Micromotor-based dual aptassay for early cost-effective diagnosis of neonatal sepsis.

Given the long-life expectancy of the newborn, research aimed at improving sepsis diagnosis and management in this population has been recognized as cost-effective, which at early stages continues to be a tremendous challenge. Despite there is not an ideal-specific biomarker, the simultaneous detection of biomarkers with different behavior during an infection such as procalcitonin (PCT) as high specificity biomarker with one of the earliest biomarkers in sepsis as interleukin-6 (IL-6) increases diagnostic performance. This is not only due to their high positive predictive value but also, since it can also help the clinician to rule out infection and thus avoid the use of antibiotics, due to their high negative predictive value. To this end, we explore a cutting-edge micromotor (MM)-based OFF-ON dual aptassay for simultaneous determination of both biomarkers in 15 min using just 2 µL of sample from low-birth-weight neonates with gestational age less than 32 weeks and birthweight below 1000 g with clinical suspicion of late-onset sepsis. The approach reached the high sensitivities demanded in the clinical scenario (LODPCT = 0.003 ng/mL, LODIL6 = 0.15 pg/mL) with excellent correlation performance (r > 0.9990, p < 0.05) of the MM-based approach with the Hospital method for both biomarkers during the analysis of diagnosed samples and reliability (Er < 6% for PCT, and Er < 4% for IL-6). The proposed approach also encompasses distinctive technical attributes in a clinical scenario since its minimal sample volume requirements and expeditious results compatible with few easy-to-obtain drops of heel stick blood samples from newborns admitted to the neonatal intensive care unit. This would enable the monitoring of both sepsis biomarkers within the initial hours after the manifestation of symptoms in high-risk neonates as a valuable tool in facilitating prompt and well-informed decisions about the initiation of antibiotic therapy.These results revealed the asset behind micromotor technology for multiplexing analysis in diagnosing neonatal sepsis, opening new avenues in low sample volume-based diagnostics.

[Estimation of prevalence of chronic Chagas disease in Brazilian municipalitiesEstimación de la prevalencia de la enfermedad de Chagas crónica en los municipios brasileños]. / Estimativa de prevalência de doença de Chagas crônica nos municípios brasileiros.

Objective: The objective of this study is to estimate the prevalence of chronic Chagas disease (CCD) in Brazil: in the general population, in women, and in women of childbearing age. Methods: A meta-analysis of the literature was conducted to extract data on the prevalence of CCD in municipalities in Brazil in the 2010-2022 period: in the general population, in women, and in women of childbearing age. Municipal-level CCD indicators available in health information systems were selected. Statistical modeling of the data extracted from the meta-analysis (based on data obtained from information systems) was applied to linear, generalized linear, and additive models. Results: The five most appropriate models were selected from a total of 549 models tested to obtain a consensus model (adjusted R2 = 54%). The most important predictor was self-reported CCD in the primary health care information system. Zero prevalence was estimated in 1 792 (32%) of Brazil's 5 570 municipalities; in the remaining 3 778 municipalities, average prevalence of the disease was estimated at 3.25% (± 2.9%). The number of carriers of CCD was estimated for the Brazilian population (~3.7 million), for women (~2.1 million) and for women of childbearing age (~590 000). The disease reproduction rate was calculated at 1.0336. All estimates refer to the 2015-2016 period. Conclusions: The estimated prevalence of CCD, especially among women of childbearing age, highlights the challenge of vertical transmission in Brazilian municipalities. Mathematical projections suggest that these estimates should be included in the national program for the elimination of vertical transmission of Chagas disease.

