Vaginal Microbiota and Proinflammatory Status in Patients with Polycystic Ovary Syndrome: An Exploratory Study.

Background/Purpose: Polycystic ovary syndrome (PCOS) is an endocrine-metabolic disease most common in patients of childbearing age. This pathology is associated with clinical, metabolic, and reproductive complications. We evaluated the diversity of the vaginal microbiota (VM), the vaginal inflammatory reaction (VIR), the proinflammatory state, and the activation of polymorphonuclear neutrophils (PMN) with the production of neutrophil extracellular traps (NETs). Methods: Thirty-three patients who attended a consultation at the Hospital UTPL-Santa Inés, Loja, Ecuador, from May to August 2023 who were diagnosed with PCOS participated in this study. Blood samples, vaginal discharge, and a survey were obtained. Results: A high number of patients, 23/33 (69.7%), presented altered microbiota in clinical variables associated with PCOS phenotypes A and B, sexual partners (>2), and oligomenorrhoea. A significant statistical association was only observed for sexually transmitted infections at sampling (p = 0.023) and insulin (p = 0.002). All eight cases studied with VIR had PMN/NETotic activity. A high frequency of proinflammatory states was observed in all vaginal microbiota states. Conclusions: These results suggest that the PCOS could trigger a proinflammatory state in the vaginal epithelium independently of the state of the vaginal microbiota. Furthermore, the presence of NETs observed in the cases studied could decrease fertility in these PCOS patients.

High Presence of NETotic Cells and Neutrophil Extracellular Traps in Vaginal Discharges of Women with Vaginitis: An Exploratory Study.

Infectious vaginitis is a microbiological syndrome of great importance in public health that affects millions of women worldwide. However, no studies have explored the phenomenon of the production of the neutrophil extracellular traps (NETs) that are released into the female reproductive tract in these pathologies. This study aimed to determine the presence of NETosis in vaginal discharges of women with bacterial vaginosis, candidiasis, and trichomoniasis by characterizing NETs. Extracellular DNA with neutrophil elastase and citrullinated histones was identified to confirm the NET components (n = 10). The concentration, phenotypes of NETs, and number of NETotic cells were determined. The results showed an increase in NETotic cells in women with Candida albicans (CA) and Trichomonas vaginalis (TV) and an increase in NETs in TV-induced vaginitis. Samples of CA- and TV-infected women showed different NET phenotypes (diffNETs, sprNETs, and aggNETs); diffNETs were found in high concentrations in samples with CA and were increased in three types of NETs in TV infections. Samples with intermediate microbiota and bacterial vaginosis showed increased NETotic cells while the intermediate microbiota presented a higher concentration of NETs. Therefore, alterations in the microbiota and the presence of fungal and parasitic infections are important stimuli for the activation and induction of NETosis, and their cytotoxic effects could enhance tissue damage.

[Factors associated with Chlamydia trachomatis in women with vaginitis attending in primary health care]. / Factores asociados a Chlamydia trachomatis en mujeres con vaginitis atendidas en atención primaria en salud.

BACKGROUND: Chlamydia trachomatis is recognized as the causative agent of one of the most common and curable sexually transmitted infections (STIs). In women, it can be a trigger for pelvic inflammatory disease, a risk factor for infertility or acquisition of other STIs. AIM: To determine CT in women with vaginitis and associate it with sociodemographic, sexual, clinical, and vaginal microbiota factors. METHODS: 121 female volunteers > 18 years of age, with a clinical diagnosis of vaginitis and abnormal vaginal discharge, attending in primary health care services (PHC) in Temuco-Chile were recluted. A vaginal discharge sample was taken, determining C. trachomatis by polymerase chain reaction and vaginal microbiota by Gram stain. Clinical, sociodemographic and sexual data were collected by survey. RESULTS: Age range was 18-61 years. The frequency of C. trachomatis was 14.9%. C. trachomatis was associated with a higher number of sexual partners in the last 5 years (p = 0.001), oral plus anal sex (p = 0.055) and sex toys (p = 0.027); there was a non-significant association with bacterial vaginosis 31.2% (p = 0.091) and initiation of sexual activity < 18 years 20.6% (p = 0.07). CONCLUSIONS: It would be beneficial to incorporate in the gynecological semiology of the women attended in PHC services, the factors found to favor C. trachomatis infection, as this could help the management of this STI.

