Rapid diagnostic tests duo as alternative to conventional serological assays for conclusive Chagas disease diagnosis

Chagas disease is caused by the parasite Trypanosoma cruzi. It affects several million people, mainly in Latin America, and severe cardiac and/or digestive complications occur in ~30% of the chronically infected patients. Disease acute stage is mostly asymptomatic and infection goes undiagnosed. In the chronic phase direct parasite detection is hampered due to its concealed presence and diagnosis is achieved by serological methods, like ELISA or indirect hemagglutination assays. Agreement in at least two tests must be obtained due to parasite wide antigenic variability. These techniques require equipped labs and trained personnel and are not available in distant regions. As a result, many infected people often remain undiagnosed until it is too late, as the two available chemotherapies show diminished efficacy in the advanced chronic stage. Easy-to-use rapid diagnostic tests have been developed to be implemented in remote areas as an alternative to conventional tests. They do not need electricity, nor cold chain, they can return results within an hour and some even work with whole blood as sample, like Chagas Stat-Pak (ChemBio Inc.) and Chagas Detect Plus (InBIOS Inc.). Nonetheless, in order to qualify a rapidly diagnosed positive patient for treatment, conventional serological confirmation is obligatory, which might risk its start. In this study two rapid tests based on distinct antigen sets were used in parallel as a way to obtain a fast and conclusive Chagas disease diagnosis using whole blood samples. Chagas Stat-Pak and Chagas Detect Plus were validated by comparison with three conventional tests yielding 100% sensitivity and 99.3% specificity over 342 patients seeking Chagas disease diagnosis in a reference centre in Sucre (Bolivia). Combined used of RDTs in distant regions could substitute laborious conventional serology, allowing immediate treatment and favouring better adhesion to it

Use of rapid diagnostic tests (RDTs) for conclusive diagnosis of chronic Chagas disease – field implementation in the Bolivian Chaco region

Chagas disease, caused by the parasite Trypanosoma cruzi, is the neglected tropical disease with a highest burden in Latin America. Its acute stage is mostly asymptomatic and goes unnoticed. Symptoms appear at the chronic stage, which is when diagnosis is usually made. This is based on the agreement of two conventional serological tests such as Enzyme-Linked Immunosorbent Assays (ELISAs). There are commercial kits with good sensitivity and specificity but their use is impractical in many highly endemic regions with poorly equipped laboratories. Luckily, several rapid diagnostic tests (RDTs) are available for the detection of anti-T. cruzi immunoglobulins. They are easy to operate, require no cold storage, provide fast turnaround of results, and some can work with a tiny volume of whole blood as sample. With the aim to field validate their use we compared an alternative algorithm based on a combination of RDTs with the standard based on ELISAs. In both cases a third test was available in case of discordance. RDTs were implemented by mobile teams in field campaigns to detect chronic T. cruzi-infections in the Chaco region of Bolivia. ELISAs were made in the reference laboratories located in the main hospitals of Yacuiba and Villa Montes, two major cities of the region. We enrolled 685 subjects who voluntarily participated in the study and had not been treated against the disease before. The agreement between the two main RDTs was 93.1% (638/685) (kappa index = 0.86; CI 95% 0.83-0.90). In comparison to the ELISAs algorithm, the combined use of the RDTs provided a sensitivity of 97.7% and a specificity of 96.1%. These results support the use of RDTs for the diagnosis of chronic Chagas disease in the studied region, and encourage their evaluation in other regions of Bolivia and other endemic countries

A strategy for scaling up access to comprehensive care in adults with Chagas disease in endemic countries: The Bolivian Chagas Platform