Objetivo: El objetivo de este estudio fue estimar la prevalencia de la enfermedad de Chagas crónica en la población brasileña en general, en las mujeres y en las mujeres en edad fértil. Métodos: Se realizó un metanálisis de la bibliografía para extraer datos sobre la prevalencia de la enfermedad de Chagas crónica en la población brasileña en general, en las mujeres y en las mujeres en edad fértil, en los municipios de Brasil durante el período 2010-2022. Se seleccionaron los indicadores relacionados con esa enfermedad disponibles en los sistemas municipales de información de salud. La modelización estadística de los datos extraídos del metanálisis, en función de los obtenidos de los sistemas de información, se aplicó a modelos lineales, lineales generalizados y aditivos. Resultados: Se seleccionaron los cinco modelos más apropiados de un total de 549 modelos evaluados, para obtener un modelo de consenso (R2 ajustado = 54%). El factor predictor más importante fue el registro de la enfermedad de Chagas crónica autodeclarada en el sistema de información de atención primaria de salud. De los 5570 municipios brasileños, en 1792 (32%) la prevalencia estimada fue nula y en los 3778 restantes la prevalencia media fue del 3,25% (± 2,9%). El número estimado de pacientes con enfermedad de Chagas crónica en la población brasileña en general, en las mujeres y en las mujeres en edad fértil fue de ~3,7 millones, ~2,1 millones y ~590 000, respectivamente. La tasa calculada de reproducción de la enfermedad fue de 1,0336. Todas las estimaciones se refieren al período 2015-2016. Conclusiones: La prevalencia estimada de la enfermedad de Chagas crónica, especialmente en las mujeres en edad fértil, pone de manifiesto el desafío que representa la transmisión vertical en los municipios brasileños. Estas estimaciones están en línea con los patrones de las proyecciones matemáticas, y sugieren la necesidad de incorporarlas al Pacto Nacional para la Eliminación de la Transmisión Vertical de la Enfermedad de Chagas.

Analysis of microsamples by miniaturized magnetic-based pipette tip microextraction: determination of free cortisol in serum and urine from very low birth weight preterm newborns.

Miniaturized magnetic-based pipette tip microextraction is presented as a sample preparation approach for microsamples. It involves quick dispersion of a diminutive amount of a magnetic sorbent material in a low-volume sample (10 µL) to entrap the target analytes. Next, the dispersion is aspirated using a (semi)automatic pipette through a pipette tip with a small cubic neodymium magnet inside, which retrieves the magnetic sorbent containing the analytes. After discarding the rest of the sample, the sorbent is properly rinsed by aspirating/dispensing deionized water, and then, the analytes are eluted by aspirating/dispensing an appropriate solvent. This approach was employed for the determination of free cortisol in serum and urine from very low birth weight preterm newborns, a vulnerable patient group who present low availability for sampling biological fluids. A magnetic immunosorbent made of a cortisol antibody was employed for the selective extraction, followed by liquid chromatography-tandem mass spectrometry. Good analytical features were obtained, such as limits of detection and quantification of 0.08 and 0.27 ng mL-1, respectively, linearity up to 50 ng mL-1 (R2 > 0.999), RSD values under 15% and relative recoveries between 91 and 111%. The cross-reactivity with other glucocorticoids (i.e., cortisone and prednisolone) was evaluated to show the selectivity of the extraction. Finally, the method applicability was demonstrated towards the determination of free cortisol in the serum and urine samples from low birth weight preterm newborns.

Predictors of subclinical atherosclerosis in HIV.

BACKGROUND: Cardiovascular disease is a major cause of morbidity and mortality in people with HIV. The detection of subclinical atherosclerosis through vascular ultrasound allows us to identify patients at an increased risk of cardiovascular disease as a primary prevention strategy; this test is not routine. Our objective is to identify predictors of subclinical atherosclerosis in a population with HIV. METHODS: People with HIV infection were selected for primary prevention and underwent carotid and femoral ultrasound to detect atheromatous plaques. Logistic regression analysis including vascular risk factors was performed to predict the presence of atherosclerosis. RESULTS: One hundred eighty-three patients were included, 54% of whom were smokers; the mean duration of HIV infection was 9.52 years, and all patients were undergoing antiretroviral treatment. Subclinical atherosclerosis was present in 62.29% of the patients; 83.32% had plaque in the carotid territory, 57.93% in the femoral territory and 25.6% in both vascular territories. Compared to those without atherosclerosis, patients with atherosclerosis were on average 5.35 years older (53.86 vs. 48.51, p < 0.001) and had a higher prevalence of smoking (63.23% vs. 39.12%, p = 0.020) and a CD4/CD8 ratio below 0.7 (44.23% vs. 29.02%, p = 0.043). A CD4/CD8 ratio lower than 0.3 was always associated with subclinical atherosclerosis (95% confidence interval (CI): 83.9-100%). The inclusion of smoking, the CD4/CD8 ratio and age in the logistic regression analysis led to a diagnostic yield of 72% measured by the area under the receiving operator characteristic (ROC) curve (95% CI: 64-80%). CONCLUSIONS: Tobacco use, age and a CD4/CD8 ratio below 0.7 allow prediction of the presence of subclinical atherosclerosis in primary prevention. A CD4/CD8 ratio below 0.3 was a diagnostic indicator of atherosclerosis in HIV patients undergoing primary prevention in our sample.

