherapy with an essential oil blend or a single essential oil were examined. All studies included a control intervention and use of a validated measurement tool. The time period under review was years 2000-2021. Due to the high level of heterogeneity and element of bias, a narrative synthesis was conducted. RESULTS: The search strategy initially retrieved 628 studies and through application of the selection criteria and the removal of duplicates, 76 studies were selected for review with a total of 6539 patients. In 42% of the RCTs, physiological measures including vital signs and/or salivary cortisol were used in addition to questionnaires. Over 70% of the studies reported a positive effect on anxiety levels in the aromatherapy intervention groups compared with the control. However, in many cases this is limited by the absence of safety data, imprecise reporting of plant species and dosage of essential oil. CONCLUSION: Inhalation aromatherapy has the potential to reduce stress and anxiety with data emerging to further support this result across a wide modality of clinical treatments. However, there is a clear need for the development of standard protocols for research in this area, generating measurable results which will create the opportunity for more rigorous evidence-based outcomes.

Incidence rate, predictors and outcomes of interruption of HIV care: nationwide results from the Belgian HIV cohort.

OBJECTIVES: We aimed to study the incidence rate, predictors and outcomes of HIV care interruption (HCI) in Belgium. METHODS: We analysed data for adult patients with at least two HIV care records in the Belgian HIV cohort between 1 January 2007 and 31 December 2016. An HCI episode was defined as 1 year without an HIV care record. The HCI incidence rate was analysed using Poisson regression, return to HIV care using a cumulative incidence function with death as a competing risk, and viral load (VL) status upon return to HIV care using logistic regression. RESULTS: We included 16 066 patients accounting for 78 625 person-years of follow-up. The incidence rate of HCI was 5.3/100 person-years [95% confidence interval (CI) 5.1-5.4/100 person-years]. The incidence of return to HIV care after HCI was estimated at 77.5% (95% CI 75.7-79.2%). Of those who returned to care, 43.7% had a VL ≤ 200 HIV-1 RNA copies/mL, suggesting care abroad or suboptimal care (without an HIV-related care record) in Belgium during the HCI, and 56.3% returned without controlled VL and were therefore considered as having experienced a real gap in HIV care; they represented 2.3/100 person-years of follow-up. Factors individually associated with HCI were no antiretroviral therapy (ART) uptake, lower age, injecting drug use, non-Belgian nationality, male gender, not being a man who has sex with men, a shorter time since HIV diagnosis, no high blood pressure and CD4 count < 350 cells/µL. CONCLUSIONS: This study highlights the need to investigate return to care and viral status at return, to better understand HCI. Identified predictors can help health care workers to target patients at higher risk of HCI for awareness and support.

Late presentation to HIV testing is overestimated when based on the consensus definition.

OBJECTIVES: In 2011, a consensus was reached defining "late presenters" (LPs) as individuals presenting for care with a CD4 count < 350 cells/µL or with an AIDS-defining event, regardless of CD4 count. However, a transient low CD4 count is not uncommon in recent infections. The objective of this study was to investigate how measurements of late presentation change if the clinical stage at the time of diagnosis is taken into account. METHODS: Case surveillance data for newly diagnosed patients in Belgium in 1998-2012 were analysed, including CD4 count at diagnosis, the presence of AIDS-defining events, and recent infections (< 6 months) as reported by clinicians in the case of acute illness or a recent negative test. First, proportions of LPs were calculated according to the consensus definition. Secondly, LPs were reclassified as "nonlate" if infections were reported as recent. RESULTS: A total of 7949 HIV diagnoses were included in the study. Recent infections were increasingly reported over time, accounting for 8.2% of new infections in 1998 and 37.5% in 2012. The consideration of clinical stage significantly modified the proportion of LPs: 18.2% of men who have sex with men (MSM) diagnosed in 2012 would be classified as LPs instead of 30.9% using the consensus definition (P < 0.001). The proportion of patients misclassified as LPs increased significantly over time: 5% in MSM in 1998 vs. 41% in 2012. CONCLUSIONS: This study suggests that low CD4 counts in recent infections may lead to overestimation of late presentation when applying the consensus definition. The impact of transient CD4 count on late presentation estimates should be assessed and, if relevant, the introduction of clinical stage in the definition of late presentation should be considered.

