Haplotype-Aware Detection of SERPINA1 Variants by Nanopore Sequencing.

α-1 Antitrypsin (AAT) is an acute-phase reactant with immunomodulatory properties that mainly inhibits neutrophil elastase. Low serum levels cause AAT deficiency (AATD), an underdiagnosed condition that predisposes to pulmonary and hepatic diseases. The SERPINA1 gene, which encodes AAT, contains >500 variants. PI∗Z and PI∗S alleles are the most diagnosed causes of AATD, but the role of the SERPINA1 haplotypes in AAT function remains unknown. SERPINA1 gene was PCR amplified from 94 patients with asthma, using primers with tails for indexing. Sequencing libraries were loaded into a MinION-Mk1C, and MinKNOW was used for basecalling and demultiplexing. Nanofilt and Minimap2 were used for filtering and mapping/alignment. Variant calling/phasing were performed with PEPPER-Margin-DeepVariant. SERPINA1 gene was 100% covered for all samples, with a minimum sequencing depth of 500×. A total of 75 single-nucleotide variants (SNVs) and 4 insertions/deletions were detected, with 45 and 2 of them highly polymorphic (minor allele frequency >0.1), respectively. Nine of the SNVs showed differences in allele frequencies when compared with the overall Spanish population. More than 90% of heterozygous SNVs were phased, yielding 91 and 58 different haplotypes for each SERPINA1 amplified region. Haplotype-based linkage disequilibrium analysis suggests that a recombination hotspot could generate variation in the SERPINA1 gene. The proposed workflow enables haplotype-aware genotyping of the SERPINA1 gene by nanopore sequencing, which will allow the development of novel AATD diagnostic strategies.

Entomo-virological surveillance of Flavivirus in mosquitoes in Yucatan State, Mexico.

The genus Flavivirus (Family: Flaviviridae) comprises arboviruses with the capacity to infect humans and animals. It also integrates insect-specific viruses. This study aimed to identify Flavivirus in mosquitoes captured in 17 municipalities in Yucatan State, Mexico. The mosquitoes were caught in households from November 2021 to May 2022. A total of 4,321 adult mosquitoes from five species were caught. The most abundant were Culex quinquefasciatus (n = 3,563) and Aedes aegypti (n = 734). For molecular investigations, 600 female mosquitoes were split into groups of 10, mostly for species and site location. Reverse transcriptase polymerase chain reaction (RT-PCR) amplified a region of the NS5 gene to find the Flavivirus ribonucleic acids (RNA). A total of 24 pools that were positive for Flavivirus were detected in Ae. aegypti specimens and subsequently subjected to sequencing using the Sanger method. A total of 12 sequences matched the established quality criteria and were subsequently employed for sequence homology analysis. We found that one sequence corresponded to the Zika virus (ZIKV), and 11 sequences had sequence similarity with Phlebotomus-associated flavivirus (PAFV), an insect-specific virus (ISF). In conclusion, we found ZIKV in the Merida municipality, Yucatan State, which suggests that the virus is silently circulating. Phlebotomus-associated flavivirus is distributed in five municipalities in Yucatan State, Mexico. Future studies could focus on isolating this virus and studying its biological role within Ae. aegypti.

Neural Basis of Number Sense in Larval Zebrafish.

Number sense, the ability to discriminate the quantity of objects, is crucial for survival. To understand how neurons work together and develop to mediate number sense, we used two-photon fluorescence light sheet microscopy to capture the activity of individual neurons throughout the brain of larval Danio rerio, while displaying a visual number stimulus to the animal. We identified number-selective neurons as early as 3 days post-fertilization and found a proportional increase of neurons tuned to larger quantities after 3 days. We used machine learning to predict the stimulus from the neuronal activity and observed that the prediction accuracy improves with age. We further tested ethanol's effect on number sense and found a decrease in number-selective neurons in the forebrain, suggesting cognitive impairment. These findings are a significant step towards understanding neural circuits devoted to discrete magnitudes and our methodology to track single-neuron activity across the whole brain is broadly applicable to other fields in neuroscience.

