Optimizing dsRNA engineering strategies and production in E. coli HT115 (DE3).

Producing double-stranded RNA (dsRNA) represents a bottleneck for the adoption of RNA interference technology in agriculture, and the main hurdles are related to increases in dsRNA yield, production efficiency, and purity. Therefore, this study aimed to optimize dsRNA production in E. coli HT115 (DE3) using an in vivo system. To this end, we designed a new vector, pCloneVR_2, which resulted in the efficient production of dsRNA in E. coli HT115 (DE3). We performed optimizations in the culture medium and expression inducer in the fermentation of E. coli HT115 (DE3) for the production of dsRNA. Notably, the variable that had the greatest effect on dsRNA yield was cultivation in TB medium, which resulted in a 118% increase in yield. Furthermore, lactose induction (6 g/L) yielded 10 times more than IPTG. Additionally, our optimized up-scaled protocol of the TRIzol™ extraction method was efficient for obtaining high-quality and pure dsRNA. Finally, our optimized protocol achieved an average yield of 53.3 µg/mL after the production and purification of different dsRNAs, reducing production costs by 72%.

Malnutrition among Children under Age Five in Panama: Results of the ENSPA 2019.

Background: Malnutrition has important short- and long-term consequences in children under age five. Malnutrition encompasses undernutrition, overnutrition, and the coexistence of both of them, known as the double burden of malnutrition (DBM). Objective: The aim of this study was to estimate the prevalence of undernutrition, overnutrition, and the DBM among these children at the national level and by living area in Panama. Methods: Data from the National Health Survey of Panama (ENSPA, Spanish acronym), a population-based, cross-sectional study carried out in 2019 were used. Stunting, wasting, overweight, and obesity were defined according to the cut-off points of the World Health Organization Growth Standards. Undernutrition was defined as being stunted only, wasted only or both; overnutrition was defined as being overweight only or obese only; and the DBM was defined as the co-occurence of stunting and overweight/obesity in the same child. Prevalence and general characteristics at the national level and by living area were weighted. Findings: The prevalence of undernutrition was 15.3% (95% confidence interval (CI) 13.4-17.3) at the national level and 36.6% (CI: 30.1-43.5) in indigenous areas. The prevalence of overnutrition was 10.2% (8.2-12.6) at the national level and 11.9% (CI: 8.5-16.3), 8.4% (CI: 6.5-10.7) and 8.7% (CI: 5.2-14.3) in urban, rural and indigenous areas, respectively. The DBM prevalence was 1.4% (CI: 1.0-2.1) at the national level and 2.7% (CI: 1.4-5.1) in indigenous areas. Conclusions: Undernutrition is still the most prevalent malnutrition condition in our country. Panama has the highest prevalence of overnutrition in Central America. The highest prevalence of undernutrition and DBM was found among children living in indigenous areas.

[Double diabetes mellitus. Double challenge for the exercise prescription. Systematic review]. / Diabetes mellitus doble. Doble desafío para la prescripción de ejercicio. Revisión sistemática.

Double diabetes (DD) refers to patients with type 1 diabetes who have developed insulin resistance. The objective of this review is to update relevant information on the prescription of physical activity, pharmacological adjustments and consumption of carbohydrates in DD. A systematic search for scientific articles was carried out in the following databases: PubMed, Cochrane, EBSCO, WoS, ScienceDirect and Medline. The evidence analyzed shows that both physical activity (PA) and physical exercise (PE) are essential to achieve metabolic control in people with DD. Physiological considerations such as: insulin adjustments, insulin injection sites, time to perform PA and PE, absolute and relative contraindications are essential to avoid complications, especially hypoglycemia.

Development of a dispersive solid phase microextraction method based on the application of MgAl, NiAl, and CoAl-layered double hydroxides for the efficient removal of α- and ß-naphthol isomers from water samples by capillary electrophoresis.

