Therapeutic Applications of Essential Oils from Native and Cultivated Ecuadorian Plants: Cutaneous Candidiasis and Dermal Anti-Inflammatory Activity.

Essential oils are a complex mixture of aromatic substances whose pharmacological actions, including antimicrobial, antioxidant, anticancer, and anti-inflammatory activities, have been widely reported. This study aimed to evaluate the anti-Candida and dermal anti-inflammatory activity of essential oils from native and cultivated Ecuadorian plants. Essential oils from Bursera graveolens, Dacryodes peruviana, Mespilodaphne quixos, and Melaleuca armillaris were isolated by hydrodistillation and were characterized physically and chemically. Its tolerance was analyzed by in vitro and in vivo studies. The antifungal activity was studied against Candida albicans, Candida glabrata, and Candida parapsilosis, whereas the anti-inflammatory effect was evaluated by a mouse ear edema model. The main compounds were limonene, α-phellandrene, (E)-methyl cinnamate, and 1,8-cineole, respectively. All essential oils showed high tolerability for skin application, antifungal activity against the three Candida strains, and anti-inflammatory efficacy by decreasing edema and overexpression of pro-inflammatory cytokines. Dacryodes peruviana essential oil showed the highest antifungal activity. On the other hand, Dacryodes peruviana and Melaleuca armillaris showed the greatest anti-inflammatory potential, decreasing edema by 53.3% and 65.25%, respectively, and inhibiting the overexpression of TNF-α, IL-8, IL-17A, and IL-23. The results suggest that these essential oils could be used as alternative therapies in the treatment of both cutaneous candidiasis and dermal inflammation.

Characterization and In Vivo Anti-Inflammatory Efficacy of Copal (Dacryodes peruviana (Loes.) H.J. Lam) Essential Oil.

Essential oils are natural aromatic substances that contain complex mixtures of many volatile compounds frequently used in pharmaceutical and cosmetic industries. Dacryodes peruviana (Loes.) H.J. Lam is a native species from Ecuador whose anti-inflammatory activity has not been previously reported, thus the aim of this study was to evaluate the anti-inflammatory activity of D. peruviana essential oil. To that end, essential oil from D. peruviana fruits was isolated by hydrodistillation and characterized physically and chemically. The tolerance of the essential oil was analyzed by cytotoxicity studies using human keratinocytes. The anti-inflammatory activity was evaluated by an arachidonic acid-induced edema model in mouse ear. The predominant compounds in D. peruviana essential oil were α-phellandrene, limonene, and α-pinene, with the three compounds reaching approximately 83% of the total composition. Tolerance studies showed high biocompatibility of this essential oil with human keratinocytes. In vivo studies demonstrated a moisturizing effect and an alleviation of several events occurred during the inflammatory process after topical treatment with D. peruviana essential oil such as decline in skin edema; reduction in leukocytic infiltrate; and decrease in inflammatory cytokines TNFα, IL-8, IL-17A, and IL-23. Therefore, this essential oil could be an attractive treatment for skin inflammation.

Comparative Study of Donepezil-Loaded Formulations for the Treatment of Alzheimer's Disease by Nasal Administration.

Alzheimer's disease is characterized by a progressive deterioration of neurons resulting in a steady loss of cognitive functions and memory. Many treatments encounter the challenge of overcoming the blood-brain barrier, thus the intranasal route is a non-invasive effective alternative that enhances the drug delivery in the target organ-the brain-and reduces the side effects associated with systemic administration. This study aimed at developing intranasal gels of donepezil as an approach to Alzheimer's disease. Three different gels were elaborated and characterized in terms of pH, morphology, gelation temperature, rheology, and swelling. An in vitro release study and an ex vivo permeation in porcine nasal mucosa were conducted on Franz diffusion cells. The tolerability of the formulations was determined by the cytotoxicity in human nasal cells RPMI 2650. Results showed that pluronic gels exhibit the higher release rate and enhanced permeation compared to chitosan gel. Moreover, the combination of Pluronic F-127 and Transcutol® P exerted a synergic effect on the permeation of donepezil through the nasal mucosa. The resulting gels showed suitable tolerance in the RPMI 2650 cell line and physicochemical characteristics for intranasal delivery, and thus gel formulations administered by nasal mucosa could be an alternative strategy to improve the bioavailability of donepezil.

