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Male infertility is a multifactorial disorder that involves different physiopathological mechanisms and multiple genes. In this sense, we analyse the role of miRNAs in this pathology. Gene expression analysis can provide relevant information to detect biomarkers, signalling pathways, pathologic mechanisms, and potential therapeutic targets for the disease. In this review, we describe four miRNA microarrays related to patients who present infertility diseases, including azoospermia, asthenozoospermia, and oligoasthenozoospermic. We selected 13 miRNAs with altered expressions in testis tissue (hsa-miR-122-5p, hsa-miR-145-5p, hsa-miR-16-5p, hsa-miR-193a-3p, hsa-miR-19a-3p, hsa-miR-23a-3p, hsa-miR-30b-5p, hsa-miR-34b-5p, hsa-miR-34c-5p, hsa-miR-374b-5p, hsa-miR-449a, hsa-miR-574-3p and hsa-miR-92a-3p), and systematically examine the mechanisms of four relevant miRNAs (hsa-miR-16-5p, hsa-miR-19a-3p, hsa-miR-92a-3p and hsa-miR-30b-5p) which we found that regulated a large number of proteins. An interaction network was generated, and its connections allowed us to identify signalling pathways and interactions between proteins associated with male infertility. In this way, we confirm that the most affected and relevant pathway is the PI3K-Akt signalling.

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BACKGROUND: Adherence to TB treatment and therefore treatment success could be improved using digital adherence technology.OBJECTIVE: To evaluate the effectiveness of a medication event reminder monitor system (MERM) on treatment success and treatment adherence in patients with drug-susceptible pulmonary TB in Perú.METHODS: This was an experimental, randomised, open-label, controlled study conducted among patients in the second phase of TB treatment. The intervention group received their medications through MERM with the support of a treatment monitor, whereas the control group used the usual strategy. Participants were followed until they completed the 54 doses of the second phase of treatment.RESULTS: The study included 53 patients in each group; four in the intervention group withdrew from the study. Treatment success was significantly more frequent in the MERM group (RR 1.15, 95% CI 1.02-1.30; P = 0.0322). There was no significant difference in the adherence outcomes; however, the percentage of patients who missed at least one dose and patients with more than 10% of total doses missed were lower in the intervention group.CONCLUSION: The use of MERM in the second phase of treatment showed a significant improvement in the treatment success rate in patients with drug-susceptible pulmonary TB.

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Platelet activity is essential in cardiovascular diseases. Therefore our objective was to evaluate the main effects of activating RAGE in platelets which are still unknown. A search for RAGE expression in different databases showed poor or a nonexistent presence in platelets. We confirmed the expression in platelets and secreted variable of RAGE (sRAGE). Platelets from elderly adults expressed in resting showed 3.2 fold more RAGE from young individuals (p < 0.01) and 3.3 fold with TRAP-6 (p < 0.001). These results could indicate that the expression of RAGE is more inducible in older adults. Then we found that activating RAGE with AGE-BSA-derived from methylglyoxal and subthreshold TRAP-6, showed a considerable increase with respect to the control in platelet aggregation and expression of P-selectin (respectively, p < 0.01). This effect was almost completely blocked by using a specific RAGE inhibitor (FSP-ZM1), confirming that RAGE is important for the function and activation platelet. Finally, we predict the region stimulated by AGE-BSA is located in region V of RAGE and 13 amino acids are critical for its binding. In conclusion, the activation of RAGE affects platelet activation and 13 amino acids are critical for its stimulation, this information is crucial for future possible treatments for CVD.

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Germline pathogenic variants in the CDH1 gene are a well-established cause of hereditary diffuse gastric cancer (HDGC) syndrome. The aim of this study was to characterize CDH1 mutations associated with HDGC from Chile, a country with one of the highest incidence and mortality rates in the world for gastric cancer (GC). Here, we prospectively include probands with family history/early onset of diffuse-type of GC. The whole coding sequence of the CDH1 gene was sequenced from genomic DNA in all patients, and a multidisciplinary team managed each family member with a pathogenic sequence variant. Thirty-six cases were included (median age 44 years/male 50%). Twenty-seven (75%) patients had diffuse-type GC at ≤50 years of age and 19 (53%) had first or second-degree family members with a history of HDGC. Two cases (5.5%) carried a non-synonymous germline sequence variant in the CDH1 gene: (a) The c.88C>A missense variant was found in a family with three diffuse-type GC cases; and (b) c.1531C>T a nonsense pathogenic variant was identified in a 22-year-old proband with no previous family history of HDGC. Of note, six family members carry the same nonsense pathogenic variant. Prophylactic gastrectomy in the proband's sister revealed stage I signet-ring cell carcinoma. The finding of 1531C>T pathogenic variant in the CDH1 in proband with no previous family history of HDGC warrants further study to uncover familial clustering of disease in CDH1 negative patients. This finding may be particularly relevant in high incidence countries, such as the case in this report.

