Global analysis of protein-RNA interactions in SARS-CoV-2-infected cells reveals key regulators of infection.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes coronavirus disease 2019 (COVID-19). SARS-CoV-2 relies on cellular RNA-binding proteins (RBPs) to replicate and spread, although which RBPs control its life cycle remains largely unknown. Here, we employ a multi-omic approach to identify systematically and comprehensively the cellular and viral RBPs that are involved in SARS-CoV-2 infection. We reveal that SARS-CoV-2 infection profoundly remodels the cellular RNA-bound proteome, which includes wide-ranging effects on RNA metabolic pathways, non-canonical RBPs, and antiviral factors. Moreover, we apply a new method to identify the proteins that directly interact with viral RNA, uncovering dozens of cellular RBPs and six viral proteins. Among them are several components of the tRNA ligase complex, which we show regulate SARS-CoV-2 infection. Furthermore, we discover that available drugs targeting host RBPs that interact with SARS-CoV-2 RNA inhibit infection. Collectively, our results uncover a new universe of host-virus interactions with potential for new antiviral therapies against COVID-19.

Reactive oxygen species activate the Drosophila TNF receptor Wengen for damage-induced regeneration.

Tumor necrosis factor receptors (TNFRs) control pleiotropic pro-inflammatory functions that range from apoptosis to cell survival. The ability to trigger a particular function will depend on the upstream cues, association with regulatory complexes, and downstream pathways. In Drosophila melanogaster, two TNFRs have been identified, Wengen (Wgn) and Grindelwald (Grnd). Although several reports associate these receptors with JNK-dependent apoptosis, it has recently been found that Wgn activates a variety of other functions. We demonstrate that Wgn is required for survival by protecting cells from apoptosis. This is mediated by dTRAF1 and results in the activation of p38 MAP kinase. Remarkably, Wgn is required for apoptosis-induced regeneration and is activated by the reactive oxygen species (ROS) produced following apoptosis. This ROS activation is exclusive for Wgn, but not for Grnd, and can occur after knocking down Eiger/TNFα. The extracellular cysteine-rich domain of Grnd is much more divergent than that of Wgn, which is more similar to TNFRs from other animals, including humans. Our results show a novel TNFR function that responds to stressors by ensuring p38-dependent regeneration.

System-wide Profiling of RNA-Binding Proteins Uncovers Key Regulators of Virus Infection.

The compendium of RNA-binding proteins (RBPs) has been greatly expanded by the development of RNA-interactome capture (RIC). However, it remained unknown if the complement of RBPs changes in response to environmental perturbations and whether these rearrangements are important. To answer these questions, we developed "comparative RIC" and applied it to cells challenged with an RNA virus called sindbis (SINV). Over 200 RBPs display differential interaction with RNA upon SINV infection. These alterations are mainly driven by the loss of cellular mRNAs and the emergence of viral RNA. RBPs stimulated by the infection redistribute to viral replication factories and regulate the capacity of the virus to infect. For example, ablation of XRN1 causes cells to be refractory to SINV, while GEMIN5 moonlights as a regulator of SINV gene expression. In summary, RNA availability controls RBP localization and function in SINV-infected cells.

Extracellular Kir2.1C122Y Mutant Upsets Kir2.1-PIP2 Bonds and Is Arrhythmogenic in Andersen-Tawil Syndrome.

