[Adherence to biological therapies in patients with rheumatoid arthritis, psoriatic arthritis and ankylosing spondylitis. (Study ADhER-1)]. / Adherencia terapéutica a fármacos biológicos en pacientes con artritis reumatoide, artritis psoriásica y espondilitis anquilosante. (Estudio ADhER-1).

BACKGROUND: To quantify adherence to biological disease-modifying anti-rheumatic drugs (DMARD) and to determine the factors that can predict adherence in patients with rheumatoid arthritis, psoriatic arthritis and ankylosing spondylitis in daily clinical practice. METHODS: An observational, descriptive, cross-sectional and single-center study was carried out. Patients with rheumatoid arthritis, ankylosing spondylitis and psoriatic arthritis who were in treatment with subcutaneous biological DMARD were included. Variables related to socioeconomic status, disease, biological therapy and safety were recorded. Adherence was calculated by using medication possession ratio, Compliance Questionnaire on Rheumatology and Morisky Medication Adherence Scale Questionnaire. RESULTS: One hundred twelve patients and 6 different biological DMARDs were included. Mean age was 56.8±13.2 years and 52.7% were women. The percentage of adherent patients was 59.3% in rheumatoid arthritis, 62.5% in psoriatic arthritis and 76.2% in ankylosing spondylitis. Lesser adherence was associated with the administration of the drug by a family member and/or caregiver (odds ratio: 9.6; 95% confidence interval: 1.5-61.8 (p <.05)). There were no differences between adherent and non-adherent patients in terms of the biological DMARD used. CONCLUSIONS: There are no differences in adherence to biological therapies among patients with chronic inflammatory arthropathies. Adherence correlates negatively with administration of biological DMARD by a family member and / or caregiver.

Gold(I) and Silver(I) Complexes with 2-Anilinopyridine-Based Heterocycles as Multitarget Drugs against Colon Cancer.

A series of gold(I) and silver(I) derivatives with N- or S-donor ligands derived from 2-anilinopyridine has been synthesized and characterized. The mononuclear structure of [Au(L1)(PPh3)](TfO) (1a) and [Au(L2)(PPh3)](TfO) (1b) was confirmed by X-ray diffraction studies, as well as the dinuclear structure in the case of [Ag(TfO)(L1)]2 (4a). Most of the complexes are cytotoxic against a model of colorectal adenocarcinoma (Caco-2 cell line) and breast adenocarcinoma cancer cell lines (MCF-7). [Au(L1)(PPh3)](TfO) (1a) was able to induce caspases 8 and 3 activation, loss of mitochondrial membrane potential, and reactive oxygen species (ROS)-dependent cell death on Caco-2 cells upon 24 h incubation. In addition, the gold complex 1a produced a significant inhibition of the redox enzyme thioredoxin reductase as well as 20S proteasome. However, the silver(I) analogue, [Ag(TfO)(L1)(PPh3)] (2a), induced cell death independent of inhibition of thioredoxin reductase and 20S proteasome, triggered ROS-independent apoptosis mediated by caspase 8 and 3 activation, and loss of mitochondrial membrane potential, which points to a different mechanism of action for both derivatives, dependent on the metal center.

Synthesis of Gold(III) Complexes with Bidentate Amino-Thiolate Ligands as Precursors of Novel Bifunctional Acyclic Diaminocarbenes.

Two neutral bis(pentafluorophenyl)thiolate gold(III) complexes with the unsymmetrical S^N ligands 2-aminothiophenol or cysteamine have been synthesized and their reactivity has been studied. Homo- and heterodinuclear compounds were obtained by their coordination to gold(I) or silver(I) derivatives through the sulfur atom. Interestingly, a tetranuclear derivative bearing short gold(I)···gold(I) and the more unusual gold(I)···gold(III) interactions has been prepared. These amino-thiolate derivatives can be used as precursors for the synthesis of novel gold(III) acyclic diaminocarbene complexes by reaction with isocyanides CNR. The nucleophilic attack of the amino group to isocyanide molecules affords the synthesis of unprecedented bidentate C^S acyclic diaminocarbene ligands. All of the complexes are air- and moisture-stable at room temperature and have been spectroscopically and structurally characterized.

Efficient Gold(I) Acyclic Diaminocarbenes for the Synthesis of Propargylamines and Indolizines.