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INTRODUCTION: Infection from different bacterial may increase the risk of thrombosis and atherosclerosis risk by production and secretion of many proinflammatory factors. Human platelets have toll-like receptor 4 (TLR4), the principal receptor for lipopolysaccharide (LPS). The activation of platelet produces Platelet-derived Microparticles (PDMPs) measuring less than 1.0 micron (that are very abundant in circulation >90%), which are associated with the development of Cardiovascular Diseases (CVDs), the leading cause of death in the world. OBJECTIVES: Experiments were designed to evaluate the generation of pro-thrombogenic microparticles in vitro on platelets via TLR4 activation. METHODS: Platelet-rich plasma and washed platelets from healthy volunteers were incubated for the generation of PDMPs. The best source for the generation of microparticles was washed platelets. Then the washed platelets were incubated for 15 minutes with ultrapure Escherichia coli LPS (0-9 µg/mL) followed by activation with ADP (1 µM, subaggregant concentration), centrifuged for 60 minutes and analyzed by flow cytometry. RESULTS: Incubating platelets with LPS (9 µg/mL) and ADP (1 µM) produced a 34-fold increase in PDMPs generation. Finally, we evaluated this protocol to detect the inhibition of PDMPs generation, washed platelets were incubated with acetylsalicylic acid (10 µM) and an inhibition of 7.7-fold in PDMPs generation for activation of TLR4 was found. CONCLUSION: A new and easy protocol for PDMPs generation and analysis by Flow Cytometry is established. In the future it could be used to determine the association of PDMPs with different pathologies.

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The main component of Alzheimer's disease (AD) is the amyloid-beta peptide (Aß), the brain of these patients is characterized by deposits in the parenchyma and cerebral blood vessels known as cerebral amyloid angiopathy (CAA). On the other hand, the platelets are the major source of the Aß peptide in circulation and once secreted can activate the platelets and endothelial cells producing the secretion of several inflammatory mediators that finally end up unchaining the CAA and later AD. In the present study we demonstrate that cAMP/PKA pathway plays key roles in the regulation of calpain activation and secretion of Aß in human platelets. We confirmed that inhibition of platelet functionality occurred when platelets were incubated with forskolin (molecule that rapidly increased cAMP levels). In this sense we found that platelets pre-incubated with forskolin (20 µM) present a complete inhibition of calpain activity and this effect is reversed using an inhibitor of protein kinase A. Consequentially, when platelets were inhibited by forskolin a reduction in the processing of the APP with the consequent decrease in the Aß peptide secretion was observed. Therefore our study provides novel insight in relation to the mechanism of processing and release of the Aß peptide from human platelets.

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BACKGROUND: Paediatric obesity and insulin resistance (IR) are potentially reversible inflammatory conditions. Long chain polyunsaturated fatty acids omega-3 (LCPUFA-ω3) show anti-inflammatory and metabolic properties, but their clinical efficacy is unclear. OBJECTIVE: The objective of this study is to evaluate whether supplementation with LCPUFA-ω3 for 3 months reduces insulin resistance and weight to adolescents with obesity. METHODS: Double-blind trial of 366 adolescents with obesity randomly assigned to 1.2-g LCPUFA-ω3 (DO3) or 1-g sunflower oil (DP) daily for 3 months; both groups received an energy-restricted diet. Children attended monthly for anthropometric, dietary, and clinical measurements. Basal and final blood samples were obtained to measure metabolic markers and erythrocytes fatty acids. Regression models were used for analysis. RESULTS: A total of 119 DO3 and 126 DP children completed follow-up. At baseline, 92% of children presented IR, 66% hypertriglyceridemia, 37% low-grade inflammation, and 32% metabolic syndrome. Despite erythrocytes LCPUFA-ω3 increased more in DO3 (Median differences = 0.984 w/w%; 95 IC = 0.47, 1.53, P < 0.001), body weight, insulin, and HOMA changed similarly in both groups at the end of intervention. Adjusting for basal values, changes in weight, insulin, and HOMA was not related with supplementation. CONCLUSIONS: Supplementation with LCPUFA-ω3 does not affect body weight or insulin in adolescents with obesity.