Concordance Between Clinical and Laboratory Diagnosis of Abnormal Vaginal Discharge in Chilean Women.

OBJECTIVE: To determine the concordance between the clinical diagnosis of women with abnormal vaginal discharge (AVD) and laboratory results using molecular detection and observation of the vaginal microbiota. METHODS: Cross-sectional study conducted in 2018 in Temuco, Chile. A total of 25 midwives from 12 health centers participated. A total of 125 women > 18 years old, volunteers, were recruited. The sample of the posterior vaginal fornix was obtained by speculoscopy. Characteristics of the discharge and of the external and internal genitalia were observed. Gram staining was used to observe vaginal microbiota, blastoconidia and pseudohyphae, and polymerase chain reaction was used for the detection of Trichomonas vaginalis and Candida albicans. The Cohen kappa coefficient was used in the concordance analysis. RESULTS: Out of a total of 125 women with AVD, 85.6% consulted spontaneously and 14.4% were diagnosed clinically during a routine check-up. Absolute concordance was significant (p = 0.0012), with an agreement of 13.6%. The relative concordance was significant, but fair for bacterial vaginosis (Kappa = 0.21; p = 0.003) and candidiasis (Kappa = 0.22; p = 0.001), and slight for trichomoniasis (Kappa = 0.14; p = 0.009). The percentage of coincidence of the diagnoses (single or mixed) by laboratory and midwives was: bacterial vaginosis 63.2% (12/19), candidiasis 36.5% (27/74), and trichomoniasis 12.5% (4/32). There was 20% coinfection. A total of 36% of the clinical diagnoses of AVD had negative laboratory tests. CONCLUSION: The vulvovaginitis conditions candidiasis and trichomoniasis appear to be overdiagnosed, and bacterial vaginosis appears to be underdiagnosed by the clinical diagnosis when compared with the laboratory diagnosis. The low concordance obtained shows the importance of complementing the clinical diagnosis with a laboratory study of AVD, particularly in women with failed treatments and/or coinfections with unspecific and varying signs and symptoms.

OBJETIVO: Determinar a concordância entre o diagnóstico clínico de mulheres com corrimento vaginal anormal (AVD) e os resultados laboratoriais por meio da detecção molecular e observação da microbiota vaginal. MéTODOS: Estudo transversal realizado em 2018 em Temuco, Chile. Participaram 25 parteiras de 12 centros de saúde. Um total de 125 mulheres > 18 anos, voluntárias, foram recrutadas. A amostra do fórnice vaginal posterior foi obtida por especuloscopia. Foram observadas características da secreção e da genitália externa e interna. A coloração de Gram foi usada para observar a microbiota vaginal, blastoconídios e pseudo-hifas, e a reação em cadeia da polimerase foi usada para a detecção de Trichomonas vaginalis e Candida albicans. O coeficiente kappa de Cohen foi usado na análise de concordância. RESULTADOS: De um total de 125 mulheres com AVD, 85,6% consultaram espontaneamente e 14,4% foram diagnosticados clinicamente durante um check-up de rotina. A concordância absoluta foi significativa (p = 0,0012), com concordância de 13,6%. A concordância relativa foi significativa, mas razoável para vaginose bacteriana (Kappa = 0,21; p = 0,003) e candidíase (Kappa = 0,22; p = 0,001), e leve para tricomoníase (Kappa = 0,14; p = 0,009). O percentual de coincidência dos diagnósticos (solteiros ou mistos) por laboratório e parteiras foi: vaginose bacteriana 63,2% (12/19), candidíase 36,5% (27/74) e tricomoníase 12,5% (4/32). Houve 20% de coinfecção. Um total de 36% dos diagnósticos clínicos de AVD tiveram exames laboratoriais negativos. CONCLUSãO: As condições de vulvovaginite candidíase e tricomoníase parecem ser sobrediagnosticadas, e a vaginose bacteriana parece ser subdiagnosticada pelo diagnóstico clínico quando comparado com o diagnóstico laboratorial. A baixa concordância obtida mostra a importância de complementar o diagnóstico clínico com estudo laboratorial de AVD, principalmente em mulheres com falha de tratamento e / ou coinfecções com sinais e sintomas inespecíficos e variáveis.

