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Maternal nutrition is critical in mammalian development, influencing the epigenetic reprogramming of gametes, embryos, and fetal programming. We evaluated the effects of different levels of sulfur (S) and cobalt (Co) in the maternal diet throughout the pre- and periconceptional periods on the biochemical and reproductive parameters of the donors and the DNA methylome of the progeny in Bos indicus cattle. The low-S/Co group differed from the control with respect to homocysteine, folic acid, B12, insulin growth factor 1, and glucose. The oocyte yield was lower in heifers from the low S/Co group than that in the control heifers. Embryos from the low-S/Co group exhibited 2320 differentially methylated regions (DMRs) across the genome compared with the control embryos. We also characterized candidate DMRs linked to the DNMT1 and DNMT3B genes in the blood and sperm cells of the adult progeny. A DMR located in DNMT1 that was identified in embryos remained differentially methylated in the sperm of the progeny from the low-S/Co group. Therefore, we associated changes in specific compounds in the maternal diet with DNA methylation modifications in the progeny. Our results help to elucidate the impact of maternal nutrition on epigenetic reprogramming in livestock, opening new avenues of research to study the effect of disturbed epigenetic patterns in early life on health and fertility in adulthood. Considering that cattle are physiologically similar to humans with respect to gestational length, our study may serve as a model for studies related to the developmental origin of health and disease in humans.
Evidence suggests that maternal prepregnancy body mass index (BMI) is associated with offspring cardiometabolic risk factors. This study was aimed at assessing the association of maternal prepregnancy BMI with offspring cardiometabolic risk factors in adolescence and adulthood. We also evaluated whether offspring BMI was a mediator in this association. The study included mother–offspring pairs from three Pelotas birth cohorts. Offspring cardiometabolic risk factors were collected in the last follow-up of each cohort [mean age (in years) 30.2, 22.6, 10.9]. Blood pressure was measured using an automatic device, cholesterol by using an enzymatic colorimetric method, and glucose from fingertip blood, using a portable glucose meter. In a pooled analysis of the cohorts, multiple linear regression was used to control for confounding. Mediation analysis was conducted using G-computation formula. In the adjusted model, mean systolic blood pressure of offspring from overweight and obese mothers was on average 1.25 (95% CI: 0.45; 2.05) and 2.13 (95% CI: 0.66; 3.59) mmHg higher than that of offspring from normal-weight mothers; for diastolic blood pressure, the means were 0.80 (95% CI: 0.26; 1.34) and 2.60 (95% CI: 1.62; 3.59) mmHg higher, respectively. Non-HDL cholesterol was positively associated with maternal BMI, whereas blood glucose was not associated. Mediation analyses showed that offspring BMI explained completely the association of maternal prepregnancy BMI with offspring systolic and diastolic blood pressure, and non-HDL cholesterol. Our findings suggest that maternal prepregnancy BMI is positively associated with offspring blood pressure, and blood lipids, and this association is explained by offspring BMI.
The role of fire in the management of degraded areas remains strongly debated. Here we experimentally compare removal and infestation of popcorn kernels (Zea mays L. – Poaceae) and açaí fruits (Euterpe oleracea Mart. – Arecaceae) in one burned and two unburned savanna habitats in the eastern Brazilian Amazon. In each habitat, a total of ten experimental units (five per seed type) were installed, each with three treatments: (1) open access, (2) vertebrate access, and (3) invertebrate access. Generalized linear models showed significant differences in both seed removal (P < 0.0001) and infestation (P < 0.0001) among seed type, habitats and access treatments. Burned savanna had the highest overall seed infestation rate (24.3%) and invertebrate access increased açaí seed infestation levels to 100% in the burned savanna. Increased levels of invertebrate seed infestation in burned savanna suggest that preparation burning may be of limited use for the management and restoration of such habitats in tropical regions.