Development of a Topical Amphotericin B and Bursera graveolens Essential Oil-Loaded Gel for the Treatment of Dermal Candidiasis.

The higher molecular weight and low solubility of amphotericin B (AmB) hinders its topical administration. The aim of this study was to incorporate Bursera graveolens essential oil into an AmB topical gel (AmB + BGEO gel) in order to promote the diffusion of the drug through the skin in the treatment of cutaneous candidiasis. AmB + BGEO gel formulation was determined using a factorial experiment. Physical and chemical parameters, stability, in vitro release profile and ex vivo permeation in human skin were evaluated. In vitro antimicrobial activity was studied using strains of C. albicans, C. glabrata and C. parapsilosis. The tolerability was evaluated using in vitro and in vivo models. AmB + BGEO gel presented appropriate characteristics for topical administration, including pH of 5.85, pseudoplastic behavior, optimal extensibility, as well as high stability and acceptable tolerability. In vitro release studies showed that the formulation releases the drug following a Boltzmann sigmoidal model. Finally, AmB + BGEO gel exhibited higher amount of drug retained inside the skin and lower Minimum Inhibitory Concentration than a formulation sans essential oil. Therefore, these results suggest that the incorporation of B. graveolens essential oil in the formulation could be used as strategy to promote a local effect in the treatment of cutaneous candidiasis.

Caracterización de Cepas de Virus Sincitial Respiratorio en el Gran Buenos Aires y Aspectos de su Diseminación / Characterization of Respiratory Syncytial Virus Strains in the Greater Buenos Aires and Spreading Aspects

INTRODUCCIÓN: El virus sincitial respiratorio (VSR) es el agente viral más frecuente de infecciones respiratorias agudas bajas (IRAB) en la primera infancia y el mayor responsable de las hospitalizaciones en el período invernal. OBJETIVOS: Describir las características de los brotes de VSR en la Zona Sanitaria VI de la provincia de Buenos Aires, establecer la diversidad de las cepas circulantes y realizar el análisis bioinformático y filogeográfico de las secuencias de la glicoproteína G. MÉTODOS: Se estudió a pacientes pediátricos internados con presentación compatible con IRAB durante dos picos epidémicos (2014-2015) en cuatro hospitales. Se recopilaron datos clínicos, demográficos y socio-sanitarios, y se detectaron patógenos virales en aspirados nasofaríngeos de estos pacientes por inmunofluorescencia (IF), obteniéndose la secuencia del gen de la proteína G en los VSR positivos. RESULTADOS: De 1296 casos estudiados, 317 fueron positivos para algún agente viral. De ellos, 266 (84%) fueron VSR positivos. Se hallaron asociaciones significativas entre las poblaciones positivas y negativas para VSR. Una tendencia al hacinamiento y vivienda precaria en los casos VSR positivos fue reflejada en los estudios filogeográficos. CONCLUSIONES: Los datos de firma molecular permitieron trazar orígenes y vías de diseminación del VSR. Esto ayuda a señalar zonas y situaciones de vulnerabilidad, estableciendo la población primaria blanco de planes de vacunación u otras medidas profilácticas.