Mononuclear gold(I) acyclic diaminocarbenes (ADCs) were prepared by the reaction of 1,2-cyclohexanediamine with the corresponding isocyanide complexes [AuCl(CNR)] (R = Cy, t Bu). The three-component coupling of aldehydes, amines, and alkynes was investigated by using these gold(I) ADC complexes. The new gold(I) metal complexes are highly efficient catalysts for the synthesis of propargylamines and indolizines in the absence of solvent and in mild conditions. This method affords the corresponding final products with excellent yields in short reaction times. Additionally, chiral gold(I) complexes with ADCs have been prepared and tried in the enantioselective synthesis of propargylamines.

Prediction of pathological response to neoadjuvant treatment in rectal cancer with a two-protein immunohistochemical score derived from stromal gene-profiling.

BACKGROUND: Preoperative chemoradiotherapy followed by surgical mesorectal resection is the standard of care for locally advanced rectal carcinomas. Yet, predicting that patients will respond to treatment remains an unmet clinical challenge. EXPERIMENTAL DESIGN: Using laser-capture microdissection we isolated RNA from stroma and tumour glands from prospective pre-treatment samples (n = 15). Transcriptomic profiles were obtained hybridising PrimeView Affymetrix arrays. We modelled a carcinoma-associated fibroblast-specific genes filtering data using GSE39396. RESULTS: The analysis of differentially expressed genes of stroma/tumour glands from responder and non-responder patients shows that most changes were associated with the stromal compartment; codifying mainly for extracellular matrix and ribosomal components. We built a carcinoma-associated fibroblast (CAF) specific classifier with genes showing changes in expression according to the tumour regression grade (FN1, COL3A1, COL1A1, MMP2 and IGFBP5). We assessed these five genes at the protein level by means of immunohistochemical staining in a patient's cohort (n = 38). For predictive purposes we used a leave-one-out cross-validated model with a positive predictive value (PPV) of 83.3%. Random Forest identified FN1 and COL3A1 as the best predictors. Rebuilding the leave-one-out cross-validated regression model improved the classification performance with a PPV of 93.3%. An independent cohort was used for classifier validation (n = 36), achieving a PPV of 88.2%. In a multivariate analysis, the two-protein classifier proved to be the only independent predictor of response. CONCLUSION: We developed a two-protein immunohistochemical classifier that performs well at predicting the non-response to neoadjuvant treatment in rectal cancer.

Adherence to capecitabine in preoperative treatment of stage II and III rectal cancer: do we need to worry?

BACKGROUND: Preoperative oral capecitabine plus radiotherapy has been progressively adopted in oncology units to provide more convenient care to patients with rectal cancer, but little is known about adherence to this therapy. PATIENTS AND METHODS: Prospective, multicentre observational study in six hospitals in metropolitan Barcelona (Spain), in patients with stage II and III rectal cancer. Assessment of adherence was based on the medical report in the clinical history, a patient questionnaire and a pill count in the pharmacy service upon finalization of treatment. Patients were considered adherent if they had taken 80%-110% of the prescribed treatment. We evaluated clinical variables, adverse effects, anxiety and depression (using the hospital anxiety depression scale [HADS]), and quality of life (EORTC QLQ-30). We analysed adherence-associated variables using a logistic regression model and concordance between adherence measures by means of the modified Kappa index. RESULTS: We included 119 participants. Adherence measures showed little concordance between the assessment methods used: adherence was 100% according to the clinical history, 83.2% according to self-report and 67.9% according to the pill count. In the multivariable analysis, the most relevant variable associated with non-adherence was anxiety prior to treatment (adjusted odds ratio [ORa] 6.96, 95% confidence interval [CI] 1.48-32.7). We did not observe any relevant association between adherence and clinical variables and baseline quality of life parameters. CONCLUSIONS: Adherence to short-term oral neoadjuvant treatment in rectal cancer may be a clinical problem, and it should be acknowledged and systematically evaluated by clinicians during treatment. The limited concordance between different measures of adherence highlights the challenges in monitoring it and the need to use different approaches to assess its impact in clinical practice.

Group 11 complexes with amino acid derivatives: Synthesis and antitumoral studies.