Factores asociados a Chlamydia trachomatis en mujeres con vaginitis atendidas en atención primaria en salud / Factors associated with Chlamydia trachomatis in women with vaginitis attending in primary health care

INTRODUCCIÓN: Chlamydia trachomatis es reconocida como el agente causal de una de las infecciones de transmisión sexual (ITS) más común y curable. En mujeres, puede ser gatillante de enfermedad inflamatoria pélvica, un factor de riesgo para infertilidad o adquisición de otras ITS. OBJETIVO: Determinar C. trachomatis en mujeres con diagnóstico clínico de vaginitis asociando factores sociodemográficos, sexuales, clínicos y microbiota vaginal. PACIENTES Y MÉTODOS: Participaron 121 mujeres voluntarias > 18 años, con diagnóstico clínico de vaginitis y descarga vaginal anormal, atendidas en servicios de atención primaria en salud (APS) en Temuco-Chile. Se tomó muestra de flujo vaginal, determinándose C. trachomatis por una reacción de polimerasa en cadena convencional y microbiota vaginal por tinción de Gram. Se colectaron datos clínicos, sociodemográficos y sexuales mediante encuesta. RESULTADOS: Rango de edad 18-61 años. La frecuencia de C. trachomatis fue 14,9%. Se asoció C. trachomatis a mayor número de parejas sexuales en los últimos 5 años (p = 0,001), relaciones sexuales oral más anal (p = 0,055) y juguetes sexuales (p = 0,027). Asociación no significativa hubo con vaginosis bacteriana 31,2% (p = 0,091) e inicio de la actividad sexual < 18 años 20,6% (p = 0,07). CONCLUSIONES: Sería beneficioso incorporar en la semiología ginecológica de las mujeres atendidas en servicios de APS, los factores encontrados como favorecedores de infección por C. trachomatis, pues ello podría ayudar al manejo de esta ITS.

BACKGROUND: Chlamydia trachomatis is recognized as the causative agent of one of the most common and curable sexually transmitted infections (STIs). In women, it can be a trigger for pelvic inflammatory disease, a risk factor for infertility or acquisition of other STIs. AIM: To determine CT in women with vaginitis and associate it with sociodemographic, sexual, clinical, and vaginal microbiota factors. Methods: 121 female volunteers > 18 years of age, with a clinical diagnosis of vaginitis and abnormal vaginal discharge, attending in primary health care services (PHC) in Temuco-Chile were recluted. A vaginal discharge sample was taken, determining C. trachomatis by polymerase chain reaction and vaginal microbiota by Gram stain. Clinical, sociodemographic and sexual data were collected by survey. RESULTS: Age range was 18-61 years. The frequency of C. trachomatis was 14.9%. C. trachomatis was associated with a higher number of sexual partners in the last 5 years (p = 0.001), oral plus anal sex (p = 0.055) and sex toys (p = 0.027); there was a non-significant association with bacterial vaginosis 31.2% (p = 0.091) and initiation of sexual activity < 18 years 20.6% (p = 0.07). CONCLUSIONS: It would be beneficial to incorporate in the gynecological semiology of the women attended in PHC services, the factors found to favor C. trachomatis infection, as this could help the management of this STI.