The association between language use and food insecurity among Hispanic adults residing in the USA depends on nativity.

OBJECTIVE: To examine the association between language use - predominantly English, English and Spanish equally and predominantly Spanish - and food insecurity among Hispanic adults residing in the USA, 1999-2018. DESIGN: Pooled cross-sectional study design. SETTING: United States. PARTICIPANTS: 15 073 Hispanic adults. RESULTS: Compared with Hispanic adults who predominantly spoke English and after adjusting for age, sex, family income-to-poverty ratio, education level and employment status, Hispanic adults who spoke English and Spanish equally (OR = 1·28, 95 % CI = 1·05, 1·56) or predominantly Spanish (OR = 1·25, 95 % CI = 1·04, 1·49) had higher odds of food insecurity. After stratifying by country of birth, language use was associated with higher odds of food insecurity only for Hispanic adults born outside of the USA, but not for Hispanic adults born in the USA. Hispanic adults born outside of the USA who spoke English and Spanish equally (OR = 1·27, 95 % CI = 1·04, 1·55) or spoke predominantly Spanish (OR = 1·24, 95 % CI = 1·04, 1·48) had higher odds of food insecurity when compared with those who predominantly spoke English. CONCLUSION: Foreign-born Hispanic adults who speak predominantly Spanish, or English and Spanish equally, have higher odds of food insecurity. Food and nutrition assistance programmes that serve Hispanic immigrants should make sure to provide linguistically and culturally appropriate services to this population.

An improved microfluidic device to enhance the enrichment factors in liquid phase microextraction: application to the simultaneous extraction of polar and non-polar acids in biological samples.

A new microfluidic device to enhance the enrichment factor in miniaturized systems is proposed. The microfluidic system was design for liquid phase microextractions, and it was applied to the simultaneous extraction of acidic compounds of a wide range of polarity (0.5 < log P < 3). The device operated under stagnant acceptor phase conditions and all the operational parameters involved were optimized. Tributyl phosphate was found to be a new highly efficient supported liquid membrane to simultaneously extract analytes of very different polarities. The optimal donor and acceptor phase were pH 2 and pH 13, respectively. The donor flow rate and the extraction time were investigated simultaneously, offering great versatility with high enrichment factors (EFs). Limits of quantitation were within 0.02 and 0.09 µg mL-1 for all compounds at 10 µL min-1 as donor flow rate and 20-min extractions, offering EFs between 11 and 18 with only 200-µL sample volume consumption. The method was successfully applied to human urine samples, observing recoveries between 47 and 90% for all compounds. This new proposed microfluidic system increases the wide range of applications, especially when the analytes are present in lower concentrations in the sample.

Prussian Blue/Chitosan Micromotors with Intrinsic Enzyme-like Activity for (bio)-Sensing Assays.

Prussian Blue (PB)/chitosan enzyme mimetic tubular micromotors are used here for on-the-fly (bio)-sensing assays. The micromotors are easily prepared by direct deposition of chitosan into the pores of a membrane template and in situ PB synthesis during hydrogel deposition. Under judicious pH control, PB micromotors display enzyme mimetic capabilities with three key functions on board: the autonomous oxygen bubble propulsion (with PB acting as a catalase mimic for hydrogen peroxide decomposition), 3,3',5,5'-tetramethylbenzidine (TMB) oxidation (with PB acting as a peroxidase mimic for analyte detection), and as a magnetic material (to simplify the (bio)-sensing steps). In connection with chitosan capabilities, these unique enzyme mimetic micromotors are further functionalized with acetylthiocholinesterase enzyme (ATChE) to be explored in fast inhibition assays (20 min) for the colorimetric determination of the nerve agent neostigmine, with excellent analytical performance in terms of quantification limit (0.30 µM) and concentration linear range (up to 500 µM), without compromising efficient micromotor propulsion. The new concept illustrated holds considerable potential for a myriad of (bio)-sensing applications, including forensics, where this conceptual approach remains to be explored. Micromotor-based tests to be used in crime scenes are also envisioned due to the reliable neostigmine determination in unpretreated samples.