Chemoinformatic Characterization of NAPROC-13: A Database for Natural Product 13C NMR Dereplication.

Natural products (NPs) are secondary metabolites of natural origin with broad applications across various human activities, particularly the discovery of bioactive compounds. Structural elucidation of new NPs entails significant cost and effort. On the other hand, the dereplication of known compounds is crucial for the early exclusion of irrelevant compounds in contemporary pharmaceutical research. NAPROC-13 stands out as a publicly accessible database, providing structural and 13C NMR spectroscopic information for over 25 000 compounds, rendering it a pivotal resource in natural product (NP) research, favoring open science. This study seeks to quantitatively analyze the chemical content, structural diversity, and chemical space coverage of NPs within NAPROC-13, compared to FDA-approved drugs and a very diverse subset of NPs, UNPD-A. Findings indicated that NPs in NAPROC-13 exhibit properties comparable to those in UNPD-A, albeit showcasing a notably diverse array of structural content, scaffolds, ring systems of pharmaceutical interest, and molecular fragments. NAPROC-13 covers a specific region of the chemical multiverse (a generalization of the chemical space from different chemical representations) regarding physicochemical properties and a region as broad as UNPD-A in terms of the structural features represented by fingerprints.

Symptoms of attention deficit hyperactivity disorder and oral health problems among children in Spain.

INTRODUCTION: The aim of this study was to explore the association between symptoms of attention deficit hyperactivity disorder (ADHD) and oral health in a representative sample of the Spanish population aged 6 to 14 years, we examined the contribution of several sociodemographic and behavioral determinants of children/adolescents and their family environment. METHODS: A cross-sectional study involving 3,402 subjects aged between 6 and 14 years from the Spanish National Health Survey. The presence of ADHD symptoms was determined using the self-reported hyperactivity/inattention subscale of the Strength and Difficulties Questionnaire. Oral health was assessed using five indicators: caries, extraction, restorations, gingival bleeding, and presence of teeth in poor condition (e.g. broken or misaligned). Logistic regressions were used to examine the association between the presence of ADHD symptoms and oral health indicators. These models were adjusted for sociodemographic factors, lifestyle variables, and oral hygiene behaviors relating to both the children and their parents. RESULTS: Compared to children without ADHD symptoms, children with ADHD symptoms had a higher risk of caries (fully adjusted odds ratio: 2.16; 95% confidence interval: 1.66-2.82), extraction (1.42; 1.09-1.85), restoration (1.47; 1.19-1.82) and gingival bleeding (1.64; 1.11-2.41). The increased risk of caries was maintained when the analyses were restricted to middle/high social class families and to children with low sugar intake, good oral hygiene behaviors and regular dental visits. CONCLUSIONS: Children with ADHD symptoms in Spain had worse oral health indicators than those without ADHD symptoms. Our results suggest that the association of ADHD symptoms with caries was independent of socioeconomic level, cariogenic diet, frequency of toothbrushing and dental visits.

Connecting high-resolution 3D chromatin maps with cell division and cell differentiation at the root apical meristem.

KEY MESSAGE: We used marker-free technologies to study chromatin at cellular resolution. Our results show asymmetric chromatin distribution, explore chromatin dynamics during mitosis, and reveal structural differences between trichoblast and atrichoblast cell. The shapes, sizes, and structural organizations of plant nuclei vary considerably among cell types, tissues, and species. This diversity is dependent on various factors, including cellular function, developmental stage, and environmental or physiological conditions. The differences in nuclear structure reflect the state of chromatin, which, in turn, controls gene expression and regulates cell fate. To examine the interrelationship between nuclear structure, cell morphology, and tissue-specific cell proliferation and differentiation processes, we conducted multiple visualizations of H3K4me1, H3K9me2, 4',6-diamidino-2-phenylindole, 5-ethynyl 2'-deoxyuridine, and SCRI Renaissance 2200, followed by subsequent quantitative analysis of individual cells and nuclei. By assigning cylindrical coordinates to the nuclei in the iRoCS toolbox, we were able to construct in situ digital three-dimensional chromatin maps for all the tissue layers of individual roots. A detailed analysis of the nuclei features of H3K4me1 and H3K9me2 in the mitotic and the elongation zones in trichoblast and atrichoblast cells at the root apical meristem revealed cell type-specific chromatin dynamics with asymmetric distribution of euchromatin and heterochromatin marks that may be associated with cell cycle and cell differentiation characteristics of specific cells. Furthermore, the spatial distribution of nuclei stained with 5-ethynyl 2'-deoxyuridine in the epidermis and cortex tissues suggests short-range coordination of cell division and nuclear migration in a linear sequence through an unknown regulatory mechanism.

