Integrated full-scale solar CPC/UV-LED-filtration system as a tertiary treatment in a conventional WWTP for agricultural reuse purposes.

Today, the emergence of increasingly restrictive treatment and reuse policies make the implementation of full-scale tertiary treatment, capable of improving the quality of water, a priority. Full-scale TiO2 photocatalysis systems are resulting in a promising option, since TiO2 is commercially available. However, questions such as how to work continuously during day/night irradiation cycle, or the removing of TiO2 in outlet flow are still unresolved. In this work, a full-scale system integrating a solar CPC/UV-LED step combined with commercial microfiltration membranes was installed in a conventional WWTP for agricultural reuse purposes. After optimization, 0.5 g/L of catalyst and combined SOLAR + UV-LED showing the highest pharmaceutical removal percentages, while a self-designed UV-LED included in the own reaction tank resulting in higher efficiencies compared with commercial lamps. Longer membrane surface area decreased fouling problems in the system. However, 60 min of irradiation time was necessary to reach the most restrictive water quality values according with (EU 2020/741). After optimization step, total costs were reduced by 45%. However, it was shown that a reduction in operating and maintenance costs, along with the development of more effective and economical commercial filtration membranes is a key factor; therefore, working on these aspects is essential in the treated water cost reduction.

Risk assessment for uptake and accumulation of pharmaceuticals by baby leaf lettuce irrigated with reclaimed water under commercial agricultural activities.

The use of reclaimed water to irrigate agricultural crops has increased in recent years as a consequence of water shortage constituting a potential risk for human health. The main objective of this study was to evaluate the impact on the soil-plant system and determining the accumulation of carbamazepine (CBZ), diclofenac (DCF), ketoprofen (KTP) and naproxen (NPX) in the edible part of lettuce under commonly used agricultural practices in commercial production. For this purpose, red oak baby lettuce (Lactuca sativa L.) was irrigated with reclaimed water fortified with different concentrations of pharmaceuticals. The study was carried out in two different scenarios: soil and tray. The tray experiments were conducted with substrate and took place at three different seasons of the year. Lettuce tissue sampled from these experiments were analysed 3 times during the lettuce growing cycle (first, second and third harvest). The practices of first harvest regrowth were also evaluated. For all experiments, CBZ showed the highest accumulation in lettuce leaves of the pharmaceuticals tested, showing a correlation between irrigation exposure time and pharmaceutical uptake. Unexpectedly, DCF was the compound with the highest uptake levels after regrowth practices. Results suggested that pharmaceuticals uptake could be directly associated with the irrigation method and possible accumulation in soil and substrates, while concentration of pharmaceuticals in substrates were 10 times higher. Based on the concentration values detected in lettuce leaves, the risk assessment suggests that no compounds imply any risk to human health, except CBZ for those on vegetarian diets in the tray scenario. Although commercial agricultural practices are usually not considered with regards to risk reduction, in this experiment we demonstrated that climatic conditions are a key factor in pharmaceuticals uptake and different agricultural practices (soil cropping and drip irrigation) can limit the presence of pharmaceutical compounds in crops.

The Bark Beetle Dendroctonus rhizophagus (Curculionidae: Scolytinae) Has Digestive Capacity to Degrade Complex Substrates: Functional Characterization and Heterologous Expression of an α-Amylase.

Dendroctonus-bark beetles are natural agents contributing to vital processes in coniferous forests, such as regeneration, succession, and material recycling, as they colonize and kill damaged, stressed, or old pine trees. These beetles spend most of their life cycle under stem and roots bark where they breed, develop, and feed on phloem. This tissue is rich in essential nutrients and complex molecules such as starch, cellulose, hemicellulose, and lignin, which apparently are not available for these beetles. We evaluated the digestive capacity of Dendroctonusrhizophagus to hydrolyze starch. Our aim was to identify α-amylases and characterize them both molecularly and biochemically. The findings showed that D. rhizophagus has an α-amylase gene (AmyDr) with a single isoform, and ORF of 1452 bp encoding a 483-amino acid protein (53.15 kDa) with a predicted signal peptide of 16 amino acids. AmyDr has a mutation in the chlorine-binding site, present in other phytophagous insects and in a marine bacterium. Docking analysis showed that AmyDr presents a higher binding affinity to amylopectin compared to amylose, and an affinity binding equally stable to calcium, chlorine, and nitrate ions. AmyDr native protein showed amylolytic activity in the head-pronotum and gut, and its recombinant protein, a polypeptide of ~53 kDa, showed conformational stability, and its activity is maintained both in the presence and absence of chlorine and nitrate ions. The AmyDr gene showed a differential expression significantly higher in the gut than the head-pronotum, indicating that starch hydrolysis occurs mainly in the midgut. An overview of the AmyDr gene expression suggests that the amylolytic activity is regulated through the developmental stages of this bark beetle and associated with starch availability in the host tree.