BACKGROUND: Bolivia has the highest prevalence of Chagas disease (CD) in the world (6.1%), with more than 607,186 people with Trypanosoma cruzi infection, most of them adults. In Bolivia CD has been declared a national priority. In 2009, the Chagas National Program (ChNP) had neither a protocol nor a clear directive for diagnosis and treatment of adults. Although programs had been implemented for congenital transmission and for acute cases, adults remained uncovered. Moreover, health professionals were not aware of treatment recommendations aimed at this population, and research on CD was limited; it was difficult to increase awareness of the disease, understand the challenges it presented, and adapt strategies to cope with it. Simultaneously, migratory flows that led Bolivian patients with CD to Spain and other European countries forced medical staff to look for solutions to an emerging problem. INTERVENTION: In this context, thanks to a Spanish international cooperation collaboration, the Bolivian platform for the comprehensive care of adults with CD was created in 2009. Based on the establishment of a vertical care system under the umbrella of ChNP general guidelines, six centres specialized in CD management were established in different epidemiological contexts. A common database, standardized clinical forms, a and a protocolized attention to adults patients, together with training activities for health professionals were essential for the model success. With the collaboration and knowledge transfer activities between endemic and non-endemic countries, the platform aims to provide care, train health professionals, and create the basis for a future expansion to the National Health System of a proven model of care for adults with CD. RESULTS: From 2010 to 2015, a total of 26,227 patients were attended by the Platform, 69% (18,316) were diagnosed with T. cruzi, 8,567 initiated anti-parasitic treatment, more than 1,616 health professionals were trained, and more than ten research projects developed. The project helped to increase the number of adults with CD diagnosed and treated, produce evidence-based clinical practice guidelines, and bring about changes in policy that will increase access to comprehensive care among adults with CD. The ChNP is now studying the Platform's health care model to adapt and implement it nationwide. CONCLUSIONS: This strategy provides a solution to unmet demands in the care of patients with CD, improving access to diagnosis and treatment. Further scaling up of diagnosis and treatment will be based on the expansion of the model of care to the NHS structures. Its sustainability will be ensured as it will build on existing local resources in Bolivia. Still human trained resources are scarce and the high staff turnover in Bolivia is a limitation of the model. Nevertheless, in a preliminary two-years-experience of scaling up this model, this limitations have been locally solved together with the health local authorities

Characterization of digestive disorders of patients with chronic Chagas disease in Cochabamba, Bolivia

Background: Chagas disease (CD) is endemic in Latin America and particularly common in Bolivia, but there is little information on the characteristics of chronic digestive involvement. Objectives: To determine the prevalence and characterize digestive manifestations in chronic CD patients in Cochabamba, Bolivia. Methods: Eighty-five T. cruzi-seropositive individuals with or without digestive symptoms (G1 group), and fifteen T. cruzi-seronegative patients with similar digestive symptoms to those seen in CD (G2 group) were included in the study. All patients underwent a detailed history including past medical history, epidemiological information, hygiene and dietary habits, a complete physical examination, two serological tests for T. cruzi, video endoscopy, barium swallow, and barium enema. Findings: We observed digestive manifestations in T. cruzi seropositive and seronegative patients. Colonic manifestations were detected in both groups, highlighting the relevance of other confounder factors in the region. Constipation was present in 52.9% of G1 patients, 62.4% presented two or more upper digestive tract symptoms, and 5.9% of them presented esophageal manifestations. Helicobacter pylori infection was detected in 58.8% of G1 patients, and all patients presented gastritis on endoscopy. Conclusions: Prevalence of digestive involvement in CD patients is higher than expected. However, digestive symptoms are not always caused by T. cruzi infection and require differential diagnoses

Molecular detection and parasite load of Trypanosoma cruzi in digestive tract tissue of Chagas disease patients affected by megacolon

Chagas disease, caused by the Trypanosoma cruzi parasite in the Americas affects similar to 7 million people, 30% with cardiac tissue damage and 10-15% with digestive disorders. In this study, we have developed a protocol to detect the presence of the parasite and estimate its load in resected dysfunctional tissue segments of chronically infected patients with digestive megacolon. We have included samples from 43 individuals, 38/5 with positive/negative serology for Chagas disease and digestive syndromes. Samples of 1.5 to 2.0 cm(2) were taken from different points of the dysfunctional digestive tract in specialized centres in Cochabamba, Bolivia. T. cruzi cultures were performed by inoculation with NNN-LIT culture medium, and genomic material was obtained from the samples for multiplex qPCR with TaqMan probes targeting satellite nuclear DNA. Cultures failed to isolate T. cruzi but qPCR reached a sensitivity of 42.1% (16/38) with all three spots and in triplicate. A new quantification methodology using synthetic satellite DNA as quantitation standard revealed parasite loads ranging from 2.2 x 10(2) to 1.0 x 10(6) satellite DNA copies/mu l. Positive samples from the distal end showed a higher parasite load. The results of the present study strengthen and add further evidence to previous findings in an experimental mouse model of chronic T. cruzi infection, providing a valuable tool to improve scientific knowledge on the relevance of the digestive tract in parasite persistence, and underline the need of a better understanding of host-pathogen interaction in digestive tissues, considering pathophysiology, disease immunology and response to treatment