The present work describes a quick, simple, and efficient method based on the use of layered double hydroxides (LDH) coupled to dispersive solid phase micro-extraction (DSPME) to remove α-naphthol (α-NAP) and ß-naphthol (ß-NAP) isomers from water samples. Three different LDHs (MgAl-LDH, NiAl-LDH, and CoAl-LDH) were used to study how the interlayer anion and molar ratio affected the removal performance. The critical factors in the DSPME procedure (pH, LDH amount, contact time) were optimized by the univariate method under the optimal conditions: pH, 4-8; LDH amount, 5 mg; and contact time, 2.5 min. The method can be successfully applied in real sample waters, removing NAP isomers even in ultra-trace concentrations. The large volume sample stacking (LVSS-CE) technique provides limits of detections (LODs) of 5.52 µg/L and 6.36 µg/L for α-naphthol and ß-naphthol, respectively. The methodology's precision was evaluated on intra- and inter-day repeatability, with %RSD less than 10% in all cases. The MgAl/Cl--LDH selectivity was tested in the presence of phenol and bisphenol A, with a removal rate of >92.80%. The elution tests suggest that the LDH MgAl/Cl--LDH could be suitable for pre-concentration of α-naphthol and ß-naphthol in future works.

LDH-Indomethacin Nanoparticles Antitumoral Action: A Possible Coadjuvant Drug for Cancer Therapy.

Indomethacin (INDO) has a mechanism of action based on inhibiting fatty acids cyclooxygenase activity within the inflammation process. The action mechanism could be correlated with possible anticancer activity, but its high toxicity in normal tissues has made therapy difficult. By the coprecipitation method, the drug carried in a layered double hydroxides (LDH) hybrid matrix would reduce its undesired effects by promoting chemotherapeutic redirection. Therefore, different samples containing INDO intercalated in LDH were synthesized at temperatures of 50, 70, and 90 °C and synthesis times of 8, 16, 24, and 48 h, seeking the best structural organization. X-ray diffraction (XRD), vibrational Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), spectrophotometric analysis in UV-VIS, and differential thermogravimetric analysis (TGA/DTA) were used for characterization. Our results indicate that higher temperatures and longer synthesis time through coprecipitation reduce the possibility of INDO intercalation. However, it was possible to establish a time of 16 h and a temperature of 50 °C as the best conditions for intercalation. In vitro results confirmed the cell viability potential and anticancer activity in the LDH-INDO sample (16 h and 50 °C) for gastric cancer (AGP01, ACP02, and ACP03), breast cancer (MDA-MB-231 and MCF-7), melanoma (SK-MEL-19), lung fibroblast (MRC-5), and non-neoplastic gastric tissue (MN01) by MTT assay. Cell proliferation was inhibited, demonstrating higher and lower toxicity against MDA-MB-231 and SK-MEL-19. Thus, a clinical redirection of INDO is suggested as an integral and adjunctive anticancer medication in chemotherapy treatment.

Inequality of opportunity in the double burden of malnutrition in Mexico.

This paper proposes a pseudo-birth-cohort approach to deal with a lack of longitudinal data to measure health inequities over time. Using Roemer's framework for inequality of opportunity, this study measures ex-ante and ex-post inequalities in malnutrition, a concept that spans both sides of the nutrition continuum. The total contribution of observed circumstances and the direct contribution of observed efforts to the variation of malnutrition are disentangled for people born between 1983 and 1988 in Mexico. Results indicate that inequality of opportunity has been persistent across this 30-year lifespan for that cohort. Some evidence suggests that a lack of opportunities has been transmitted from parents to children and that people's circumstances account for most of the explained variation in the double burden of malnutrition. However, stratifying the analysis by sex shows that efforts account for more of the explained variation of inequality of opportunity for women in their middle adulthood than for men in most of the outcomes analyzed.

Toxicological effects of solvent-extracted organic matter associated with PM2.5 on human bronchial epithelial cell line NL-20.