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Cardiovascular diseases are one of the main public health problems, and many of them, their pathophysiology involves alterations in platelet activity. Platelet activation is an essential event that is regulated by the intracellular concentrations of Ca2+ and cAMP. Interestingly, it has been shown that the activation of adenosine A2A receptors increases cAMP levels and produces the inhibition of platelet aggregation, which appears as a potential target for regulation of platelet activity. Therefore, we tried to activate A2A receptors using Indiplon, a drug developed for the treatment of insomnia, and analyze its effect on platelet activity in vitro. Our results indicate that Indiplon is able to interact in silico with the adenosine A2A receptor (ΔGbind of -73.321 kcal/mol, similar to that obtained with adenosine), which is involved in the regulation of platelet cAMP levels. In functional studies using PRP, a reduction in platelet aggregation induced by ADP was observed in the presence of Indiplon at 500 µM with a percentage of inhibition 70%, where the use of specific inhibitors (ZM241385 and MSX-2) of the A2A receptor also blocked these effects reducing the percentage of inhibition to 41% and 34.1%, respectively. Also, the use of Indiplon produced a decrease in the expression in the membrane of P-selectin. Thus, Indiplon acts as an A2A receptor agonist and whose activation results in inhibition of platelet aggregation and activation, showing a possible cardiovascular protective role.

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Tubulointerstitial nephritis (TIN) is the main renal involvement associated with primary Sjögren syndrome (pSS). TIN can manifest as distal renal tubular acidosis (RTA), nephrogenic diabetes insipidus, proximal tubular dysfunction, and others. We present a 31-year-old female with hypokalemic paralysis due to distal RTA (dRTA). She received symptomatic treatment and hydroxychloroquine with a good response. There is insufficient information on whether to perform a kidney biopsy in these patients or not. The evidence suggests that there is an inflammatory background and therefore a potential serious affection to these patients, such as hypokalemic paralysis. We found 52 cases of hypokalemic paralysis due to dRTA in pSS patients. The majority of those patients were treated only with symptomatic medication. Patients who received corticosteroids had stable evolution even though they did not have another symptomatology. With such heterogeneous information, prospective studies are needed to assess the value of adding corticosteroids as a standardized treatment of this manifestation.

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The objectives of this study were to describe alterations that age and dietary inclusion of direct-fed microbial (DFM) Bacillus subtilis (BS) and a specific essential oil (EO) blend (carvacrol, cinnamaldehyde, cineol, and pepper extract) causes in the activity of digestive enzymes (maltase: MALT; aminopeptidase-N: APN; intestinal alkaline phosphate: IAP) and expression patterns of genes related to transport (oligopeptide transporter gene: SLC15A1; Na+-dependent glucose and galactose transporter gene: SLC5A1; Na+-independent glucose, galactose, and fructose transporter gene: SLC2A2; ATPase, Na+/K+ transporting gene: ATP1A1) and digestion (aminopeptidase-N gene: ANPEP; maltase-glucoamylase gene: MGAM; Sucrase-isomaltase gene: SI) of carbohydrates and proteins in the small intestine of broilers. Also, the objective was to analyze if growth performance of broilers is affected by supplementation (BS and EO blend). Day-old male broiler chicks (n = 1,320) were assigned to 5 treatments. Diets included a basal diet (BD) as a negative control (CON); experimental diets were BD + BS; BD + BS + EO; BD + EO; BD + antibiotic growth promoter (AGP) avilamycin was the positive control. Performance was evaluated between 1 to 42 d. Transcript abundance of transport-related genes and digestion-related genes were assayed by RT-qPCR and determined at d 7, 21, and 42. MALT-, APN-, and IAP-specific activities were determined at d 7, 21, and 42. Broilers fed BS had greater SLC15A1 mRNA abundance compared to CON, while EO and AGP were related to higher activities of IAP and APN. Analysis over time revealed higher abundance of MGAM, SLC2A2, SLC15A1, SLC5A1 and SI mRNA at d 42 when compared to d 7. Activity of IAP decreased after d 7 and activity of MALT increased with age. The current study suggests that age had effect over carbohydrate and protein transport and carbohydrate digestion. The supplementation of BS DFM hade evident effect over protein transport and that the use of EO in the diet enhanced the activities of carbohydrate and protein digestion, reflecting improvement in digestive and transport physiology of birds. Changes performed by BS DFM and EO did not favor performance.

