Exploring HLA-C methylation patterns and nutritional status in Kichwa mothers and infants from Tena, Ecuador.

Environment and lifestyle can affect the epigenome passed down from generation to generation. A mother's nutrition can impact the methylation levels of her offspring's epigenome, but it's unclear which genes may be affected by malnutrition during gestation or early development. In this study, we examined the levels of methylated GC in the promoter region of HLA-C in mothers and infants from the Kichwa community in Ecuador. To do this, we analyzed saliva samples using bisulfite DNA sequencing. While we did not observe any significant differences in the mean methylation percentages in exon 1 of HLA-C between mothers and their infants after the first two years of lactation and life, respectively, we did find that infants tended to increase their methylation level during the first two years of life, while mothers tended to decrease it after the first two years of breastfeeding. When we compared methylation levels between mothers and infants using an ANOVA/posthoc Tukey test, we found that the average methylation for the entire population was less than 3% at T1 and T2. Although there was a tendency for infants to have higher methylation levels during their first two years of life and for mothers to have lower methylation levels after the first two years of breastfeeding, the mean values were not significantly different. However, we found a significant difference when we contrasted the data using a Kruskal-Wallis test at 0.05 for T1 AND T2 (p-value: 0.0148). Specifically, mothers had an average of X̅ = 2.06% and sons had X̅ = 1.57% at T2 (p-value: 0.7227), while the average for mothers was X̅ = 1.83% and for sons X̅ =1.77%. Finally, we identified three CpG motif nucleotide positions (32-33, 43-44, and 96-97) along the 122 bp analysis of HLA-C exon one, which was found to retain methylation patterns over time and is inherited from mother to offspring. Finally, our small pilot study did not reveal significant correlations between maternal and offspring nutritional status and DNA methylation levels of HLA-C exon one.

Surveillance of COVID-19 vaccines: A comprehensive analysis of the first immunization drive in Ecuador.

The initial phase of the COVID-19 vaccination in Ecuador occurred between April and November 2021. Initially, it focused on priority populations, including health professionals and other front-line workers. During this period, there was limited knowledge about the vaccine's adverse effects. A non-probability, observational study was conducted among university staff in Guayaquil, Ecuador, who received the AstraZeneca vaccine (n = 423) between April and November 2021. This study aimed to compare the acute adverse reactions by doses and to report the incidence of long-term adverse reactions within the AstraZeneca group. As a result, comparing acute adverse reactions between doses, the odds ratio for local pain, headache, muscle pain, fever, and chills are statistically higher after the first dose than the second dose. Survival curves indicated these symptoms appeared mainly within the first 6 h post-vaccination. This is the first pharmacovigilance study from Ecuador that analyzes survival probabilities for the AstraZeneca vaccine's adverse effects.

Essential oils: a systematic review on revolutionizing health, nutrition, and omics for optimal well-being.

Purpose: Essential oils from various plants have diverse therapeutic properties and are researched extensively. They have applications in medicine, aromatherapy, microbiology, agriculture, livestock, and the food industry, benefiting the population. Methods: This systematic review followed the PRISMA verification protocol. The study focused on the anti-inflammatory effects, nutraceutical properties, antioxidant and antibacterial activity of essential oils in lemon, orange, cumin, cinnamon, coriander, rosemary, thyme, and parsley. We also looked at their presence in the diet, their effect, their mechanism of action on health, and the most important active compounds. The search was conducted in the PubMed database for the last 12 years of publications, including in vitro, in vivo, and online cell model tests. Results: Essential oils have been shown to have multiple health benefits, primarily due to their antimicrobial and anti-inflammatory effects. The mechanism of action of cinnamon oil alters bacterial membranes, modifies lipid profiles, and inhibits cell division, giving a potential benefit in protection against colitis. On the other hand, a significant improvement was observed in the diastolic pressure of patients with metabolic syndrome when supplementing them with cumin essential oil. The antimicrobial properties of coriander essential oil, especially its application in seafood like tilapia, demonstrate efficacy in improving health and resistance to bacterial infections. Cumin essential oil treats inflammation. Parsley essential oil is an antioxidant. Orange peel oil is antibacterial, antifungal, antiparasitic, and pro-oxidative. Lemon essential oil affects mouse intestinal microbiota. Thyme essential oil protects the colon against damage and DNA methylation. Carnosic acid in rosemary oil can reduce prostate cancer cell viability by modifying the endoplasmic reticulum function. Conclusion and discussion: Essential oils have many therapeutic and antiparasitic properties. They are beneficial to human health in many ways. However, to understand their potential benefits, more research is needed regarding essential oils such as coriander, parsley, rosemary, cumin, and thyme. These research gaps are relevant since they restrict understanding of the possible benefits of these crucial oils for health-related contexts.