Novel clinical and molecular findings in Spanish patients with naevoid basal cell carcinoma syndrome.

BACKGROUND: Naevoid basal cell carcinoma syndrome (NBCCS) is an autosomal dominant disorder characterized by developmental alterations and multiple basal cell carcinomas. Mutations in PTCH1, which encodes a membrane receptor for Sonic Hedgehog, are associated with the development of the disease. Most of them produce a truncated protein, which is unable to suppress Smoothened protein and continuously activates the downstream pathway. OBJECTIVES: We aimed to characterize 22 unrelated Spanish patients with NBCCS, the largest cohort with Gorlin syndrome reported to date in Spain. METHODS: Genomic analysis of PTCH1 was performed in patients with NBCCS and controls, and mutations were analysed using bioinformatics tools. RESULTS: We report for the first time two young patients, one each with uterus didelphys and ganglioneuroma, within the context of NBCCS. One patient showing a severe phenotype of the disease had developed basal cell carcinomas since childhood. Sanger sequencing of PTCH1 in this cohort identified 17 novel truncating mutations (11 frameshift, five nonsense and one mutation affecting an exon-intron splice site) and two novel missense mutations that were predicted to be pathogenic. The patients showed great clinical variability and inconsistent genotype-phenotype correlation, as seen in relatives carrying similar mutations. CONCLUSIONS: This study contributes to increase the pool of clinical manifestations of NBCCS, as well as increasing the number of pathogenic mutations identified in PTCH1 predisposing to the condition. The inconsistencies found between phenotype and genotype suggest the involvement of other modifying factors, genetic, epigenetic or environmental.

Practical approach to the evaluation of industrial wastewater treatment by the application of advanced microbiological techniques.

In cork industry, the operation of boiling raw cork generates large volumes of wastewater named Cork Boiling Wastewater (CBW). The main characteristics are the low biodegradability and medium to low acute toxicity, resulting in the necessity of designing advanced biological treatments by possible conventional activated sludge adaptation. In order to evaluate the variation of bacterial population along that process, a study based on optical microscopy, plate count, DNA extraction, qPCR and massive sequencing techniques was performed. Results showed a diminution of the total and volatile solids (TSS and VSS), jointly with a decrease in DNA concentration, general bacteria (16 S) and ammonia-oxidizing bacteria (AOB). After a few hours of testing, diverse microbiological species died while others showed a possible adaptation of the biological system, accompained by a dissolved organic carbon (DOC) reduction. In addition, toxicity tests based on activated sludge showed the development of chronic toxicity through the contact time. Combination of classical and advanced microbiological techniques, such as quantitative real time Polymerase Chain Reaction (qPCR) and metagenomics, was essential to predict the variation of species during the experiment and to conclude if effective biological adaptation could be finally attained for the target complex wastewater.

Heat recovery during treatment of highly concentrated wastewater: economic evaluation and influencing factors.

This paper assesses the economics of heat recovery from biological wastewater treatment plants (WWTPs) treating concentrated wastewater, as higher concentrations result in higher heat generation in the treatment basin. A heat balance model has been applied to calculate the amount of recoverable heat from the system and the effect of the heat extraction capacity on the economics of a heat pump installation, evaluated using the internal rate of return. A sensitivity analysis has been performed to evaluate the effect of several parameters on the economics of heat recovery in this type of WWTP: the electricity price, the price of the fuel substituted by heating savings, the investment costs, the coefficient of performance (COP) and the amount of heat extracted from the system. It was calculated that the heat pump capacity has to be high enough to recover a significant amount of heat, but low enough to improve the economics of the system. The economic performance of the system is very dependent on the energy prices of both electrical power to run the heat pump and the fuel (heat) cost substituted by the heat pump.