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A high degree of specialization between host and parasite is a well-known outcome of a long history of coevolution, and it is strikingly illustrated in a coordination of their life cycles. In some cases, the arms race ensued at the establishment of a symbiotic relationship results in the adoption of manipulative strategies by the parasite. We have already learned that Steinina ctenocephali, a gregarine living in the alimentary canal of cat flea, Ctenocephalides felis follows its phenology and metamorphosis. Despite these findings the outcome of their symbiotic partnership (mutualist, parasitic or commensal) remains unclear. To address this important question, we measured life history parameters of the flea in the presence of varying infection intensities of gregarine oocysts in laboratory conditions. We found that neither the emergence nor survival rate of fleas was affected by harbouring the gregarines. More surprisingly, our results show that flea larvae infected with gregarines developed faster and emerged earlier than the control group. This gregarine therefore joins the selected group of protists that can modify physiological host traits and provides not only new model taxa to be explored in an evolutionary scenario, but also potential development of control strategies of cat flea.

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Salmonella enterica subsp. enterica serovar Gallinarum biovar Gallinarum (SG) causes fowl typhoid (FT), a septicaemic disease which can result in high mortality in poultry flocks. The absence of flagella in SG is thought to favour systemic invasion, since bacterial recognition via Toll-like receptor (TLR)-5 does not take place during the early stages of FT. In the present study, chicks susceptible to FT were inoculated with a wild type SG (SG) or its flagellated motile derivative (SG Fla(+)). In experiment 1, mortality and clinical signs were assessed, whereas in experiment 2, gross pathology, histopathology, systemic invasion and immune responses were evaluated. SG Fla(+) infection resulted in later development of clinical signs, lower mortality, lower bacterial numbers in the liver and spleen, and less severe pathological changes compared to SG. The CD8(+) T lymphocyte population was higher in the livers of chicks infected with SG at 4 days post-inoculation (dpi). Chicks infected with SG had increased expression of interleukin (IL)-6 mRNA in the caecal tonsil at 1 dpi and increased expression of IL-18 mRNA in the spleen at 4 dpi. In contrast, the CD4(+) T lymphocyte population was higher at 6 dpi in the livers of birds infected with SG Fla(+). Therefore, flagella appeared to modulate the chicken immune response towards a CD4(+) T profile, resulting in more efficient bacterial clearance from systemic sites and milder infection.

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Two experiments were conducted to test the hypothesis that when using similar protein/amino acid diets and environment temperature conditions, the performance and carbon turnover in muscle and liver tissues, as measured by the incorporation of stable isotopes ((13)C/(12)C), must be different between fast-growing Cobb 500® and slow-growing Label Rouge broilers. For both experiments (Cobb and Label Rouge), 21-d-old birds were distributed in a completely randomised, 3 × 3 factorial design; three environmental temperatures (cyclic heat stress ad libitum, 22°C ad libitum, and 22°C restricted) and three crude protein concentrations (189.1, 210 and 220 g/kg CP) were used. The Cobb 500® had better performance with higher concentrations of crude protein. Cyclic heat stress (a temperature factor), negatively affected this genetic strain's performance. For the Label Rouge birds, the crude protein concentrations in the diet presented inconsistent results and cyclic heat stress did not affect the performance. The carbon turnover rate was affected in the Cobb 500® strain, with a high protein content reducing carbon turnover in the evaluated tissues (liver and muscles). Feed intake had a greater impact on carbon turnover rates than cyclic heat stress. The Label Rouge birds were not affected by the evaluated factors, suggesting that genetic improvement has a leading role on tissue carbon turnover. There is a genetic influence on carbon turnover in the liver and muscle tissues of broiler chickens. In addition, genetically fast-growing broilers are more susceptible to variations in diet composition and environmental temperature than less rapidly growing animals.