Metabolic syndrome: Nutri-epigenetic cause or consequence?

Metabolic syndrome is a cluster of conditions that results from the interplay of genetic and environmental factors, which increase the comorbidity risk of obesity, hyperglycemia, dyslipidemia, arterial hypertension, stroke, and cardiovascular disease. In this article, we review various high-impact studies which link epigenetics with metabolic syndrome by comparing each study population, methylation effects, and strengths and weaknesses of each research. We also discuss world statistical data on metabolic syndrome incidence in developing countries where the metabolic syndrome is common condition that has significant public health implications.

"HLA-C: evolution, epigenetics, and pathological implications in the major histocompatibility complex".

HLA-C, a gene located within the major histocompatibility complex, has emerged as a prominent target in biomedical research due to its involvement in various diseases, including cancer and autoimmune disorders; even though its recent addition to the MHC, the interaction between HLA-C and KIR is crucial for immune responses, particularly in viral infections. This review provides an overview of the structure, origin, function, and pathological implications of HLA-C in the major histocompatibility complex. In the last decade, we systematically reviewed original publications from Pubmed, ScienceDirect, Scopus, and Google Scholar. Our findings reveal that genetic variations in HLA-C can determine susceptibility or resistance to certain diseases. However, the first four exons of HLA-C are particularly susceptible to epigenetic modifications, which can lead to gene silencing and alterations in immune function. These alterations can manifest in diseases such as alopecia areata and psoriasis and can also impact susceptibility to cancer and the effectiveness of cancer treatments. By comprehending the intricate interplay between genetic and epigenetic factors that regulate HLA-C expression, researchers may develop novel strategies for preventing and treating diseases associated with HLA-C dysregulation.

A report on SARS-CoV-2 first wave in Ecuador: drug consumption dynamics.

Introduction: The first COVID-19 wave in Ecuador started in March 2020 and extended until November. Several types of drugs have been proposed as a potential treatment during this period, and some affected people have self-medicated. Method: A retrospective study was conducted with 10,175 individuals who underwent RT-PCR tests for SARS-CoV-2 from July to November 2020. We compared the number of positive and negative cases in Ecuador with symptoms and drug consumption. The Chi-square test of independence compared clinical and demographic data and PCR test results. Odds ratios analyzed drug consumption dynamics. Results: Of 10,175 cases, 570 were positive for COVID-19, while 9,605 were negative. In positive cases, there was no association between the RT-PCR result and sex, age, or comorbidities. When considering demographic data, Cotopaxi and Napo had the highest rates of positive cases (25.7% and 18.8%, respectively). Manabí, Santa Elena, and Guayas regions had fewer than 10% positive cases. The Drug consumption dynamic analysis showed that negative COVID-19 cases presented higher drug consumption than positive cases. In both groups, the most consumed medication was acetaminophen. Acetaminophen and Antihistamines had higher odds of consumption in positive PCR cases than in negative. Symptoms like fever and cough were more related to positive RT-PCR results. Conclusion: The first COVID-19 wave in Ecuador has affected the provinces differently. At a national level, the consumption of drugs has been highly associated with self-medication.

Anti-inflammatory potential of processing Vernonanthura patens (Kunth) H. Rob. leaves aqueous extract.