Deconvolution of complex differential scanning calorimetry profiles for protein transitions under kinetic control.

A frequent outcome in differential scanning calorimetry (DSC) experiments carried out with large proteins is the irreversibility of the observed endothermic effects. In these cases, DSC profiles are analyzed according to methods developed for temperature-induced denaturation transitions occurring under kinetic control. In the one-step irreversible model (native → denatured) the characteristics of the observed single-peaked endotherm depend on the denaturation enthalpy and the temperature dependence of the reaction rate constant, k. Several procedures have been devised to obtain the parameters that determine the variation of k with temperature. Here, we have elaborated on one of these procedures in order to analyze more complex DSC profiles. Synthetic data for a heat capacity curve were generated according to a model with two sequential reactions; the temperature dependence of each of the two rate constants involved was determined, according to the Eyring's equation, by two fixed parameters. It was then shown that our deconvolution procedure, by making use of heat capacity data alone, permits to extract the parameter values that were initially used. Finally, experimental DSC traces showing two and three maxima were analyzed and reproduced with relative success according to two- and four-step sequential models.

A Biological Approach for the Synthesis of Bismuth Nanoparticles: Using Thiolated M13 Phage as Scaffold.

We report the synthesis of Bi nanoparticles (Bi NPs) using the M13 phage as scaffold. The p8 protein of the phage is functionalized with thiol groups of different lengths, and these thiolated regions act as nucleation centers for Bi(3+) ions. The size distribution, shape, and resilience to oxidation of the Bi NPs depend on the length of the thiol group used. The NPs are characterized by high resolution transmission electron microscopy, Raman, and IR spectroscopies, matrix assisted laser desorption/ionization, and optical absorption. These results show that the nanoparticles are crystalline and have a typical diameter of ∼3.0 nm. The method of preparation presented here is reproducible and implies "greener" conditions than those reported elsewhere. To the best of our knowledge, this is the first report of bismuth nanoparticles synthesized by a biomineralization method.

Cardiac arrhythmias in stroke unit patients. Evaluation of the cardiac monitoring data. / Arritmias cardiacas en la unidad de ictus: análisis de los datos de la monitorización cardiaca.

INTRODUCTION: Cardiac arrhythmias are frequent in acute stroke. Stroke units are widely equipped with cardiac monitoring systems. Pre-existing heart diseases and heart-brain interactions may be implicated in causing cardiac arrhythmias in acute stroke. This article analyses cardiac arrhythmias detected in patients hospitalised in a stroke unit. METHOD: Prospective observational study of consecutive patients admitted to a stroke unit with cardiac monitoring. We collected clinical data from patients and the characteristics of their cardiac arrhythmias over a 1-year period (2013). Time of arrhythmia onset, associated predisposing factors, and the therapeutic decisions made after detection of arrhythmia were examined. All patients underwent continuous cardiac monitoring during no less than 48hours. RESULTS: Of a total of 332 patients admitted, significant cardiac arrhythmias occurred in 98 patients (29.5%) during their stay in the stroke unit. Tachyarrhythmia (ventricular tachyarrhythmias, supraventricular tachyarrhythmias, complex ventricular ectopy) was present in 90 patients (27.1%); bradyarrhythmia was present in 13 patients (3.91%). Arrhythmias were independently associated with larger size of brain lesion and older age. In 10% of the patient total, therapeutic actions were taken after detection of significant cardiac arrhythmias. Most events occurred within the first 48hours after stroke unit admission. CONCLUSIONS: Systematic cardiac monitoring in patients with acute stroke is useful for detecting clinically relevant cardiac arrhythmias. Incidence of arrhythmia is higher in the first 48hours after stroke unit admission. Age and lesion size were predicted appearance of arrhythmias. Detection of cardiac arrhythmias in a stroke unit has important implications for treatment.

Detailed characterization of MLH1 p.D41H and p.N710D variants coexisting in a Lynch syndrome family with conserved MLH1 expression tumors.