The heterogeneity and complexity of solvent-extracted organic matter associated with PM2.5 (SEOM-PM2.5) is well known; however, there is scarce information on its biological effects in human cells. This work aimed to evaluate the effect of SEOM-PM2.5 collected in northern Mexico City during the cold-dry season (November 2017) on NL-20 cells, a human bronchial epithelial cell line. The SEOM obtained accounted for 15.5% of the PM2.5 mass and contained 21 polycyclic aromatic hydrocarbons (PAHs). The cell viability decreased following exposure to SEOM-PM2.5, and there were noticeable morphological changes such as increased cell size and the presence of cytoplasmic vesicles in cells treated with 5-40 µg/mL SEOM-PM2.5. Exposure to 5 µg/mL SEOM-PM2.5 led to several alterations compared with the control cells, including the induction of double-stranded DNA breaks based (p < 0.001); nuclear fragmentation and an increased mitotic index (p < 0.05); 53BP1 staining, a marker of DNA repair by non-homologous end-joining (p < 0.001); increased BiP protein expression; and reduced ATF6, IRE1α, and PERK gene expression. Conversely, when exposed to 40 µg/mL SEOM-PM2.5, the cells showed an increase in reactive oxygen species formation (p < 0.001), BiP protein expression (p < 0.05), and PERK gene expression (p < 0.05), indicating endoplasmic reticulum stress. Our data suggest concentration-dependent toxicological effects of SEOM-PM2.5 on NL-20 cells, including genotoxicity, genomic instability, and endoplasmic reticulum stress.

Morphological Characteristics of the Double Mental Foramen and Its Relevance in Clinical Practice: An Observational Study.

The mental foramen (MF) is an opening found bilaterally on the anterolateral aspect of the mandible; it can be round or oval and have different diameters. One of the anatomical variants of the jaw is the presence of an accessory mental foramen (AMF). These are usually smaller than the MF and can be located above, below, or to the sides of the main MF. The objective of this study was to recognize the presence of AMF in dry jaws of the Chilean population and collect information about its clinical relevance reported in the literature. In this descriptive observational study, we have collected dried jaws obtained from three higher education institutions in Santiago de Chile, from the Department of Morphology of the Andrés Bello University, the Normal Human Anatomy Unit of the University of Santiago, and the Human Anatomy pavilion from the Faculty of Medicine of the Finis Terrae University. The samples for this research were obtained by convenience, and the observation of the jaws was carried out in the human anatomy laboratories of each institution by three evaluators independently, and a fourth evaluator was included to validate that each evaluation was correct. The sample for this research came from 260 dry jaws, showing the following findings from the total jaws studied, and to classify as an accessory MF, it will be examined and measured so that it complies with what is declared in the literature as the presence of AMF, which is between 0.74 mm. and 0.89 mm. There were 17 studies included with a sample that fluctuated between 1 and 4000, with a cumulative total of 7946 and an average number of jaws analyzed from the studies of 467.4, showing statistically significant differences between the means with the sample analyzed in this study; p = 0.095. For the cumulative prevalence of the presence of AMF, this was 3.07 in this study, and in the compared studies, the average of AMF was 8.01%, which did not present a statistically significant difference; p = 0.158. Regarding the presence of variants of unilateral AMF, this occurred in five jaws, which is equivalent to 1.84% in the sample of this study, while in previous studies, it was 7.5%, being higher on the left side than on the right. The presence of AMF is a variant with high prevalence if we compare it with other variants of the jaw. Knowledge of the anatomy and position of the AMF is crucial to analyze different scenarios in the face of surgical procedures or conservative treatments of the lower anterior dental region.

Prevalence of the Double Burden of Malnutrition among Adolescents: Associations with Lifestyle Behaviors and Clusters of Social Determinants.

The double burden of malnutrition (DBM) is a condition in which malnutrition coexists with overweight, reflecting a new layer of malnutrition. Our objectives were to assess prevalence; test associations between DBM and 24-hour movement behaviors; and investigate whether DBM is associated with clusters of social determinants. Methods: This multicenter cross-sectional study included 1152 adolescents (12 to 17 years old) from four Brazilian cities. Body mass index (BMI, kg/m2) was used to estimate overweight, and the adopted cutoff points took into account the curves established for age and sex: Z-score > 1 and ≤2 (overweight) and Z-score > 2 (obesity). The serum concentration of 25-hydroxyvitamin D [25(OH)D] was stratified into three levels: vitamin D deficiency ≤ 20 ng/mL; vitamin D insufficiency = 21-29 ng/mL; optimal vitamin D ≥ 30 ng/mL. We used multilevel Poisson regression models to estimate prevalence ratios (PRs) and their respective 95% confidence intervals (95%CI) and to analyze the association between DBM and covariates. A significance level of p < 0.05 was considered. Cluster analyses were performed by applying a combination of hierarchical and non-hierarchical methods. Results: A population prevalence of DBM of 7.3% (95% CI: 5.9-8.9) was revealed. A percentage of 19.2% (95% CI: 17.0-21.6) of adolescents were overweight, and 8.3% (95% CI: 6.8-10.1) were obese. A total of 41.5% (95% CI: 38.7-44.4) had vitamin D deficiency, and 25.8% (95% CI: 23.4-28.4) had vitamin D insufficiency. However, 24-hour movement behaviors were not associated with DBM. Adolescents living in the southern region of the country, from public schools whose mothers have higher education, have a 1.94 [PR = 2.94 (95% CI: 1.20-7.23)] times greater chance of developing DBM. These results highlight the importance of specific factors to improve the nutritional health of adolescents, considering the specific social determinants identified in this study.