Transmission of HIV Drug Resistance and the Predicted Effect on Current First-line Regimens in Europe.

BACKGROUND: Numerous studies have shown that baseline drug resistance patterns may influence the outcome of antiretroviral therapy. Therefore, guidelines recommend drug resistance testing to guide the choice of initial regimen. In addition to optimizing individual patient management, these baseline resistance data enable transmitted drug resistance (TDR) to be surveyed for public health purposes. The SPREAD program systematically collects data to gain insight into TDR occurring in Europe since 2001. METHODS: Demographic, clinical, and virological data from 4140 antiretroviral-naive human immunodeficiency virus (HIV)-infected individuals from 26 countries who were newly diagnosed between 2008 and 2010 were analyzed. Evidence of TDR was defined using the WHO list for surveillance of drug resistance mutations. Prevalence of TDR was assessed over time by comparing the results to SPREAD data from 2002 to 2007. Baseline susceptibility to antiretroviral drugs was predicted using the Stanford HIVdb program version 7.0. RESULTS: The overall prevalence of TDR did not change significantly over time and was 8.3% (95% confidence interval, 7.2%-9.5%) in 2008-2010. The most frequent indicators of TDR were nucleoside reverse transcriptase inhibitor (NRTI) mutations (4.5%), followed by nonnucleoside reverse transcriptase inhibitor (NNRTI) mutations (2.9%) and protease inhibitor mutations (2.0%). Baseline mutations were most predictive of reduced susceptibility to initial NNRTI-based regimens: 4.5% and 6.5% of patient isolates were predicted to have resistance to regimens containing efavirenz or rilpivirine, respectively, independent of current NRTI backbones. CONCLUSIONS: Although TDR was highest for NRTIs, the impact of baseline drug resistance patterns on susceptibility was largest for NNRTIs. The prevalence of TDR assessed by epidemiological surveys does not clearly indicate to what degree susceptibility to different drug classes is affected.

Good continuum of HIV care in Belgium despite weaknesses in retention and linkage to care among migrants.

BACKGROUND: The Belgian HIV epidemic is largely concentrated among men who have sex with men and Sub-Saharan Africans. We studied the continuum of HIV care of those diagnosed with HIV living in Belgium and its associated factors. METHODS: Data on new HIV diagnoses 2007-2010 and HIV-infected patients in care in 2010-2011 were analysed. Proportions were estimated for each sequential stage of the continuum of HIV care and factors associated with attrition at each stage were studied. RESULTS: Of all HIV diagnosed patients living in Belgium in 2011, an estimated 98.2% were linked to HIV care, 90.8% were retained in care, 83.3% received antiretroviral therapy and 69.5% had an undetectable viral load (<50 copies/ml). After adjustment for sex, age at diagnosis, nationality and mode of transmission, we found lower entry into care in non-Belgians and after preoperative HIV diagnoses; lower retention in non-Belgians and injecting drug users; higher retention in men who have sex with men and among those on ART. Younger patients had lower antiretroviral therapy uptake and less viral suppression; those with longer time from diagnosis had higher ART uptake and more viral suppression; Sub-Saharan Africans on ART had slightly less viral suppression. CONCLUSIONS: The continuum of HIV care in Belgium presents low attrition rates over all stages. The undiagnosed HIV-infected population, although not precisely estimated, but probably close to 20% based on available survey and surveillance results, could be the weakest stage of the continuum of HIV care. Its identification is a priority along with improving the HIV care continuum of migrants.

HIV-1 resistance and trend analysis in newly diagnosed patients in Belgium.

