Low-pass whole genome sequencing as a cost-effective alternative to chromosomal microarray analysis for low- and middle-income countries.

Low-pass whole genome sequencing (LP-WGS) has been applied as alternative method to detect copy number variants (CNVs) in the clinical setting. Compared with chromosomal microarray analysis (CMA), the sequencing-based approach provides a similar resolution of CNV detection at a lower cost. In this study, we assessed the efficiency and reliability of LP-WGS as a more affordable alternative to CMA. A total of 1363 patients with unexplained neurodevelopmental delay/intellectual disability, autism spectrum disorders, and/or multiple congenital anomalies were enrolled. Those patients were referred from 15 nonprofit organizations and university centers located in different states in Brazil. The analysis of LP-WGS at 1x coverage (>50kb) revealed a positive testing result in 22% of the cases (304/1363), in which 219 and 85 correspond to pathogenic/likely pathogenic (P/LP) CNVs and variants of uncertain significance (VUS), respectively. The 16% (219/1363) diagnostic yield observed in our cohort is comparable to the 15%-20% reported for CMA in the literature. The use of commercial software, as demonstrated in this study, simplifies the implementation of the test in clinical settings. Particularly for countries like Brazil, where the cost of CMA presents a substantial barrier to most of the population, LP-WGS emerges as a cost-effective alternative for investigating copy number changes in cytogenetics.

A retrospective clinical study to compare the ability of cone-beam computed tomographic images and periapical radiographs to reveal cracked teeth, split teeth, and teeth with vertical root fractures.

BACKGROUND: This retrospective clinical study aimed to compare the sensitivity of cone-beam computed tomographic (CBCT) images and periapical (PA) radiographs to reveal cracked teeth, split teeth, and teeth with vertical root fractures (VRFs). METHODS: The authors included 98 patients (98 teeth) diagnosed with a longitudinal tooth fracture (LTF) (cracked tooth, split tooth, VRF) through direct visualization after extraction and with comprehensive clinical and radiographic records. They collected demographic, clinical, and radiographic data. The authors evaluated PA radiographs and CBCT images to identify fractures, fracture lines, and the different patterns of bone loss associated with these teeth. They used the McNemar test to compare PA radiographs and CBCT scans when assessing bone loss. They used the Fisher test to determine statistical relationships between fracture types and demographic, clinical, and radiologic traits. They used an analysis of variance test to compare patient age with fracture types. RESULTS: CBCT images were significantly more effective (P < .05) in detecting bone loss patterns associated with LTFs than with PA radiographs, with 71% of cases detected via CBCT images compared with 42% via radiographs. Mean age was significantly greater (P < .05) in patients with teeth with VRFs than in patients with split teeth. A significant relationship was observed between the type of fracture and the following variables: root canal treatment (split, VRF, P = .002), deep probing depth (≥ 5 mm) (VRF, P = .026), and having more than 8 teeth extracted from the mouth (VRF, P = .032). Overall, there was a significant difference (P < .001) between the visualization of fracture lines (45% on PA radiographs, 65% on CBCT images). CONCLUSIONS: CBCT scans provided more information on LTFs than PA radiographs, particularly in the identification of periradicular bone changes. PRACTICAL IMPLICATIONS: CBCT imaging can assist in making the clinical diagnosis of LTFs through observation of bone loss patterns, providing more information than PA radiographs.

Effect of SARS-CoV-2 S protein on the proteolytic cleavage of the epithelial Na+ channel ENaC.

Severe cases of COVID-19 are characterized by development of acute respiratory distress syndrome (ARDS). Water accumulation in the lungs is thought to occur as consequence of an exaggerated inflammatory response. A possible mechanism could involve decreased activity of the epithelial Na+ channel, ENaC, expressed in type II pneumocytes. Reduced transepithelial Na+ reabsorption could contribute to lung edema due to reduced alveolar fluid clearance. This hypothesis is based on the observation of the presence of a novel furin cleavage site in the S protein of SARS-CoV-2 that is identical to the furin cleavage site present in the alpha subunit of ENaC. Proteolytic processing of αENaC by furin-like proteases is essential for channel activity. Thus, competition between S protein and αENaC for furin-mediated cleavage in SARS-CoV-2-infected cells may negatively affect channel activity. Here we present experimental evidence showing that coexpression of the S protein with ENaC in a cellular model reduces channel activity. In addition, we show that bidirectional competition for cleavage by furin-like proteases occurs between 〈ENaC and S protein. In transgenic mice sensitive to lethal SARS-CoV-2, however, a significant decrease in gamma ENaC expression was not observed by immunostaining of lungs infected as shown by SARS-CoV2 nucleoprotein staining.