Lynch syndrome (LS) is an autosomal dominant cancer-susceptibility disease caused by inactivating germline mutations in mismatch repair (MMR) genes. Variants of unknown significance (VUS) are often detected in mutational analysis of MMR genes. Here we describe a large family fulfilling Amsterdam I criteria carrying two rare VUS in the MLH1 gene: c.121G > C (p.D41H) and c.2128A > G (p.N710D). Collection of clinico-pathological data, multifactorial analysis, in silico predictions, and functional analyses were used to elucidate the clinical significance of the identified MLH1 VUS. Only the c.121G > C variant cosegregated with LS-associated tumors in the family. Diagnosed colorectal tumors were microsatellite unstable although immunohistochemical staining revealed no loss of MMR proteins expression. Multifactorial likelihood analysis classified c.2128A > G as a non-pathogenic variant and c.121G > C as pathogenic. In vitro functional tests revealed impaired MMR activity and diminished expression of c.121G > C. Accordingly, the N710 residue is located in the unconserved MLH1 C-terminal domain, whereas D41 is highly conserved and located in the ATPase domain. The obtained results will enable adequate genetic counseling of c.121G > C and c.2128A > G variant carriers and their families. Furthermore, they exemplify how cumulative data and comprehensive analyses are mandatory to refine the classification of MMR variants.

Using the M13 Phage as a Biotemplate to Create Mesoporous Structures Decorated with Gold and Platinum Nanoparticles.

By taking advantage of the physical and chemical properties of the M13 bacteriophage, we have used this virus to synthesize mesoporous silica structures. Major coat protein p8 was chemically modified by attaching thiol groups. As we show, the resulting thiolated phage can be used as a biotemplate able to direct the formation of mesoporous silica materials. Simultaneously, this thiol functionality acts as an anchor for binding metal ions, such as Au(3+) and Pt(4+), forming reactive M13-metal ionic complexes which evolve into metal nanoparticles (NPs) trapped in the mesoporous network. Interestingly, Au(3+) ions are reduced to Au(0) NPs by the protein residues without requiring an external reducing agent. Likewise, silica mesostructures decorated with Au and Pt NPs are prepared in a one-pot synthesis and characterized using different techniques. The obtained results allow us to propose a mechanism of formation. In addition, gold-containing mesoporous structures are tested for the reduction of 4-nitrophenol (4-NP) and methylene blue (MB) in the presence of NaBH4. Although all of the gold-containing catalysts exhibit catalytic activity, those obtained with thiolated phages present a better performance than that obtained with M13 alone. This behavior is ascribed to the position of the Au NPs, which are partially embedded in the wall of the final mesostructures.

Biomimetic sol-gel synthesis of TiO2 and SiO2 nanostructures.

We report the heptapeptide-mediated biomineralization of titanium dioxide nanoparticles from titanium alkoxides. We evaluated the influence of pH on the biomineralized products and found that nanostructured TiO2 was formed in the absence of external ions (water only) at pH ~ 6.5. Several variants (mutants) of the peptides with different properties (i.e., different charges, isoelectric points (pIs), and sequences) were designed and tested in biomineralization experiments. Acid-catalyzed experiments were run using the H1 (HKKPSKS) peptide at room temperature, which produced anatase nanoparticles (~5 nm in size) for the first time via a heptapeptide and sol-gel approach. In addition, the peptide H1 was used to synthesize SiO2 nanoparticles. The influence of the pH and the added ions were monitored: at higher pH levels (8-9), SiO2 nanoparticles (20-30 nm in size) were obtained. In addition, whereas borate and Tris ions allowed the formation of colloidal systems, phosphate ions were unable to produce sols. The results presented here demonstrate that biomineralization depends on the sequence and charge of the peptide, and ions in solution can optimize the formation of nanostructures.

Gut transcriptome of two bark beetle species stimulated with the same kairomones reveals molecular differences in detoxification pathways.