BACKGROUND: Andersen-Tawil syndrome type 1 is a rare heritable disease caused by mutations in the gene coding the strong inwardly rectifying K+ channel Kir2.1. The extracellular Cys (cysteine)122-to-Cys154 disulfide bond in the channel structure is crucial for proper folding but has not been associated with correct channel function at the membrane. We evaluated whether a human mutation at the Cys122-to-Cys154 disulfide bridge leads to Kir2.1 channel dysfunction and arrhythmias by reorganizing the overall Kir2.1 channel structure and destabilizing its open state. METHODS: We identified a Kir2.1 loss-of-function mutation (c.366 A>T; p.Cys122Tyr) in an ATS1 family. To investigate its pathophysiological implications, we generated an AAV9-mediated cardiac-specific mouse model expressing the Kir2.1C122Y variant. We employed a multidisciplinary approach, integrating patch clamping and intracardiac stimulation, molecular biology techniques, molecular dynamics, and bioluminescence resonance energy transfer experiments. RESULTS: Kir2.1C122Y mice recapitulated the ECG features of ATS1 independently of sex, including corrected QT prolongation, conduction defects, and increased arrhythmia susceptibility. Isolated Kir2.1C122Y cardiomyocytes showed significantly reduced inwardly rectifier K+ (IK1) and inward Na+ (INa) current densities independently of normal trafficking. Molecular dynamics predicted that the C122Y mutation provoked a conformational change over the 2000-ns simulation, characterized by a greater loss of hydrogen bonds between Kir2.1 and phosphatidylinositol 4,5-bisphosphate than wild type (WT). Therefore, the phosphatidylinositol 4,5-bisphosphate-binding pocket was destabilized, resulting in a lower conductance state compared with WT. Accordingly, on inside-out patch clamping, the C122Y mutation significantly blunted Kir2.1 sensitivity to increasing phosphatidylinositol 4,5-bisphosphate concentrations. In addition, the Kir2.1C122Y mutation resulted in channelosome degradation, demonstrating temporal instability of both Kir2.1 and NaV1.5 proteins. CONCLUSIONS: The extracellular Cys122-to-Cys154 disulfide bond in the tridimensional Kir2.1 channel structure is essential for the channel function. We demonstrate that breaking disulfide bonds in the extracellular domain disrupts phosphatidylinositol 4,5-bisphosphate-dependent regulation, leading to channel dysfunction and defects in Kir2.1 energetic stability. The mutation also alters functional expression of the NaV1.5 channel and ultimately leads to conduction disturbances and life-threatening arrhythmia characteristic of Andersen-Tawil syndrome type 1.

GLP-1 Receptor Agonists Promote Weight Loss Among People with HIV.

BACKGROUND: Weight gain and associated metabolic complications are increasingly prevalent among people with HIV (PWH). Glucagon-like peptide-1 receptor agonists (GLP-1RAs) are incretin-based therapies for diabetes and weight management that have been shown to result in substantial weight loss; however, studies of their effects in PWH are limited. METHODS: A retrospective single-center cohort study was conducted among PWH who were taking GLP-1RAs at UC San Diego Owen Clinic between 2/1/2021 to 2/1/2023. Baseline clinical data were collected and changes in weight, body mass index (BMI), and hemoglobin A1C (A1C) before starting GLP-1RAs compared to the most recent clinic visit were calculated (with a minimum of 3 months follow-up time required). Logistic regression was performed to identify variables associated with >5% of total body weight loss. RESULTS: A total of 225 patients received on average 13 months of GLP-1RA therapy, with 85 (37.8%) achieving the maximum GLP-1RA dose. GLP-1RA therapy resulted, on average, in a loss of 5.4 kg, decrease in BMI by 1.8 kg/m2, and decrease in A1C by 0.6%. In the multivariable analysis, higher baseline BMI [OR 1.10 (1.03-1.16)], treatment duration of GLP-1RA therapy greater than 6 months [OR 3.12 (1.49-6.49], and use of tirzepatide [OR 5.46 (1.44-20.76)] were significantly more likely to be associated with >5% weight loss. CONCLUSIONS: Use of GLP-1RAs led to declines in weight, BMI, and hemoglobin A1C among PWH and offers an additional strategy to address weight gain and diabetes.

Exploring Attentional Bias toward Alcohol Content: Insights from Eye-Movement Activity.