Evaluation of microplastic pollution using bee colonies: An exploration of various sampling methodologies.

Recent research has highlighted the potential of honeybees and bee products as biological samplers for monitoring xenobiotic pollutants. However, the effectiveness of these biological samplers in tracking microplastics (MPs) has not yet been explored. This study evaluates several methods of sampling MPs, using honeybees, pollen, and a novel in-hive passive sampler named the APITrap. The collected samples were characterized using a stereomicroscopy to count and categorise MPs by morphology, colour, and type. To chemical identification, a micro-Fourier transform-infrared (FTIR) spectroscopy was employed to determine the polymer types. The study was conducted across four consecutive surveillance programmes, in five different apiaries in Denmark. Our findings indicated that APITrap demonstrated better reproducibility, with a lower variation in results of 39%, compared to 111% for honeybee samples and 97% for pollen samples. Furthermore, the use of APITrap has no negative impact on bees and can be easily applied in successive samplings. The average number of MPs detected in the four monitoring studies ranged from 39 to 67 in the APITrap, 6 to 9 in honeybee samples, and 6 to 11 in pollen samples. Fibres were the most frequently found, accounting for an average of 91% of the total MPs detected in the APITrap, and similar values for fragments (5%) and films (4%). The MPs were predominantly coloured black, blue, green and red. Spectroscopy analysis confirmed the presence of up to five different synthetic polymers. Polyethylene terephthalate (PET) was the most common in case of fibres and similarly to polypropylene (PP), polyethylene (PE), polyacrylonitrile (PAN) and polyamide (PA) in non fibrous MPs. This study, based on citizen science and supported by beekeepers, highlights the potential of MPs to accumulate in beehives. It also shows that the APITrap provides a highly reliable and comprehensive approach for sampling in large-scale monitoring studies.

GJB2 c.35del variant up-regulates GJA1 gene expression and affects differentiation of human stem cells.

Pathogenic DNA alterations in GJB2 are present in nearly half of non-syndromic hearing loss cases with autosomal recessive inheritance. The most frequent variant in GJB2 causing non-syndromic hearing loss is the frameshifting c.35del. GJB2 encodes Cx26, a protein of the connexin family that assembles hemichannels and gap junctions. The expression of paralogous proteins is believed to compensate for the loss of function of specific connexins. As Cx26 has been involved in cell differentiation in distinct tissues, we employed stem cells derived from human exfoliated deciduous teeth (SHEDs), homozygous for the c.35del variant, to assess GJB2 roles in stem cell differentiation and the relationship between its loss of function and the expression of paralogous genes. Primary SHED cultures from patients and control individuals were compared. SHEDs from patients had significantly less GJB2 mRNA and increased amount of GJA1 (Cx43), but not GJB6 (Cx30) or GJB3 (Cx31) mRNA. In addition, they presented higher induced differentiation to adipocytes and osteocytes but lower chondrocyte differentiation. Our results suggest that GJA1 increased expression may be involved in functional compensation for GJB2 loss of function in human stem cells, and it may explain changes in differentiation properties observed in SHEDs with and without the c.35del variant.

Diverse projected climate change scenarios affect the physiology of broccoli plants to different extents.