Physiological breeding for yield improvement in soybean: solar radiation interception-conversion, and harvest index.

KEY MESSAGE: Efficiency of light interception, Radiation use efficiency and harvest index can be used as targets to improve grain yield potential in soybean. Grain yield (GY) production can be expressed as the result of three main efficiencies: light interception (Ei), radiation use (RUE), and harvest index (HI). Although dissecting GY through these three efficiencies is not entirely new, there is a lack of knowledge about the phenotypic variation, the genetic architecture, and the relative contribution of these three efficiencies on GY in soybean. This knowledge gap coupled with laborious phenotyping prevents the active consideration of these efficiencies into breeding programs. This study aims to reveal the phenotypic variation, heritability, genetic relationships, genetic architecture, and genomic prediction for Ei, RUE, and HI in soybean. We evaluated a maturity control panel of 383 Recombinant Inbred Lines (RILs) selected from the soybean nested association mapping (SoyNAM) population. Dry matter ground measured along with canopy coverage (CC) from UAS imagery were collected in three environments. Light interception was modeled through a logistic curve using CC as a proxy. The total above-ground biomass collected during the growing season and its respective cumulative light intercepted were used to derive RUE through linear models fitting. Additive-genetic correlations, genome-wide association (GWA) and whole-genome regressions (WGR) were performed to evaluate the relationship between traits, their association with genomic regions, and the feasibility of predicting these efficiencies with genomic information. Correlation analyses considered three groups: the entire data set, and the high- and low-yielding RILs to determine association as a function of the GY. Our results revealed moderate to high phenotypic variation for Ei, RUE, and HI with ranges of 8.5%, 1.1 g MJ-1, and 0.2, respectively. Additive-genetic correlation revealed a strong relationship of GY with HI and moderate with RUE and Ei when whole data set was considered, but negligible contribution of HI on GY when just the top 100 was analyzed. The GWA analyses showed that Ei is associated with three SNPs; two of them located on chromosome 7 and one on chromosome 11 with no previous quantitative trait loci (QTLs) reported for these regions. RUE is associated with four SNPs on chromosomes 1, 7, 11, and 18. Some of these QTLs are novel, while others are previously documented for plant architecture and chlorophyll content. Two SNPs positioned on chromosome 13 and 15 with previous QTLs reported for plant height and seed set, weight and abortion were associated with HI. WGR showed high predictive ability for Ei, RUE, and HI with maximum correlation ranging between 0.75 and 0.80. Future improvements in GY can be expected through strategies prioritizing Ei for short-term results when using high yielding germplasm and RUE for medium- and long-term outcomes. This work is a pioneer attempt to integrate traditional physiological traits into the breeding process in the context of physiological breeding.

Bioaccumulation and biochemical responses in the peppery furrow shell Scrobicularia plana exposed to a pharmaceutical cocktail at sub-lethal concentrations.

Pharmaceutical drugs in the aquatic medium may pose significant risk to non-target organisms. In this study, the potential toxicity of a mixture of three compounds commonly detected in marine waters (ibuprofen, ciprofloxacin and flumequine) was assessed, by studying bioaccumulation, oxidative stress and neurotoxicity parameters (catalase CAT, superoxide dismutase SOD, glutathione reductase GR, glutathione S-transferase GST, lipid peroxidation LPO, glutathione peroxidase GPX, metallothionein MT and acetylcholinesterase AChE) in the clam Scrobicularia plana. Temporal evolution of selected endpoints was evaluated throughout an exposure period (1, 7 and 21 days) followed by a depuration phase. The accumulation of all drugs was fast, however clams showed the ability to control the internal content of drugs, keeping their concentration constant throughout the exposure and reducing their content after 7 days of depuration. The induction of biochemical alterations (SOD, CAT, LPO, MT, AChE) was observed in gills and digestive gland probably related to an imbalance in the redox state of clams as a consequence of the exposure to the drug mixture. These alterations were also maintained at the end of the depuration week when the high levels of SOD, CAT, GST and LPO indicated the persistence of oxidative stress and damage to lipids despite the fact that clams were no longer exposed to the mixture.