Folk medicine uses decoctions of Vernonanthura patens (Kunth) H. Rob. leaves for healing wounds, and moderate pains. In this study, anti-inflammatory activity of decocted aqueous extract and its fractions is discussed. The fractions were obtained by liquid-liquid extraction in a separating funnel with solvents of increasing polarity: hexane, chloroform, and ethyl acetate. Antioxidant capacity, COX1, and COX2 cyclooxygenase inhibitory activities of aqueous extract (A1), aqueous (A2), and ethyl acetate (A3) fractions were assessed. A3 revealed the highest flavonoid content, and DPPH (2,2-diphenyl-1-picrylhydrazyl) scavenging activity. Nevertheless, no significance differences were observed between IC50 values of A1 and A2, and A1 showed anti-inflammatory activity with potential selectivity against COX2 enzyme, but intermediate COX1 inhibition. Further experiments are required to complement the remarkable anti-inflammatory effect of assessed aqueous extract. These results support the medicinal use of this plant species and indicate that A1 can be used as raw material for prospective nutraceutical products.

Experimental Biology 2021 Meeting Abstracts.

According to WHO's definition, self-medication is the use of drugs to treat self-diagnosed disorders or symptoms. There is a high risk of self-medication on the general population, mainly in South America. This phenomenon has been increasing with the SARS-CoV-2 outbreak, generating concerns, and anxieties in people, especially during the period of confinement. The present study used an online survey to determine the standard self-medication practices, risk, and medication-severity-index of the Ecuadorian confinement population. This study is a quantitative descriptive analysis survey designed by pharmacists and medical professionals for the self-medication during the confinement. A total of 400 Ecuadorian adults were recruited, and 380 adults accepted the digital informed consent, regardless of whether or not they had had symptoms of COVID19 during the quarantine. The survey platform used was Google Forms, and the digital social networks (WhatsApp, Facebook) were its broadcast medium. The inclusion criteria were adults who respect the confinement conditions. We excluded health professionals or who were already taking long-term medication. The data were analyzed by R, and the Hazard-Risk-Assessment-Score (HRAS) used the analysis of clinically relevant questions. There is a high risk of drug-related problems, despite the high level of education of the surveyed population (63.1% third level and 26.72% 4th level). The 55% of the surveyed Ecuadorian population had an extreme risk of self-medication (HRAS25 points), 35% prefer a drug therapy recommendation of TV, Social networks, and influencer, instead of Health professional (HRAS 20 points, moderate risk), and 39% ends up their medication before recommendations of a health professional (doctor or pharmaceutical), meaning in a moderate risk of self-medication (HRAS 12 points). There is a high risk of the Ecuadorian population to self-medication regardless of the level of education acquired. Even though the majority of the people answered requesting a prescription for the acquisition and consumption of drugs, they presented drug-related problems.

Acute oral toxicity of a novel functional drink based on Ilex guayusa, Vernonanthura patens, and cocoa husk.

A novel functional drink with nutraceutical properties was formulated from the aqueous extracts of Ilex guayusa, and Vernonanthura patens leaves, and cocoa husks. This juice contains various bioactive compounds, such as phenolic compounds and methylxanthines, with antioxidant and stimulant properties of pharmacological interest. However, it is known whether herbal extracts' interaction may have adverse toxic effects on human health. To evaluate this functional drink's innocuity, we estimated the acute oral toxicity (AOT) in experimental mice. This paper presents the AOT evaluation of two formulations of a functional drink (pre-formulation and microencapsulation) at a single dose of 2000 mg/kg of body weight (b.w.). No signs of adverse toxicity and mortality were observed after a single oral dose of 2000 mg/kg b.w. Likewise, no significant body and organ weight changes, food and water consumption behavior, and no histopathological changes were observed in the main organs evaluated. In conclusion, this functional drink can be categorized as low toxicity " according to the Globally Harmonized Classification System (GHS), making it a potential beverage with high nutritional and pharmacological value.

Reducing unnecessary antibiotic prescription through implementation of a clinical guideline on self-limiting respiratory tract infections.