Climate change caused by global warming involves crucial plant growth factors such as atmospheric CO2 concentration, ambient temperature or water availability. These stressors usually co-occur, causing intricate alterations in plant physiology and development. This work focuses on how elevated atmospheric CO2 levels, together with the concomitant high temperature, would affect the physiology of a relevant crop, such as broccoli. Particular attention has been paid to those defence mechanisms that contribute to plant fitness under abiotic stress. Results show that both photosynthesis and leaf transpiration were reduced in plants grown under climate change environments compared to those grown under current climate conditions. Furthermore, an induction of carbohydrate catabolism pointed to a redistribution from primary to secondary metabolism. This result could be related to a reinforcement of cell walls, as well as to an increase in the pool of antioxidants in the leaves. Broccoli plants, a C3 crop, grown under an intermediate condition showed activation of those adaptive mechanisms, which would contribute to coping with abiotic stress, as confirmed by reduced levels of lipid peroxidation relative to current climate conditions. On the contrary, the most severe climate change scenario exceeded the adaptive capacity of broccoli plants, as shown by the inhibition of growth and reduced vigour of plants. In conclusion, only a moderate increase in atmospheric CO2 concentration and temperature would not have a negative impact on broccoli crop yields.

Association between fatigue and MDS-UPDRS in individuals with Parkinson's disease: cross-sectional study.

BACKGROUND: Fatigue is significant in the context of Parkinson's disease (PD), considering that one-third of patients classify it as the most restricting symptom in their daily life activities (DLAs). The objective was to verify the relationship (association) between fatigue and non-motor and motor symptoms of PD. METHODS: A cross-sectional study which included 100 individuals with PD. Initially, demographic and clinical data (modified Hoehn and Yahr scale-HY, anxiety, and depression) were collected. To assess the non-motor and motor symptoms of PD, the Movement Disorders Society-Unified Parkinson's Disease Rating Scale (MDS-UPDRS) was applied. Fatigue was evaluated using the Parkinson Fatigue Scale. RESULTS: A higher HY score, greater severity of non-motor aspects of DLAs and motor aspects of DLAs, more motor complications, and higher levels of anxiety as well as depression were observed in the "fatigue" group. Fatigue was associated with a lower daily equivalent levodopa dose (LEDD), a higher body mass index (BMI), anxiety, depression, and the presence of non-motor symptoms. CONCLUSION: Non-motor symptoms are more determining factors for fatigue than the motor condition itself, with an association between fatigue and higher BMI scores, increased anxiety and depression, lower LEDD, and greater severity of non-motor aspects of DLAs. Individuals in the "fatigue" group had higher HY scores, anxiety, and depression, worse non-motor and motor symptoms related to experiences of daily life, as well as motor complications.

Cuidados al final de la vida en unidades de cuidado intensivo neonatal de Iberoamérica: una mirada desde la perspectiva de enfermería / End-of-life care in neonatal intensive care units in Iberoamerica: A look from the nursing perspective

Introducción: Los cuidados al final de la vida (CFV) son una parte fundamental de la asistencia al recién nacido ingresado en una unidad de cuidados intensivos neonatal (UCIN). El objetivo de este estudio fue explorar cómo se implementan los CFV en UCIN de Iberoamérica. Métodos: Estudio transversal realizado en 2022 mediante encuesta vía correo electrónico y redes oficiales de SIBEN a profesionales de enfermería de UCIN de países de Iberoamérica, sobre aspectos relacionados con los CFV. Recolección y análisis de datos con REDCap y STATA 14.0. Resultados: Obtuvimos 400 respuestas de enfermeros/as de 11 países, el 73% procedentes de hospitales terciarios. El 86% de los respondedores eran responsables directos de brindar CFV, si bien un 48% dijeron no haber recibido formación. Solo 67 (17%) afirmaron que la UCIN en la que trabajan cuenta con un protocolo. Las acciones que se implementan durante los CFV fueron infrecuentes (≤50%) en todos los ítems explorados y muy poco frecuentes (<20%) en permitir el acceso libre de familiares, contar con privacidad, brindar asistencia psicológica, registrar el proceso en la historia clínica, asistir con los procesos burocráticos u otorgar un plan de seguimiento para el duelo. Conclusión: La mayoría de los profesionales de enfermería encuestados eran responsables directos de proveer CFV, pero no contaban con protocolos, no habían recibido capacitación y consideraron que los CFV podrían mejorarse significativamente. Las estrategias para los CFV en la región iberoamericana requieren ser optimizadas.(AU)