INTRODUCTION: Attentional bias (AB) is an implicit selective attention toward processing disorder-significant information while neglecting other environmental cues. Considerable empirical evidence highlights the clinical implication of AB in the onset and maintenance of substance use disorder. An innovative method to explore direct measures of AB relies on the eye-movement activity using technologies like eye-tracking (ET). Despite the growing interest regarding the clinical relevance of AB in the spectrum of alcohol consumption, more research is needed to fully determine the AB patterns and its transfer from experimental to clinical applications. The current study consisted of three consecutive experiments. The first experiment aimed to design an ad-hoc visual attention task (VAT) consisting of alcohol-related and neutral images using a nonclinical sample (n = 15). The objective of the second and third experiments was to analyze whether the effect of type of image (alcohol-related vs. neutral images) on AB toward alcohol content using the VAT developed in the first experiment was different for type of drinker (light vs. heavy drinker in the second experiment [n = 30], and occasional social drinkers versus alcohol use disorder (AUD) patients in the third experiment [n = 48]). METHODS: Areas of interest (AOIs) within each type of image (neutral and alcohol-related) were designed and raw ET-based data were subsequently extracted through specific software analyses. For experiment 1, attention maps were created and processed for each image. For experiments 2 and 3, data on ET variables were gathered and subsequently analyzed through a two-way ANOVA with the aim of examining the effects of the type of image and drinker on eye-movement activity. RESULTS: There was a statistically significant interaction effect between type of image and type of drinker (light vs. heavy drinker in experiment 2, F(1, 56) = 13.578, p < 0.001, partial η2 = 0.195, and occasional social drinker versus AUD patients in the experiment 3, F(1, 92) = 35.806, p < 0.001, partial η2 = 0.280) for "first fixation" with large effect sizes, but not for "number of fixations" and "dwell time." The simple main effect of type of image on mean "first fixation" score for AUD patients was not statistically significant. CONCLUSION: The data derived from the experiments indicated the importance of AB in sub-clinical populations: heavy drinkers displayed an implicit preference for alcohol-related images compared to light drinkers. Nevertheless, AB fluctuations in patients with AUD compared to the control group were found. AUD patients displayed an early interest in alcohol images, followed by an avoidance attentional processing of alcohol-related images. The results are discussed in light of recent literature in the field.

The importance of virion-incorporated cellular RNA-Binding Proteins in viral particle assembly and infectivity.

RNA is a central molecule in RNA virus biology due to its dual function as messenger and genome. However, the small number of proteins encoded by viral genomes is insufficient to enable virus infection. Hence, viruses hijack cellular RNA-binding proteins (RBPs) to aid replication and spread. In this review we discuss the 'knowns' and 'unknowns' regarding the contribution of host RBPs to the formation of viral particles and the initial steps of infection in the newly infected cell. Through comparison of the virion proteomes of ten different human RNA viruses, we confirm that a pool of cellular RBPs are typically incorporated into viral particles. We describe here illustrative examples supporting the important functions of these RBPs in viral particle formation and infectivity and we propose that the role of host RBPs in these steps can be broader than previously anticipated. Understanding how cellular RBPs regulate virus infection can lead to the discovery of novel therapeutic targets against viruses.

An approach to the evaluation of the potency of cannabis resin in Madrid: A health hazard? / Aproximación a la evaluación de la potencia de la resina de cannabis en Madrid: ¿Un riesgo para la salud?

The present study investigates the concentration of Delta (9)-tetrahidrocannabinol (THC), cannabidiol (CBD) and cannabinol (CBN) in 60 samples of cannabis resin acquired on the streets of Madrid region and its potential danger to consumers' health. Additionally, we study the possible correlation between the potency of samples and their organoleptic characteristics. The analysis of cannabinoids was carried out using a high performance liquid chromatography (RP-HPLC-UV). To classify samples, a strength scale based on THC content was established. THC content in 76.7% of the samples was higher than 15%. This potency allows these samples to be classified as Schedule I or drugs with "unacceptable risk" for human health. THC content in 36.7% of the samples was 28.8% on average, which means very high potency. The mean CBD content was 5%, while the correlation between the CBD/THC ratio and potency was negative. The mean content of CBN was 1.74% and the CBN/THC ratio also showed a negative correlation in respect to potency. When investigating the possible correlation between sample potency and organoleptic characteristics, those samples which simultaneously presented sticky texture, high elasticity and light brown colour had very high potency, with an average THC content of 28.7%. Our study shows that the THC content of most of the cannabis that can be purchased in Madrid region is over 15% and poses a health hazard. Additionally, we demonstrate for the first time that only those samples with very high potency can be directly associated with certain organoleptic characteristics.