Development, synthesis, and application of magnetic layered double hydroxides (Fe3O4@SiO-LDH/DS-) as an efficient adsorbent for the removal of tetracyclines from milk samples.

This work presents the development, synthesis, and application of a layered double hydroxide (LDH) coupled to magnetic particles for the removal of antibiotics as tetracyclines (TC´s): tetracycline (TC), chlortetracycline (CT), oxytetracycline (OT), and doxycycline (DT) from milk samples. The LDH synthesis conditions, reaction time (30-90 min), molar ratios Mg2+/Al3+ (7:1-1:7), interlayer anion (NO3-, Cl-, CO32-, and dodecyl sulphate (DS-)) were evaluated. Under synthesis conditions (reaction time of 30 min, Mg2+/Al3+ molar ratio of 7:1, and DS- as interlayer anion), the LDH was coupled in a magnetic solid phase microextraction (MSPµE) methodology. At the optimal extraction conditions (pH 6, 5 min of contact time, 10 mg of adsorbent), a removal percentage of 99.0 % was obtained for each tetracycline. FTIR, TGA, SEM, and adsorption isotherms were employed to characterize the optimal adsorbent. Each experiment was corroborated by large-volume sample stacking capillary electrophoresis (LVSS-CE). The adsorbent was applied directly to positive milk samples (previously tested) for TC´s removal.

"Social learning, innovation, and sustainability: The search for directions beyond a systematic literature review".

The literature frames social learning as a critical concept when searching for sustainability and innovation. Its prominent position in various studies has raised questions about the role played by such a theory and the possibility of new research perspectives. Therefore, this paper analyzed future research directions in social learning, sustainability, and innovation through a systematic literature review. By using four guiding questions, this qualitative study conducted a systematic literature review in the Web of Science, Scopus, and Scientific Electronic Library Online (SciELO) databases using the search terms "social learning," "sustain*," and "innovation." to identify, evaluate, and interpret relevant available studies. For the analysis, hermeneutic units were created to construct relationships between papers of different perspectives and subjects, namely social learning for sustainability and innovation. The results demonstrated that instrumentalizing social learning theory has been essential to promoting sustainability and innovation. Regarding innovation, social learning creates opportunities for innovation because it motivates change and questions rules and norms. Studies on social learning for sustainability were the most representative although few studies have focused on what individuals become after an intervention based on social learning; they only applied the concept and demonstrated the processes of carrying out interventions, who participates, and how and why it happens. Additionally, the highlighted changes are not ontological, but epistemological, even though social learning and innovation seem to intertwine to change and promote sustainability. This study's contribution lies in the theoretical perspective since a kind of taxonomy was produced with the results from the databases, enabling advances for future research. The uniform treatment attributed to innovation was a limitation, refraining from differentiating its various nuances. Nevertheless, the originality of this article is the intersection between two theoretical perspectives - social learning and innovation - in the common and complex field of sustainability.

Comparison of ddRADseq and EUChip60K SNP genotyping systems for population genetics and genomic selection in Eucalyptus dunnii (Maiden).