BACKGROUND: Clinical guidelines (CG) are used to reduce variability in practice when the scientific evidence is sparse or when multiple therapies are available. The development and implementation of evidence-based CG is intended to organize and provide the best available evidence to support clinical decision making in order to improve quality of care. Upper respiratory tract infections (URTI) are the leading cause of misuse of antibiotics and a CG may reduce the unnecessary antibiotic prescription. METHODS: The aim of this quasi-experimental, before-after study was to analyze the short- and long-term effects of the implementation of a CG to decrease the rate of antibiotic prescription in URTI cases in the emergency department of a third level private hospital in Quito, Ecuador. The study included 444 patients with a main diagnosis of URTI. They were distributed in three groups: a baseline cohort 2011 (n = 114), a first post-implementation cohort 2011 (n = 114), and a later post-implementation cohort 2018 (n = 216). The implementation strategy consisted of five key steps: acceptance of the need for implementation of the CG, dissemination of the CG, an educational campaign, constant feedback, and sustainability of the strategy through continuous training. RESULTS: The results of this study show a 42.90% of antibiotic prescription rate before the CG implementation. After the implementation of the CG, the prescription rate of antibiotics was significantly reduced by 24.5% (42.9% vs 18.4%, p<0.0001) and the appropriate antibiotic prescription rate was significantly increased by 44.2% (22.4% vs 66.6%, p<0.0001) in the first post-implementation cohort 2011. There was not a significant difference in antibiotic prescription rate and appropriate antibiotic prescription rate between two post-implementation cohorts: 18.4% vs 25.9% (p = 0.125) and 66.6% vs 50% (p = 0.191), respectively. CONCLUSIONS: The implementation of CGs decreases the rate of antibiotic prescription in URTI cases. The results are remarkable after early implementation, but the effect persists over time. The emphasis must shift from guideline development to strategy implementation.

Knowledge, Attitudes and Practices in Antibiotic Use in Family Medicine Students.

BACKGROUND: Inappropriate prescriptions of antibiotics lead to ineffective and unsafe treatments and worsening of diseases. Medical students may have deficiencies in their prescription skills and they may need further training in the use of antibiotics for their practice. Medical skills in prescribing antibiotics can be improved through continuous medical education. The aim of this study was to assess the current levels of knowledge, attitudes, and practices (KAP) in antibiotic prescription in upper respiratory tract infections (URTI) among postgraduate family medicine students in Ecuador. METHODS: A cross-sectional study with an on-line survey, based on micro-curricular contents, to evaluate KAPs regarding antibiotic prescription in URTI among postgraduate family medicine students in 5 provinces of Ecuador. RESULTS: Two hundred and seventy-three physicians responded (94.1%). Most physicians treated between 1 and 5 URTI cases per day. The odds for inadequate knowledge and inappropriate practices in URTI among postgraduate family medicine students were 8.74 (95%CI, 4.94-15.46, P < .001) and 5.99, (IC95%, 2.66-13.50, P < .001) in physicians who were students of the first half of the study program. CONCLUSION: The knowledge in URTI was limited among physicians. Nonetheless, they expressed a positive attitude toward not using antibiotics in URTI. A postgraduate program can significantly improve the knowledge and practices related to antibiotic prescriptions in URTI.

Análisis molecular de Helicobacter pylori (genes de patogenicidad) en biopsias gástricas de pacientes de la Sierra y Oriente Ecuatorianos / Molecular analysis of Helicobacter pylori (pathogenicity genes) in gastric biopsies of patients from the Ecuadorian Sierra and Oriente