Dendroctonus bark beetles are the most destructive agents in coniferous forests. These beetles come into contact with the toxic compounds of their host's chemical defenses throughout their life cycle, some of which are also used by the insects as kairomones to select their host trees during the colonization process. However, little is known about the molecular mechanisms by which the insects counteract the toxicity of these compounds. Here, two sibling species of bark beetles, D. valens and D. rhizophagus, were stimulated with vapors of a blend of their main kairomones (α-pinene, ß-pinene and 3-carene), in order to compare the transcriptional response of their gut. A total of 48 180 unigenes were identified in D. valens and 43 704 in D. rhizophagus, in response to kairomones blend. The analysis of differential gene expression showed a transcriptional response in D. valens (739 unigenes, 0.58-10.36 Log2FC) related to digestive process and in D. rhizophagus (322 unigenes 0.87-13.08 Log2FC) related to xenobiotics metabolism. The expression profiles of detoxification genes mainly evidenced the up-regulation of COEs and GSTs in D. valens, and the up-regulation of P450s in D. rhizophagus. Results suggest that terpenes metabolism comes accompanied by an integral hormetic response, result of compensatory mechanisms, including the activation of other metabolic pathways, to ensure the supply of energy and the survival of organisms which is specific for each species, according to its life history and ecological strategy.

Selection of patients for percutaneous closure in nonlacunar cryptogenic stroke associated with patent foramen ovale. Data from the NORDICTUS cooperative registry.

INTRODUCTION: There is an extending use of percutaneous closure of patent foramen ovale (PFO) as therapy for PFO-associated cryptogenic strokes. The aim of our study was to investigate the clinical practice of percutaneous closure of PFO and to analyse the variables for decision-making on the selection of patients for this procedure. METHOD: A prospective observational multicentric survey was conducted using all the cases of cryptogenic stroke/transient ischaemic attack associated with PFO recorded in the NORDICTUS hospital registry during the period 2018-2021. Clinical data, radiological patterns, echocardiogram data and factors related to PFO-associated stroke (thromboembolic disease and paradoxical embolism criteria) were recorded. The indication for closure was analysed according to age (≤/> 60 years) and the characteristics of the PFO. RESULTS: In the group ≤ 60 years (n = 488), 143 patients (29.3%) underwent PFO closure. The most influential variables for this therapy were detection of a high-risk PFO (OR 4.11; IC 2.6-6.5, P < .001), criteria for paradoxical embolism (OR 2.61; IC 1.28-5.28; P = .008) and previous use of antithrombotics (OR 2.67; IC 1.38-5.18; P = .009). In the > 60 years group (n = 124), 24 patients had PFO closure (19%). The variables related to this option were history of pulmonary thromboembolism, predisposition to thromboembolic disease, paradoxical embolism criteria, and high-risk PFO. CONCLUSIONS: The detection of a high-risk PFO (large shunt, shunt with associated aneurysm) is the main criterion for a percutaneous closure-based therapy. Other conditions to consider in the eligibility of patients are the history of thromboembolic disease, paradoxical embolism criteria or the previous use of antithrombotics.

Microbial dynamics during the ripening of a mixed cow and goat milk cheese manufactured using frozen goat milk curd.

To overcome the seasonal shortage of goat milk in mixed milk cheese manufacture, pasteurized goat milk curd and high-pressure-treated raw goat milk curd manufactured in the spring were held at -24 °C for 4 mo, thawed, and mixed with fresh cow milk curd for the manufacture of experimental cheeses. Control cheeses were made from a mixture of pasteurized cow and goat milk. The microbiota of experimental and control cheeses was studied using culture-dependent and culture-independent techniques. Bacterial enumeration by classical methods showed lactic acid bacteria to be the dominant population in both control and experimental cheeses. In total, 681 isolates were grouped by partial amplified rDNA restriction analysis (ARDRA) into 4 groups and identified by 16S rRNA gene sequencing as Lactococcus lactis ssp. lactis (563 isolates), Leuconostoc pseudomesenteroides (72 isolates), Lactobacillus spp. (34 isolates), and Lc. lactis ssp. cremoris (12 isolates). Temporal temperature gradient gel electrophoresis (TTGE) analysis of cheese showed (1) the predominance of Lc. lactis in all cheeses; (2) the presence of Leu. pseudomesenteroides population in all cheeses from d 15 onward; (3) the presence of a Lactobacillus plantarum population in control cheese until d 15 and in experimental cheeses throughout the ripening period. Due to the most diverse and complete set of peptidases present in the genus Lactobacillus, the prevalence of this population in experimental cheeses could give rise to differences in cheese flavor between experimental and control cheeses.

Mass and heat balances for biological nitrogen removal in an activated sludge process: to couple or not to couple?