El presente estudio investiga la concentración de Delta(9)-tetrahidrocannabinol (THC), cannabidiol (CBD) y cannabinol (CBN) en 60 muestras de resina de cannabis adquiridas en las calles de Madrid y su potencial riesgo para la salud del consumidor. Adicionalmente, estudiamos la posible asociación entre la potencia de las muestras y sus características organolépticas. El análisis de cannabinoides se llevó a cabo mediante cromatografía líquida de alta resolución (RP-HPLC-UV). Atendiendo al contenido en THC se estableció una escala de potencia para clasificar las muestras. El 76,7% de las muestras tenía un contenido en THC superior al 15%, esta potencia las cataloga como drogas de Grado I con "riesgo inaceptable" para la salud. El 36,7% de las muestras presentaron un contenido medio en THC del 28,8% (potencia muy alta). El contenido medio en CBD fue del 5% y el de CBN 1,74%; ambas ratios, CBD/THC y CBN/THC, mostraron una correlación negativa con la potencia. Al investigar la posible asociación entra potencia y características organolépticas, se observó que las muestras que presentaban a la vez una textura pegajosa, una elasticidad alta y un color marrón claro, tenían una potencia muy alta, con un contenido medio en THC del 28.7%. Nuestro estudio muestra que el contenido en THC de la mayoría de la resina de cannabis que puede adquirirse en Madrid es superior al 15% y supone un elevado riesgo para la salud. Adicionalmente, demostramos por primera vez que solo aquellas muestras con una potencia muy alta pueden asociarse directamente con ciertas características organolépticas.

Empirical rescue treatment of Helicobacter pylori infection in third and subsequent lines: 8-year experience in 2144 patients from the European Registry on H. pylori management (Hp-EuReg).

OBJECTIVE: To evaluate the use, effectiveness and safety of Helicobacter pylori empirical rescue therapy in third and subsequent treatment lines in Europe. DESIGN: International, prospective, non-interventional registry of the clinical practice of European gastroenterologists. Data were collected and quality reviewed until October 2021 at Asociación Española de Gastroenterología-Research Electronic Data Capture. All cases with three or more empirical eradication attempts were assessed for effectiveness by modified intention-to-treat and per-protocol analysis. RESULTS: Overall, 2144 treatments were included: 1519, 439, 145 and 41 cases from third, fourth, fifth and sixth treatment lines, respectively. Sixty different therapies were used; the 15 most frequently prescribed encompassed >90% of cases. Overall effectiveness remained <90% in all therapies. Optimised treatments achieved a higher eradication rate than non-optimised (78% vs 67%, p<0.0001). From 2017 to 2021, only 44% of treatments other than 10-day single-capsule therapy used high proton-pump inhibitor doses and lasted ≥14 days. Quadruple therapy containing metronidazole, tetracycline and bismuth achieved optimal eradication rates only when prescribed as third-line treatment, either as 10-day single-capsule therapy (87%) or as 14-day traditional therapy with tetracycline hydrochloride (95%). Triple amoxicillin-levofloxacin therapy achieved 90% effectiveness in Eastern Europe only or when optimised. The overall incidence of adverse events was 31%. CONCLUSION: Empirical rescue treatment in third and subsequent lines achieved suboptimal effectiveness in most European regions. Only quadruple bismuth-metronidazole-tetracycline (10-day single-capsule or 14-day traditional scheme) and triple amoxicillin-levofloxacin therapies reached acceptable outcomes in some settings. Compliance with empirical therapy optimisation principles is still poor 5 years after clinical practice guidelines update. TRIAL REGISTRATION NUMBER: NCT02328131.

Assessment of dissolved mercury by diffusive gradients in thin films devices in abandoned ponds impacted by small scale gold mining.

In order to fulfil the Minamata Convention on Mercury, it is necessary to monitor the Hg contamination in freshwater ecosystems nearby artisanal and small scale gold mining (ASGM) areas. Since most of these ASGM communities are located in remote areas, a convenient method for sampling, preserving and transporting samples is needed. In this study we evaluated the feasibility of the diffusive gradient in thin-films (DGT) technique to detect and quantify the labile fraction of Hg and other metals (Pb, Cu, Zn, Cd, Ni, Mn and Cr) in a hard-to-reach gold mining district in the state of Chocó, Colombia. We deployed DGT at sampling sites along the Atrato river and abandoned mining ponds (AMPs) which were deserted in different periods since 1997 to 2019 (6-15 years). In average, the labile THg concentrations in AMPs (148.9 ± 43.2 ng L-1) were a 50% higher than in the river water (99.9 ± 37.4 ng L-1). In the ponds, no significant differences were found in labile Hg with respect abandonment period. Labile Ni (0.9-493.1), Mn (1.33-11.48), Cu (0.030-2.233), and Zn (0.67-10.29) (in µg L-1) were found in higher amounts than for the rest of metals. Labile concentrations of metals are related with their downstream proximity to gold mining activities, being higher in devices deployed close to ASGM sites. Moreover, this study demonstrates the feasibility of the DGT technique to sample, transport, storage, and preserve labile Hg from hard-to-reach ASGM areas.