INTRODUCTION: The respiratory syncytial virus (RSV) is the most frequent viral agent associated to acute lower respiratory infections (ALRIs) in early childhood, being the main responsible for hospitalizations during winter. OBJECTIVES: To describe the characteristics of RSV outbreaks in the Health Area VI of Buenos Aires Province, to establish the diversity of circulating strains and to perform a bioinformatic and phylogeographic analysis of glycoprotein G sequences. METHODS: Pediatric inpatients with ALRI-compatible x|presentation during two epidemic peaks (2014-2015) were studied in four hospitals. Clinical, demographic and socio-sanitary data were collected, viral pathogens were detected by immunofluorescence (IF), and the sequence of the G protein gene was obtained in the positive RSVs. RESULTS: From 1296 cases, 317 were positive for some viral agent and 266 (84%) out of these were RSV positive. Significant associations were found among the positive and negative populations for RSV. A trend towards overcrowding and precarious housing in positive RSV cases was reflected in phylogeographic studies. CONCLUSIONS: The molecular signature data allowed tracing origins and routes of RSV dissemination. This helps identify areas and situations of vulnerability, establishing the primary target population for vaccination plans or other prophylactic measures.

Unravelling respiratory syncytial virus outbreaks in Buenos Aires, Argentina: Molecular basis of the spatio-temporal transmission.

Respiratory syncytial virus (RSV) is the main viral cause of hospitalization due to acute lower respiratory tract infections in infants worldwide. Several vaccines against RSV are under research and development, which are about to be approved. We evaluated transmission patterns in different settings to determine age-specific vaccination targets from a viral perspective. We sequenced the G glycoprotein's ectodomain of a constant clinical sampling between two epidemic outbreaks in a limited geographical region and performed phylogeographic analyses. We described a spatio-temporal transmission between local strains, which were originated in the center of the analyzed area and then spread to others. Interestingly, that central area reported the highest population density of the region and also showed overcrowding. This information should be considered by public health systems to evaluate vaccination at all ages in those areas to decrease viral transmission and in lower density populations only susceptible children should be vaccinated.

Frecuencia de la mutación 185delAG en el gen BRCA1 en mujeres chilenas sanas con antecedentes familiares de cáncer de mama / Frequency of the 185delAG mutation in the BRCA1 gene in chilean healthy women with family history of breast cancer

Background: Breast cancer is the most common malignancy among women, and is the second cause of cancer mortality among Chilean women. Female mortality due to breast cancer in Chile has shown a steady increase from 9.5 deaths per 100.000 women in 1985 to 12.8 deaths per 100.000 in 1995. A family history of breast cancer is one of the main risk factors for the development of the disease. BRCA1 and BRCA2 are two major hereditary breast cancer susceptibility genes. Mutations in these genes are associated to inherited breast cancer; 664 predisposing mutations have been described, but in specific populations only some of them, such as 185delAG have been found to be associated with susceptibility to breast cancer. Aim: To establish the frequency of the 185delAG mutation in the BRCA1 gene in Chilean healthy women with a family history of breast cancer. Patients and Methods: The 185delAG mutation was studied by mismatch polymerase chain (PCR) reaction in 382 Chilean healthy women with at least two relatives affected with breast cancer. The PCR products were digested with the restriction enzyme HinfI. Digestion of the normal allele (170 pb fragment) produces a 150 pb fragment; the PCR product for the mutant allele does not contain a site for HinfI and therefore remains as a 170 bp fragment after digestion. Results: One of the 382 healthy women presented the fragment of 170 pb after digestion with HinfI suggesting that she was heterozygous carrier for this mutation. The mutant patient had a mammography without suspicion of cancer. Conclusions: The frequency of the 185delAG mutation in BRCA1 was 0.26 percent (1/382) in Chilean healthy women with a family history of breast cancer

Síndrome de X frágil: análisis clínico en 300 pacientes con retardo mental inespecífico en la población chilena / Fragile X syndrome: clinical analysis of 300 Chilean patients with unspecific mental retardation