A total of 1055 nucleotide sequences obtained from HIV patients diagnosed in 2008 and 2009 in Belgium were included in this prevalence study. The study population is a group of patients whose visit was considered by the clinician as the first contact with a Belgian AIDS reference centre or with another clinical centre experienced in HIV care. Prevalences of surveillance drug resistance mutations (SDRM) of 11·7% (47/394) and 11·0% (73/661) were observed in 2008 and 2009, respectively. The highest level of SDRM was observed towards nucleoside reverse transcriptase inhibitors (NRTIs) (7·8%), followed by the non-nucleoside reverse transcriptase inhibitors (NNRTIs) (4·2%) and Protease inhibitors (PIs) (2·3%). A potential clinical impact of the SDRM was demonstrated when using the current first-line therapy. A particularly high prevalence of SDRM was observed among intravenous drug users (IDUs) (29·4%). Reanalysis and comparing the data from previous Belgian studies using similar interpretation algorithms could not reveal a significant trend in SDRM prevalence over the last 5 years.

A national cohort of HIV-infected patients in Belgium: design and main characteristics.

In Belgium, individual laboratory and treatment data of all HIV-infected patients seen in the 9 AIDS Reference Centres and 7 AIDS Reference Laboratories are collected prospectively since 2006. We present here an analysis of patients recorded in the cohort database between 1st of January 2006 and 31st of December 2008. During that period, 11982 patients were under medical follow-up in Belgium. Sixty-one percent of the patients were male and the median age was 39.8 at the time of first recorded viral load. Among the patients whose nationality or probable mode of transmission was recorded, nearly half (48.0%) were Belgian and 38.3% originated from Sub-Saharan Africa; heterosexual contacts were reported in the majority of cases (56.0%) followed by homosexual contacts (35.3%). A total of 145 deaths were reported. Around three quarters of the patients were on ART. The median CD4 cell count rose from 470 cells/mm3 in 2006 to 501 cells/mm3 in 2008. This cohort enabled us to obtain comprehensive information on the numbers and characteristics of HIV-infected patients currently being followed up in Belgium, and on trends in antiretroviral therapy and biological results. This will serve for planning purposes, evaluation of access to care and as a source of information for further studies.

Retinal astrocytomas: long-term follow-up.

BACKGROUND: Retinal astrocytomas are exceedingly rare benign tumours of the retina. Their occurrence can be solitary or multiple, uni- or bilateral, isolated or in association with a phakomatosis such as tuberous sclerosis or neurofibromatosis type 1. PATIENTS AND METHODS: We report the long-term follow-up in three patients with retinal astrocytomas. RESULTS: Over many years of follow-up all astrocytomas showed very little progression and no deterioration of visual function. Subtle changes occurred inside the lesions. CONCLUSIONS: Even after long-term follow-up the natural course of retinal astrocytic hamartomas seems to be favourable, with visual loss and significant growth being unlikely to occur. A thorough ophthalmological and general evaluation, in order to rule out an underlying systemic disease and to document the ocular status, are needed initially. Thereafter eye examinations can be scheduled in long intervals.

Lymphogranuloma venereum in Europe, 2003-2008.

Lymphogranuloma venereum, caused by the L serovars of Chlamydia trachomatis, emerged in Europe in 2003 and a series of outbreaks were reported in different countries. The infection presents as a severe proctitis in men who have sex with men, many of whom are co-infected with HIV and other sexually transmitted infections. This paper reviews the number of cases reported over a five year period, from 2003 to 2008, from countries that were part of the European Surveillance of Sexually Transmitted Infections (ESSTI) network. Reports were received from Belgium, Denmark, France, Germany, the Netherlands, Portugal, Spain, Sweden, and the United Kingdom. It appears that after five years the characteristics of the patients infected has overall remained unchanged, although the total number of cases has increased and more countries in Europe have now identified cases of LGV.

HIV infections and STI co-infections in men who have sex with men in Belgium: sustained increase in HIV diagnoses.