Antecedentes: Helicobacter pylori es una bacteria Gram negativa, reconocida como la causa de la úlcera péptica (UP) y cáncer el gástrico (CG). Se han identificado genes de virulencia asociados con la patogenicidad del H. pylori incluyendo la isla de patogenicidad cagA y la citotoxina vacuolizante A (vacA). La frecuencia de los genes de patogenicidad se ha asociado con la localización geográfica y condiciones de vida de las personas. Pocos estudios en el Ecuador, han demostrado la relación entre los genes de patogenicidad de H. pylori y regiones geográficas de diferente altitud. Este estudio analizó los genes de patogenicidad de biopsias gástricas dos parroquias del Ecuador: una ubicada en la altura, Zumbahua (Sierra Central) y otra a nivel del mar, Shushufindi (Amazonía). Métodos: Se obtuvieron 127 muestras de biopsias gástricas embebidas en parafina de sujetos provenientes de Zumbahua (n = 90) y Shushufindi (n = 37). Mediante un análisis histopatológico se determinó la presencia de la infección y alteraciones patológicas tisulares. Se seleccionaron las muestras de los pacientes con mayor índice de infección por H. pylori (++ y +++ en el examen histopatológico) para el análisis molecular del H. pylori; se aisló su ADN y se evaluaron los genes de patogenicidad por PCR. Resultados: Se determinó la presencia de 5 casos de cáncer gástrico en la parroquia de Zumbahua, con mayor frecuencia en hombres que en mujeres. En la parroquia de Sushufindi hubo mayor prevalencia de infección por H. pylori comparada con Zumbahua. El análisis molecular de los genes de patogenicidad determinó que hubo una mayor expresión de estos en las muestras provenientes de la parroquia de Zumbahua; el 20% de las muestras amplificaron para vacAm1, 8.57% para vacAs1 y el 20% para vacAs2; mientras que para Shushufindi, únicamente el 8.0% amplificó para el gen vacAm1. Conclusiones: La prevalencia de infección por H. pylori en las muestras de las parroquias estudiadas es alta. Los genes de patogenicidad asociados con mayor virulencia provinieron de Zumbahua así como también las muestras con cáncer. Por otro lado, en las muestras de Shushufindi los genes de patogenicidad fueron menos virulentos y no hubo casos de malignidad. Es necesario establecer sistemas de tamizaje tanto para detectar cepas de H. pylori con genes de virulencia como para la detección temprana del cáncer gástrico.

Rab7b participation on the TLR4 (Toll-like receptor) endocytic pathway in Shiga toxin-associated Hemolytic Uremic Syndrome (HUS).

BACKGROUND: The inflammatory response of the host to Shiga toxin and/or lipopolysaccharide (LPS) of Escherichia coli (E. coli) is included in (HUS). The TLR4-LPS complex is internalized and TLR4 induced inflammatory signaling is stopped by targeting the complex for degradation. Rab7b, a small guanosine triphosphatase (GTPase) expressed in monocytes, regulates the later stages of the endocytic pathway. OBJECTIVE: we studied the Rab7b participation on the TLR4 endocytic pathway and its effect on monocyte cytokine production along the acute course of pediatric Shiga toxin-associated HUS. METHODS AND RESULTS: Monocytes were identified according to their positivity in CD14 expression. Surface TLR4 expression in monocytes from 18 HUS patients significantly increased by day 1 to 6, showing the highest increase on day 4 compared to monocytes of 10 healthy children. Significant higher surface TLR4 expression was accompanied by increased proinflammatory intracellular cytokines, tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6). In contrast, after these time points, surface TLR4 expression and intracellular TNF-α levels, returned to near control levels after 10 days. Furthermore, confocal immunofluorescence microscopy proved colocalization of increased intracellular TLR4/Rab7b determined by Pearson's coefficient in monocytes from HUS patients from day 1 on the highest colocalization of both proteins by day 4. Decreased TLR4/Rab7b colocalization was shown 10 days after HUS onset. CONCLUSION: The colocalization of TLR4 and Rab7b allows us to suggest Rab7b participation in the control of the TLR4 endocytic pathway in HUS patient monocytes. A consequential fall in cytokine production throughout the early follow up of HUS is demonstrated.

Review: Prevalence and dynamics of Helicobacter pylori infection during childhood.