Introduction: End-of-life care (ELC) represents a quality milestone in neonatal intensive care units (NICU). The objective of this study was to explore how ELC are carried out in NICUs in Iberoamerica. Methods: Cross-sectional study, carried out in 2022 through the administration of an anonymous survey sent to neonatal nursing professionals. The survey included general data and work activity data; existence and contents of ELC protocols in the NICU and training received. The survey was distributed by email and published on official SIBEN social networks. REDCap and STATA 14.0 software were used for data collection and analysis. Results: We obtained 400 responses from nurses from 11 countries, 73% working in third level hospitals. A percentage of 86 of the respondents are directly responsible for providing ELC, although 48% of them said they had not received training on this subject. Only 67 (17%) state that the NICU in which they work has a protocol that establishes a strategy for performing the ELC. Finally, the actions that are implemented during the ELC are globally infrequent (≤50%) in all the items explored and very infrequent (<20%) in relation to allowing free access to family members, having privacy, providing psychological assistance, register the process in the medical record, assist with bureaucratic processes or grant a follow-up plan for grief. Conclusion: Most of the nursing professionals surveyed are directly responsible for this care, do not have protocols, have not received training, and consider that the ELC could be significantly improved. Strategies for ELCs in the Ibero-American region need to be optimized.(AU)

KS-WNK1 is required for the renal response to extreme changes in potassium intake.

Kidney-specific with-no-lysine kinase 1 (KS-WNK1) is an isoform of WNK1 kinase that is predominantly found in the distal convoluted tubule of the kidney. The precise physiological function of KS-WNK1 remains unclear. Some studies have suggested that it could play a role in regulating potassium renal excretion by modulating the activity of the Na+-Cl- cotransporter (NCC). However, changes in the potassium diet from normal to high failed to reveal a role for KS-WNK1, but under a normal-potassium diet, the expression of KS-WNK1 is negligible. It is only detectable when mice are exposed to a low-potassium diet. In this study, we investigated the role of KS-WNK1 in regulating potassium excretion under extreme changes in potassium intake. After following a zero-potassium diet (0KD) for 10 days, KS-WNK1-/- mice had lower plasma levels of K+ and Cl- while exhibiting higher urinary excretion of Na+, Cl-, and K+ compared with KS-WNK1+/+ mice. After 10 days of 0KD or normal-potassium diet (NKD), all mice were challenged with a high-potassium diet (HKD). Plasma K+ levels markedly increased after the HKD challenge only in mice previously fed with 0KD, regardless of genotype. KSWNK1+/+ mice adapt better to HKD challenge than KS-WNK1-/- mice after a potassium-retaining state. The difference in the phosphorylated NCC-to-NCC ratio between KS-WNK1+/+ and KS-WNK1-/- mice after 0KD and HKD indicates a role for KS-WNK1 in both NCC phosphorylation and dephosphorylation. These observations show that KS-WNK1 helps the distal convoluted tubule to respond to extreme changes in potassium intake, such as those occurring in wildlife.NEW & NOTEWORTHY The findings of this study demonstrate that kidney-specific with-no-lysine kinase 1 plays a role in regulating urinary electrolyte excretion during extreme changes in potassium intake, such as those occurring in wildlife. .

Monitoring SpO2: The Basics of Retinopathy of Prematurity (Back to Basics) and Targeting Oxygen Saturation.

Oxygen (O2) is a drug frequently used in newborn care. Adverse effects of hypoxia are well known but the damaging effects of excess oxygen administration and oxidative stress have only been studied in the last 2 decades. Many negative effects have been described, including retinopathy of prematurity . Noninvasive pulse oximetry (SpO2) is useful to detect hypoxemia but requires careful evaluation and understanding of the frequently changing relationship between O2 and hemoglobin to prevent hyperoxemia. Intention to treat SpO2 ranges should be individualized for every newborn receiving supplemental O2, according to gestational age, post-natal age, and clinical condition.

Evaluation of the sorption/desorption processes of pesticides in biodegradable mulch films used in agriculture.