The objective of this study was to analyse the dynamics of spatial dispersion of the coronavirus disease 2019 (COVID-19) in Brazil by correlating them to socioeconomic indicators. This is an ecological study of COVID-19 cases and deaths between 26 February and 31 July 2020. All Brazilian counties were used as units of analysis. The incidence, mortality, Bayesian incidence and mortality rates, global and local Moran indices were calculated. A geographic weighted regression analysis was conducted to assess the relationship between incidence and mortality due to COVID-19 and socioeconomic indicators (independent variables). There were confirmed 2 662 485 cases of COVID-19 reported in Brazil from February to July 2020 with higher rates of incidence in the north and northeast. The Moran global index of incidence rate (0.50, P = 0.01) and mortality (0.45 with P = 0.01) indicate a positive spatial autocorrelation with high standards in the north, northeast and in the largest urban centres between cities in the southeast region. In the same period, there were 92 475 deaths from COVID-19, with higher mortality rates in the northern states of Brazil, mainly Amazonas, Pará and Amapá. The results show that there is a geospatial correlation of COVID-19 in large urban centres and regions with the lowest human development index in the country. In the geographic weighted regression, it was possible to identify that the percentage of people living in residences with density higher than 2 per dormitory, the municipality human development index (MHDI) and the social vulnerability index were the indicators that most contributed to explaining incidence, social development index and the municipality human development index contributed the most to the mortality model. We hope that the findings will contribute to reorienting public health responses to combat COVID-19 in Brazil, the new epicentre of the disease in South America, as well as in other countries that have similar epidemiological and health characteristics to those in Brazil.
To quantify the change in the risk of food insecurity and maternal mental disorder (MMD) before and during the coronavirus disease 2019 (COVID-19) pandemic.
Repeated cross-sectional survey. Between 17 July and 10 September 2020, mother–child pairs who were enrolled in a population-based survey in 2017 were re-contacted by telephone for consent and to complete a telephonic COVID-19 survey. We used the Brazilian Food Insecurity Scale to assess food security and the Self Reporting Questionnaire-20 to assess MMD. McNemar’s test for paired data that also accounted for clustering was used. Logistic regression was used to assess the relationship of unemployment and receipt of government assistance with food insecurity and MMD in 2020.
Five hundred and seventy-seven mother–child pairs completed the 2017 and 2020 surveys. At the time of the 2020 interview, the child cohort was 36–108 months of age.
The proportion of mothers reporting food insecurity was 15·5 % higher (95 % CI 5·9, 25·1, P value < 0·001) during the pandemic in July–August 2020 as compared with November 2017, while the prevalence of MMD was 40·2 % higher during the pandemic (95 % CI 32·6, 47·8, P value < 0·001). Loss of formal employment was associated with increased risk of food insecurity, but not with the risk of MMD.
The risk of food insecurity and MMD in Ceará increased during the COVID-19 pandemic. These findings highlight the need for policies and interventions to reduce the impact of the COVID-19 pandemic on maternal and child health, nutrition and well-being in Brazil.
This study aimed to (1) evaluate the effects of flint maize processing methods on the estimation of the readily soluble fraction (a), the potentially degradable fraction (b) and the rate of degradation of b (c) for dry matter (DM), organic matter (OM) and starch in the rumen; and (2) verify whether two different applications of in situ technique can be used to estimate in vivo DM, OM and starch digestibilities. Five ruminally cannulated Nellore bulls (265 ± 18.2 kg; 8 ± 1.0 mo) were distributed in a 5 × 5 Latin square. Three experimental diets were composed of 0.30 whole-plant maize silage, 0.10 supplement and 0.60 of one of the following processing methods: dry ground maize grain (DMG); high-moisture maize (HMM); reconstituted maize grain silage (RMG). Two additional diets were composed of 0.10 supplement, 0.80 snaplage and 0.10 stalklage (SNAP-80); or 0.10 supplement and 0.90 snaplage (SNAP-90). Digestibilities were estimated using in vivo procedure or predicted from in situ technique using a single 24 h incubation point or an equation proposed in previous literature. Diets based on ensiled grains presented greater (P < 0.05) fraction a and c and lower (P < 0.05) fraction b of DM, OM and starch compared to DMG. Both alternative use of in situ technique accurately estimated (P > 0.05) in vivo DM, OM and starch digestibilities. The results suggest that ensilage process may increase the availability of nutrients. The two different applications of in situ technique showed precision and accuracy to estimate in vivo digestibility.
To systematically review studies that used indexes to assess feeding practices of children under 2 years.
Seven databases were searched with no limit on language or publication date.
The reviewed studies included thirteen Asian, ten Latin American, four European, four North American, three Oceanian and three African.
Children under 2 years.