Gold(I), gold(III), silver(I) and copper(I) complexes with modified amino acid esters and phosphine ligands have been prepared in order to test their cytotoxic activity. Two different phosphine fragments, PPh3 and PPh2py (py=pyridine), have been used. The amino acid esters have been modified by introducing an aromatic amine as pyridine that coordinates metal fragments through the nitrogen atom, giving complexes of the type [M(L)(PR3)](+) or [AuCl3(L)] (L=l-valine-N-(4-pyridylcarbonyl) methyl ester (L1), l-alanine-N-(4-pyridylcarbonyl) methyl ester (L2), l-phenylalanine-N-(4-pyridylcarbonyl) methyl-ester) (L3); M=Au(I), Ag(I), Cu(I), PR3=PPh3, PPh2py). The in vitro cytotoxic activity of metal complexes was tested against four tumor human cell lines and one tumor mouse cell line. A metabolic activity test (3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide, MTT) was used and IC50 values were compared with those obtained for cisplatin. Several complexes displayed significant cytotoxic activities. In order to determine whether antiproliferation and cell death are associated with apoptosis, NIH-3T3 cells were exposed to five selected complexes (Annexin V+ FITC, PI) and analyzed by flow cytometry. These experiments showed that the mechanism by which the complexes inhibit cell proliferation inducing cell death in NIH-3T3 cells is mainly apoptotic.

First-line single-agent panitumumab in frail elderly patients with wild-type KRAS metastatic colorectal cancer and poor prognostic factors: A phase II study of the Spanish Cooperative Group for the Treatment of Digestive Tumours.

BACKGROUND: Frail elderly patients with metastatic colorectal cancer (mCRC) are not candidates for chemotherapy. Monotherapy with anti-epidermal growth factor receptor (EGFR) monoclonal antibodies may be an option for these patients with few systemic toxic effects. PATIENTS AND METHODS: Single-arm, multicentre, phase II trial including patients ⩾ 70y ears with wild-type (WT) KRAS (exon 2) mCRC, Eastern Cooperative Oncology Group (ECOG) status ⩽ 3, KPC (Köhne Prognostic Classification)--defined intermediate or high risk status, frailty and/or ineligibility for chemotherapy. Patients received panitumumab until progression or unacceptable toxicity. The primary end-point was progression free survival (PFS) rate at 6 months. RESULTS: The study included 33 patients (intention-to-treat (ITT) population). Median age: 81 years; sex: 66.7% male; high-risk KPC status: 45.4%. Median treatment duration was 14 weeks and 6-month PFS rate was 36.4% (95% confidence interval (CI): 20.0-52.8). The objective response rate: 9.1% (95% CI: 0-18.9) (all partial responses), and there were 18 stable diseases (54.5%). Median PFS was 4.3 months (95% CI: 2.8-6.4) and median overall survival (OS) was 7.1 months (95% CI: 5.0-12.3). There were no deaths or grade 4-5 adverse events (AEs) related to panitumumab and the most common grade 3-related AE was rash acneiform (15.2%). A significant association between clinical response and RAS status was observed (P=0.037). In the WT RAS subgroup (WT exons 2, 3, and 4 of KRAS and NRAS, N = 15), 6-month PFS rate was 53.3% (95% CI: 30.1-75.2) and median PFS and OS were 7.9 and 12.3 months, respectively. CONCLUSIONS: Single-agent panitumumab is active and well tolerated and may be a therapeutic option for high-risk frail elderly patients with WT RAS tumours considered not candidates for chemotherapy (clinicaltrials.gov identifier NCT01126112).

The fluxional amine gold(III) complex as an excellent catalyst and precursor of biologically active acyclic carbenes.

A new amine gold(III) complex [Au(C6F5)2(DPA)]ClO4 with the di-(2-picolyl)amine (DPA) ligand has been synthesised. In the solid state the complex has a chiral amine nitrogen because the ligand coordinates to the gold centre through one nitrogen atom from a pyridine and through the NH moiety, whereas in solution it shows a fluxional behaviour with a rapid exchange between the pyridine sites. This complex can be used as an excellent synton to prepare new gold(III) carbene complexes by the reaction with isocyanide CNR. The resulting gold(III) derivatives have unprecedented bidentate C^N acyclic carbene ligands. All the complexes have been spectroscopically and structurally characterized. Taking advantage of the fluxional behaviour of the amine complex, its catalytic properties have been tested in several reactions with the formation of C-C and C-N bonds. The complex showed excellent activity with total conversion, without the presence of a co-catalyst, and with a catalyst loading as low as 0.1%. These complexes also present biological properties, and cytotoxicity studies have been performed in vitro against three tumour human cell lines, Jurkat (T-cell leukaemia), MiaPaca2 (pancreatic carcinoma) and A549 (lung carcinoma). Some of them showed excellent cytotoxic activity compared with the reference cisplatin.