[Continuous glucose monitoring devices in people over 65 years: literature review]. / Dispositivos de monitorización continua de glucosa en mayores de 65 años: revisión bibliográfica.

Background: The use of continuous glucose monitoring (CGM) in elderly patients with diabetes is not well documented. The present study search to know the current status of use of CGM devices in patients with diabetes over 65 years of age. Methods: A systematic literature search was performed in PubMed and Scopus databases in November 2023. The following descriptors were used, linked using the Boolean operators AND and OR: Diabetes, Continuous glucose monitoring, Flash glucose monitoring, Glycemia, Glycemic control, Controlled diabetes, Diabetes care, Metrics, Older and Elder. The CASPe critical reading system and the PRISMA methodology were applied to document the review. Results: Fourteen selected studies published between 2019 and 2023 were reviewed. Publications from European countries used small sample sizes and were intervention studies. The duration of the interventions was generally short: 7 were ≤14 days in duration and only 3 were 6-12 months in duration. Overall, the selected articles found that CGM improved glycemic outcomes due to a decrease in glycosylated hemoglobin value and an improvement in CGM metrics, and diabetes-associated complications in elderly patients with diabetes. Conclusion: CGM seems to be a useful and effective tool to improving glycemic outcomes of people with diabetes over age 65. Further studies under real-world conditions and of longer duration are needed to expand and improve the evidence.

Drought risk in Moldova under global warming and possible crop adaptation strategies.

This study analyzes the relationship between drought processes and crop yields in Moldova, together with the effects of possible future climate change on crops. The severity of drought is analyzed over time in Moldova using the Standard Precipitation Index, the Standardized Precipitation Evapotranspiration Index, and their relationship with crop yields. In addition, rainfall variability and its relationship with crop yields are examined using spectral analysis and squared wavelet coherence. Observed station data (1950-2020 and 1850-2020), ERA5 reanalysis data (1950-2020), and climate model simulations (period 1970-2100) are used. Crop yield data (maize, sunflower, grape), data from experimental plots (wheat), and the Enhanced Vegetation Index from Moderate Resolution Imaging Spectroradiometer satellites were also used. Results show that although the severity of meteorological droughts has decreased in the last 170 years, the impact of precipitation deficits on different crop yields has increased, concurrent with a sharp increase in temperature, which negatively affected crop yields. Annual crops are now more vulnerable to natural rainfall variability and, in years characterized by rainfall deficits, the possibility of reductions in crop yield increases due to sharp increases in temperature. Projections reveal a pessimistic outlook in the absence of adaptation, highlighting the urgency of developing new agricultural management strategies.

[Detection of people at risk of having diabetes in the community pharmacy with the Findrisc test 2014-2021]. / Detección de personas en riesgo de padecer diabetes en la farmacia comunitaria con el test de Findrisc en los años 2014-2021.