Eucalyptus dunnii is one of the most important Eucalyptus species for short-fiber pulp production in regions where other species of the genus are affected by poor soil and climatic conditions. In this context, E. dunnii holds promise as a resource to address and adapt to the challenges of climate change. Despite its rapid growth and favorable wood properties for solid wood products, the advancement of its improvement remains in its early stages. In this work, we evaluated the performance of two single nucleotide polymorphism, (SNP), genotyping methods for population genetics analysis and Genomic Selection in E. dunnii. Double digest restriction-site associated DNA sequencing (ddRADseq) was compared with the EUChip60K array in 308 individuals from a provenance-progeny trial. The compared SNP set included 8,011 and 19,008 informative SNPs distributed along the 11 chromosomes, respectively. Although the two datasets differed in the percentage of missing data, genome coverage, minor allele frequency and estimated genetic diversity parameters, they revealed a similar genetic structure, showing two subpopulations with little differentiation between them, and low linkage disequilibrium. GS analyses were performed for eleven traits using Genomic Best Linear Unbiased Prediction (GBLUP) and a conventional pedigree-based model (ABLUP). Regardless of the SNP dataset, the predictive ability (PA) of GBLUP was better than that of ABLUP for six traits (Cellulose content, Total and Ethanolic extractives, Total and Klason lignin content and Syringyl and Guaiacyl lignin monomer ratio). When contrasting the SNP datasets used to estimate PAs, the GBLUP-EUChip60K model gave higher and significant PA values for six traits, meanwhile, the values estimated using ddRADseq gave higher values for three other traits. The PAs correlated positively with narrow sense heritabilities, with the highest correlations shown by the ABLUP and GBLUP-EUChip60K. The two genotyping methods, ddRADseq and EUChip60K, are generally comparable for population genetics and genomic prediction, demonstrating the utility of the former when subjected to rigorous SNP filtering. The results of this study provide a basis for future whole-genome studies using ddRADseq in non-model forest species for which SNP arrays have not yet been developed.

Mechanically Excellent, Notch-Insensitive, and Highly Conductive Double-Network Hydrogel for Flexible Strain Sensor.

A novel double-network conductive hydrogel based on lithium acetate/gelatin/polyacrylamide (PAAM) was synthesized by heating-cooling and subsequent γ-ray radiation-induced polymerization and cross-linking. Owing to the hydrogen bonding interaction between lithium acetate, physical cross-linked gelatin, and chemical cross-linked PAAM, the resultant hydrogel exhibited high tensile strength (1260 kPa), high ionic conductivity (35.2 mS cm-1), notch-insensitivity (tensile strength 415 kPa, elongation at break 872% with transverse notch), and extensive strain monitoring range (0.15-800%) under optimum conditions. The lithium acetate/gelatin/polyacrylamide hydrogel strain sensor attached to the skin can sensitively monitor the subtle movements of the human body. The strain sensor based on the resultant hydrogel with transverse notch can still work for 1200 cycles, due to that the covalent-cross-linked PAAm chain bridges the cracks and stabilizes the deformation, while the physical-cross-linked gelatin was unzipped to make the blunting of notch. The conductive hydrogel with high-sensitivity and high stability is expected to be used as materials for the preparation of flexible strain sensors in the future.

Full spectrum cannabidiol-rich extract reduced propofol dosage required for anesthetic induction in dogs-a pilot study.

Introduction: Cannabinoids show great therapeutic potential, but their effect on anesthesia still remains unclear. Use of chronic recreational Cannabis in humans undergoing anesthetic procedures tends to require a higher dose when compared to non-users. On the other hand, studies on rodents and dogs have shown that cannabinoid agonists may potentiate certain anesthetics. This contrast of effects possibly occurs due to different time lengths of administration of different phytocannabinoids at different doses, and their distinct effects on the Endocannabinoid System, which is also affected by anesthetics such as propofol and isoflurane. Methods: Twenty-seven healthy male dogs, client-owned, ranging from 1 to 7 years, and from 5 to 35 kg were selected, mean weight 15.03±7.39 kg, with owners volunteering their animals to participate in the research performed in the Federal University of Santa Catarina (UFSC). Dogs were randomized into 3 groups. The Control Group (CON, n = 9), receiving only Extra Virgin Olive Oil, the same oil-base used in the treatment groups. Group 2 (G2, n = 9) received 2 mg/kg of total phytocannabinoids, and Group 3 (G3, n = 9) received 6 mg/kg of total phytocannabinoids. All groups received their treatments transmucosally, 75 min before their induction with propofol. Heart and respiratory rate, blood pressure, temperature and sedation were evaluated prior to, and at 30, 60, and 75 min after administration of the fsCBD-rich extract or Placebo extract. Preanesthetic medication protocol was also included across all treatment groups, 15 min before induction. Parametric data was analyzed with one-way ANOVA, followed by Student-Newman-Keuls (SNK) if significant statistical differences were found. Non-parametric data was analyzed using Friedman's test, followed by Dunn test for comparisons between all timepoints in the same group. Kruskal-Wallis followed by Dunn was utilized for between groups comparisons. Propofol dose necessary for induction was analyzed through One-way ANOVA followed by Tukey's Multiple Comparisons Test, using Instat by Graphpad, and differences were considered statistically significant when p < 0.05. Our analysis assessed if statistical significance was present between time points in the same group, and between groups in the same time points. Results: In our study, 6 mg/kg of total phytocannabinoids were able to reduce the dose of propofol necessary for induction by 23% when compared to the control group. The fsCBD-rich extract did not produce significant sedation within or between groups, although statistically significant differences in heart rate and systolic blood pressure were found. Discussion: Our findings indicate that phytocannabinoids could be an adjunct option in anesthesia, although further research is necessary to better confirm this data. Additionally, further research is needed to determine the best dosage, delivery method, time for administration, ideal molecular profile for desired effects, safety, drug-drug interactions, and transurgical effects.