A human health risk assessment of methylmercury, arsenic and metals in a tropical river basin impacted by gold mining in the Colombian Pacific region.

The Atrato River basin is one of the most biodiverse areas worldwide, and paradoxically, it is one of the sites in Colombia with the highest environmental impact from gold mining. This study assessed the distribution of Hg, As, Pb, and Cd in 47 fish species (n = 1372) and the accumulative human health risk in inhabitants (n = 2325) from 13 municipalities located along the Atrato River basin. The results revealed that Hg and As in fish present a high potential human health risk based on their mean concentrations. Estimated daily intake (EDI) calculations showed that humans could present detrimental health effects, while that target hazard quotient (THQ) above 1 showed that the exposed population might experience noncarcinogenic health risks, mainly from the accumulative effects of Hg (80.4%) and As (18.2%). The species that would most affect the health of the inhabitants are carnivorous H. malabaricus, A. pardalis, P. schultzi, R. quelen, and C. kraussii, which are among the fourteen most consumed in the region. These species had values of estimated weekly intake (EWI) above the provisional tolerable weekly intake thresholds for MeHg (PTWI of 1.6 and 3.2 µg/kg bw/week for adults and children, respectively) in 7 of the 13 localities evaluated. According to the surveys, the calculated weekly allowable fish amount (MFW) showed that carnivorous fish may generate adverse effects on the consumers because the allowed MeHg is about 2 times higher than the upper reference limit. Other results indicate a significant carcinogenic health risk, mainly from As, in 8 of the 13 localities evaluated. Due to the high rates of unsatisfied basic needs and the monetary poverty in the region, the possibility that inhabitants can replace fish as the principal source of protein is low. Therefore, a food guidance is required to avoid risks, obtain nutritional benefits, and sustain fish populations.

A viral RNA motif involved in signaling the initiation of translation on non-AUG codons.

Noncanonical translation, and particularly initiation on non-AUG codons, are frequently used by viral and cellular mRNAs during virus infection and disease. The Sindbis virus (SINV) subgenomic mRNA (sgRNA) constitutes a unique model system to analyze the translation of a capped viral mRNA without the participation of several initiation factors. Moreover, sgRNA can initiate translation even when the AUG initiation codon is replaced by other codons. Using SINV replicons, we examined the efficacy of different codons in place of AUG to direct the synthesis of the SINV capsid protein. The substitution of AUG by CUG was particularly efficient in promoting the incorporation of leucine or methionine in similar percentages at the amino terminus of the capsid protein. Additionally, valine could initiate translation when the AUG is replaced by GUG. The ability of sgRNA to initiate translation on non-AUG codons was dependent on the integrity of a downstream stable hairpin (DSH) structure located in the coding region. The structural requirements of this hairpin to signal the initiation site on the sgRNA were examined in detail. Of interest, a virus bearing CUG in place of AUG in the sgRNA was able to infect cells and synthesize significant amounts of capsid protein. This virus infects the human haploid cell line HAP1 and the double knockout variant that lacks eIF2A and eIF2D. Collectively, these findings indicate that leucine-tRNA or valine-tRNA can participate in the initiation of translation of sgRNA by a mechanism dependent on the DSH. This mechanism does not involve the action of eIF2, eIF2A, or eIF2D.

In vitro anti-Acanthamoeba activity of flavonoid glycosides isolated from Delphinium gracile, D. staphisagria, Consolida oliveriana and Aconitum napellus.

Acanthamoeba spp. are widely distributed in the environment and cause serious infections in humans. Treatment of Acanthamoeba infections is very challenging and not always effective which requires the development of more efficient drugs against Acanthamoeba spp. The purpose of the present study was to test medicinal plants that may be useful in the treatment of Acanthamoeba spp. Here we evaluated the trophozoital and cysticidal activity of 13 flavonoid glycosides isolated from Delphinium gracile, D. staphisagria, Consolida oliveriana and from Aconitum napellus subsp. Lusitanicum against the amoeba Acanthamoeba castellanii. AlamarBlue Assay Reagent® was used to determine the activity against trophozoites of A. castellanii, and cytotoxic using Vero cells. Cysticidal activity was assessed on treated cysts by light microscopy using a Neubauer chamber to quantify cysts and trophozoites. Flavonoids 1, 2, 3 and 4 showed higher trophozoital activity and selectivity indexes than the reference drug chlorhexidine digluconate. In addition, flavonoid 2 showed 100% cysticidal activity at a concentration of 50 µm, lower than those of the reference drug and flavonoid 3 (100 µm). These results suggest that flavonoids 2 and 3 might be used for the development of novel therapeutic approaches against Acanthamoeba infections after satisfactory in vivo evaluations.