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OBJECTIVE: To evaluate the usefulness of serial determinations of asymmetric dimethylarginine (ADMA) and homocysteine (Hcy) concentrations during pregnancy to predict pre-eclampsia, taking into account maternal obesity and B vitamin status. DESIGN: Longitudinal study. SETTING: Two obstetric referral hospitals. SAMPLE: Two hundred and fifty-two of 411 women invited to participate in the study. METHODS: The women made monthly visits from ≤20 weeks of gestation until delivery for measurements of plasma ADMA, Hcy, and vitamins B6 , B12, and folic acid, and for the recording of clinical information. MAIN OUTCOME MEASURE: Early elevations in plasma ADMA and Hcy related to the development of pre-eclampsia. RESULTS: Of the 252 women who completed the study, 179 had no complications, 49 developed pre-eclampsia, and 24 presented with complications other than pre-eclampsia. ADMA and Hcy increased gradually throughout pregnancy in the pre-eclampsia group, independent of maternal B-vitamin status and obesity, but remained constant in women with no complications. Relative to the preceding month, ADMA and Hcy levels increased 1 month prior to the onset of pre-eclampsia: 124 ± 27 nmol (P < 0.001) and 1177 ± 278 nmol (P = 0.001), respectively, in the pre-eclampsia group. The group of women with no complications did not show any significant changes. Increases of 80 nmol ADMA and 1000 nmol Hcy at 1 month prior to the onset of pre-eclampsia demonstrated the best potential for prediction. CONCLUSIONS: Increased ADMA and Hcy levels precede clinical manifestations of pre-eclampsia. Therefore, serial determinations of their concentrations may be helpful in identifying women at risk. TWEETABLE ABSTRACT: Increased ADMA and Hcy precede clinical pre-eclampsia and may identify women at risk.

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The present assay attempts to evaluate the feasibility of using embryo transfer in small community farmers by in vivo study and by modelling the results obtained. From the total of 59 donor cows, 62.7% responded to treatment, with a significant difference (p = 0.002) in the percentage of the response between breeds, being 90.5% (19/21) in Holstein and 47.4% (18/38) in Brahman. A total of 283 embryos were graded as transferable, while 141 as non-transferable, without difference in the percentage of transferable embryo by breed (p = 0.18). The mean of transferable embryos graded as class I and II was not different between Holstein and Brahman (p = 0.96 and p = 0.92, respectively); besides, no differences were observed in the other grades (non-transferable). The highest difference in costs, regardless of its quality by breed, was seen in the lower levels of probable fertility of the embryo transferred, even reaching several hundred dollars. When modelling the expected costs for embryo produced and transferred, values can reach nearly $2000.00 when the probable fertility is only 10%. However, when the probable fertility was 60%, embryo cost was close to $300.00. This technology seems to be viable on average or high-scale systems, having a superovulatory response between 60 and 80% with 4-6 transferrable embryos. Yet, in small-scale farming, due to the reduced number of donors and/or recipients, the costs surpass the economical feasibility of the technique.

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1. An experiment was conducted to study the effect of two different diets on zootechnical performance and the major bacterial groups in association with the host mucosa and dispersed in the lumen contents of the small intestine of broiler chickens. 2. The two experimental diets were maize or sorghum-based. In addition to the total bacteria, bacterial groups belonging to the Enterobacteriaceae (Enterococcus and Lactobacillus) were quantified by real-time PCR. 3. There were no differences in body weight gain and feed intake, but feed conversion ratio increased for sorghum-fed broilers at 21 and 42 d of age. 4. The Enterococcus group decreased in all gut segments from 7 to 42 d, while the Lactobacillus group increased in both ecosystems. In the ileal mucosa, the enterobacterial counts decreased from 7 to 42 d in the maize-based diet, but remained stable in the sorghum-based diet. 5. The results shed light on the spatial and temporal distribution of bacterial groups that play important physiological roles in the small intestine of chickens. Specifically, the increased Enterobacteria population in the ileum is consistent with the relatively poor feed conversion in sorghum-fed broilers.