INTRODUCTION: Long-term persistent Helicobacter pylori infection has been associated with ulceropeptic disease and gastric cancer. Although H. pylori is predominantly acquired early in life, a clear understanding of infection dynamics during childhood has been obfuscated by the diversity of populations evaluated, study designs, and methods used. AIM: Update understanding of true prevalence of H. pylori infection during childhood, based on a critical analysis of the literature published in the past 5 years. METHODS: Comprehensive review and meta-analysis of original studies published from 2011 to 2016. RESULTS: A MEDLINE® /PubMed® search on May 1, 2016, using the terms pylori and children, and subsequent exclusion, based on abstract review using predefined criteria, resulted in 261 citations. An Embase® search with the same criteria added an additional 8 citations. In healthy children, meta-analysis estimated an overall seroprevalence rate of 33% (95% CI: 27%-38%). Seven healthy cohort studies using noninvasive direct detection methods showed infection prevalence estimates ranging from 20% to 50% in children ≤5 and 38% to 79% in children >5 years. The probability of infection persistence after a first positive sample ranged from 49% to 95%. Model estimates of cross-sectional direct detection studies in asymptomatic children indicated a prevalence of 37% (95% CI: 30%-44%). Seroprevalence, but not direct detection rates increased with age; both decreased with increasing income. The model estimate based on cross-sectional studies in symptomatic children was 39% (95% CI: 35%-43%). CONCLUSIONS: The prevalence of H. pylori infection varied widely in the studies included here; nevertheless, model estimates by detection type were similar, suggesting that overall, one-third of children worldwide are or have been infected. The few cohort and longitudinal studies available show variability, but most studies, show infection rates over 30%. Rather surprisingly, overall infection prevalence in symptomatic children was only slightly higher, around 40%. Studies including only one positive stool sample should be interpreted with caution as spontaneous clearance can occur.

Helicobacter pylori Infection Is Associated with Decreased Expression of SLC5A8, a Cancer Suppressor Gene, in Young Children.

Background:Helicobacter pylori infects half of the world's population and causes gastric cancer in a subset of infected adults. Previous blood microarray findings showed that apparently healthy children, persistently infected with H. pylori have differential gene expression compared to age-matched, non-infected children. SLC5A8, a cancer suppressor gene with decreased expression among infected children, was chosen for further study based on bioinformatics analysis. Methods: A pilot study was conducted using specific qRT-PCR amplification of SLC5A8 in blood samples from H. pylori infected and non-infected children, followed by a larger, blinded, case-control study. We then analyzed gastric tissue from H. pylori infected and non-infected children undergoing endoscopy for clinical purposes. Results: Demographics, clinical findings, and family history were similar between groups. SLC5A8 expression was decreased in infected vs. non-infected children in blood, 0.12 (IQR: 0-0.89) vs. 1.86 (IQR: 0-8.94, P = 0.002), and in gastric tissue, 0.08 (IQR: 0.04-0.15) vs. 1.88 (IQR: 0.55-2.56; P = 0.001). Children who were both stool positive and seropositive for H. pylori had the lowest SLC5A8 expression levels. Conclusions:H. pylori infection is associated with suppression of SCL5A8, a cancer suppressor gene, in both blood and tissue samples from young children. Key Points: Young children, persistently infected with Helicobacter pylori show decreased expression of SLC5A8 mRNA in both blood and tissue samples as compared to non-infected children.

Small structural changes on a hydroquinone scaffold determine the complex I inhibition or uncoupling of tumoral oxidative phosphorylation.

Mitochondria participate in several distinctiveness of cancer cell, being a promising target for the design of anti-cancer compounds. Previously, we described that ortho-carbonyl hydroquinone scaffold 14 inhibits the complex I-dependent respiration with selective anti-proliferative effect on mouse mammary adenocarcinoma TA3/Ha cancer cells; however, the structural requirements of this hydroquinone scaffold to affect the oxidative phosphorylation (OXPHOS) of cancer cells have not been studied in detail. Here, we characterize the mitochondrial metabolism of TA3/Ha cancer cells, which exhibit a high oxidative metabolism, and evaluate the effect of small structural changes of the hydroquinone scaffold 14 on the respiration of this cell line. Our results indicate that these structural changes modify the effect on OXPHOS, obtaining compounds with three alternative actions: inhibitors of complex I-dependent respiration, uncoupler of OXPHOS and compounds with both actions. To confirm this, the effect of a bicyclic hydroquinone (9) was evaluated in isolated mitochondria. Hydroquinone 9 increased mitochondrial respiration in state 4o without effects on the ADP-stimulated respiration (state 3ADP), decreasing the complexes I and II-dependent respiratory control ratio. The effect on mitochondrial respiration was reversed by 6-ketocholestanol addition, indicating that this hydroquinone is a protonophoric uncoupling agent. In intact TA3/Ha cells, hydroquinone 9 caused mitochondrial depolarization, decreasing intracellular ATP and NAD(P)H levels and GSH/GSSG ratio, and slightly increasing the ROS levels. Moreover, it exhibited selective NAD(P)H availability-dependent anti-proliferative effect on cancer cells. Therefore, our results indicate that the ortho-carbonyl hydroquinone scaffold offers the possibility to design compounds with specific actions on OXPHOS of cancer cells.