We analysed thirty-six studies: twenty-two presenting original indexes and fourteen using adapted indexes. Among the original indexes, thirteen assess breast-feeding, fourteen food consumption, ten food groups, and ten other feeding practices. Original indexes were mainly adapted to fit the data available in the study, to update for current nutritional recommendations or to add components not present in the original indexes. Seven studies evaluated the associations between the indexes and nutrient intake or nutritional status. The main limitations cited by the authors were: flaws in the definition of the index components, criteria for cut-off points and weighting of the evaluated index components.
The assessment of feeding practices for young children and its comparison across countries remains a challenge, especially due to the lack of consensus on the construction of indexes and regional differences in dietary recommendations and practices. Lack of validation for some indexes also makes it difficult to choose the most appropriate index for a given objective. Adapting existing indexes is a viable option. We point out relevant recommendations that may contribute to future research. Validation and longitudinal studies in diverse populations are favourable to qualify the assessment of feeding practices in this group.
The consumption of fructose has increased in children and adolescents and is partially responsible for the high incidence of metabolic diseases. The lifestyle during postnatal development can result in altered metabolic programming, thereby impairing the reproductive system and fertility during adulthood. Therefore, the aim of this study was to evaluate the effect of a high-fructose diet in the male reproductive system of pubertal and adult rats. Male Wistar rats (30 d old) were assigned to four different groups: Fr30, which received fructose (20%) in water for 30 d and were euthanized at postnatal day (PND) 60; Re-Fr30, which received fructose (20%) for 30 d and were euthanized at PND 120; and two control groups C30 and Re-C30, which received water ad libitum and were euthanized at PND 60 and 120, respectively. Fructose induced an increase in abnormal seminiferous tubules with epithelial vacuoles, degeneration, and immature cells in the lumen. Moreover, Fr30 rats showed altered spermatogenesis and daily sperm production (DSP), as well as increased serum testosterone concentrations. After discontinuing high-fructose consumption, DSP and sperm number decreased significantly. We observed tissue remodeling in the epididymis, with a reduction in stromal and epithelial compartments that might have influenced sperm motility. Therefore, we concluded that fructose intake in peripubertal rats led to changes in the reproductive system observed both during puberty and adulthood.
To investigate the timing and routes of contamination of the rooms of patients newly admitted to the hospital.
Observational cohort study and simulations of pathogen transfer.
A Veterans’ Affairs hospital.
Patients newly admitted to the hospital with no known carriage of healthcare-associated pathogens.
Interactions between the participants and personnel or portable equipment were observed, and cultures of high-touch surfaces, floors, bedding, and patients’ socks and skin were collected for up to 4 days. Cultures were processed for Clostridioides difﬁcile, methicillin-resistant Staphylococcus aureus (MRSA), and vancomycin-resistant enterococci (VRE). Simulations were conducted with bacteriophage MS2 to assess plausibility of transfer from contaminated floors to high-touch surfaces and to assess the effectiveness of wearing slippers in reducing transfer.
Environmental cultures became positive for at least 1 pathogen in 10 (59%) of the 17 rooms, with cultures positive for MRSA, C. difficile, and VRE in the rooms of 10 (59%), 2 (12%), and 2 (12%) participants, respectively. For all 14 instances of pathogen detection, the initial site of recovery was the floor followed in a subset of patients by detection on sock bottoms, bedding, and high-touch surfaces. In simulations, wearing slippers over hospital socks dramatically reduced transfer of bacteriophage MS2 from the floor to hands and to high-touch surfaces.
Floors may be an underappreciated source of pathogen dissemination in healthcare facilities. Simple interventions such as having patients wear slippers could potentially reduce the risk for transfer of pathogens from floors to hands and high-touch surfaces.
We aimed at evaluating the association of maternal pre-pregnancy nutritional status with offspring anthropometry and body composition. We also evaluated whether these associations were modified by gender, diet and physical activity and mediated by birth weight.
Birth cohort study.
Waist circumference was measured with an inextensible tape, and fat and lean mass were measured using dual-energy X-ray absorptiometry. Multiple linear regression was used to adjust for possible confounders and allele score of BMI. We carried out mediation analysis using G-formula.
In 1982, 1993 and 2004, all maternity hospitals in Pelotas (South Brazil) were visited daily and all live births whose families lived in the urban area of the city were evaluated. These subjects have been followed up at different ages.