Belgium is currently experiencing an upward trend in the number of new HIV diagnoses characterised by a continuous increase in the number of cases among men who have sex with men (MSM). Based on surveillance data, in the past decade the yearly number of newly diagnosed HIV cases in MSM increased more than threefold, from 101 cases diagnosed in 1999 to 332 cases in 2008. During this period, the majority of new HIV infections in MSM were diagnosed among Belgian citizens (72%), followed by other European nationalities (13%). The increase in HIV diagnoses does not reflect an increase in HIV testing since the number of tests performed nationwide remained remarkably stable over time. The steady increase in the number of newly diagnosed HIV cases among MSM, and the high proportion of MSM among HIV-positive patients co-infected with other sexually transmitted infections (STI) (95.6% in 2008) indicate increases in unsafe sex practices in this group. Development of behavioural surveillance and more qualitative research on reasons for unsafe sex are needed in order to develop more effective prevention strategies.

[Incidence of Neisseria gonorrhoeae infections in Belgium: trends 2000-2006]. / Incidence des infections à Neisseria gonorrhoeae en Belgique: tendances 2000-2006.

In Belgium, three registration systems collect epidemiological information on N. gonorrhoeae infections. The descriptive analysis of the data presented in this article allows describing the epidemiology of N. gonorrhoeae infections in Belgium in terms of trends in time, describing the characteristics of the patients, and providing information on resistance to antibiotics. The results on the incidence of N. gonorrhoeae infections show an important increase since the year 2000, and this increase is even more pronounced between 2005 and 2006. The majority of the patients reside in big cities, mainly in the district of Antwerp and in the Brussels-Capital region. Among the N. gonorrhoeae specimens that were sent to the reference laboratory, the proportion of specimens resistant to ciprofloxacine increases each year; this proportion reaches 61.4% in 2006. The increase in the incidence of N. gonorrhoeae infections and in antimicrobial resistance is also observed in other European countries. The increase in incidence may be partly related to the important increase of resistance to ciprofloxacine. It is very important to continue the surveillance of antimicrobial resistance, to adapt treatment in function of the recent evolutions and to inform physicians at a regular basis. The results show that homo- and bisexual men are most at risk for N. gonorrhoeae infections. The prevention campaigns for sexually transmitted infections and screening policy have to be reinforced, particularly among homo- and bisexual men.

Effect of natural and synthetic antioxidants on the oxidative stability of cooked, frozen pork patties.

The effect of grape seed extract (GS; 0.02%), oleoresin rosemary (OR; 0.02%), water-soluble oregano extract (WS; 0.02%), propyl gallate (PG; 0.02% of fat), butylated hydroxyanisole (BHA; 0.02% of fat), and butylated hydroxytoluene (BHT; 0.02% of fat) on the oxidative and color stability of precooked pork patties stored at -18 degrees C for up to 6 mo were determined. Pork lean and trim were ground and mixed (30% fat). Antioxidants mixed with salt (2%) were added. Patties were formed, cooked to 71 degrees C, over wrapped in PVC, and stored at -18 degrees C. Lipid oxidation was determined using thiobarbituric acid-reactive substances (TBARS) and descriptive sensory evaluation. Color was determined instrumentally and visually. Samples were evaluated after 0, 1, 2, 3, 4, 5, and 6 mo of frozen storage. Based upon TBARS values, PG (0.21 mg MDA/kg) and GS extract (0.23) had more antioxidant activity over the storage period than did WS, OR, BHA, and BHT. GS had no effect on a* or b* values. Grape seed extract (0.02%) has the potential to inhibit oxidative rancidity as well as current synthetic antioxidants.

Chemoimmunotherapy versus chemotherapy for metastatic malignant melanoma.