(Aminophosphane)gold(I) and silver(I) complexes as antibacterial agents.

This manuscript describes the synthesis of new Au(I) and Ag(I) complexes with aminophosphane ligands and a study of their antibacterial activity against Gram-negative Salmonella enterica serovar typhimurium and Escherichia coli and Gram-positive Listeria monocytogenes and Staphylococcus aureus. The bactericidal assays revealed the effectiveness of these compounds on paradigm Gram-negative and Gram-positive pathogens, showing a moderate antimicrobial activity, comparable with the antibiotics of reference, for all gold(I) complexes and the silver(I) complexes without coordinated PPh3 groups. For those complexes that were found to show inhibitory activity, serial dilutions in liquid broth method were performed for determination of MIC (minimum inhibitory concentration) and MBC (minimum bactericidal concentration).

Clinical management of regorafenib in the treatment of patients with advanced colorectal cancer

Colorectal cancer is one of the most common tumors worldwide and at least 50 % of patients with this disease develop metastases. In this setting, additional treatment options are needed for patients presenting disease progression after exhausting all standard therapies. Regorafenib is an orally administered multikinase inhibitor which has been shown to provide survival benefits to patients with metastatic colorectal cancer (mCRC). Although most adverse events (AEs) associated with regorafenib may resolve within the first 8 weeks of treatment, some of them may require dose reduction or treatment interruption. Overall, while remaining aware of the safety profile of regorafenib and how to manage the most common toxicities related to its use, this drug should be considered a new standard of care for patients with pretreated mCRC. This review addresses practical aspects of its use, such as dosing, patient monitoring, and management of the most common regorafenib-related AEs (AU)

No disponible

Clinical management of regorafenib in the treatment of patients with advanced colorectal cancer.

Colorectal cancer is one of the most common tumors worldwide and at least 50 % of patients with this disease develop metastases. In this setting, additional treatment options are needed for patients presenting disease progression after exhausting all standard therapies. Regorafenib is an orally administered multikinase inhibitor which has been shown to provide survival benefits to patients with metastatic colorectal cancer (mCRC). Although most adverse events (AEs) associated with regorafenib may resolve within the first 8 weeks of treatment, some of them may require dose reduction or treatment interruption. Overall, while remaining aware of the safety profile of regorafenib and how to manage the most common toxicities related to its use, this drug should be considered a new standard of care for patients with pretreated mCRC. This review addresses practical aspects of its use, such as dosing, patient monitoring, and management of the most common regorafenib-related AEs.

A comparative study of structural patterns and luminescent properties of silver-DAFO complexes with carborane- versus "classical"-diphosphanes.

New complexes with the DAFO (4,5-diazafluoren-9-one) ligand of stoichiometry [Ag(DAFO)(P-P)]OTf [P-P = dppe, 1,2-bis(diphenylphosphane)ethane; dppp, 1,3-bis(diphenylphosphane)propane; dppph, ortho-bis(diphenylphosphane)benzene; dppcc, 1,2-bis(diphenylphosphane)-1,2-dicarba-closo-dodecaborane; dipcc, 1,2-bis(diisopropylphosphane)-1,2-dicarba-nido-dodecaborane], [Ag(DAFO)(P-P*)] [P-P* = dppnc, 7,8-bis(diphenylphosphane)-7,8-dicarba-nido-undecaborate(-1); dipnc, 7,8-bis(diisopropylphosphane)-7,8-dicarba-closo-undecaborate(-1)] and [Ag(DAFO)(OTf)L] [L = PPh3; dpccMe, 1-diphenylphosphane-2-methyl-1,2-dicarba-closo-dodecaborane] are reported. The structures of the complexes depend on the skeleton of the diphosphane. Most of these compounds are luminescent and their emissions seem to have originated from IL (DAFO) transitions, modified upon coordination to silver.