Biotechnological Advances to Improve Abiotic Stress Tolerance in Crops.

The major challenges that agriculture is facing in the twenty-first century are increasing droughts, water scarcity, flooding, poorer soils, and extreme temperatures due to climate change. However, most crops are not tolerant to extreme climatic environments. The aim in the near future, in a world with hunger and an increasing population, is to breed and/or engineer crops to tolerate abiotic stress with a higher yield. Some crop varieties display a certain degree of tolerance, which has been exploited by plant breeders to develop varieties that thrive under stress conditions. Moreover, a long list of genes involved in abiotic stress tolerance have been identified and characterized by molecular techniques and overexpressed individually in plant transformation experiments. Nevertheless, stress tolerance phenotypes are polygenetic traits, which current genomic tools are dissecting to exploit their use by accelerating genetic introgression using molecular markers or site-directed mutagenesis such as CRISPR-Cas9. In this review, we describe plant mechanisms to sense and tolerate adverse climate conditions and examine and discuss classic and new molecular tools to select and improve abiotic stress tolerance in major crops.

Glioblastoma Treatment: State-of-the-Art and Future Perspectives.

(1) Background: Glioblastoma is the most frequent and lethal primary tumor of the central nervous system. Through many years, research has brought various advances in glioblastoma treatment. At this time, glioblastoma management is based on maximal safe surgical resection, radiotherapy, and chemotherapy with temozolomide. Recently, bevacizumab has been added to the treatment arsenal for the recurrent scenario. Nevertheless, patients with glioblastoma still have a poor prognosis. Therefore, many efforts are being made in different clinical research areas to find a new alternative to improve overall survival, free-progression survival, and life quality in glioblastoma patients. (2) Methods: Our objective is to recap the actual state-of-the-art in glioblastoma treatment, resume the actual research and future perspectives on immunotherapy, as well as the new synthetic molecules and natural compounds that represent potential future therapies at preclinical stages. (3) Conclusions: Despite the great efforts in therapeutic research, glioblastoma management has suffered minimal changes, and the prognosis remains poor. Combined therapeutic strategies and delivery methods, including immunotherapy, synthetic molecules, natural compounds, and glioblastoma stem cell inhibition, may potentiate the standard of care therapy and represent the next step in glioblastoma management research.

Pseudocrossidium replicatum (Taylor) R.H. Zander is a fully desiccation-tolerant moss that expresses an inducible molecular mechanism in response to severe abiotic stress.

KEY MESSAGE: The moss Pseudocrossidium replicatum is a desiccation-tolerant species that uses an inducible system to withstand severe abiotic stress in both protonemal and gametophore tissues. Desiccation tolerance (DT) is the ability of cells to recover from an air-dried state. Here, the moss Pseudocrossidium replicatum was identified as a fully desiccation-tolerant (FDT) species. Its gametophores rapidly lost more than 90% of their water content when exposed to a low-humidity atmosphere [23% relative humidity (RH)], but abscisic acid (ABA) pretreatment diminished the final water loss after equilibrium was reached. P. replicatum gametophores maintained good maximum photosystem II (PSII) efficiency (Fv/Fm) for up to two hours during slow dehydration; however, ABA pretreatment induced a faster decrease in the Fv/Fm. ABA also induced a faster recovery of the Fv/Fm after rehydration. Protein synthesis inhibitor treatment before dehydration hampered the recovery of the Fv/Fm when the gametophores were rehydrated after desiccation, suggesting the presence of an inducible protective mechanism that is activated in response to abiotic stress. This observation was also supported by accumulation of soluble sugars in gametophores exposed to ABA or NaCl. Exogenous ABA treatment delayed the germination of P. replicatum spores and induced morphological changes in protonemal cells that resembled brachycytes. Transcriptome analyses revealed the presence of an inducible molecular mechanism in P. replicatum protonemata that was activated in response to dehydration. This study is the first RNA-Seq study of the protonemal tissues of an FDT moss. Our results suggest that P. replicatum is an FDT moss equipped with an inducible molecular response that prepares this species for severe abiotic stress and that ABA plays an important role in this response.