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Evaluar y comparar de forma clínica, tomográfica e histológica el proceso de reparación ósea y la conservación del reborde alveolar en dos alveolos post-extracción de paredes intactas, con y sin la utilización de un material sustituto óseo regenerativo a base de sulfato de calcio hemihidratado como relleno y barrera, antes de la colocación de implantes a cuatro meses.

To evaluate and compare clinically, tomographic and histological the bone repair process and the preservation of the alveolar walls of two sockets intact, with and without the use of a bone substitute material based regenerative calcium sulfate hemihydrate as a filler and barrier before implant placement to four months.

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The aim of the present study was to comparatively evaluate hemogram, blood serum components and anatomopathologic alterations in commercial layers experimentally challenged with an attenuated vaccine candidate strain (SG∆cobS∆cbiA) and other two pathogenic strains (SGDcobS and SGNalr) of Gallinarum (SG). In total, 280 commercial layers were randomly divided into 4 groups (G1, G2, G3 and G4). At five days of age, birds from groups G1 received approximately 107 colony forming units (CFU) of SGDcobS; meanwhile birds from group G2 and G3 received the same dose of SGNalr and SG∆cobS∆cbiA, respectively. Birds from G4 were not infected. At 24 hours before (DBI) and 24 hours after (1 DAI), and three (3 DAI), five (5 DAI), seven (7 DAI) ten (10 DAI), and fifteen (15 DAI) days after the infection, 10 birds of each group were humanely killed and blood samples collected to hematological and serum tests. Samples of liver, spleen, thymus, bursa of Fabricius, kidney and heart were also collected for the histological examination. Birds inoculated with SGDcobS and SGNalr showed similar alterations in hemogram, blood serum components and anatomopathologic exams. On the other hand, the exams of birds inoculated with SG∆cobS∆cbiA strain were similar to those of the uninfected birds. However, changes could be noticed in levels of uric acid and cholesterol during the course of the infection of birds from G3. Decrease in levels of light IgG 3 DAI was also observed in birds from this group. Pyknosis in kidney cells was a microscopic alteration found in birds from G3. Further studies must be done to verify if these alterations will not interfere in the performance of the vaccinate birds with SG∆cobS∆cbiA strain.(AU)

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Background: ConQoL questionnaire assesses health related quality of life among children with congenital heart diseases. It has a version for children aged 8 to 11 years and anotherfor children aged 12 to 16years. Aim: To validate ConQol questionnaire for Chilean children with a congenital heart disease. Material and Methods: Using a multicentric cross sectional design, 334 children from four hospitals (54% males), were surveyed. Among them 45% were aged 8 to 11 years and 55%, 12 to 16 years. The study involved three stages: cross cultural adjustment of the original questionnaire, pre-test study, and estimation of its psychometric properties. Content, construct and criterion validity and internal consistency with Cronbach's alpha, were assessed. Results: The version for children aged 8 to 11 years and comprised by three domains (symptoms, activity and relationships), obtained and α ≥ 0.60. In the questionnaire for children aged 12 to 16years, there is one more domain called coping, which obtained an α of 0.53, that was different to the other three domains that obtained an α > 0.70. The correlation between Health Quality of Life and Perception of Health Quality of Life was statistically significant for both groups. The association between Health Quality ofLife and health capability was only significant among children aged 12 to 16years (p < 0.01). Conclusions: The adapted ConQol questionnaire matched properly with the original one. The adapted questionnaire is valid and reliable to assess Health Quality ofLife among Chilean children with congenital heart diseases.

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Experimental studies have shown that hatching rate depends, among other factors, on the main physical characteristics of the eggs. The physical parameters used in our work were egg weight, eggshell thickness, egg sphericity, and yolk per albumen ratio. The relationships of these parameters in the incubation process were modeled by Fuzzy logic. The rules of the Fuzzy modeling were based on the analysis of the physical characteristics of the hatching eggs and the respective hatching rate using a commercial hatchery by applying a trapezoidal membership function into the modeling process. The implementations were performed in software. Aiming to compare the Fuzzy with a statistical modeling, the same data obtained in the commercial hatchery were analyzed using multiple linear regression. The estimated parameters of multiple linear regressions were based on a backward selection procedure. The results showed that the determination coefficient and the mean square error were higher using the Fuzzy method when compared with the statistical modeling. Furthermore, the predicted hatchability rates by Fuzzy Logic agreed with hatching rates obtained in the commercial hatchery.

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