Structural and photophysical study on heterobimetallic complexes with d8-d10 interactions supported by carborane ligands: theoretical analysis of the emissive behaviour.

Heterobimetallic complexes of formula [M{(PPh2)2C2B9H10}(S2C2B10 H10)M'(PPh3)] (M=Pd, Pt; M'=Au, Ag, Cu) and [Ni{(PPh2)2C2B9H10}(S2C2B10H10)Au(PPh3)] were obtained from the reaction of [M{(PPh2)2C2B10H10}(S2C2B10H10)] (M=Pd, Pt) with [M'(PPh3)](+) (M'=Au, Ag, Cu) or by one-pot synthesis from [(SH2C2B10H10], (PPh2 )2C2B10H10, NiCl2 ⋅6 H2 O, and [Au(PPh3)](+). They display d(8)-d(10) intermetallic interactions and emit red light in the solid state at 77 K. Theoretical studies on [M{(PPh2)2C2B9H10}(S2C2B10H10)Au(PPh3)] (M=Pd, Pt, Ni) attribute the luminescence to ligand (thiolate, L)-to-"P2-M-S2" (ML') charge-transfer (LML'CT) transitions for M=Pt and to metal (M)-to-"P2-M-S2" (ML') charge-transfer (MML'CT) transitions for M=Ni, Pd.

Strong inhibition of thioredoxin reductase by highly cytotoxic gold(I) complexes. DNA binding studies.

Biological properties of a series of aminophosphine-thiolate gold(I) complexes [Au(SR)(PPh2NHpy)] [Ph2PNHpy=2-(diphenylphosphinoamino)pyridine; HSR=2-mercaptopyridine (2-HSpy) (3), 2-mercaptonicotinic acid (2-H2-mna) (4), 2-thiouracil (2-HTU) (5) or 2-thiocytosine (2-HTC) (6)] and [Au(SR){PPh2NH(Htrz)}] [Ph2PNH(Htrz)=3-(diphenylphosphinoamino)-1,2,4-triazole]; HSR=2-mercaptopyridine (2-HSpy) (7), 2-thiocytosine (2-HTC) (8) or 6-thioguanine (6-HTG) (9) have been studied. Their antitumor properties have been tested in vitro against two tumor human cell lines, HeLa (derived from cervical cancer) and MCF-7 (derived from breast cancer), using a metabolic activity test (3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide, MTT). Some of them showed excellent cytotoxic activity. With the aim to obtain more information about the mechanisms of action of these derivatives, the interactions of complexes 3, 5, 7 and 9 with thioredoxin reductase in HeLa cells were studied. They showed a potent inhibition of thioredoxin reductase activity. In order to complete this study, interactions of the complexes with calf thymus (CT-) DNA and with different bacterial DNAs, namely the plasmid pEMBL9 and the promoter region of the furA (ferric uptake regulator A) gene from Anabaena sp. PCC 7120 were investigated. Although interactions of complexes with CT-DNA have been verified, none of them cause significant changes in its structure.

Synthesis of new gold(I) thiolates containing amino acid moieties with potential biological interest.

The reaction of the gold(I) complex [Au(SpyCOOH)(PPh3)], which contains nicotinic acid thiolate, with several amino acid esters such as glycine methyl ester or the enantiomerically pure L isomers of alanine methyl ester, phenylalanine methyl ester, valine methyl ester, methionine methyl ester, and proline methyl ester produces the gold(I) derivatives with the new thiolate containing amino acid ester ligands [Au{SpyCONHCH(R)COOMe}(PPh3)]. The reaction of these amino acid ester derivatives with LiOH in methanol and acidification with KHSO4 until pH 3-4 afford the corresponding acids, which are water-soluble species. These amino acid compounds can be further coupled with other amines, such as, for example, isopropylamine, to give the corresponding amide derivatives. The species with glycine methyl ester and valine methyl ester have been characterized by X-ray crystallography, showing, in the second case, only one of the enantiomers, which proves that retention of the configuration after reaction occurs.

Influence of the group 11 metal on the emissive properties of complexes [M{(PR2)2C2B9H10}L].