Systematics of the Osteocephalusbuckleyi species complex (Anura, Hylidae) from Ecuador and Peru.

We present a new phylogeny, based on DNA sequences of mitochondrial and nuclear genes, for frogs of the genus Osteocephalus with emphasis in the Osteocephalus buckleyi species complex. Genetic, morphologic, and advertisement call data are combined to define species boundaries and describe new species. The phylogeny shows strong support for: (1) a basal position of Osteocephalus taurinus + Osteocephalus oophagus, (2) a clade containing phytotelmata breeding species, and (3) a clade that corresponds to the Osteocephalus buckleyi species complex. Our results document a large proportion of hidden diversity within a set of populations that were previously treated as a single, widely distributed species, Osteocephalus buckleyi. Individuals assignable to Osteocephalus buckleyi formed a paraphyletic group relative to Osteocephalus verruciger and Osteocephalus cabrerai and contained four species, one of which is Osteocephalus buckleyisensu stricto and three are new. Two of the new species are shared between Ecuador and Peru (Osteocephalus vilmaesp. n. and Osteocephalus cannatellaisp. n.) and one is distributed in the Amazon region of southern Peru (Osteocephalus germanisp. n.) We discuss the difficulties of using morphological characters to define species boundaries and propose a hypothesis to explain them.

Efecto antiinflamatorio y antimicótico del extracto alcohólico y composición química del aceite de hojas de Conyza bonariensis (L) Cronquist (canilla de venado) / Anti-inflammatory and antimitotic effect of the alcoholic extract and chemical composition of the oil from Conyza bonariensis (L) Cronquist (deer shinbone) leaves

Introducción: la especie Conyza bonariensis (L.) Cronquist, comúnmente llamada canilla de venado, es de origen americano. Objetivos: contribuir al estudio farmacológico y toxicológico de los extractos alcohólicos y a la evaluación química del aceite de hojas de C bonariensis de Ecuador. Métodos: se emplearon 2 tinturas a 50 por ciento con etanol a 70 y 90 por ciento de hojas secas. Se realizó un estudio fitoquímico a través del sistema acoplado de cromatografía gaseosa-espectrometría de masas, se determinó el efecto antimicótico mediante inhibición de dilución en placa con siembra en superficie por diseminación, y antiinflamatorio en el modelo de edema de la oreja inducido por aceite de Croton en ratones albinos de la línea OF1. Resultados: el tamizaje fitoquímico mostró presencia de aceites esenciales, triterpenos-esteroides, compuestos fenólicos, flavonoides, quinonas, antocianidinas, saponinas y compuestos reductores. Se determinó estructuras a 32 componentes del aceite esencial extraído de las hojas y el sesquiterpeno: trans-b-farneseno resultó el componente mayoritario con 70,82 por ciento, este compuesto se informa por primera vez en esta especie. El extracto hidroalcohólico (25 µL a cada lado de las orejas tratadas) inhibió significativamente la inflamación comparado con bencidamina. Conclusiones: los extractos hidroalcohólicos mostraron efecto antiinflamatorio en las condiciones experimentales del estudio y se determinó la composición química del aceite de hojas de C. bonariensis(AU)