Microplastics from mulch films can be a source of chemical contamination to agricultural soils. In this context, biodegradable films have been widely positioned as a greener choice. However, their sorption/desorption capabilities, in contrast to the conventional plastic types remain understudied. It is for this reason that objective evaluation of their interactions with residual agricultural contaminants becomes important. Our findings reveal that polyethylene (PE) mulch films retained lower amounts of pesticide residues and demonstrated a higher desorption/release [median desorption = 71.86 µg/L or about 50%], while polybutylene adipate terephthalate (PBAT) mulch films retained higher amounts of pesticide residues onto their surface and demonstrated a much lower desorption [median desorption = 24.27 µg/L or about 17%] after a spraying event. A higher ambient temperature had no significant effect on final desorption amounts in both PE [median = 65.27 µg/L at 20 °C and 74.23 µg/L at 40 °C] and PBAT [median = 24.26 µg/L at 20 °C and 24.78 µg/L at 40 °C] mulch films. However, it did favour a faster desorption pace in PE films. Desorption in PBAT and PE plastic types was correlated with the log Kow value [Spearman's correlation: 0.857 and 0.837 respectively, p < 0.05]. However, only a moderate correlation with pKa was observed in PBAT [Spearman's correlation: 0.478, p < 0.05], while none for PE plastic type. Sorption of pesticides onto biodegradable PBAT microplastics were best explained by Elovich [R2: 0.937-0.959] and pseudo-second order kinetics [R2: 0.942-0.987], suggesting the presence of chemisorption. Furthermore, Weber Morris plots suggested the presence of a multi-step process and Boyd plots indicated that film diffusion or chemical bond formation was the rate-limiting step governing this phenomenon.

A biomechanical analysis of turning during gait in individuals with different subtypes of Parkinson's disease.

BACKGROUND: Turning while walking is a complex component of locomotor capacity, which can be challenging in the daily lives of people with Parkinson's disease. The aim of the study was to compare biomechanical strategies during turning of gait in individuals with Parkinson's disease and its different clinical subtypes. METHODS: A cross-sectional study, comprising of 43 individuals with idiopathic Parkinson's disease, divided in subgroups: akineto-rigid, dominant tremor and mixed. Motor impairment was assessed using the Unified Parkinson's Disease Rating Scale. The gait biomechanical parameters (number of steps, step length, cadence, amplitude, velocity and radius of the turn) were analyzed during turning in a kinematics laboratory. In the statistical analysis, a comparison was made between subgroups, and correlations between biomechanical parameters. FINDINGS: There was no statistically significant difference between the subgroups. In the correlation analysis, notable correlations were found between the anticipatory step length and the following variables: number of steps (r = -0.418), step length while turning (r = 0.805), step length after turning (r = 0.644), average velocity (r = 0.830), average velocity while turning (r = 0.755), and maximum velocity (rho = 0.835). INTERPRETATION: The difficulties primarily occur during the anticipatory phase of the turn, which affects the entire task. The greater the length of the anticipatory step, the greater the length of the step taken to turn as well as the step taken after turning. And the greater the velocity, the greater the step length, and to fewer steps taken to perform the task.

End-of-life care in neonatal intensive care units in Iberoamerica: A look from the nursing perspective.

INTRODUCTION: End-of-life care (ELC) represents a quality milestone in neonatal intensive care units (NICU). The objective of this study was to explore how ELC are carried out in NICUs in Iberoamerica. METHODS: Cross-sectional study, through the administration of an anonymous survey sent to neonatal nursing professionals. The survey included general data and work activity data; existence and contents of ELC protocols in the NICU and training received. The survey was distributed by email and published on official SIBEN social networks. REDCap and STATA 14.0 software were used for data collection and analysis. RESULTS: We obtained 400 responses from nurses from 11 countries in the Ibero-American region. 86% of the respondents are directly responsible for providing ELC, although 48% of them said they had not received training on this subject. Only 67 (17%) state that the NICU in which they work has a protocol that establishes a strategy for performing the ELC. Finally, the actions that are implemented during the ELC are globally infrequent (≤50%) in all the items explored and very infrequent (<20%) in relation to allowing free access to family members, having privacy, providing psychological assistance, register the process in the medical record, assist with bureaucratic processes or grant a follow-up plan for grief. CONCLUSION: Most of the nursing professionals surveyed are directly responsible for this care, do not have protocols, have not received training, and consider that the ELC could be significantly improved. Strategies for ELCs in the Ibero-American region need to be optimized.