The Internet of Cooperative Agents Architecture (X-IoCA) for Robots, Hybrid Sensor Networks, and MEC Centers in Complex Environments: A Search and Rescue Case Study.

Cloud robotics and advanced communications can foster a step-change in cooperative robots and hybrid wireless sensor networks (H-WSN) for demanding environments (e.g., disaster response, mining, demolition, and nuclear sites) by enabling the timely sharing of data and computational resources between robot and human teams. However, the operational complexity of such multi-agent systems requires defining effective architectures, coping with implementation details, and testing in realistic deployments. This article proposes X-IoCA, an Internet of robotic things (IoRT) and communication architecture consisting of a hybrid and heterogeneous network of wireless transceivers (H2WTN), based on LoRa and BLE technologies, and a robot operating system (ROS) network. The IoRT is connected to a feedback information system (FIS) distributed among multi-access edge computing (MEC) centers. Furthermore, we present SAR-IoCA, an implementation of the architecture for search and rescue (SAR) integrated into a 5G network. The FIS for this application consists of an SAR-FIS (including a path planner for UGVs considering risks detected by a LoRa H-WSN) and an ROS-FIS (for real-time monitoring and processing of information published throughout the ROS network). Moreover, we discuss lessons learned from using SAR-IoCA in a realistic exercise where three UGVs, a UAV, and responders collaborated to rescue victims from a tunnel accessible through rough terrain.

Acute pancreatitis in SARS-CoV-2 infection. Beyond respiratory distress.

We read with great interest the two letters published in November regarding SARS-CoV-2 infection and acute pancreatitis (AP). We report our only case of AP related to such infection.

Acute pancreatitis related to SARS-CoV-2 infection. Disclaimer.

Regarding our article "Daño pancreático: pancreatitis aguda en pacientes COVID-19", we would like to clarify that the case previously described met the diagnostic criteria for acute pancreatitis, defined in the Atlanta classification and mentioned in several guidelines.

Assessing the effectiveness of AS-48 in experimental mice models of Chagas' disease.

OBJECTIVES: We report the in vivo trypanocidal activity of the bacteriocin AS-48 (lacking toxicity), which is produced by Enterococcus faecalis, against the flagellated protozoan Trypanosoma cruzi, the aetiological agent of Chagas' disease. METHODS: We determined the in vivo activity of AS-48 against the T. cruzi Arequipa strain in BALB/c mice (in both acute and chronic phases of Chagas' disease). We evaluated the parasitaemia, the reactivation of parasitaemia after immunosuppression and the nested parasites in the chronic phase by PCR in target tissues. RESULTS: AS-48 reduced the parasitaemia profile in acute infection and showed a noteworthy reduction in the parasitic load in chronic infection after immunosuppression according to the results obtained by PCR (double-checking to demonstrate cure). CONCLUSIONS: AS-48 is a promising alternative that provides a step forward in the development of a new therapy against Chagas' disease.

Bismuth quadruple regimen with tetracycline or doxycycline versus three-in-one single capsule as third-line rescue therapy for Helicobacter pylori infection: Spanish data of the European Helicobacter pylori Registry (Hp-EuReg).