HIF-transcribed p53 chaperones HIF-1α

Chronic hypoxia is associated with a variety of physiological conditions such as rheumatoid arthritis, ischemia/reperfusion injury, stroke, diabetic vasculopathy, epilepsy and cancer. At the molecular level, hypoxia manifests its effects via activation of HIF-dependent transcription. On the other hand, an important transcription factor p53, which controls a myriad of biological functions, is rendered transcriptionally inactive under hypoxic conditions. p53 and HIF-1α are known to share a mysterious relationship and play an ambiguous role in the regulation of hypoxia-induced cellular changes. Here we demonstrate a novel pathway where HIF-1α transcriptionally upregulates both WT and MT p53 by binding to five response elements in p53 promoter. In hypoxic cells, this HIF-1α-induced p53 is transcriptionally inefficient but is abundantly available for protein-protein interactions. Further, both WT and MT p53 proteins bind and chaperone HIF-1α to stabilize its binding at its downstream DNA response elements. This p53-induced chaperoning of HIF-1α increases synthesis of HIF-regulated genes and thus the efficiency of hypoxia-induced molecular changes. This basic biology finding has important implications not only in the design of anti-cancer strategies but also for other physiological conditions where hypoxia results in disease manifestation

Long-term postharvest aroma evolution of tomatoes with the alcobaça (alc) mutation