DNA barcode analysis of the endangered green turtle (Chelonia mydas) in Mexico1.

Technological and analytical advances to study evolutionary biology, ecology, and conservation of green turtles (Chelonia mydas) are realized through molecular approaches including DNA barcoding. We characterized the usefulness of COI DNA barcodes in green turtles in Mexico to better understand genetic divergence and other genetic parameters of this species. We analyzed 63 sequences, including 25 from green turtle field specimens collected from the Gulf of Mexico and from the Mexican Pacific and 38 already present in the Barcode of Life Data Systems (BOLD). A total of 13 haplotypes were identified with four novel haplotypes from the Pacific Ocean and three novel haplotypes from the Atlantic Ocean. Intraspecific distance values among COI gene sequences by two different models were 0.01, demonstrating that there is not a subdivision for green turtle species. Otherwise, the interspecific distance interval ranged from 0.07 to 0.13, supporting a clear subdivision among all sea turtle species. Haplotype and total nucleotide diversity values of the COI gene reflect a medium genetic diversity average. Green turtles of the Mexican Pacific showed common haplotypes to some Australian and Chinese turtles, but different from the haplotypes of the Mexican Atlantic. COI analysis revealed new haplotypes and confirmed that DNA barcodes were useful for evaluation of the population diversity of green turtles in Mexico.

Assessment of pharmaceutical mixture (ibuprofen, ciprofloxacin and flumequine) effects to the crayfish Procambarus clarkii: A multilevel analysis (biochemical, transcriptional and proteomic approaches).

The knowledge about the effects of pharmaceuticals on aquatic organisms has been increasing in the last decade. However, due to the variety of compounds presents in the aquatic medium, exposure scenarios and exposed organisms, there are still many gaps in the knowledge on how mixtures of such bioactive compounds affect exposed non target organisms. The crayfish Procambarus clarkii was used to analyze the toxicity effects of mixtures of ciprofloxacin, flumequine and ibuprofen at low and high concentrations (10 and 100 µg/L) over 21 days of exposure and to assess the recovery capacity of the organism after a depuration phase following exposure during additional 7 days in clean water. The crayfish accumulated the three compounds throughout the entire exposure in the hepatopancreas. The exposure to the mixture altered the abundance of proteins associated with different cells functions such as biotransformation and detoxification processes (i.e. catalase and glutathione transferase), carbohydrate metabolism and immune responses. Additionally changes in expression of genes encoding antioxidant enzymes and in activity of the corresponding enzymes (i.e. superoxide dismutase, glutathione peroxidase and glutathione transferase) were reported. Alterations at different levels of biological organization did not run in parallel under all circumstances and can be related to changes in the redox status of the target tissue. No differences were observed between control and exposed organisms for most of selected endpoints after a week of depuration, indicating that exposure to the drug mixture did not produce permanent damage in the hepatopancreas of P. clarkii.

Monitoring of pharmaceuticals in aquatic biota (Procambarus clarkii) of the Doñana National Park (Spain).