Offspring of obese mothers had on average higher BMI, waist circumference and fat mass index than those of normal weight mothers, and these differences were higher among daughters. The magnitudes of the association were similar in the cohorts, except for height, where the association pattern was not clear. In the 1982 cohort, further adjustment for a BMI allele score had no material influence on the magnitude of the associations. Mediation analyses showed that birth weight captured part of this association.
Our findings suggest that maternal pre-pregnancy nutritional status is positively associated with offspring BMI and adiposity in offspring. And this association is higher among daughters whose mother was overweight or obese and, birth weight explains part of this association.
Austenitic stainless steel is used in several industrial branches due to its mechanical and thermal properties, and to its good corrosion resistance. With low cost and biocompatibility, it is used to manufacture prostheses and devices for bone fixation. However, direct contact with body fluids may cause corrosion. Thin films of FeAlCr intermetallic alloy can be used to increase service life of prostheses and avoid replacement surgeries. The aim of this work was to cover the austenitic stainless steel to study the effect of target–substrate distance on the film characteristics. Coatings were performed using the magnetron sputtering technique with the substrate positioned at different distances from the target. The influence on film thickness, morphology, roughness, and adhesion to the substrate was investigated. The thin films of FeAlCr (160 nm thick deposited at 100 mm far from the substrate) were formed by smaller particles (11.2 nm long), densely packed (551,000 particles/mm2), with flat and regular appearance, and greater adherence to the substrate.
Trypanosoma cruzi has three biochemically and morphologically distinct developmental stages that are programmed to rapidly respond to environmental changes the parasite faces during its life cycle. Unlike other eukaryotes, Trypanosomatid genomes contain protein coding genes that are transcribed into polycistronic pre-mRNAs and have their expression controlled by post-transcriptional mechanisms. Transcriptome analyses comparing three stages of the T. cruzi life cycle revealed changes in gene expression that reflect the parasite adaptation to distinct environments. Several genes encoding RNA binding proteins (RBPs), known to act as key post-transcriptional regulatory factors, were also differentially expressed. We characterized one T. cruzi RBP, named TcZH3H12, which contains a zinc finger domain and is up-regulated in epimastigotes compared to trypomastigotes and amastigotes. TcZC3H12 knockout (KO) epimastigotes showed decreased growth rates and increased capacity to differentiate into metacyclic trypomastigotes. Transcriptome analyses comparing wild type and TcZC3H12 KOs revealed a TcZC3H12-dependent expression of epimastigote-specific genes such as genes encoding amino acid transporters and proteins associated with differentiation (PADs). RNA immunoprecipitation assays showed that transcripts from the PAD family interact with TcZC3H12. Taken together, these findings suggest that TcZC3H12 positively regulates the expression of genes involved in epimastigote proliferation and also acts as a negative regulator of metacyclogenesis.
This research communication addresses the hypothesis that, during the summer in the subtropics, natural tree shade helps to improve milk functional characteristics such as stability and acidity. Sixteen Holstein lactating cows were enrolled. The study consisted of three periods (pre-stress, heat stress and post-stress) based on allocating grazing cows into two treatments (with and without access to shade during the Heat Stress period). Overall THI during the trial was (mean ± se) 76.0 ± 3.4. Access to shade prevented the heat stress-related decrease in milk stability both in the ethanol and in the coagulation time test, as well as maintained milk acidity within an acceptable range (14 to 18°D).
The transgenic Liberty Link® (LL) soybean is tolerant to glufosinate, conferred by the enzyme phosphinothricin acetyltransferase (PAT), which is encoded by the pat gene from Streptomyces viridochromogenes. Because symptoms of injury can be observed in soybean [Glycine max (L.) Merr.] plants in some situations, this study evaluated the effects of rates of glufosinate on agronomic performance; quality of LL soybean seeds; and the ammonia, glufosinate, and N-acetyl-l-glufosinate concentration (NAG) in soybeans with and without the pat gene after application of increasing glufosinate rates. Field and greenhouse experiments were conducted; the first evaluated the selectivity of glufosinate in LL soybeans, and the second evaluated the metabolic changes in soybeans with (LL) and without (RR2) the pat gene, after application of glufosinate. For fieldwork, application of glufosinate at rates up to four times the maximum recommended caused initial injury symptoms (up to 38.5%) in LL soybean plants. However, no negative effect was found on seed quality and agronomic performance of LL plants, including yield. This shows the selectivity of glufosinate promoted by pat gene insertion for application in POST (V4), in LL soybean. For the greenhouse experiment, it was concluded that the LL soybean plants presented high glufosinate metabolism, lower ammonia concentration, and no reduction in dry matter, in comparison with RR2 soybean, after application of high rates of glufosinate.