Arginine vasopressin regulates the renal Na+-Cl- and Na+-K+-Cl- cotransporters through with-no-lysine kinase 4 and inhibitor 1 phosphorylation.

Vasopressin regulates water homeostasis via the V2 receptor in the kidney at least in part through protein kinase A (PKA) activation. Vasopressin, through an unknown pathway, upregulates the activity and phosphorylation of Na+-Cl- cotransporter (NCC) and Na+-K+-2Cl- cotransporter 2 (NKCC2) by Ste20-related proline/alanine-rich kinase (SPAK) and oxidative stress-responsive kinase 1 (OSR1), which are regulated by the with-no-lysine kinase (WNK) family. Phosphorylation of WNK4 at PKA consensus motifs may be involved. Inhibitor 1 (I1), a protein phosphatase 1 (PP1) inhibitor, may also play a role. In human embryonic kidney (HEK)-293 cells, we assessed the phosphorylation of WNK4, SPAK, NCC, or NKCC2 in response to forskolin or desmopressin. WNK4 and cotransporter phosphorylation were studied in desmopressin-infused WNK4-/- mice and in tubule suspensions. In HEK-293 cells, only wild-type WNK4 but not WNK1, WNK3, or a WNK4 mutant lacking PKA phosphorylation motifs could upregulate SPAK or cotransporter phosphorylation in response to forskolin or desmopressin. I1 transfection maximized SPAK phosphorylation in response to forskolin in the presence of WNK4 but not of mutant WNK4 lacking PP1 regulation. We observed direct PP1 regulation of NKCC2 dephosphorylation but not of NCC or SPAK in the absence of WNK4. WNK4-/- mice with desmopressin treatment did not increase SPAK/OSR1, NCC, or NKCC2 phosphorylation. In stimulated tubule suspensions from WNK4-/- mice, upregulation of pNKCC2 was reduced, whereas upregulation of SPAK phosphorylation was absent. These findings suggest that WNK4 is a central node in which kinase and phosphatase signaling converge to connect cAMP signaling to the SPAK/OSR1-NCC/NKCC2 pathway.NEW & NOTEWORTHY With-no-lysine kinases regulate the phosphorylation and activity of the Na+-Cl- and Na+-K+-2Cl- cotransporters. This pathway is modulated by arginine vasopressin (AVP). However, the link between AVP and WNK signaling remains unknown. Here, we show that AVP activates WNK4 through increased phosphorylation at putative protein kinase A-regulated sites and decreases its dephosphorylation by protein phosphatase 1. This work increases our understanding of the signaling pathways mediating AVP actions in the kidney.

Dysregulation of the WNK4-SPAK/OSR1 pathway has a minor effect on baseline NKCC2 phosphorylation.