BACKGROUND: Malignant melanoma, one of the most aggressive of all skin cancers, is increasing in incidence throughout the world. Surgery remains the cornerstone of curative treatment in earlier stages. Metastatic disease is incurable in most affected people, because melanoma does not respond to most systemic treatments. A number of novel approaches are under evaluation and have shown promising results, but they are usually associated with increased toxicity and cost. The combination of chemotherapy and immunotherapy has been reported to improve treatment results, but it is still unclear whether evidence exists to support this choice, compared with chemotherapy alone. No language restrictions were imposed. OBJECTIVES: To compare the effects of therapy with chemotherapy and immunotherapy (chemoimmunotherapy) versus chemotherapy alone in people with metastatic malignant melanoma. SEARCH STRATEGY: We searched the Cochrane Skin Group Specialised Register (14 February 2006), the Cochrane Central Register of Controlled Trials (The Cochrane Library Issue 3, 2005), MEDLINE (2003 to 30 January 2006 ), EMBASE (2003 to 20 July 2005) and LILACS (1982 to 20 February 2006). References, conference proceedings, and databases of ongoing trials were also used to locate trials. SELECTION CRITERIA: All randomised controlled trials that compared the use of chemotherapy versus chemoimmunotherapy on people of any age, diagnosed with metastatic melanoma. DATA COLLECTION AND ANALYSIS: Two authors independently assessed each study to determine whether it met the pre-defined selection criteria, with differences being resolved through discussion with the review team. Two authors independently extracted the data from the articles using data extraction forms. Quality assessment included an evaluation of various components associated with biased estimates of treatment effect. Whenever possible, a meta-analysis was performed on the extracted data, in order to calculate a weighed treatment effect across trials. MAIN RESULTS: Eighteen studies met our criteria and were included in the meta-analysis, with a total of 2625 participants. We found evidence of an increase of objective response rates in people treated with chemoimmunotherapy, in comparison with people treated with chemotherapy. Nevertheless, the impact of these increased response rates was not translated into a survival benefit. We found no difference in survival to support the addition of immunotherapy to chemotherapy in the systemic treatment of metastatic melanoma, with a hazard ratio of 0.89 (95% CI 0.72 to 1.11, p=0.31). Additionally, we found increased hematological and non-hematological toxicities in people treated with chemoimmunotherapy. AUTHORS' CONCLUSIONS: We failed to find any clear evidence that the addition of immunotherapy to chemotherapy increases survival of people with metastatic melanoma. Further use of combined immunotherapy and chemotherapy should only be done in the context of clinical trials.

Intimal flap in a common carotid artery in a patient with Marfan's syndrome.

Dissection of the common carotid artery is a rare but important complication of Marfan's syndrome. The following case describes a patient with formation of an intimal flap of the common carotid artery who had suffered from an aortic dissection years before.

Unexpected pathogenic mechanism of a novel mutation in the coding sequence of SPG4 (spastin).

The authors report a nucleotide substitution (c.1216A>G) in SPG4 (spastin) causing hereditary spastic paraplegia. This apparent missense mutation in the ATPase domain confers aberrant, in-frame splicing and results in destabilization of mutated transcript. Mutated protein is deficient in microtubule-severing activity but, unlike neighboring mutations, shows regular subcellular localization. The authors' data point to haploinsufficiency rather than a dominant negative effect as the disease-causing mechanism for this mutation.

High HIV prevalence among patients choosing anonymous and free testing in Belgium, 1990-2002.

From 1990 through 2002, 25,250 anonymous and free HIV tests were performed at a testing site, which carried out the majority (85%) of anonymous testing in Belgium. During the same period, approximately 7.3 million confidential tests were registered nationwide. The rate of new HIV infections diagnosed at the anonymous testing site was 11.1/1000 tests; it was significantly higher than the rate observed among confidential tests (relative risk = 7.41; P < 0.0001). New HIV cases diagnosed through anonymous testing include a higher proportion of young adults (42.0% versus 32.5% in confidential testing; P < 0.001) and a higher proportion of men who have sex with men (32.7% vs. 25.9% in confidential testing; P < 0.02). Anonymous and free HIV testing was particularly sought by persons with higher infection risk, and efficiently contributed to HIV diagnosis in this population. Anonymous and free testing should be and remain an accessible alternative integrated in HIV testing policies.

