Reverse engineering synthetic antiviral amyloids

Human amyloids have been shown to interact with viruses and interfere with viral replication. Based on this observation, we employed a synthetic biology approach in which we engineered virus-specific amyloids against influenza A and Zika proteins. Each amyloid shares a homologous aggregation-prone fragment with a specific viral target protein. For influenza we demonstrate that a designer amyloid against PB2 accumulates in influenza A-infected tissue in vivo. Moreover, this amyloid acts specifically against influenza A and its common PB2 polymorphisms, but not influenza B, which lacks the homologous fragment. Our model amyloid demonstrates that the sequence specificity of amyloid interactions has the capacity to tune amyloid-virus interactions while allowing for the flexibility to maintain activity on evolutionary diverging variants. Some human amyloid proteins have been shown to interact with viral proteins, suggesting that they may have potential as therapeutic agents. Here the authors design synthetic amyloids specific for influenza A and Zika virus proteins, respectively, and show that they can inhibit viral replication

The prion-like RNA-processing protein HNRPDL forms inherently toxic amyloid-like inclusion bodies in bacteria

BACKGROUND: The formation of protein inclusions is connected to the onset of many human diseases. Human RNA binding proteins containing intrinsically disordered regions with an amino acid composition resembling those of yeast prion domains, like TDP-43 or FUS, are being found to aggregate in different neurodegenerative disorders. The structure of the intracellular inclusions formed by these proteins is still unclear and whether these deposits have an amyloid nature or not is a matter of debate. Recently, the aggregation of TDP-43 has been modelled in bacteria, showing that TDP-43 inclusion bodies (IBs) are amorphous but intrinsically neurotoxic. This observation raises the question of whether it is indeed the lack of an ordered structure in these human prion-like protein aggregates the underlying cause of their toxicity in different pathological states. RESULTS: Here we characterize the IBs formed by the human prion-like RNA-processing protein HNRPDL. HNRPDL is linked to the development of limb-girdle muscular dystrophy 1G and shares domain architecture with TDP-43. We show that HNRPDL IBs display characteristic amyloid hallmarks, since these aggregates bind to amyloid dyes in vitro and inside the cell, they are enriched in intermolecular β-sheet conformation and contain inner amyloid-like fibrillar structure. In addition, despite their ordered structure, HNRPDL IBs are highly neurotoxic. CONCLUSIONS: Our results suggest that at least some of the disorders caused by the aggregation of human prion-like proteins would rely on the formation of classical amyloid assemblies rather than being caused by amorphous aggregates. They also illustrate the power of microbial cell factories to model amyloid aggregation. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12934-015-0284-7) contains supplementary material, which is available to authorized users

Phage Therapy and Photodynamic Therapy: Low Environmental Impact Approaches to Inactivate Microorganisms in Fish Farming Plants

Owing to the increasing importance of aquaculture to compensate for the progressive worldwide reduction of natural fish and to the fact that several fish farming plants often suffer from heavy financial losses due to the development of infections caused by microbial pathogens, including multidrug resistant bacteria, more environmentally-friendly strategies to control fish infections are urgently needed to make the aquaculture industry more sustainable. The aim of this review is to briefly present the typical fish farming diseases and their threats and discuss the present state of chemotherapy to inactivate microorganisms in fish farming plants as well as to examine the new environmentally friendly approaches to control fish infection namely phage therapy and photodynamic antimicrobial therapy

Phage therapy as an approach to prevent Vibrio anguillarum infections in fish larvae production

Fish larvae in aquaculture have high mortality rates due to pathogenic bacteria, especially the Vibrio species, and ineffective prophylactic strategies. Vaccination is not feasible in larvae and antibiotics have reduced efficacy against multidrug resistant bacteria. A novel approach to controlling Vibrio infections in aquaculture is needed. The potential of phage therapy to combat vibriosis in fish larvae production has not yet been examined. We describe the isolation and characterization of two bacteriophages capable of infecting pathogenic Vibrio and their application to prevent bacterial infection in fish larvae. Two groups of zebrafish larvae were infected with V. anguillarum (∼106 CFU mL-1) and one was later treated with a phage lysate (∼108 PFU mL-1). A third group was only added with phages. A fourth group received neither bacteria nor phages (fish control). Larvae mortality, after 72 h, in the infected and treated group was similar to normal levels and significantly lower than that of the infected but not treated group, indicating that phage treatment was effective. Thus, directly supplying phages to the culture water could be an effective and inexpensive approach toward reducing the negative impact of vibriosis in larviculture