The with-no-lysine kinase 4 (WNK4)-sterile 20/SPS-1-related proline/alanine-rich kinase (SPAK)/oxidative stress-responsive kinase 1 (OSR1) pathway mediates activating phosphorylation of the furosemide-sensitive Na+-K+-2Cl- cotransporter (NKCC2) and the thiazide-sensitive NaCl cotransporter (NCC). The commonly used pT96/pT101-pNKCC2 antibody cross-reacts with pT53-NCC in mice on the C57BL/6 background due to a five amino acid deletion. We generated a new C57BL/6-specific pNKCC2 antibody (anti-pT96-NKCC2) and tested the hypothesis that the WNK4-SPAK/OSR1 pathway strongly regulates the phosphorylation of NCC but not NKCC2. In C57BL/6 mice, anti-pT96-NKCC2 detected pNKCC2 and did not cross-react with NCC. Abundances of pT96-NKCC2 and pT53-NCC were evaluated in Wnk4-/-, Osr1-/-, Spak-/-, and Osr1-/-/Spak-/- mice and in several models of the disease familial hyperkalemic hypertension (FHHt) in which the CUL3-KLHL3 ubiquitin ligase complex that promotes WNK4 degradation is dysregulated (Cul3+/-/Δ9, Klhl3-/-, and Klhl3R528H/R528H). All mice were on the C57BL/6 background. In Wnk4-/- mice, pT53-NCC was almost absent but pT96-NKCC2 was only slightly lower. pT53-NCC was almost absent in Spak-/- and Osr1-/-/Spak-/- mice, but pT96-NKCC2 abundance did not differ from controls. pT96-NKCC2/total NKCC2 was slightly lower in Osr1-/- and Osr1-/-/Spak-/- mice. WNK4 expression colocalized not only with NCC but also with NKCC2 in Klhl3-/- mice, but pT96-NKCC2 abundance was unchanged. Consistent with this, furosemide-induced urinary Na+ excretion following thiazide treatment was similar between Klhl3-/- and controls. pT96-NKCC2 abundance was also unchanged in the other FHHt mouse models. Our data show that disruption of the WNK4-SPAK/OSR1 pathway only mildly affects NKCC2 phosphorylation, suggesting a role for other kinases in NKCC2 activation. In FHHt models NKCC2 phosphorylation is unchanged despite higher WNK4 abundance, explaining the thiazide sensitivity of FHHt.NEW & NOTEWORTHY The renal cation cotransporters NCC and NKCC2 are activated following phosphorylation mediated by the WNK4-SPAK/OSR1 pathway. While disruption of this pathway strongly affects NCC activity, effects on NKCC2 activity are unclear since the commonly used phospho-NKCC2 antibody was recently reported to cross-react with phospho-NCC in mice on the C57BL/6 background. Using a new phospho-NKCC2 antibody specific for C57BL/6, we show that inhibition or activation of the WNK4-SPAK/OSR1 pathway in mice only mildly affects NKCC2 phosphorylation.

Optimal population-specific HLA imputation with dimension reduction.

Human genomics has quickly evolved, powering genome-wide association studies (GWASs). SNP-based GWASs cannot capture the intense polymorphism of HLA genes, highly associated with disease susceptibility. There are methods to statistically impute HLA genotypes from SNP-genotypes data, but lack of diversity in reference panels hinders their performance. We evaluated the accuracy of the 1000 Genomes data as a reference panel for imputing HLA from admixed individuals of African and European ancestries, focusing on (a) the full dataset, (b) 10 replications from 6 populations, and (c) 19 conditions for the custom reference panels. The full dataset outperformed smaller models, with a good F1-score of 0.66 for HLA-B. However, custom models outperformed the multiethnic or population models of similar size (F1-scores up to 0.53, against up to 0.42). We demonstrated the importance of using genetically specific models for imputing populations, which are currently underrepresented in public datasets, opening the door to HLA imputation for every genetic population.

Distribution of PD-L1, TROP2 and HER2- "lowness" in early triple-negative breast cancer: an opportunity for treatment de-escalation.

BACKGROUND: HER2, TROP2 and PD-L1 are novel targets in triple-negative breast cancer (TNBC). The combined expression status of these targets, and whether they can define prognostic subgroups, is currently undefined. METHODS: Immunohistochemistry was used to determine HER2, TROP2 and PD-L1 levels in 459 TNBC cases, that received in the adjuvant/neoadjuvant setting active surveillance, CMF, anthracycline-, anthracycline plus taxane-, or carboplatin-containing regimes. RESULTS: HER2-low patients with PD-L1 > 1 CPS (double-positive, herein "DP") had a mean PFS of 4768 days (95% CI: 4267-5268) versus 3522 days (95% CI: 3184-3861) for non-DP patients (P = 0.002). Regarding the received adjuvant treatment, DP patients (versus non-DP) receiving anthracyclines plus taxanes exhibited a mean PFS time of 4726 (95% CI: 4022-5430) versus 3302 (95% CI: 2818-3785) days (P = 0.039). Finally, 100% of DP patients that received a carboplatin-based regimen were long-term disease-free. CONCLUSIONS: Early HER2-low, PD-L1-positive TNBC patients have a very good prognosis, particularly if treated with anthracycline/taxane- or carboplatin-containing regimes.