Aim: To analyze diabetes risk screening using the Findrisc questionnaire, performed in Spanish community pharmacies (CP) since 2014. Methods: Descriptive cumulative study of the results of campaigns from Global Diabetes Day, in 2014, 2016-2018, 2020 and 2021. Subjects: Users ≥18 years not diagnosed with diabetes with signed consent. Variables: Findrisc test score, mean (m) and standard deviation (SD) and subjects at risk intervals, n (%). Demographic (sex, age) and anthropometric variables, body mass index (BMI) (kg/m2), waist circumference (cm), capillary glycaemia (SD) (mg/dL). Results: A total of 1146 pharmacists took part; 12,402 users. A total of 8799 (70.9%) had BMI ≥25 kg/m2; 7366 (59.4%) were taking anti-hypertensives, 6047 (48.8%) with excessive abdominal circumference. In total, 5962 (48.0%) had a family history of diabetes.Average risk (Findrisc score) was 11.3 (4.6), without any sex differences (P>0.05). The number of subjects with high/very high risk (F≥15) was 3107 (25.0%) without any sex differences (P>0.05). The high/very high risk increased with age, from 282 (15.1%) people aged 45 to 54 up to 1695 (40.1%) people aged >64. A total of 1762 (14.2%) were referred to the doctor. There are no data on the result.Average interview time: 10.3 (5.3) minutes, no differences between sexes (P>0.05). Conclusions: One quarter of those surveyed had a high/very high risk and one in seven were referred to the doctor.The most prevalent risk factors were BMI, hypertension, abdominal circumference and family history of diabetes.Interprofessional communication should be improved as no result was obtained from referrals to the doctor.

Outpatient ATG-free hematopoietic transplantation for aplastic anemia in limited-resource environments offers excellent results: Data from a single LATAM center.

OBJECTIVE: To document the results of outpatient hematopoietic stem cell transplantation (HSCT) from the peripheral blood (PB) of sibling donors without anti-thymocyte globulin (ATG) in the conditioning regimen. MATERIAL AND METHODS: Patients from a low-income population with severe AA who received a PB, unmanipulated sibling HLA-identical HSCT between 2000 and 2020 at a single institution were studied. Survival was the primary outcome. RESULTS: Forty-one transplants were performed. Time between diagnosis and transplant was five months (1-104). Median age was 37 (range, 4-61) years; 25 (61 %) recipients were males and 32 (78 %) had treatment failure, 9 (22 %) have not received treatment. ATG was administered in 5 (12.2 %) cases; the graft source was PB in 38 (92.7 %) transplants. Twenty-six (63.4 %) transplants were carried out in the outpatient setting. Infections developed in 14 (34.1 %) patients. Primary graft failure (GF) occurred in 3 (7.3 %) patients. The 15-year OS was 81 %, EFS was 77.4 %. Patients with high pre-HSCT transfusion burden had lower OS (p = 0.035) and EFS (p = 0.026). Previous treatment failure and age were not associated with lower OS (p = 0.115, p = 0.069) or EFS (p = 0.088, p = 0.5, respectively). CONCLUSIONS: HLA-identical T-cell replete outpatient HSCT from the PB of sibling donors for AA patients using ATG-free conditioning offers excellent long-term survival.

Performance Enhancement of Biopolyester Blends by Reactive Compatibilization with Maleic Anhydride-Grafted Poly(butylene succinate-co-adipate).

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) is a very promising biodegradable copolyester of high interest in food packaging. Its inherent brittleness and narrow processing window make it necessary to blend it with flexible biopolyesters, such as poly(butylene succinate-co-adipate) (PBSA). However, the resultant biopolyester blends are thermodynamically immiscible, which impairs their performance and limits their applications. This study is the first to explore the use of poly(butylene succinate-co-adipate) grafted with maleic anhydride (PBS-g-MAH) as a novel reactive additive to compatibilize PHBV/PBSA blends. The compatibilizer was prepared by a reactive melt-mixing process of PBSA and maleic anhydride (MAH) using dicumyl peroxide (DCP) as an organic radical initiator, achieving a grafting degree (Gd) of 5.4%. Biopolyester blend films were thereafter prepared via cast extrusion and their morphological, thermal, mechanical, and barrier properties were characterized. Compatibilization by PBSA-g-MAH was confirmed by observing an improved phase interaction and lower dispersed domain sizes in the blends with 15 wt% PBSA. These compatibilized PHBV/PBSA blends were thermally stable up to 285 °C, showed enhanced ductility and toughness, as well as providing an improved barrier against water and limonene vapors and oxygen. These findings suggest that the use of MAH-grafted biopolyesters can represent an effective strategy to improve the properties of biopolyester blends and open up new opportunities for the application of PHBV-based formulations for food packaging.