Concordance Between Clinical and Laboratory Diagnosis of Abnormal Vaginal Discharge in Chilean Women / Concordância entre o diagnóstico clínico e laboratorial de corrimento vaginal anormal em mulheres chilenas

Abstract Objective To determine the concordance between the clinical diagnosis of women with abnormal vaginal discharge (AVD) and laboratory results using molecular detection and observation of the vaginal microbiota. Methods Cross-sectional study conducted in 2018 in Temuco, Chile. A total of 25 midwives from 12 health centers participated. A total of 125 women>18 years old, volunteers, were recruited. The sample of the posterior vaginal fornix was obtained by speculoscopy. Characteristics of the discharge and of the external and internal genitalia were observed. Gram staining was used to observe vaginal microbiota, blastoconidia and pseudohyphae, and polymerase chain reaction was used for the detection of Trichomonas vaginalis and Candida albicans. The Cohen kappa coefficient was used in the concordance analysis. Results Out of a total of 125 women with AVD, 85.6% consulted spontaneously and 14.4% were diagnosed clinically during a routine check-up. Absolute concordance was significant (p=0.0012), with an agreement of 13.6%. The relative concordance was significant, but fair for bacterial vaginosis (Kappa=0.21; p=0.003) and candidiasis (Kappa=0.22; p=0.001), and slight for trichomoniasis (Kappa=0.14; p=0.009). The percentage of coincidence of the diagnoses (single or mixed) by laboratory and midwives was: bacterial vaginosis 63.2% (12/19), candidiasis 36.5% (27/74), and trichomoniasis 12.5% (4/32). There was 20% coinfection. A total of 36% of the clinical diagnoses of AVD had negative laboratory tests. Conclusion The vulvovaginitis conditions candidiasis and trichomoniasis appear to be overdiagnosed, and bacterial vaginosis appears to be underdiagnosed by the clinical diagnosis when compared with the laboratory diagnosis. The low concordance obtained shows the importance of complementing the clinical diagnosis with a laboratory study of AVD, particularly in women with failed treatments and/or coinfections with unspecific and varying signs and symptoms.

Resumo Objetivo Determinar a concordância entre o diagnóstico clínico de mulheres com corrimento vaginal anormal (AVD) e os resultados laboratoriais por meio da detecção molecular e observação da microbiota vaginal. Métodos Estudo transversal realizado em 2018 em Temuco, Chile. Participaram 25 parteiras de 12 centros de saúde. Um total de 125 mulheres>18 anos, voluntárias, foramrecrutadas. A amostra do fórnice vaginal posterior foi obtida por especuloscopia. Foram observadas características da secreção e da genitália externa e interna. A coloração de Gram foi usada para observar a microbiota vaginal, blastoconídios e pseudo-hifas, e a reação em cadeia da polimerase foi usada para a detecção de Trichomonas vaginalis e Candida albicans. O coeficiente kappa de Cohen foi usado na análise de concordância. Resultados De um total de 125 mulheres com AVD, 85,6% consultaram espontaneamente e 14,4% foram diagnosticados clinicamente durante um check-up de rotina. A concordância absoluta foi significativa (p=0,0012), com concordância de 13,6%. A concordância relativa foi significativa, mas razoável para vaginose bacteriana (Kappa =0,21; p=0,003) e candidíase (Kappa=0,22; p=0,001), e leve para tricomoníase (Kappa=0,14; p=0,009). O percentual de coincidência dos diagnósticos (solteiros ou mistos) por laboratório e parteiras foi: vaginose bacteriana 63,2% (12/19), candidíase 36,5% (27/74) e tricomoníase 12,5% (4/32). Houve 20% de coinfecção. Umtotal de 36% dos diagnósticos clínicos de AVD tiveram exames laboratoriais negativos. Conclusão As condições de vulvovaginite candidíase e tricomoníase parecem ser sobrediagnosticadas, e a vaginose bacteriana parece ser subdiagnosticada pelo diagnóstico clínico quando comparado com o diagnóstico laboratorial. A baixa concordância obtida mostra a importância de complementar o diagnóstico clínico comestudo laboratorial de AVD, principalmente emmulheres com falha de tratamento e / ou coinfecções com sinais e sintomas inespecíficos e variáveis.

First Molecular Characterization of Trichostrongylus colubriformis Infection in Rural Patients from Chile.