STI surveillance by a sentinel network of physicians in Belgium.

The STI sentinel surveillance network by physicians in Belgium started in 2000. During 4 months a year, from October until January, STI patients were registered using a standardized protocol. The main goal is to determine STI incidence trends by comparing the results of the analyses using the data of the physicians that registered in all registration periods (Oct. 2000-Jan. 2001; Oct. 2001-Jan. 2002; Oct. 2002-Jan. 2003; Oct. 2003-Jan. 2004). Between the registration periods 2000-2001 and 2002-2003, there was a significant increase in the number of syphilis diagnoses (p<0.01), largely attributable to infections in men who have sex with men (MSM). A high proportion of MSM with syphilis were HIV positive. 83% of HIV positive MSM were already aware of their HIV positive status. The proportion of STI patients with HIV co-infection increased significantly throughout the different periods (from 7.4% in 2000-2001 to 18.1% in 2003-2004; p<0.01). These findings emphasize the importance of the proposal of a HIV test in a STI patient and call for intensification of prevention measures, particularly in MSM and people living with HIV.

Colony stimulating factors for prevention of myelosupressive therapy induced febrile neutropenia in children with acute lymphoblastic leukaemia.

BACKGROUND: Acute lymphoblastic leukaemia (ALL) is the most common cancer in childhood and febrile neutropenia is a potentially life-threatening side effect of its treatment. Current treatment consists of supportive care plus antibiotics. Clinical trials have attempted to evaluate the use of colony-stimulating factors (CSF) as additional therapy to prevent febrile neutropenia in children with ALL. The individual trials do not show whether there is significant benefit or not. Systematic review provides the most reliable assessment and the best recommendations for practice. OBJECTIVES: To evaluate the safety and effectiveness of the addition of G-CSF or GM-CSF to myelosuppressive chemotherapy in children with ALL, in an effort to prevent the development of febrile neutropenia. Evaluation of number of febrile neutropenia episodes, length to neutrophil count recovery, incidence and length of hospitalisation, number of infectious disease episodes, incidence and length of treatment delays, side effects (flu-like syndrome, bone pain and allergic reaction), relapse and overall mortality (death). SEARCH STRATEGY: The search covered the Cochrane Central Register of Controlled Trials, MEDLINE, EMBASE, CANCERLIT, LILACS, and SciElo. We manually searched records of conference proceedings of ASCO and ASH from 1985 to 2003 as well as databases of ongoing trials. We consulted experts and scanned references from the relevant articles. SELECTION CRITERIA: We looked for randomised controlled trials (RCTs) comparing CSF with placebo or no treatment as primary or secondary prophylaxis to prevent febrile neutropenia in children with ALL. DATA COLLECTION AND ANALYSIS: Two authors independently selected, critically appraised studies and extracted relevant data. The end points of interest were:* Primary end points: number of febrile neutropenia episodes and overall mortality (death) * Secondary end points: time to neutrophil count recovery, incidence and length of hospitalisation, number of infectious diseases episodes, incidence and length of treatment delays, side effects (flu-like syndrome, bone pain and allergic reaction) and relapse. We conducted a meta-analysis of these end points and expressed the results as Peto odds ratios. For continuous outcomes we calculated a weighted mean difference and a standardised mean difference. For count data, meta-analysis of the logarithms of the rate ratios using generic inverse variance was employed. MAIN RESULTS: We scanned more than 5500 citations and included six studies with a total of 332 participants in the analysis. There were insufficient data to assess the effect on survival. The use of CSF significantly reduced the number of episodes of febrile neutropenia episodes (Rate Ratio = 0.63; 95% confidence interval (CI) 0.46 to 0.85; p =0.003, with substantial heterogeneity), the length of hospitalisation (weighted mean difference (WMD) = -1.58; 95% CI -3.00 to -0.15; p = 0.03), and number of infectious diseases episodes (Rate Ratio=0.44; 95%CI 0.24 to 0.80; p=0.002). In spite of these results, CSF did not influence the length of episodes of neutropenia (WMD = -1.11; 95% CI -3.55 to 1.32; p = 0.4) or delays in chemotherapy courses (Rate Ratio=0.77; 95%CI 0.49 to 1,23; p=0.28) . AUTHORS' CONCLUSIONS: Children with ALL treated with CSF benefit from shorter hospitalisation and fewer infections. However, there was no evidence for a shortened duration of neutropenia nor fewer treatment delays, and no useful information about survival. The role of CSF regarding febrile neutropenia episodes is still uncertain. Although current data shows statistical benefit for CSF use, substantial heterogeneity between included trials does not allow this conclusion.