The postharvest evolution of Penjar tomatoes has been studied in four accessions representative of the variability of the varietal type. The long-term shelf life of these materials, which carry the alc allele, was confirmed with 31.2-59.1% of commercial fruits after 6 months of effective conservation at room temperature and a limited loss of weight (21.1-27.9%). Aroma in Penjar tomatoes is differentiated from other tomato varieties by a characteristic 'sharp-floral' aroma descriptor. The evolution of the 'sharp-floral' aroma during postharvest showed a peak of intensity at 2 months of postharvest, though in one accession a delay of 2 months in this response was detected. Out of 25 volatiles analysed, including main and background notes, a reverse iPLS variable selection revealed that the main candidates behind this aromatic behaviour are ¿-terpineol, trans-2-hexenal, 6-methyl-5-hepten-2-one, trans-2-octenal, ¿-pinene, ß-ionone, 2 + 3-methylbutanol and phenylacetaldehyde. Between harvest and 2 months postharvest, most compounds reduced considerably their concentration, while the intensity of the 'sharp-floral' descriptor increased, which means that probably there is a rearrangement of the relative concentrations among volatiles that may lead to masking/unmasking processes. © 2011 Springer-Verlag.This work was supported by grants from the Conselleria de Agricultura, Pesca y Alimentacio de la Comunidad Valenciana, the Fundacion de la Comunidad Valenciana para la Investigacion Agroalimentaria (AGROALIMED) and from the Departament d'Agricultura, Alimentacio i Accio Rural (DAR) de la Generalitat de Catalunya.Casals Missio, J.; Cebolla Cornejo, J.; Rosello Ripolles, S.; Beltran Arandes, J.; Casanas, F.; Nuez Viñals, F. (2011). Long-term postharvest aroma evolution of tomatoes with the alcobaça (alc) mutation. European Food Research and Technology. 233(2):331-342. doi:10.1007/s00217-011-1517-6S3313422332Petro-Turza M (1987) Flavor of tomato and tomato products. Food Rev Int 2:309–351Butterry RG (1993) Quantitative and sensory aspects of flavor of tomato and other vegetables and fruits. In: Acree TE, Teranishi R (eds) Flavor science: sensible principles and techniques. American Chemical Society, WashingtonGoff SA, Klee HJ (2006) Plant volatile compounds: sensory cues for health and nutritional value? Science 311:815–819Tieman DM, Zeigler M, Schmelz EA, Taylor MG, Bliss P, Kirst M, Klee MJ (2006) Identification of loci affecting flavour volatile emissions in tomato fruits. J Exp Bot 57:887–896Zanor MI, Rambla JL, Chaïb J, Steppa A, Medina A, Granell A, Fernie AR, Causse M (2009) Metabolic characterization of loci affecting sensory attributes allows an assessment of the influence of the levels of primary metabolites and volatile organic contents. J Exp Bot 60:2139–2154Ortiz-Serrano P, Gil JV (2010) Quantitative comparison of free and bound volatiles of two commercial tomato cultivars (Solanum lycopersicum L.) during ripening. J Agric Food Chem 58:1106–1114Boukobza F, Taylor AJ (2002) Effect of postharvest treatment on flavour volatiles of tomatoes. Postharvest Biol Technol 25:321–331Vrebalov J, Ruezinsky D, Padmanabhan V, White R, Medrano D, Drake R, Schuch W, Giovannoni J (2002) A MADS-box gene necessary for fruit ripening at the tomato ripening-inhibitor (rin) locus. Science 296:343–346Giovannoni JJ, Tanksley SD, Vrebalov J, Noensie E (2004) NOR gene for use in manipulation of fruit quality and ethylene response. US Patent No 5,234,834 issued 13 July 2004McGlasson WB, Last JH, Shaw KJ, Meldrum SK (1987) Influence of the non-ripening mutants rin and nor on the aroma of tomato fruit. HortScience 22:632–634Baldwin EA, Scott JW, Shewmaker CK, Schuch W (2000) Flavor trivia and tomato aroma: biochemistry and possible mechanisms for control of important aroma components. HortScience 35:1013–1022Kovács K, Rupert CF, Tikunov Y, Graham N, Bradley G, Seymour GB, Bovy AG, Grierson D (2009) Effect of pleiotropic ripening mutations on flavour volatile biosynthesis. Phytochemistry 70:1003–1008Gao HY, Zhu BZ, Zhu HL, Zhang YL, Xie YH, Li YC, Luo YB (2007) Effect of suppression of ethylene biosynthesis on flavour products in tomato fruits. Russ J Plant Physiol 54:80–88Lewinsohn E, Sitrit Y, Bar E, Azulay Y, Meir A, Zamir D, Tadmor Y (2005) Carotenoid pigmentation affects the volatile composition of tomato and watermelon fruits, as revealed by comparative genetic analyses. J Agric Food Chem 53:3142–3148Kopeliovitch E, Mizrahi Y, Rabinowitch D, Kedar N (1980) Physiology of the mutant alcobaca. Physiol Plant 48:307–311Casals J, Pacual L, Cañizares J, Cebolla-Cornejo J, Casañas F, Nuez F (2011) Genetic basis of long shelf life and variability into Penjar tomato. Genet Resour Crop Evol. doi: 10.1007/s10722-011-9677-6Kuzyomenskii AV (2007) Effect of cumulative polymery of tomato keeping life genes. Cytol Genet 41:268–275Paran I, van der Knaap E (2007) Genetic and molecular regulation of fruit and plant domestication traits in tomato and pepper. J Exp Bot 58:3841–3852Moretti CL, Baldwin EA, Sargent SA, Huber DJ (2002) Internal bruising alters aroma volatile profiles in tomato fruit tisúes. HortScience 37:378–382Buttery RG, Teranishi R, Ling LC (1987) Fresh tomato aroma volatiles: a qualitative study. J Agric Food Chem 35:540–544Romero del Castillo R, Valero J, Casañas F, Costell E (2008) Training validation and maintenance of a panel to evaluate the texture of dry beans (Phaseolus vulgaris L.). J Sens Stud 23:303–319Beltran J, Serrano E, López FJ, Peruga A, Valcárcel M, Roselló S (2006) Comparison of two quantitative GC-MS methods for analysis of tomato aroma based on purge-and-trap and on solid-phase microextraction. Anal Bioanal Chem 385:1255–1264Martens H, Naes T (1989) Multivariate Calibration. Wiley, New YorkWise BM, Gallagher NB, Bro R, Shaver JM, Windig W, Koch RS (2006) Chemometrics tutorial for PLS_Toolbox and Solo. Eigenvector Research, WenatcheeHongsoongnern P, Chambers E (2008) A lexicon for texture and flavor characteristics of fresh and processed tomatoes. J Sens Stud 23:583–599Norgaard L, Saudland A, Wagner J, Nielsen JP, Munck L, Engelsen SB (2000) Interval partial least-squares regression (iPLS): A comparative chemometric study with an example from near-infrared spectroscopy. Appl Spectrosc 54:413–419Javanmardi J, Kubota C (2006) Variation of lycopene, antioxidant activity, total soluble solids and weight loss of tomato during postharvest storage. Postharvest Biol Technol 41:151–155Kader AA (1986) Effects of postharvest handling procedures on tomato quality. Acta Hort 190:209–222Maul F, Sargent SA, Sims CA, Baldwin EA, Balaban MO, Huber DJ (2000) Tomato flavor and aroma quality as affected by storage temperature. J Food Sci 65:1228–1237Krumbein A, Auerswald H (1998) Characterization of aroma volatiles in tomatoes by sensory analyses. Nahrung 6:S395–S399Tandon KS, Baldwin EA, Shewfelt RL (2000) Aroma perception of individual volatile compounds in fresh tomatoes (Lycopersicon esculentum Mill.) as affected by the medium of evaluation. Postharvest Biol Technol 20:261–268Cebolla-Cornejo J, Roselló S, Valcárcel M, Serrano E, Beltran J, Nuez F (2011) Evaluation of genotype and environment effects on taste and aroma flavour components of Spanish fresh tomato varieties. J Agric Food Chem 59:2440–2450Carbonell-Barrachina AA, Agustí A, Ruiz JJ (2006) Analysis of flavor volatile compounds by dynamic headspace in traditional and hybrid cultivars of Spanish tomatoes. Eur Food Res Technol 222:536–542Alonso A, Vázquez-Araújo L, García-Martínez S, Ruiz JJ, Carbonell Barrachina AA (2009) Volatile compounds of traditional and virus-resistant breeding lines of Muchamiel tomatoes. Eur Food Res Technol 230:315–323Liggett E, Drake MA, Delwiche JF (2008) Impact of flavor attributes on consumer liking of Swiss cheese. J Dairy Sci 91:466–476Ortiz-Serrano P, Gil JV (2007) Quantitation of free and glycosidically bound volatiles in and effect of glycosidase addition on three tomato varieties (Solanum lycopersicum L.). J Agric Food Chem 55:9170–9176Xu Y, Barringer S (2010) Comparison of tomatillo and tomato volatile compounds in the headspace by selected ion flow tube mass spectrometry (SIFT-MS). J Food Sci 75:C268–C273Berna AZ, Lammertyn J, Saevels S, Di Natale C, Nicolai BM (2004) Electronic nose systems to study shelf life and cultivar effect on tomato aroma profile. Sens Actuators B Chem 97:324–333Baldwin EA, Scott JW, Einstein MA, Malundo TMM, Carr BT, Shewfelt RL, Tandon KS (1998) Relationship between sensory and instrumental analysis for tomato flavor. J Am Soc Hortic Sci 12:906–915Krumbein A, Peters P, Brückner B (2004) Flavour compounds and a quantitative descriptive analysis of tomatoes (Lycopersicon esculentum Mill.) of different cultivars in short-term storage. Postharvest Biol Technol 32:15–2