Botulinum Toxin Type A (BoNT-A) Use for Post-Stroke Spasticity: A Multicenter Study Using Natural Language Processing and Machine Learning.

We conducted a multicenter and retrospective study to describe the use of botulinum toxin type A (BoNT-A) to treat post-stroke spasticity (PSS). Data were extracted from free-text in electronic health records (EHRs) in five Spanish hospitals. We included adults diagnosed with PSS between January 2015 and December 2019, stratified into BoNT-A-treated and untreated groups. We used EHRead® technology, which incorporates natural language processing and machine learning, as well as SNOMED CT terminology. We analyzed demographic data, stroke characteristics, BoNT-A use patterns, and other treatments. We reviewed the EHRs of 1,233,929 patients and identified 2190 people with PSS with a median age of 69 years; in total, 52.1% were men, 70.7% had cardiovascular risk factors, and 63.2% had suffered an ischemic stroke. Among the PSS patients, 25.5% received BoNT-A at least once. The median time from stroke to spasticity onset was 205 days, and the time from stroke to the first BoNT-A injection was 364 days. The primary goal of BoNT-A treatment was pain control. Among the study cohort, rehabilitation was the most common non-pharmacological treatment (95.5%). Only 3.3% had recorded monitoring scales. In conclusion, a quarter of patients with PSS received BoNT-A mainly for pain relief, typically one year after the stroke. Early treatment, disease monitoring, and better data documentation in EHRs are crucial to improve PSS patients' care.

Ascorbate, plant hormones and their interactions during plant responses to biotic stress.

Plants can experience a variety of environmental stresses that significantly impact their fitness and survival. Additionally, biotic stress can harm agriculture, leading to reduced crop yields and economic losses worldwide. As a result, plants have developed defense strategies to combat potential invaders. These strategies involve regulating redox homeostasis. Several studies have documented the positive role of plant antioxidants, including Ascorbate (Asc), under biotic stress conditions. Asc is a multifaceted antioxidant that scavenges ROS, acts as a co-factor for different enzymes, regulates gene expression, and facilitates iron transport. However, little attention has been given to Asc and its transport, regulatory effects, interplay with phytohormones, and involvement in defense processes under biotic stress. Asc interacts with other components of the redox system and phytohormones to activate various defense responses that reduce the growth of plant pathogens and promote plant growth and development under biotic stress conditions. Scientific reports indicate that Asc can significantly contribute to plant resistance against biotic stress through mutual interactions with components of the redox and hormonal systems. This review focuses on the role of Asc in enhancing plant resistance against pathogens. Further research is necessary to gain a more comprehensive understanding of the molecular and cellular regulatory processes involved.

Pd-imidate@SBA-15: a multifunctional heterogeneous catalyst for the aqueous room temperature hydrogenation of CO2 to formic acid.

The incorporation of hydrophilic and basic sites from phosphotriazaadamantane and saccharine in a water-soluble Pd complex and the subsequent confinement into mesoporous silica support increases the activity and stability of the palladium catalytic species for the room-temperature aqueous hydrogenation of either bicarbonate or CO2 into formic acid. The use of low Pd loadings (<0.1mmolPd·g-1) of the well-dispersed complex on SBA-15 mesoporous silica allows performing the reaction under room temperature, and aqueous conditions, exhibiting TON of ca. 40-100, for the CO2 or bicarbonate hydrogenation, respectively, which is one order of magnitude higher than the homogeneous case, allowing the easy isolation and recycling of the solid catalyst.

Influence of Education on Dental Anxiety and Fear in Mental Disorders after Viewing a Molar Extraction in Three Types of Mental Disorder Compared to Persons with No Mental Disorder.