The global diet quality score as an indicator of adequate nutrient intake and dietary quality - a nation-wide representative study.

BACKGROUND: The Global Diet Quality Score (GDQS) was developed to be a simple, timely and cost-effective tool to track, simultaneously, nutritional deficiency and non-communicable disease risks from diet in diverse settings. The objective was to investigate the performance of GDQS as an indicator of adequate nutrient intake and dietary quality in a national-representative sample of the Brazilian population. METHODS: Nationally-representative data from 44,744 men and non-pregnant and non-lactating women aging ≥ 10 years, from the Brazilian National Dietary Survey were used. Dietary data were collected through two 24-h recalls (24HR). The GDQS was calculated and compared to a proxy indicator of nutrient adequate intake (the Minimum Dietary Diversity for Women-MDD-W) and to an indicator of high-risk diet for non-communicable diseases (caloric contribution from ultra-processed foods-UPF). To estimate the odds for overall nutrient inadequacy across MDD-W and GDQS quintiles, a multiple logistic regression was applied, and the two metrics' performances were compared using Wald's post-test. RESULTS: The mean GDQS for Brazilians was 14.5 (0-49 possible range), and only 1% of the population had a low-risk diet (GDQS ≥ 23). The GDQS mean was higher in women, elderly individuals and in higher-income households. An inverse correlation was found between the GDQS and UPF (rho (95% CI) = -0.20(-0.21;-0.19)). The odds for nutrient inadequacy were lower as quintiles of GDQS and MDD-W were higher (p-trend < 0.001), and MDD-W had a slightly better performance than GDQS (p-diff < 0.001). Having a low-risk GDQS (≥ 23) lowered the odds for nutrient inadequacy by 74% (95% CI:63%-81%). CONCLUSION: The GDQS is a good indicator of overall nutrient adequacy, and correlates well with UPF in a nationally representative sample of Brazil. Future studies must investigate the relationship between the GDQS and clinical endpoints, strengthening the recommendation to use this metric to surveillance dietary risks.

Double vitrification of embryos adversely affects clinical outcomes.

OBJECTIVE: To evaluate the impact of double embryo vitrification on clinical outcomes. METHODS: This retrospective cohort study included data from January 2013 to March 2021. The study group included women aged 33.3±5.7 years with double-vitrified embryos (n=381), while the control group included women aged 32.1±6.7 years with embryos vitrified once (n=780), all transferred at the blastocyst stage. The primary endpoint was live birth rate (LBR), and secondary endpoints included percent positive ßHCG test, clinical/ongoing pregnancy rates, miscarriage/biochemical pregnancy rates and birthweight. RESULTS: LBR was significantly lower in double-vitrified embryos (30.2%) than in embryos vitrified once (45.6%, p<.05). Similarly, double-vitrified embryos were associated with significantly lower positive ßHCG tests (46% vs. 63.3%, p<.05) and clinical (34.9% vs. 52.2%, p<.05) and ongoing pregnancy (31.3% vs. 47.3%, p<.05) rates compared to embryos vitrified once. However, biochemical pregnancy (double vitrified: 24.1% vs. vitrified once: 17.9%, p>.05) and miscarriage rates (double vitrified: 10.2% vs. vitrified once: 9.4%, p>.05), as well as mean birthweight (double-vitrified embryos: 2950g vs. embryos vitrified once: 2837g, p>.05) did not differ significantly between two groups. On a secondary comparison, amongst double-vitrified embryos, the subgroup that was cultured for more than 24 hours between warming and second vitrification achieved significantly higher positive ßHCG tests (49%) and clinical pregnancy (38%) rates, compared to embryos re-vitrified on the same day of warming (31.8% and 20.5%, respectively, p<.05). Nevertheless, LBR did not differ significantly amongst these study-group embryos (embryos that remained in culture for more than 24 hours: 32.2% vs. embryos that were re-vitrified on warming day: 20.5%, p>.05). CONCLUSIONS: Double vitrification of embryos adversely affects clinical outcomes. However, it represents a valuable option concerning embryo wastage, with acceptable success rates.