In this work the presence of different pharmaceuticals at Doñana National Park (Spain) and their main entry sources (input source or entry points) have been stated over the 2011-2016 years period. Twenty-three selected pharmaceuticals (corresponding to eight therapeutic families) were evaluated in crayfish and water samples from Doñana National Park (Spain) (six sampling points selected in order to cover different possible pollution sources into and surrounding the Park). The multiresidue determination was carried out using enzymatic-microwave assisted extraction prior to high performance liquid chromatography mass spectrometry detection. Sulphonamides (sulfadiazine, sulfamerazine, sulfamethazine, and sulfamethoxazole); trimethoprim, an antibiotic that is frequently co-administered with sulfamethoxazole; amphenicols (chloramphenicol, florfenicol and thiamphenicol); fluoroquinolones (ciprofloxacin, enrofloxacin, flumequine, danofloxacin, gatifloxacin, norfloxacin, marbofloxacin and grepafloxacin); penicillins (amoxicillin); tetracyclines (chlortetracycline and oxytetracycline); non-steroidal anti-inflammatory drugs (salicylic acid and ibuprofen); beta-blocker drugs (atenolol); and antiepileptics (carbamazepine) were analysed. Ciprofloxacin, ibuprofen, salicylic acid, flumequine, and carbamazepine were detected and/or quantified at some of the selected sampling points. A clear ecotoxicological risk to the ecosystem was demonstrated from the occurrence of ciprofloxacin in samples obtained after the punctual and massive presence of people inside the Park. Furthermore, flumequine and carbamazepine have been detected in Procambarus clarkii specimens in concentrations around 30 ng g-1 and 14 ng g-1, respectively, and their occurrence in the specimens could indicate the persistence of the discharge sources. The main source of pharmaceuticals into the Park might be the livestock farming activities, and the influence of urban wastewaters from surrounding villages does not seem to be very important.

Electrochemical Microfluidic Micromotors-Based Immunoassay for C-Reactive Protein Determination in Preterm Neonatal Samples with Sepsis Suspicion.

Online coupling of a micromotor-based immunoassay and a microfluidic electrochemical detection was explored as a new approach for C-reactive protein (CRP) determination in serum and preterm neonatal plasma samples with sepsis suspicion. The approach combines the advantages of micromotors (self-fluid mixing capabilities leading to a faster assay in low sample volumes) and electrochemical microfluidic (flow-controlled ultraminiaturized electrochemical detection, high sensitivity, and low-cost) technologies. Both technologies elegantly meet the point of care testing or bed side device requirements such as low analysis times, miniaturization and simplification, and single use. Anti-CRP functionalized micromotors (anti-CRP-rGO(reduced graphene oxide)/Ni/PtNPs (platinum nanoparticles))-based immunoassay coupled to thin layer Au-based electrochemical microfluidics operating at -0.20 V under controlled fluidic detection operations (30 µL min-1) allowed the sensitive (LOD = 0.54 µg/mL) and accurate CRP determination using very low volume preterm neonatal clinical samples (<10 µL) in just 8 min of total assay time. These excellent analytical characteristics obtained linked to the full automatization of the immunoassay allowed the fast and accurate determination of CRP in hardly available clinical samples as those coming from preterm infants with suspected sepsis. These results demonstrated the usefulness of the approach which meets the clinical requirements as a future point-of-care device for clinical analysis.

Genome-wide transcriptional changes triggered by water deficit on a drought-tolerant common bean cultivar.

BACKGROUND: Common bean (Phaseolus vulgaris L.) is a relevant crop cultivated over the world, largely in water insufficiency vulnerable areas. Since drought is the main environmental factor restraining worldwide crop production, efforts have been invested to amend drought tolerance in commercial common bean varieties. However, scarce molecular data are available for those cultivars of P. vulgaris with drought tolerance attributes. RESULTS: As a first approach, Pinto Saltillo (PS), Azufrado Higuera (AH), and Negro Jamapa Plus (NP) were assessed phenotypically and physiologically to determine the outcome in response to drought on these common bean cultivars. Based on this, a Next-generation sequencing approach was applied to PS, which was the most drought-tolerant cultivar to determine the molecular changes at the transcriptional level. The RNA-Seq analysis revealed that numerous PS genes are dynamically modulated by drought. In brief, 1005 differentially expressed genes (DEGs) were identified, from which 645 genes were up-regulated by drought stress, whereas 360 genes were down-regulated. Further analysis showed that the enriched categories of the up-regulated genes in response to drought fit to processes related to carbohydrate metabolism (polysaccharide metabolic processes), particularly genes encoding proteins located within the cell periphery (cell wall dynamics). In the case of down-regulated genes, heat shock-responsive genes, mainly associated with protein folding, chloroplast, and oxidation-reduction processes were identified. CONCLUSIONS: Our findings suggest that secondary cell wall (SCW) properties contribute to P. vulgaris L. drought tolerance through alleviation or mitigation of drought-induced osmotic disturbances, making cultivars more adaptable to such stress. Altogether, the knowledge derived from this study is significant for a forthcoming understanding of the molecular mechanisms involved in drought tolerance on common bean, especially for drought-tolerant cultivars such as PS.