Background: Fragile X syndrome is the most important cause of sex linked mental retardation and the second of chromosomal origin, after Down syndrome. Aim: To apply the modified Hagerman score to patients with mental retardation and to relate clinical findings with cytogenetic and molecular diagnosis. Patients and methods: The modified Hagerman score was applied to 214 male and 86 female patients with mental retardation. The clinical variables in non fragile X and fragile X cases, determined by molecular and cytogenetic methods, were compared. Results: The score in 210 non fragile X males was 10.5 + 3.7 (range 3 23), compared to 21.4 + 2.1 (range 19 to 23) in the four fragile X patients. All fragile X patients had mental retardation, attention deficits, hyperactivity disorders, hand biting and poor visual contact. Hand biting, flapping and persevering speech were observed in a significantly higher number of fragile X males. Only one of 86 females had fragile X syndrome. Her most relevant findings were a long face and high forehead, an attention deficit, hyperactivity and poor visual contact. No clinical differences with other mentally retarded females were found. Condusions: Approximately 5 percent of institutionalized males with mental retardation have a fragile X syndrome

Pesquisa molecular y clínica del síndrome de X-frágil en 300 pacientes con retardo mental inespecífico / Clinical and metabolic screening for fragile X syndrome in 300 patients with unspecific mental retardation

Background: Fragile X syndrome is the most freqent cause of mental retardation linked to the X chromosome. In the majority of cases, the mutation responsible for the syndrome is an expansion of the trinucleotide repeat (CGG)n, present in the 5' region of exon 1 of the gene for mental retardation associated with fragile X syndrome (FMR-1). Aim: To report the results of a fragile X screening in patients with mental retardation. Patients and methods: Fragile X screening using polymerase chain reaction methods was done in 386 X chromosomes from 300 patients (214 male), aged 4 to 26 years old. The modified Hagerman test was applied to male patients. Hybridization techniques were applied in a subgroup of 51 patients. Results: (CGG)n 30 was the allele found with the highest frequency in 50.2percent of patients. (CGG)n 29 was found in 29percent of patients. One subject had an allele with 46 CGG repeats, which corresponds to the gray zone. Hybridization studies were highly concordant with PCR, detecting four males with fragile X syndrome and a carrier female. The average clinical score of mental retardation not due to fragile X syndrome was 10.3 ñ 3.4 (range 3 to 23), and 97percent of males had a score below 19. The concordance between scores over 20 and molecular genotype was 98percent. Conclusions: The distribution of (CGG)n repeats, observed in this study, was significantly different to that previously reported for a normal Chilean population. The dispersion of molecular status and clinical score was lower than previously described using cytogenetic techniques

La variación genética de MSX1 presenta un dimorfismo sexual en la fisura labiopalatina no sindrómica en la población chilena / Genetic variation of MSX1 has a sexual dimorphism in non syndromic cleft palate in the Chilean population

Background: Recent studies in mice have demonstrated that the Msx-1 homebox gene is implicated in cleft palate. Thus, it has been suggested that its human homologue, MSX1 (HOX-7), located in chromosome 4 could be involved in the etiology of non syndromic cleft lip palate. Aim: To study the linkage between non syndromic cleft palate and variations of MSX1 gene. Patients and methods: Seventy three patients with non syndromic cleft lip palate (34 simplex and 37 multiplex), 127 unaffected relatives of the cases (61 relatives of simplex cases and 66 relatives of multiplex cases) and 77 controls were studied. DNA was extracted from leukocytes and the intragenic microsatellite sequence was amplified by PCR. Results: A polymorphism of four alleles was observed, 1 (175 bp), 2 (173 bp), 3 (171 bp) and 4 (169 bp). Alleles 2 and 4 showed a joint variation in males with multiplex cleft lip palate and in their respective unaffected male relatives, that was significant when compared with male controls. Instead, the joint variation of alleles 1 and 4 of unaffected female relatives had significant differences with female controls. Females with multiplex cleft lip palate differed from female controls only in allele 1. Conclusions: These results support the hypothesis of a genetic heterogeneity in the etiology of non syndromic cleft lip palate

Asociación entre la variación genética de MSX1 (Hox-7) y la fisura labiopalatina no sindrómica en población chilena / Association between the genetic variation of MSX1 (Hox-7) and non syndromic cleft lip palate in chilean subjects