A minimally invasive approach for efficient gene delivery to rodent hearts.

Transcoronary gene delivery represents a desirable option to achieve global myocardial transgene expression but still requires aggressive surgical preparation in rodents. We therefore developed a catheter-based approach for cardiac gene transfer in the closed chest rat. A double-lumen balloon catheter was used to create aortic occlusion for specific infusion of adenoviral vectors carrying a beta-galactosidase transgene (1 x 10(11) PFU) into the coronaries. Simultaneously, venous return was obstructed by a second balloon catheter in the right atrium. To prolong viral incubation time, we induced a transient cardiac arrest (2 and 5 min) by a combination of acetylcholine and the beta-receptor antagonist, esmolol. At 72 h after transfection, the hearts showed a homogeneous and widespread beta-galactosidase expression, and the transduction efficiency increased and up to about 43% of cardiac myocytes (histochemistry) with a 400-fold increase of beta-galactosidase activity (luminescence assay) compared to sham-operated hearts. Pharmacological treatment aimed at increasing vascular permeability (SNAP and histamine) did not bring about synergistic effects on transfection efficiency. In addition, the method using high intracoronary pressure delivery (>300 mmHg) in a single-pass manner resulted in rather sparse beta-galactosidase expression in the myocardium (3-5% of cardiac myocytes). Therefore, the percutaneous gene delivery system described here provides a simple and minimally invasive procedure that represents a novel strategy for a homogeneous and highly efficient in vivo gene transfer to rodent hearts. Our results also suggest that prolongation of viral incubation time is an effective means for achieving highly efficient myocardial gene transduction.

Validation of the CALUX bioassay for PCDD/F analyses in human blood plasma and comparison with GC-HRMS.

Following the dioxin crisis of 1999, several studies were conducted to assess the impact of this crisis on the dioxin body burden in the Belgian population. The Scientific Institute of Public Health identified a population from whom plasma samples were available and from whom, during the follow up survey, plasma samples were obtained in 2000. In total, 496 samples were collected for GC-HRMS and CALUX analyses to verify statistical assessment conclusions. This study was seen as an opportunity to validate the CALUX bioassay for biological sample analysis and to compare toxic equivalency (TEQ) values obtained by the reference GC-HRMS technique and by the screening method. This article focuses on the validation results of the CALUX bioassay for the analyses of the dioxin fractions of blood plasma. The sample preparation is based on a liquid-liquid extraction, followed by an acid silica in series with an activated carbon clean-up. A good recovery (82%) and reproducibility (coefficient of variation less than 25%) were found for this method. Based on 341 plasma samples, a significant correlation was established between the bioassay and chemical method (R = 0.64). However, a proportional systematic error was observed when the results obtained with the CALUX bioassay were regressed with the results from the GC-HRMS analyses. The limit of quantification (LOQ) used to calculate TEQ values from the GC-HRMS determinations, the use of the relative potency values instead of the toxic equivalent factor and the potential of CALUX bioassay to measure all compounds with affinity for the AhR may partly explain this proportional systematic error. Nevertheless, the present results suggest that the CALUX bioassay could be a promising valid screening method for human blood plasma analyses.