BACKGROUND: Different bismuth quadruple therapies containing proton-pump inhibitors, bismuth salts, metronidazole, and a tetracycline have been recommended as third-line Helicobacter pylori eradication treatment after failure with clarithromycin and levofloxacin. AIM: To evaluate the efficacy and safety of third-line treatments with bismuth, metronidazole, and either tetracycline or doxycycline. METHODS: Sub-study with Spanish data of the "European Registry on H pylori Management" (Hp-EuReg), international multicenter prospective non-interventional Registry of the routine clinical practice of gastroenterologists. After previous failure with clarithromycin- and levofloxacin-containing therapies, patients receiving a third-line regimen with 10/14-day bismuth salts, metronidazole, and either tetracycline (BQT-Tet) or doxycycline (BQT-Dox), or single capsule (BQT-three-in-one) were included. Data were registered at AEG-REDCap database. Univariate and multivariate analyses were performed. RESULTS: Four-hundred and fifty-four patients have been treated so far: 85 with BQT-Tet, 94 with BQT-Dox, and 275 with BQT-three-in-one. Average age was 53 years, 68% were women. Overall modified intention-to-treat and per-protocol eradication rates were 81% (BQT-Dox: 65%, BQT-Tet: 76%, BQT-three-in-one: 88%) and 82% (BQT-Dox: 66%, BQT-Tet: 77%, BQT-three-in-one: 88%), respectively. By logistic regression, higher eradication rates were associated with compliance (OR = 2.96; 95% CI = 1.01-8.84) and no prior metronidazole use (OR = 1.96; 95% CI = 1.15-3.33); BQT-three-in-one was superior to BQT-Dox (OR = 4.46; 95% CI = 2.51-8.27), and BQT-Tet was marginally superior to BQT-Dox (OR = 1.67; 95% CI = 0.85-3.29). CONCLUSION: Third-line H pylori eradication with bismuth quadruple treatment (after failure with clarithromycin and levofloxacin) offers acceptable efficacy and safety. Highest efficacy was found in compliant patients and those taking 10-day BQT-three-in-one or 14-day BQT-Tet. Doxycycline seems to be less effective and therefore should not be recommended.

In Vivo Biological Evaluation of a Synthetic Royleanone Derivative as a Promising Fast-Acting Trypanocidal Agent by Inducing Mitochondrial-Dependent Necrosis.

The life-long and life-threatening Chagas disease is one of the most neglected tropical diseases caused by the protozoan parasite Trypanosoma cruzi. It is a major public health problem in Latin America, as six to seven million people are infected, being the principal cause of mortality in many endemic regions. Moreover, Chagas disease has become widespread due to migrant populations. Additionally, there are no vaccines nor effective treatments to fight the disease because of its long-term nature and complex pathology. Therefore, these facts emphasize how crucial the international effort for the development of new treatments against Chagas disease is. Here, we present the in vitro and in vivo trypanocidal activity of some oxygenated abietane diterpenoids and related compounds. The 1,4-benzoquinone 15, not yet reported, was identified as a fast-acting trypanocidal drug with efficacy against different strains in vitro and higher activity and lower toxicity than benznidazole in both phases of murine Chagas disease. The mode of action was also evaluated, suggesting that quinone 15 kills T. cruzi by inducing mitochondrion-dependent necrosis through a bioenergetics collapse caused by a mitochondrial membrane depolarization and iron-containing superoxide dismutase inhibition. Therefore, the abietane 1,4-benzoquinone 15 can be considered as a new candidate molecule for the development of an appropriate and commercially accessible anti-Chagas drug.

Repositioning of leishmanicidal [1,2,3]Triazolo[1,5-a]pyridinium salts for Chagas disease treatment: Trypanosoma cruzi cell death involving mitochondrial membrane depolarisation and Fe-SOD inhibition.

Trypanosomatidae is a family of unicellular parasites belonging to the phylum Euglenozoa, which are causative agents in high impact human diseases such as Leishmaniasis, Chagas disease and African sleeping sickness. The impact on human health and local economies, together with a lack of satisfactory chemotherapeutic treatments and effective vaccines, justifies stringent research efforts to search for new disease therapies. Here, we present in vitro trypanocidal activity data and mode of action data, repositioning leishmanicidal [1,2,3]Triazolo[1,5-a]pyridinium salts against Trypanosoma cruzi, the aetiological agent of Chagas disease. This disease is one of the most neglected tropical diseases and is a major public health issue in Central and South America. The disease affects approximately 6-7 million people and is widespread due to increased migratory movements. We screened a suite of leishmanicidal [1,2,3]Triazolo[1,5-a]pyridinium salt compounds, of which compounds 13, 20 and 21 were identified as trypanocidal drugs. These compounds caused cell death in a mitochondrion-dependent manner through a bioenergetic collapse. Moreover, compounds 13 and 20 showed a remarkable inhibition of iron superoxide dismutase activity of T. cruzi, a key enzyme in the protection from the damage produced by oxidative stress.