PURPOSE: The aim was to characterize the infection by Trichostrongylus spp. in patients from Chile using a combination of molecular detection techniques and phylogenetic analysis relating the findings to clinical and epidemiological reports of the patients METHODS: Strongylid eggs were detected in seven patients by coproparasitological techniques. From each sample a fragment of the ITS-2 ribosomal gene was amplified by PCR, sequenced and analyzed by the Neighbor-Joining method. RESULTS: All the sequences and phylogenetic clusters corresponded to T. colubriformis. Two samples presented a single nucleotide polymorphism showing two possible haplotypes. Six patients presented gastrointestinal symptoms. All of them lived on farms and used sheep manure as fertilizer. CONCLUSION: T. colubriformis was the strongylid involved in the infections of these Chilean patients associated with the presence of livestock and agricultural practices that favor infection by this type of nematode.

[Frequency of cervical cytological alterations and human papilloma virus in a sample of university students in Temuco, Chile]. / Frecuencia de alteraciones citológicas cervicales y virus papiloma humano en una muestra de estudiantes universitarias en Temuco, Chile.

BACKGROUND: The early onset of sexual activity can promote the development of cervical alterations and sexually transmitted infections, especially the human papillomavirus (HPV) very common in adolescents and young people. AIM: The condition of the cervix, HPV and sexual behavior in young women under 25 years of age were analyzed. METHODS: 182 university students, healthy, sexually active, 18-24 years old, without vaccine for HPV participated. Papanicolaou (Pap) test and classification of high and low risk HPV (HR and LR) were performed by real time polymerase chain reaction. The sexual behaviors were consulted in private. RESULTS: The 46.9% of Pap presented cytological alterations, non-specific inflammation/hemorrhagic (29.4%) and atypical smear (10.2%) being de most frequent. The overall frequency of HPV-HR was 24.3%, of these 67.4% presented an altered Pap. There was an association between cytological alterations and HPV (p < 0.0001) and years of sexual activity and atypical smear or cervical intraepithelial neoplasia grade I (CIN I) (p = 0.009). 11.9% of young women (21/177) presented atypical smear or CIN I, with 66.7% of cases HPV-HR. CONCLUSIONS: These findings alert the vulnerability of these young women who would have a potential risk of viral persistence, CIN and eventually cancer. It is important to emphasize counseling and prevention prior to the regular age of admission to the screening program for cervical cancer. This study was financed by the Universidad de La Frontera through Projects DI15-0047 and DI17-0123.

Frecuencia de alteraciones citológicas cervicales y virus papiloma humano en una muestra de estudiantes universitarias en Temuco, Chile / Frequency of cervical cytological alterations and human papilloma virus in a sample of university students in Temuco, Chile

Resumen Introducción: El inicio precoz de actividad sexual puede favorecer el desarrollo de alteraciones cervicales y de infecciones de transmisión sexual, en especial del virus papiloma humano (VPH) muy frecuente en adolescentes y jóvenes. Objetivo: Analizar el estado del cuello uterino, presencia del VPH y conductas sexuales en mujeres menores de 25 años. Material y Métodos: Participaron 182 estudiantes universitarias de 18-24 años, sanas, sexualmente activas y no vacunadas para VPH. Se realizó Papanicolaou (Pap) y clasificación del VPH en alto y bajo riesgo (AR y BR) mediante reacción de polimerasa en cadena en tiempo real. Las conductas sexuales fueron consultadas privadamente. Resultados: El 46,9% de los Pap presentaron alteraciones citológicas (inflamación inespecífica/hemorrágico: 29,4% y frotis atípicos (FA):10,2%). La frecuencia de los VPH-AR fue 24,3%; de éstos, 67,4% presentó un Pap alterado. Hubo asociación entre alteraciones citológicas y presencia de VPH (p < 0,0001) y años de actividad sexual y FA o neoplasia intraepitelial grado I (NIE I) (p = 0,009). El 11,9% de las jóvenes estudiadas (21/177) presentó FA o NIE I, con 66,7% de casos VPH-AR. Conclusiones: Estos hallazgos alertan la vulnerabilidad de estas jóvenes que tendrían un riesgo potencial de persistencia viral, NIE y eventualmente cáncer. Es importante enfatizar consejería y prevención previo a la edad normada de ingreso al programa de cribado para cáncer cérvico uterino en Chile.