Background: Based on data obtained from hospitals in the city of Belo Horizonte (population ~3,000,000), we evaluated relevant factors such as death, age, duration of surgery, potential for contamination and surgical site infection, plastic surgery, and craniotomy. The possibility of predicting surgical site infection (SSI) was then analyzed using pattern recognition algorithms based on MLP (multilayer perceptron). Methods: Data were collected by the hospital infection control committees (CCIHs) in hospitals in Belo Horizonte between 2016 and 2018. The noisy records were filtered, and the occurrences were analyzed. Finally, the predictive power of SSI of 5 types MLP was evaluated experimentally: momentum, backpropagation standard, weight decay, resilient propagation, and quick propagation. The model used 3, 5, 7, and 10 neurons in the occult layer and with resamples varied the number of records for testing (65% and 75%) and for validation (35% and 25%). Comparisons were made by measuring the AUC (area under the curve (range, 0–1). Results: From 1,096 records of craniotomy, 289 were usable for analysis. Moreover, 16% died; averaged age was 56 years (range, 40–65); mean time of surgery was 186 minutes (range, 95–250 minutes); the number of hospitalizations ranged from 1 (90.6%) to 8 (0.3%). Contamination among these cases was rated as follows: 2.7% contaminated, 23.5% potentially contaminated, 72.3% clean. The SSI rate reached 4%. The prediction process in AUCs ranged from 0.7 to 0.994. In plastic surgery, from 3,693 records, 1,099 were intact, with only 1 case of SSI and no deaths. The average age for plastic surgery was 41 years (range, 16–91); the average time of surgery was 218.5 minutes (range, 19–580 minutes); the number of hospitalizations ranged from 1 (77.4%) to 6 times (0.001%). Contamination among these cases was rated as follows: 27.90% potential contamination, 1.67% contaminated, and 0.84% infected. The prediction process ranged in AUCs from 0.2 to 0.4. Conclusions: We identified a high noise index in both surgeries due to subjectivity at the time of data collection. The profiles of each surgery in the statistical analyses were different, which was reflected in the analyzed structures. The MLP for craniotomy surgery demonstrated relevant predictive power and can guide intelligent monitoring software (available in www.sacihweb.com). However, for plastic surgeries, MLPs need more SSI samples to optimize outcomes. To optimize data collection and to enable other hospitals to use the SSI prediction tool, a mobile application was developed.
Background: Healthcare services are increasingly shifting from inpatient to outpatient settings. Outpatient settings such as emergency departments (EDs), oncology clinics, dialysis clinics, and day surgery often involve invasive procedures with the risk of acquiring healthcare-associated infections (HAIs). As a leading cause of HAI, Clostridioides difficile infection (CDI) in outpatient settings has not been sufficiently described in Canada. The Canadian Nosocomial Infection Surveillance Program (CNISP) aims to describe the epidemiology, molecular characterization, and antimicrobial susceptibility of outpatient CDI across Canada. Methods: Epidemiologic data were collected from patients diagnosed with CDI from a network of 47 adult and pediatric CNISP hospitals. Patients presenting to an outpatient setting such as the ED or outpatient clinics were considered as outpatient CDI. Cases were considered HAIs if the patient had had a healthcare intervention within the previous 4 weeks, and they were considered community-associated if there was no history of hospitalization within the previous 12 weeks. Clostridioides difficile isolates were submitted to the National Microbiology Laboratory for testing during an annual 2-month targeted surveillance period. National and regional rates of CDI were stratified by outpatient location. Results: Between January 1, 2015, and June 30, 2019, 2,691 cases of outpatient-CDI were reported, and 348 isolates were available for testing. Most cases (1,475 of 2,691, 54.8%) were identified in outpatient clinics, and 72.8% (1,960 of 2,691) were classified as community associated. CDI cases per 100,000 ED visits were highest in 2015, at 10.3, and decreased to 8.1 in 2018. Rates from outpatient clinics decreased from 3.5 in 2016 to 2.7 in 2018 (Fig. 1). Regionally, CDI rates in the ED declined in Central Canada and increased in the West after 2016. Rates in outpatient clinics were >2 times higher in the West compared to other regions. RT027 associated with NAP1 was most common among ED patients (26 of 195, 13.3%), whereas RT106 associated with NAP11 was predominant in outpatient clinics (22 of 189, 11.6%). Overall, 10.4% of isolates were resistant to moxifloxacin, 0.5% were resistant to rifampin, and 24.2% were resistant to clindamycin. No resistance was observed for metronidazole, vancomycin, or tigecycline. Compared to CNISP inpatient CDI data, outpatients with CDI were younger (51.8 ± 23.3 vs 64.2 ± 21.6; P < .001), included more females (56.4% vs 50.9%; P < .001), and were more often treated with metronidazole (63.0% vs 56.1%; P < .001). Conclusions: For the first time, CDI cases identified in outpatient settings were characterized in a Canadian context. Outpatient CDI rates are decreasing overall, but they vary by region. Predominant ribotypes vary based on outpatient location. Outpatients with CDI are younger and are more likely female than inpatients with CDI.