Richness and metallo-tolerance of cultivable fungi recovered from three high altitude glaciers from Citlaltépetl and Iztaccíhuatl volcanoes (Mexico).

In Mexico little is known about high-altitude glacial psychrotolerant or psychrophilic fungal species, with most glacial fungi isolated from polar environments or Alpine glaciers. It has been documented that some of these species may play an important role in bioremediation of contaminated environments with heavy metals. In the present study, 75 fungi were isolated from glaciers in Citlaltépetl (5675 masl) and Iztaccíhuatl (5286 masl) volcanoes. Combining morphological characteristics and molecular methods, based on ITS rDNA, 38 fungi were partially identified to genus level, 35 belonging to Ascomycota and three to Mucoromycota. The most abundant genera were Cladosporium, followed by Alternaria and Sordariomycetes order. All isolated fungi were psychrotolerant, pigmented and resistant to different concentrations of Cr(III) and Pb(II), while none tolerated Hg(II). Fungi most tolerant to Cr(III) and Pb(II) belong to the genera Stemphylium, Cladosporium and Penicillium and to a lesser extent Aureobasidium and Sordariomycetes. To our knowledge, this is the first report on cultivable mycobiota richness and their Cr and Pb tolerance. The results open new research possibilities about fungal diversity and heavy metals myco-remediation. Extremophilic fungal communities should be further investigated before global warming causes permanent changes and we miss the opportunity to describe these sites in Mexico.

An on-chip microfluidic-based electrochemical magneto-immunoassay for the determination of procalcitonin in plasma obtained from sepsis diagnosed preterm neonates.

A novel on-chip electrochemical magneto-immunoassay for the determination of procalcitonin (PCT) has been proposed. The strategy involved the on-line performing of the biorecognition event and detection on the thin-film microfluidic gold electrode chamber operating at E = -0.20 V (vs. Au). The complete assay was performed in less than 15 minutes using only 25 µL of the sample, covering the entire range of clinically relevant PCT concentrations in sepsis diagnosis with a limit of detection and quantification of 0.02 ng mL-1 and 0.05 ng mL-1, respectively (the sepsis diagnosis threshold: 0.5 ng mL-1). The on-chip electrochemical magneto-immunoassay provided excellent results in the analysis of very unique samples obtained from preterm neonates admitted with suspected sepsis, in which the sample volume is hardly available. These characteristics fulfill the POCT requirements for PCT determination in the whole clinically relevant concentration range. Because of the high clinical relevance and the important role of PCT in sepsis, this approach opens new perspectives for sepsis diagnosis and therapy guidance using low volume samples.

Direct capillary electrophoresis analysis of basic and acidic drugs from microliter volume of human body fluids after liquid-phase microextraction through nano-fibrous membrane.

In the present work, a disposable microextraction device with a polyamide 6 nano-fibrous supported liquid membrane (SLM) is employed for the pretreatment of minute volumes of biological fluids. The device is placed in a sample vial for an at-line coupling to a commercial capillary electrophoresis instrument with UV-Vis detection (CE-UV) and injections are performed fully automatically from the free acceptor solution above the SLM with no contact between the capillary and the membrane. Up to 4-fold enrichment of model basic (nortriptyline, haloperidol, loperamide, and papaverine) and acidic (ibuprofen, naproxen, ketoprofen, and diclofenac) drugs is achieved by optimizing the ratio of the donor to the acceptor solution volumes (16 to 4 µL, respectively). The actual setup enables SLM extractions from less than a drop of sample and is suitable for pretreatment of scarce human body fluids. Two unique methods are reported for efficient clean-up and enrichment of the basic and acidic drugs from capillary blood (formed as dried blood spot), serum, and urine samples, which enable their determination at therapeutic and/or toxic levels. The hyphenation of the SLM extraction with CE-UV analysis provides good repeatability (RSD, 2.4-14.9%), linearity (r2, 0.988-1.000), sensitivity (LOD, 0.017-0.22 mg L-1), and extraction recovery (ER, 20-106%) at short extraction times (10 min) and with minimum consumption of samples and reagents. Graphical abstract.