Background: Homeotic genes have regulatory functions during development. It has been postulated that the human Msx-1 homeotic gene can be involved in the etiology of non syndromic cleft lip palate, since its homologous Msx-1 is involved in cleft palate of mice. Aim: To perform an association analysis between the genetic variation of Msx-1 and non syndromic cleft lip palate in Chilean subjects. Patients and methods: Seventy patients with non syndromic cleft lip palate, 136 healthy relatives of these patients and 69 non related normal individuals were studied. CA microsatellite in Msx- gene, that was amplified with PCR, was studied. Results: No differences in the genetic frequencies of Msx-1 alleles, were observed in the three groups studied. Allelic heterogeneity for allele 2 seems to be related to cases of non syndromic cleft lip palate from multiplex families and heterogeneity for allele 3 is related with simplex families cases. Conclusions: These results seem to support the hypothesis of genetic heterogeneity in the etiology of non syndromic cleft lip palate

Aspectos genético moleculares del síndrome de X-frágil / Molecular and genetic features of fragile X syndrome

The fragile X syndrome is the most frequent cause of sexlinked mental retardation. In the majority of the cases the mutation responsible for the Martin Bell syndrome is produced when an expansion of the (CGG)n repetition is present in the region 5' of the exón 1 of the gene for X-fragile mental retardation 1 (FMR1), together with a hipermethylation in the CpG promoter region of the gene. The result of this situation is the absence of the FMRP protein coded by the gene. The correlation between length of the (CGG)n sequences and the X-fragile phenotype has permitted a more precise diagnosis of affected and carrier individuals by means of direct DNA analysis. Neverthless the molecular genetic basis of the instability and expansion of the (CGG)n sequences represents a problem not yet resolved. Two morphologic microsatellite (AC)n repetitions, FRAXAC1 and FRAXAC2 that flanck the FMR-1 gene have been recently described. It has been suggested that some haplotypes of FRAXAC1 and FRAXAC2 could be associated to long (CGG)n repetitions and thet these haplotypes would confer more instability to the repeated fragment thus increasing the probability of expansion. It has also been described that the (CGG)n repetition of the FMR-1 gene is interrupted by AGG trinucleotides and that the loss of one AGG would be an important mutational event in the generation of predisposing unstable alleles of the X-fragile syndrome

Tiempos de la erupción dentaria temporal en pacientes con síndrome de Down / Time of temporal dental eruption in patients with Down's syndrome

Se examinaron 255 pacientes con síndrome de Down que asisten a escuelas especiales de Santiago, Chile, con el propósito de describir los tiempos de erupción para la dentición decidual y compararlos con los de la población normal. En los niños con síndrome de Down se observa retraso significativo en la erupción de los siguientes dientes: el incisivo central derecho (15,27 ñ 5,515 meses) y los incisivos laterales derecho e izquierdo (18,44 ñ 9,652 y 18,15 ñ 11,82 meses) y los caninos derecho e izquierdo (25,87 ñ 7,667 y 26,65 ñ 7,431 meses, respectivamente) en el maxilar inferior. Las niñas con síndrome de Down presentan retraso significativo en la erupción de los incisivos laterales derecho e izquierdo (17,31 ñ 14,42 y 17,31 ñ 14,42 meses, respectivamente), los caninos derecho e izquierdo (30,70 ñ 6,454 y 30,60 ñ 7,249 meses, respectivamente, y el primer molar izquierdo 25,87 ñ 14,34 meses) en el maxilar superior; el incisivo central izquierdo (12,02 ñ 7,286 meses), los incisivos laterales derecho e izquierdo (27,59 ñ 10,01 y 24,66 ñ 23,86 meses, respectivamente), los caninos derecho e izquierdo (27,83 ñ 11,25 y 28,80 ñ 10,60 meses, respectivamente) y el segundo molar derecho (28,83 ñ 3,454 meses) en el maxilar inferior. La secuencia de erupción en los niños con s. de Down fue similar a la observada en los normales. Las edades de erupción mostraron una distribución gaussiana y las varianzas existentes en el grupo con s. de Down fueron significativamente superiores a las de los normales