Objective: The goal of this study is to validate the psychometric properties of the Modified Dental Anxiety Scale (MDAS) and the Dental Fear Scale (DFS) at three different times: seven days before, immediately after, and seven days after watching a video of surgical extraction of a lower third molar in a sample with four strata: anxiety disorder, adjustment disorder, mixed anxiety-depressive disorder, and no mental disorder ages 18-70 in a psychiatric clinic in Malaga. Methods: The Research Ethics Committee of the Virgen Macarena-Virgen del Rocío Hospitals approved the study. After being interviewed by a psychiatrist and subsequently completing the Hamilton Rating Scales for Anxiety and Depression, the participants were divided into 4 strata (60 persons in each). The influence of education level was then analyzed. Results: The scales demonstrated good psychometric properties, with higher MDAS and DFS scores for lower levels of education and mixed anxiety-depressive disorder. Conclusions: Patients who show higher levels of dental anxiety and dental fear will be those with lower education levels, as well as those who suffer from mixed anxiety-depressive disorder.

Coastal Dune Vegetation Dynamism and Anthropogenic-Induced Transitions in the Mexican Caribbean during the Last Decade.

In the Mexican Caribbean, environmental changes, hydrometeorological events, and anthropogenic activities promote dynamism in the coastal vegetation cover associated with the dune; however, their pace and magnitude remain uncertain. Using Landsat 7 imagery, spatial and temporal changes in coastal dune vegetation were estimated for the 2011-2020 period in the Sian Ka'an Biosphere Reserve. The SAVI index revealed cover changes at different magnitudes and paces at the biannual, seasonal, and monthly timeframes. Climatic seasons had a significant influence on vegetation cover, with increases in cover during northerlies (SAVI: p = 0.000), while the topographic profile of the dune was relevant for structure. Distance-based multiple regressions and redundancy analysis showed that temperature had a significant effect (p < 0.05) on SAVI patterns, whereas precipitation showed little influence (p > 0.05). The Mann-Kendall tendency test indicated high dynamism in vegetation loss and recovery with no defined patterns, mostly associated with anthropogenic disturbance. High-density vegetation such as mangroves, palm trees, and shrubs was the most drastically affected, although a reduction in bare soil was also recorded. This study demonstrated that hydrometeorological events and climate variability in the long term have little influence on vegetation dynamism. Lastly, it was observed that anthropogenic activities promoted vegetation loss and transitions; however, the latter were also linked to recoveries in areas with pristine environments, relevant for tourism.

Single-point and kinetics of peripheral residual disease by mass spectrometry to predict outcome in patients with high risk smoldering multiple myeloma included in the GEM-CESAR trial.

The value of quantitative immunoprecipitation mass spectrometry (QIP-MS) to identify the M-protein is being investigated in patients with monoclonal gammopathies but no data are yet available in high-risk smoldering myeloma (HRsMM). We have therefore investigated QIP-MS to monitor peripheral residual disease (PRD) in 62 HRsMM patients enrolled in the GEM-CESAR trial. After 24 cycles of maintenance, detecting the M-protein by MS or clonal plasma cells by NGF identified cases with a significantly shorter median PFS (mPFS; MS: not reached vs 1,4 years, p=0.001; NGF: not reached vs 2 years, p=0.0002) but reaching CR+sCR did not discriminate patients with different outcome. With NGF as a reference, the combined results of NGF and MS showed a high negative predictive value (NPV) of MS: 81% overall and 73% at treatment completion. When sequential results were considered, sustained negativity by MS or NGF was associated with a very favorable outcome with a mPFS not yet reached vs 1.66 years and 2.18 years in cases never attaining PRD or minimal residual disease (MRD) negativity, respectively. We can thus conclude that 1) the standard response categories of the IMWG do not seem to be useful for treatment monitoring in HRsMM patients, 2) MS could be used as a non-invasive, clinical valuable tool with the capacity of guiding timely bone marrow evaluations (based on its high NPV with NGF as a reference) and 3) similarly to NGF, sequential results of MS are able identify a subgroup of HRsMM patients with long-term disease control. This study was registered at www.clinicaltrials.gov (ClinicalTrials.gov identifier: NCT02415413).