Models adapt constantly, usually increasing the degree of detail describing physical phenomena. In water resource recovery facilities, models based on mass and/or heat balances have been used to describe and improve operation. While both mass and heat balances have proven their worth individually, the question arises to which extent their coupling, which entails increased model complexity, warrants the supposedly more precise simulation results. In order to answer this question, the need for and effects of coupling mass and heat balances in modelling studies were evaluated in this work for a biological nitrogen removal process treating highly concentrated wastewater. This evaluation consisted on assessing the effect of the coupling of mass and heat balances on the prediction of: (1) nitrogen removal efficiency; (2) temperature; (3) heat recovery. In general, mass balances are sufficient for evaluating nitrogen removal efficiency and effluent nitrogen concentrations. If one desires to evaluate the effect of temperature changes (e.g. daily, weekly, seasonally) on nitrogen removal efficiency, the use of temperature profiles as an input variable to a mass balance-based model is recommended over the coupling of mass and heat balances. In terms of temperature prediction, considering a constant biological heat generation term in the heat balance model provides sufficient information - i.e. without the coupling of mass and heat balances. Also, for evaluating the heat recovery potential of the system, constant biological heat generation values provide valuable information, at least under normal operating conditions, i.e. when the solids retention time is large enough to maintain nitrification.

SEOM clinical guideline on hereditary colorectal cancer (2019).

In the last 2 decades, clinical genetics on hereditary colorectal syndromes has shifted from just a molecular characterization of the different syndromes to the estimation of the individual risk of cancer and appropriate risk reduction strategies. In the last years, new specific therapies for some subgroups of patients have emerged as very effective alternatives. At the same time, germline multigene panel testing by next-generation sequencing (NGS) technology has become the new gold standard for molecular genetics.

Surgical management of hereditary colorectal cancer: surgery based on molecular analysis and family history.

The importance of colorectal cancer (CRC) is increasing. A proportion show a hereditary component, as in Lynch syndrome and Familial Adenomatous Polyposis, and a recently defined entity as well, namely, Familial Colorectal Cancer type X. The high probability to develop CRC in these groups may, at the time of recognition, change surgical management, including its timing or even the surgical technique. In some cases prophylactic surgery can play an important role. The possibility of using tools that allow recognition of the aforementioned syndromes, including microsatellite instability, immunohistochemistry for DNA mismatch repair system proteins, and especially their mutations, is on the basis of therapeutic strategies that differ from those employed in sporadic CRC cases.

TiBALDH as a precursor for biomimetic TiO2 synthesis: stability aspects in aqueous media.

Titanium(iv) bis(ammonium lactate)dihydroxide (TiBALDH) is a commercially available reagent frequently used to synthesize TiO2. Particularly, for the biomimetic synthesis of TiO2, TiBALDH is the preferred precursor because it can be mixed in aqueous solutions with no apparent hydrolysis or condensation reactions. Thus, proteins or other biomolecules can be used as a template in aqueous systems for the synthesis of TiO2 from TiBALDH. Nevertheless, there is evidence that TiBALDH is in equilibrium with TiO2, and even, the principal structure of the complex has been suggested as [Ti4O4(lactate)8]8-. Since that chemical equilibrium depends on the polarity of the solvent, in this work we explored a diversity of media to test the chemical stability of TiBALDH and its equilibrium with TiO2 at room temperature. TiBALDH (2.078 M) contains particles of 18.6 ± 7.3 nm in size, if it is diluted with deionized water, the particles reach a size of 5.2 ± 1.7 nm, which suggest that intermolecular interactions form polymers of titanium lactate complexes reversibly, reaching equilibrium after 10 hours. Typical buffer systems were tested; TiBALDH reacted rapidly only with phosphate groups, even if the source came from DNA. Therefore, phosphate buffer must be avoided in biomineralization TiO2 synthesis. In solutions of TiBALDH at basic pH, condensation reactions are promoted to form a gel containing anatase nanoparticles, but if the solutions are acidic, monodisperse anatase nanoparticles of ∼5 nm were observed. The results show that the commercial reagent TiBALDH contains many species of titanium lactate complexes in equilibrium with TiO2, and it is affected by the concentration, time, pH, and several ions. This peculiar behavior must be taken into account when this precursor is used and it could be useful to develop novel synthesis routes of macrostructures with biomolecules in aqueous systems.