Disclosures: Susy Hota reports contract research for Finch Therapeutics.
The chronic kidney disease (CKD) and diabetes mellitus (DM) are considered a serious public health problem. The objective was investigating the association of DM with the anthropometric measures, biochemical profile and dietary intake in patients with CKD. Is a cross-sectional study done in 2017, with 51 patients previously diagnosed with CKD. We collect socio-demographic, lifestyle variables, anthropometric measurements, biochemical profile and dietary intake. We using the Kolmogorov–Smirnov test, followed by Pearson's χ2 test and Student's t test. Data were analysed using several multivariable logistic regression models, including the socio-demographic, anthropometric, dietary intake and biochemical variable. Variables with P ≤ 0⋅20 in the univariate analyses were selected and kept in the block in the simple and multiple logistic regression analysis, to determine the differences between the categories and the factors associated with the presence of DM or not, remaining in the model final, only the significant variables (P ≤ 0⋅05). Each variable was adjusted for all other variables included in the univariate analysis. The strength of the association was assessed by the odds ratio and 95% confidence intervals (CI). The multivariate logistic regression analysis evidenced that the increase of 1 cm in waist circumference and 1 mg/dl in VLDL-c values increases the chance of DM, respectively, by 8⋅4% (OR 1⋅076; P 0⋅05) and 8⋅8% (OR 1⋅102; P 0⋅01). In contrast, an increase of 1 mg/dl in total cholesterol decreases the chance of developing DM by 3⋅1% (OR 0⋅965; P 0⋅01), that is, it becomes a protective factor. The present study identified the associations between overweight, dietary intake and biochemical tests.
To conduct international comparisons of self-reports, collateral reports, and cross-informant agreement regarding older adult psychopathology.
We compared self-ratings of problems (e.g. I cry a lot) and personal strengths (e.g. I like to help others) for 10,686 adults aged 60–102 years from 19 societies and collateral ratings for 7,065 of these adults from 12 societies.
Data were obtained via the Older Adult Self-Report (OASR) and the Older Adult Behavior Checklist (OABCL; Achenbach et al., 2004).
Cronbach’s alphas were .76 (OASR) and .80 (OABCL) averaged across societies. Across societies, 27 of the 30 problem items with the highest mean ratings and 28 of the 30 items with the lowest mean ratings were the same on the OASR and the OABCL. Q correlations between the means of the 0–1–2 ratings for the 113 problem items averaged across all pairs of societies yielded means of .77 (OASR) and .78 (OABCL). For the OASR and OABCL, respectively, analyses of variance (ANOVAs) yielded effect sizes (ESs) for society of 15% and 18% for Total Problems and 42% and 31% for Personal Strengths, respectively. For 5,584 cross-informant dyads in 12 societies, cross-informant correlations averaged across societies were .68 for Total Problems and .58 for Personal Strengths. Mixed-model ANOVAs yielded large effects for society on both Total Problems (ES = 17%) and Personal Strengths (ES = 36%).
The OASR and OABCL are efficient, low-cost, easily administered mental health assessments that can be used internationally to screen for many problems and strengths.