Introduction: the Conyza bonariensis (L) Cronquist species, usually called deer shinbone is of American origin. Objectives: to contribute to the pharmacological and toxicological study of alcoholic extracts and to the chemical assessment of C. bonariensis leaf oil from Ecuador. Methods: two 50 percent tinctures of 70 percent and 90 percent ethanol content in dry leaves were used. A phytochemical study was conducted by an combined system of mass spectrometry and gas chromatography; the amniotic effect was determined using plate dilution inhibition with surface culturing by dissemination, and the anti-inflammatory effect in the Croton oil induced ear edema model in OF1 line albino mice. Results: the phytochemical screening showed the presence of essential oils, triterpene-steroids, phenolic compounds, flavonoids, quinones, anthocyanidines, saponins and reducing compounds. The structures of 32 essential oil components extracted from the leaves were identified along with and the sesquiterpene trans-b-farnesene that was the major component (70.82 percent), this compound is reported for the first time in this species. The hydroalcoholic extract (25 µL on each side of the treated ears) significantly inhibited inflammation compared to benzidamine. Conclusions: hydroalcoholic extracts showed anti-inflammatory effect in the experimental conditions of the study and the chemical composition of C. bonariensis leaf oil was identified(AU)

Efecto antiinflamatorio y antimicótico del extracto alcohólico y composición química del aceite de hojas de Conyza bonariensis (L) Cronquist (canilla de venado) / Anti-inflammatory and antimitotic effect of the alcoholic extract and chemical composition of the oil from Conyza bonariensis (L) Cronquist (deer shinbone) leaves

Introducción: la especie Conyza bonariensis (L.) Cronquist, comúnmente llamada canilla de venado, es de origen americano. Objetivos: contribuir al estudio farmacológico y toxicológico de los extractos alcohólicos y a la evaluación química del aceite de hojas de C bonariensis de Ecuador. Métodos: se emplearon 2 tinturas a 50 por ciento con etanol a 70 y 90 por ciento de hojas secas. Se realizó un estudio fitoquímico a través del sistema acoplado de cromatografía gaseosa-espectrometría de masas, se determinó el efecto antimicótico mediante inhibición de dilución en placa con siembra en superficie por diseminación, y antiinflamatorio en el modelo de edema de la oreja inducido por aceite de Croton en ratones albinos de la línea OF1. Resultados: el tamizaje fitoquímico mostró presencia de aceites esenciales, triterpenos-esteroides, compuestos fenólicos, flavonoides, quinonas, antocianidinas, saponinas y compuestos reductores. Se determinó estructuras a 32 componentes del aceite esencial extraído de las hojas y el sesquiterpeno: trans-b-farneseno resultó el componente mayoritario con 70,82 por ciento, este compuesto se informa por primera vez en esta especie. El extracto hidroalcohólico (25 µL a cada lado de las orejas tratadas) inhibió significativamente la inflamación comparado con bencidamina. Conclusiones: los extractos hidroalcohólicos mostraron efecto antiinflamatorio en las condiciones experimentales del estudio y se determinó la composición química del aceite de hojas de C. bonariensis

Introduction: the Conyza bonariensis (L) Cronquist species, usually called deer shinbone is of American origin. Objectives: to contribute to the pharmacological and toxicological study of alcoholic extracts and to the chemical assessment of C. bonariensis leaf oil from Ecuador. Methods: two 50 percent tinctures of 70 percent and 90 percent ethanol content in dry leaves were used. A phytochemical study was conducted by an combined system of mass spectrometry and gas chromatography; the amniotic effect was determined using plate dilution inhibition with surface culturing by dissemination, and the anti-inflammatory effect in the Croton oil induced ear edema model in OF1 line albino mice. Results: the phytochemical screening showed the presence of essential oils, triterpene-steroids, phenolic compounds, flavonoids, quinones, anthocyanidines, saponins and reducing compounds. The structures of 32 essential oil components extracted from the leaves were identified along with and the sesquiterpene trans-b-farnesene that was the major component (70.82 percent), this compound is reported for the first time in this species. The hydroalcoholic extract (25 µL on each side of the treated ears) significantly inhibited inflammation compared to benzidamine. Conclusions: hydroalcoholic extracts showed anti-inflammatory effect in the experimental conditions of the study and the chemical composition of C. bonariensis leaf oil was identified