The metal atoms in group 11 complexes [M{(PR2)2C2B9H10}L] [R = Ph, (i)Pr; L = tertiary phosphane; M = Au, Ag, Cu] play an important role in the emissive properties of these compounds. The influence of the metal follows the order Au ≫ Ag ≥ Cu. The three-coordinated complexes are obtained from the reaction of [AuClL], [Ag(OTf)L], or [Cu(NO3)(PPh3)2] with the closo carborane diphosphane in refluxing ethanol. For L = PPh2NHPy, cleavage of the P-N bond and the formation of the monophosphane PPh2OEt are observed, depending on the metal and the nido carborane diphosphane substituent ((i)Pr or Ph).

Clinicopathological risk factors of Stage II colon cancer: results of a prospective study.

AIM: Adjuvant 5-fluorouracil based chemotherapy has demonstrated benefit in Stage III colon cancer but still remains controversial in Stage II. The aim of this study was to analyse the prognostic impact of clinicopathological factors that may help guide treatment decisions in Stage II colon cancer. METHOD: Between 1996 and 2006 data from patients diagnosed with colorectal cancer at Hospital Universitari Bellvitge and its referral comprehensive cancer centre Institut Català d'Oncologia/L'Hospitalet were prospectively included in a database. We identified 432 patients with Stage II colon cancer operated on at Hospital Universitari Bellvitge. The 5-year relapse-free survival (RFS) and colon-cancer-specific survival (CCSS) were determined. RESULTS: The 5-year RFS and CCSS were 83% and 88%, respectively. Lymphovascular or perineural invasion was associated with RFS [hazard ratio (HR) 1.84; 95% CI 1.01-3.35]. Gender (women, HR 0.48; 95% CI 0.23-1) and lymphovascular or perineural invasion (HR 3.51; 95% CI 1.86-6.64) together with pT4 (HR 2.79; 95% CI 1.44-5.41) influenced CCSS. In multivariate analysis pT4 and lymphovascular or perineural invasion remained significantly associated with CCSS. We performed a risk index with these factors with prognostic impact. Patients with pT4 tumours and lymphovascular or perineural invasion had a 5-year CCSS of 61%vs the 93% (HR 5.87; 95 CI 2.46-13.97) of those without any of these factors. CONCLUSION: pT4 and lymphatic, venous or perineural invasion are confirmed as significant prognostic factors in Stage II colon cancer and should be taken into account in the clinical validation process of new molecular prognostic factors.

Synthesis of gold-silver luminescent honeycomb aggregates by both solvent-based and solvent-free methods.

Heterometallic clusters with strong luminescence have been synthesized (see picture: Au(C≡CPh)(2)yellow-red, Ag(2)blue, Ored) from the metalloligand unit [Au(C≡CPh)PPh(3) ] (yellow/red bars) by using both standard solvent-based and solvent-free reactions. The aggregates are stabilized only by acetylide-metal or metal-metal interactions, and their nuclearity is controlled through the addition of different donor ligands.

Evaluation of an immunochromatographic dip strip test for simultaneous detection of Cryptosporidium spp, Giardia duodenalis, and Entamoeba histolytica antigens in human faecal samples.

Immunochromatographic (IC) tests may play an important role in the future diagnosis of parasitic diseases because of their speed and simplicity of use. A recently developed test to detect Cryptosporidium spp, Giardia duodenalis and Entamoeba histolytica was evaluated. Microscopy and PCR were the "gold standard" reference techniques and the results of this IC test were compared with those obtained with ELISA and IC single test for the three parasites. One hundred sixty stool samples were assayed. Using microscopy, 22 samples were diagnosed as positive for Cryptosporidium spp., 31 for Giardia duodenalis, 41 for Entamoeba histolytica/dispar, and 68 had a negative diagnosis for the three parasites. Results of IC tests show sensitivities of 70-72% for Cryptosporidium, 90-97% for Giardia and 62.5% for Entamoeba histolytica. Specificities were of 93.6-94.9%, >99% and 96.1%, respectively. In all diagnoses, agreement with microscopy and PCR was over 90%, except in the triple test and microscopy in E. histolytica detection that was 76.3%, due to the inability of microscopy to differentiate E. histolytica from nonpathogenic species such as E. dispar or E. moshkovskii. The triple stool immunoassays provide adequate sensitivities and specificities for use in outbreak situations, for screening proposals and for massive assays in endemic areas where a large number of samples must be analysed or as complementary test for individual diagnosis.