Clinical effect analysis of different treatment schemes for children with ulnar and radial double fractures.

Objective: The objective of the study is to evaluate the safety and efficacy of three different treatment methods for pediatric ulnar and radial double fractures. Methods: 120 children with ulnar and radial double fractures were included in the study. According to the different treatment plans, children were divided into three groups: manual reduction, splint external fixation, double elastic intramedullary fixation, and double plate fixation. Surgical indicators, radiological results, clinical efficacy, and complications were evaluated and compared among the groups. Results: The average hospital stay and operation time were significantly longer in the double plate internal fixation group compared to the other two groups. The double elastic intramedullary nailing group showed a higher fracture healing rate at 3 months compared to the other groups. There were no significant differences in clinical efficacy among the three groups. Complications were observed in all groups but did not show significant statistical differences. Conclusion: Double elastic intramedullary nailing fixation demonstrated favorable outcomes in terms of surgical indicators and fracture healing rates for pediatric ulnar and radial double fractures.

Objetivo: Evaluar la seguridad y eficacia de tres métodos de tratamiento diferentes para las fracturas dobles cubital y radial pediátricas. Métodos: Se incluyeron en el estudio 120 niños con fracturas dobles de cúbito y radio. Según los diferentes planes de tratamiento, los niños se dividieron en tres grupos: reducción manual, fijación externa con férula, fijación intramedular doble elástica y fijación con doble placa. Se evaluaron y compararon entre los grupos indicadores quirúrgicos, resultados radiológicos, eficacia clínica y complicaciones. Resultados: La estancia hospitalaria promedio y el tiempo de operación fueron significativamente más prolongados en el grupo de fijación interna con doble placa en comparación con los otros dos grupos. El grupo de clavo intramedular elástico doble mostró una mayor tasa de curación de la fractura a los 3 meses en comparación con los otros grupos. No hubo diferencias significativas en la eficacia clínica entre los tres grupos. Se observaron complicaciones en todos los grupos pero no mostraron diferencias estadísticas significativas. Conclusión: La fijación con clavo intramedular elástico doble demostró resultados favorables en términos de indicadores quirúrgicos y tasas de curación de fracturas pediátricas dobles cubital y radial.

Dynamics of the double burden of malnutrition in Guatemala: a secondary data analysis of the demographic and health surveys from 1998-2015.

OBJECTIVE: We estimated the prevalence and time trends of the double burden of malnutrition (DBM) in Guatemala and explored its occurrence based on socio-demographic factors. STUDY DESIGN: This was a secondary data analysis using information from four Demographic and Health Surveys covering the period 1998-2015. METHODS: The unit of analysis was the household within which information was gathered from women 18-49 years and their children, 6-59 months. The main outcome was the prevalence of any DBM in the household (co-existence of undernutrition and overnutrition in a woman, her children or both). We estimated the prevalence of any DBM by survey and analysed time trends. Stepwise logistic regression was used to explore the occurrence of DBM and socio-demographic factors. RESULTS: We analysed 39,749 households across all surveys. The prevalence of any DBM was 25.3% (95%CI: 22.1-28.7) in 1998-99, 23.8% (22.0-25.8) in 2002, 25.9% (24.3-27.5) in 2008-09 and 24.2% (22.9-25.5) in 2014-15, with no significant change over time (P = 0.782). Characteristics associated with lower odds of any DBM were rural residence, female-headed household, wealth and women's secondary education. Higher odds were seen for households with electricity, women >25y, indigenous and with >2 children. CONCLUSION: Our findings revealed that a quarter of Guatemala's households suffer from DBM, which has remained unchanged for 17 years. Interventions should prioritise urban areas, households of lower socio-economic status and those less educated. To increase awareness of policymakers of this pressing public health concern, further research on DBM could be strengthened by prospective study designs, integrating all household members and expanding the types of malnutrition.