Síndrome X frágil: estudio clínico citogenético y molecular en una familia chilena / Fragile X syndrome: cytogenetical clinic and molecular study in a chilean family

El síndrome del cromosoma X-frágil es la causa más frecuente de retardo mental hereditario en el varón y se asocia a un marcador citogenético ubicado en la banda Xq 27.3 (FRAXA) del cromosoma X. Durante muchos años el diagnostico estuvo basado en la identificación citogenética del marcador; sin embargo, este método no ha resultado confiable para el diagnóstico prenatal y la detección de portadores masculinos o femeninos clínicamente normales. El descubrimiento de las bases moleculares de este síndrome ha permitido desarrollar sondas que posibilitan la identificación confiable de los afectados y los portadores por análisis directo del ADN. Se describen los resultados de un estudio clínico, citogenético y molecular en la familia de una probando X frágil. En el afectado un varón de 2 años 4 meses con retraso mental y dismorfias se comprobó un sitio X frágil, pero su madre y una tía no tenían signos clínicos y sitio frágil. El análisis molecular mostró, en el paciente, una banda de 6,5 Kb, que indicaba la presencia de la mutación causal, mientras la madre exhibía dos bandas, una de 5,2 y otra de 6,0 Kb, que la identifican como heterocigota. En contraste, la tía presentó una sola banda de 5,2 Kb característica de los individuos normales

Cronología de la erupción dentaria permanente en pacientes con síndrome de Down / Chronology of the eruption of the permanent dentition in patients with Down's syndrome

Se examinaron 240 pacientes con síndrome de Down, que asisten a escuelas especiales de Santiago de Chile, con el propósito de determinar la cronología de erupción de la dentición de definitiva y compararla con el patrón de erupción de la población chilena normal. Los niños con síndrome de Down presentan en el maxilar superior e inferior un retraso significativo de la erupción de algunos dientes; éstos son los siguientes: en el maxilar superior, el primer molar derecho (85,35 ñ 20,03 meses) e izquierdo (87,41 ñ 22,37 meses), primer y segundo premolar izquierdo (161,60 ñ 60,43 y 172,10 ñ 79,57 meses, respectivamente) y canino izquierdo (163,72 ñ 81,55 meses). En el maxilar inferior, el primer molar derecho (90,98 ñ 24,52 meses), el incisivo central derecho e izquierdo (84,26 ñ 21,38 y 84,59 ñ 17,72 meses, respectivamente), el incisivo lateral derecho e izquierdo (101,89 ñ 23,79 y 117,53 ñ 83,02 meses, respectivamente) y el canino izquierdo (147,57 ñ 41,54 meses). Las niñas con síndrome de Down presentan en el maxilar superior e inferior retraso significativo en la erupción de los siguientes dientes: en el maxilar superior, los segundos molares derecho e izquierdo (172,71 ñ 83,02 y 191,88 ñ 55,90 meses, respectivamente) y el primer molar izquierdo (84,48 ñ 19,65 meses). En el maxilar inferior, segundo molar y primer premolar derechos (163,30 ñ 59,31 y 131,07 ñ 21,94 meses, respectivamente) y ambos incisivos laterales derecho e izquierdo (102,27 ñ 52,86 y 112,87 ñ 73,47 meses, respectivamente). La secuencia de la erupción dentaria superior e inferior en niños con síndrome de Down comparada con la de la población normal presenta una gran asincronía que afecta a veinte dientes (71,4%). En las niñas con síndrome de Down esta secuencia asincrónica afecta sólo a 15 dientes (53,6%). Una de las principales diferencias entre los individuos Down y normales se debe a la mayor varianza observada en la muestra Down