Effects of whole-body ELECTROMYOSTIMULATION on health and performance: a systematic review

Abstract Background Whole-body electrical myostimulation (WB-EMS) is a relatively recent training methodology that has been extraordinarily used in recent years. However, there is a lack of consensus on the effectiveness of WB-EMS in the situations in which its use has been largely popularized. The objective of this systematic review was to determine the effects produced by WB-EMS. Methods A search of PubMed, Web of Science, Scopus and Cochrane was performed to identify all the studies that have applied electrical stimulation in lower and upper limbs simultaneously and that have clearly presented their protocols for the training and application of the stimulation. The last search was performed on September 9, 2018. Studies written in English or German were included. Results A total of 21 articles met the inclusion criteria and were analyzed following the guidelines of the Cochrane Guide for Systematic Reviews. Nineteen studies analyzed the chronic effects of WB-EMS, and 2 analyzed acute effects with a total of 505 subjects (310 men and 195 women). In total, 35% were moderately trained, and 65% were sedentary subjects. Different dependent variables were studied, such as anthropometric parameters, strength parameters, energy expenditure, psychophysiological parameters and blood parameters. There is a lack of randomized controlled studies, and the studies included exhibit a moderate to high level of risk of bias. Conclusions Given the limited number of available studies on WB-EMS, the scarce amount of scientific evidence found does not allow definitive conclusions about its effects; therefore, future studies about WB-EMS are necessary