Quantification of nanoplastic uptake and distribution in the root, stem and leaves of the edible herb Lepidum sativum.

This study confirms the uptake, translocation and bioaccumulation of 100 nm polystyrene nanoplastics in the root, stem and leaves of the plant Lepidum sativum at exposure concentrations ranging from environmentally realistic 10 µg/L up to a high of 100 mg/L. Accumulation in plant tissues was characterised by aggregation in the intercellular spaces and heterogeneous distribution. Nanoplastic presence was confirmed in the root tips, root surface and stele, lateral roots, root hairs, stem vascular bundles, leaf veins and mesophyll, as well as leaf epidermis including stomatal sites. Quantification results show that majority of the particles were retained in the root and accumulation in stem and leaves was only 13 to 18 % of the median value in roots. There was a reduction of 38.89 ± 9.62 % in the germination rate, 55 % in plant fresh weight, as well as in root weight (> 80 %), root length (> 60 %), shoot weight (51 to 78 %) and number of lateral roots (> 28 %) at exposure concentrations at and above 50 mg/L. However, lower, environmentally probable exposure concentrations did not affect the plant health significantly. Our results highlight the urgent need for further exploration of this issue from the point of view of food safety and security. STATEMENT OF ENVIRONMENTAL IMPLICATION: Micro and nanoplastics have been reported in agricultural environments across the globe and reports regarding their hazardous effects over agricultural and plant health call for an urgent exploration of this issue. This work demonstrates the uptake, bioaccumulation and distribution of nanoplastics in an edible plant at an environmentally realistic concentration and raises serious concerns regarding the possible implications for food safety and security. It presents a novel approach which addresses the quantification of nanoplastic accumulation in plant tissues and helps identify the mechanism and trends behind this phenomenon which has been a challenge up until now.

An evolutionary perspective on complex neuropsychiatric disease.

The forces of evolution-mutation, selection, migration, and genetic drift-shape the genetic architecture of human traits, including the genetic architecture of complex neuropsychiatric illnesses. Studying these illnesses in populations that are diverse in genetic ancestry, historical demography, and cultural history can reveal how evolutionary forces have guided adaptation over time and place. A fundamental truth of shared human biology is that an allele responsible for a disease in anyone, anywhere, reveals a gene critical to the normal biology underlying that condition in everyone, everywhere. Understanding the genetic causes of neuropsychiatric disease in the widest possible range of human populations thus yields the greatest possible range of insight into genes critical to human brain development. In this perspective, we explore some of the relationships between genes, adaptation, and history that can be illuminated by an evolutionary perspective on studies of complex neuropsychiatric disease in diverse populations.

Resección hepática como tratamiento de lesión de vía biliar compleja / Liver resection for the treatment of complex bile duct injury

RESUMEN La lesión de la vía biliar constituye un problema de salud importante, ya que su incidencia se ha duplicado desde el advenimiento de la colecistectomía laparoscópica. En casos de lesiones complejas con afectación de la confluencia, asociación a lesiones vasculares, atrofia hepática, colangitis o intentos fallidos de reparación, el tratamiento suele ser un desafío. Presentamos el caso de una paciente con lesión compleja por compromiso vascular del pedículo hepático derecho que desarrolló una atrofia del parénquima hepático. Ante la presencia de complicaciones sépticas debió realizarse una hepatectomía asociada a la reparación definitiva de la vía biliar, con buenos resultados.

ABSTRACT Bile duct injury represents a serious health problem, with its incidence doubling since the advent of laparoscopic cholecystectomy. In cases of complex lesions affecting the hepatic confluence, association with vascular injuries, hepatic atrophy, cholangitis or failed repair, treatment is often challenging. We report the case of a female patient with a complex bile duct injury due to vascular involvement of the right hepatic pedicle who developed right liver atrophy. In the presence of septic complications, hepatectomy was necessary, along with a definitive repair of the biliary tract, resulting in positive outcomes.