La hiperglucemia como factor de mal pronóstico en el síndrome coronario agudo

Objetivo: La hiperglucemia es una observación frecuente en el síndrome coronario agudo (SCA). Hemos analizado la relación existente entre la hiperglucemia al ingreso y el pronóstico de los pacientes que ha sufrido un SCA. Material y métodos: Estudio prospectivo de 455 pacientes con SCA con y sin elevación del segmento ST de alto riesgo (criterios de la ACA/AHA). Dividimos la muestra según la mediana de la glucemia al ingreso en < 139 mg/dl y ≥ 139 mg/dl, y observamos variables analíticas, electrocardiográficas, ecocardiográficas y epidemiológicas. Mediante un análisis de riesgos proporcionales de Cox se analizó su relación con la mortalidad como variable principal en los siguientes seis meses al evento. Resultados: La edad media fue de 64,3 ± 12,7 años, el 80,4% eran varones y el 21,8% habían sido diagnosticados de diabetes. La glucemia media al ingreso fue de 163 ± 72 mg/dL. Un total de 47 pacientes fallecieron (10,3%). La glucemia media de los que fallecieron fue de 190 ± 79 mg/dl frente a 160 ± 70 mg/dl en los supervivientes (p = 0,003). Aquellos que presentaban al ingreso hiperglucemia (≥ 139 mg/dL) tuvieron una mayor mortalidad (Hazard ratio [HR] = 2,98; intervalo de confianza [IC] 95%: 1,06-8,4; p = 0,039). La edad avanzada, el sexo masculino, la disfunción ventricular y el descenso inicial de la presión arterial también mostraron relación independiente con la mortalidad. Conclusiones: La hiperglucemia al ingreso ≥ 139 mg/dl en pacientes con SCA se asocia a un mayor riesgo de fallecer en los próximos seis meses, independientemente del antecedente de diabetes y otros factores de riesgo conocidos. Objective: Hyperglycemia is a frequent observation in the acute coronary syndrome. We analyzed the relationship between hyperglycemia on admission and patients with acute coronary syndrome. Material and methods: Prospective study of 455 patients with acute coronary syndrome with and without elevation of ST segment with high risk according to ACA/AHA criteria. We divided the sample according to the median glycemia on admission into < 139 mg/dl and ≥ 139 mg/dl. We studied the analytic, electrocardiography, echocardiography and epidemiologic variables. Using the Cox Proportional Hazard Model, we analyzed their relationship with the mortality as principal variable during a six-month period after the acute coronary syndrome. Results: Mean age was 64.3 ± 12.7 years, 80.4% were male and 21.8% had been diagnosed with diabetes. Mean glycemia on admission was 163.3 ± 71.8 mg/dl. Forty-seven patients died (10.3%), Mean glycemia of those who had died was 189.8 ± 78.8 mg/dl compared to 160.3 ± 70.4 mg/dl in the survival group (P = 0.003). Patients with hyperglycemia on admission ≥ 139 mg/dl had higher mortality, hazard ratio (HR) =2.98 (confidence interval [CI 95%]: 1.06-8.4; P = 0.039). Elderly patients, being a male, having ventricular dysfunction and initial decrease of blood pressure also showed an independent relationship with mortality. Conclusions: Hyperglycemia on admission ≥ 139 mg/dl in acute coronary syndrome patients is associated with a higher risk of death in the following six months, independently of diabetes or other risk factors known

Spontaneous Dissection of Internal Carotid Artery Masquerading as Angioedema

Spontaneous dissection of the internal carotid artery usually presents with stroke-like symptoms secondary to ischemia in its vascular territory, as well as local signs and symptoms, which may include head, face or neck pain, Horner’s syndrome, pulsatile tinnitus, and cranial nerve palsies. We report a case of a 44-year-old healthy white male who presented with tongue swelling mimicking angioedema as an unusual manifestation of spontaneous dissection of the internal carotid artery. Two weeks after the initial presentation, the patient returned with similar symptoms and slurred speech. Upon physical examination, he was noted to have isolated left-sided hypoglossal nerve palsy. Subsequent diagnostic imaging revealed segmental narrowing of the left internal carotid artery. The appearance was consistent with the presence of a spontaneous internal carotid artery dissection with associated pseudoaneurysm formation

Effects of Toasting Time on Digestive Hydrolysis of Soluble and Insoluble 00-Rapeseed Meal Proteins

Thermal damage to proteins can reduce their nutritional value. The effects of toasting time on the kinetics of hydrolysis, the resulting molecular weight distribution of 00-rapeseed meal (RSM) and the soluble and insoluble protein fractions separated from the RSM were studied. Hydrolysis was performed with pancreatic proteases to represent in vitro protein digestibility. Increasing the toasting time of RSM linearly decreased the rate of protein hydrolysis of RSM and the insoluble protein fractions. The extent of hydrolysis was, on average, 44% higher for the insoluble compared with the soluble protein fraction. In contrast, the rate of protein hydrolysis of the soluble protein fraction was 3–9-fold higher than that of the insoluble protein fraction. The rate of hydrolysis of the insoluble protein fraction linearly decreased by more than 60% when comparing the untoasted to the 120 min toasted RSM. Increasing the toasting time elicited the formation of Maillard reaction products (furosine, N (ε)-carboxymethyl-lysine and N (ε)-carboxyethyl-lysine) and disulfide bonds in the insoluble protein fraction, which is proposed to explain the reduction in the hydrolysis rate of this fraction. Overall, longer toasting times increased the size of the peptides resulting after hydrolysis of the RSM and the insoluble protein fraction. The hydrolysis kinetics of the soluble and insoluble protein fractions and the proportion of soluble:insoluble proteins in the RSM explain the reduction in the rate of protein hydrolysis observed in the RSM with increasing toasting time

Reverse engineering synthetic antiviral amyloids

Human amyloids have been shown to interact with viruses and interfere with viral replication. Based on this observation, we employed a synthetic biology approach in which we engineered virus-specific amyloids against influenza A and Zika proteins. Each amyloid shares a homologous aggregation-prone fragment with a specific viral target protein. For influenza we demonstrate that a designer amyloid against PB2 accumulates in influenza A-infected tissue in vivo. Moreover, this amyloid acts specifically against influenza A and its common PB2 polymorphisms, but not influenza B, which lacks the homologous fragment. Our model amyloid demonstrates that the sequence specificity of amyloid interactions has the capacity to tune amyloid-virus interactions while allowing for the flexibility to maintain activity on evolutionary diverging variants.status: publishe