Prevalent role of homologous recombination in the repair of specific double-strand breaks in Rhizobium etli.

Double-strand breaks (DSBs) are the most dangerous injuries for a genome. When unrepaired, death quickly ensues. In most bacterial systems, DSBs are repaired through homologous recombination. Nearly one-quarter of bacterial species harbor a second system, allowing direct ligation of broken ends, known as Non-Homologous End Joining (NHEJ). The relative role of both systems in DSBs repair in bacteria has been explored only in a few cases. To evaluate this in the bacterium Rhizobium etli, we used a modified version of the symbiotic plasmid (264 kb), containing a single copy of the nifH gene. In this plasmid, we inserted an integrative plasmid harboring a modified nifH gene fragment containing an I-SceI site. DSBs were easily inflicted in vivo by conjugating a small, replicative plasmid that expresses the I-SceI nuclease into the appropriate strains. Repair of a DSB may be achieved through homologous recombination (either between adjacent or distant repeats) or NHEJ. Characterization of the derivatives that repaired DSB in different configurations, revealed that in most cases (74%), homologous recombination was the prevalent mechanism responsible for repair, with a relatively minor contribution of NHEJ (23%). Inactivation of the I-SceI gene was detected in 3% of the cases. Sequence analysis of repaired derivatives showed the operation of NHEJ. To enhance the number of derivatives repaired through NHEJ, we repeated these experiments in a recA mutant background. Derivatives showing NHEJ were readily obtained when the DSB occurred on a small, artificial plasmid in a recA mutant. However, attempts to deliver a DSB on the symbiotic plasmid in a recA background failed, due to the accumulation of mutations that inactivated the I-SceI gene. This result, coupled with the absence of derivatives that lost the nonessential symbiotic plasmid, may be due to an unusual stability of the symbiotic plasmid, possibly caused by the presence of multiple toxin-antitoxin modules.

Outcomes of Aortomitral Continuity Reconstruction During Concomitant Aortic and Mitral Valve Replacement.

OBJECTIVES: To describe outcomes of reconstruction of the aortomitral continuity (AMC) during concomitant aortic and mitral valve replacement (ie, the "Commando" procedure). DESIGN: A retrospective study of consecutive cardiac surgeries from 2010 to 2022. SETTING: At a single institution. PARTICIPANTS: All patients undergoing double aortic and mitral valve replacement. INTERVENTIONS: Patients were dichotomized by the performance (or not) of AMC reconstruction. MEASUREMENTS AND MAIN RESULTS: A total of 331 patients underwent double-valve replacement, of whom 21 patients (6.3%) had a Commando procedure. The Commando group was more likely to have had a previous aortic valve replacement (AVR) or mitral valve replacement (MVR) (66.7% v 27.4%, p < 0.001), redo cardiac surgery (71.4% v 31.3%, p < 0.001), and emergent/salvage surgery (14.3% v 1.61%, p = 0.001), whereas surgery was more often performed for endocarditis in the Commando group (52.4% v 22.9%, p = 0.003). The Commando group had higher operative mortality (28.6% v 10.7%, p = 0.014), more prolonged ventilation (61.9% v 31.9%, p = 0.005), longer cardiopulmonary bypass time (312 ± 118 v 218 ± 85 minutes, p < 0.001), and longer ischemic time (252 ± 90 v 176 ± 66 minutes, p < 0.001). Despite increased short-term morbidity in the Commando group, Kaplan-Meier survival estimation showed no difference in long-term survival between each group (p = 0.386, log-rank). On multivariate Cox analysis, the Commando procedure was not associated with an increased hazard of death, compared to MVR + AVR (hazard ratio 1.29, 95% CI: 0.65-2.59, p = 0.496). CONCLUSIONS: Although short-term postoperative morbidity and mortality were found to be higher for patients undergoing the Commando procedure, AMC reconstruction may be equally durable in the long term.