Background: The early onset of sexual activity can promote the development of cervical alterations and sexually transmitted infections, especially the human papillomavirus (HPV) very common in adolescents and young people. Aim: The condition of the cervix, HPV and sexual behavior in young women under 25 years of age were analyzed. Methods: 182 university students, healthy, sexually active, 18-24 years old, without vaccine for HPV participated. Papanicolaou (Pap) test and classification of high and low risk HPV (HR and LR) were performed by real time polymerase chain reaction. The sexual behaviors were consulted in private. Results: The 46.9% of Pap presented cytological alterations, non-specific inflammation/hemorrhagic (29.4%) and atypical smear (10.2%) being de most frequent. The overall frequency of HPV-HR was 24.3%, of these 67.4% presented an altered Pap. There was an association between cytological alterations and HPV (p < 0.0001) and years of sexual activity and atypical smear or cervical intraepithelial neoplasia grade I (CIN I) (p = 0.009). 11.9% of young women (21/177) presented atypical smear or CIN I, with 66.7% of cases HPV-HR. Conclusions: These findings alert the vulnerability of these young women who would have a potential risk of viral persistence, CIN and eventually cancer. It is important to emphasize counseling and prevention prior to the regular age of admission to the screening program for cervical cancer. This study was financed by the Universidad de La Frontera through Projects DI15-0047 and DI17-0123.

Trichinella spiralis Infecting Wild Boars in Southern Chile: Evidence of an Underrated Risk.

Trichinellosis in Chilean wild animals has scarcely been documented. The introduction of wild boars into the wild environment represents a viable new host with a potential risk of infection for human health. The aim of this study was to determine the presence and prevalence of Trichinella in wild boars. Two hundred seventy eight wild boars from of the Southern Chile were examined by compression and artificial digestion techniques. The larvae in the positive samples were collected for taxonomic analysis through polymerase chain reaction-inter-simple sequence repeats and to calculate the parasitic burden. A prevalence of 1.8% (5/278) of infected animals and an average parasitic burden of 6.8 ± 2.1 larvae per gram were estimated. The only species identified by molecular techniques was Trichinella spiralis. Prevalence of T. spiralis in wild boars was similar to those described around the world. T. spiralis infection rate and parasite burden detected in Chilean wild boars represent a certain food-borne risk for human population.

A PCR-RFLP assay for discrimination of Echinococcus granulosus sensu stricto and Taenia spp. in dogs stool.

In order to develop a method of identification and discrimination of Echinococcus granulosus sensu stricto from faecal samples of dogs infected with taeniid eggs (Echinococcus spp., Taenia spp.), a combined strategy of Polymerase Chain Reaction (PCR) and Restriction Fragments of Lenght Polymorphisms (RFLP) was proposed. Initially, a pair of primers was designed to amplify a fragment of the 12 Subunit of ribosomal RNA gene (12SrRNA) from mitochondrial DNA. The amplified product was digested by SspI restriction enzyme, which in E. granulosus kept the intact fragment of 160 basis pairs (bp), while in Taenia spp. produced two fragments (62 bp and another of 98 bp). The method was tested using positive controls of DNA, in faecal samples experimentally contaminated with eggs of E. granulosus and Taenia spp. and in dogs naturally infected. In all of them, reproducible results were obtained and the primers were specific to amplify only Taeniidae DNA. The sensitivity of the technique was tested, achieving amplification of DNA extractions with a single egg. In conclusion, the technique developed was optimal and easy to identify patent infections by E. granulosus s.s., constituting a possible alternative for epidemiological studies in dogs, especially in endemic areas where this infection occurs.

DNA extraction in Echinococcus granulosus and Taenia spp. eggs in dogs stool samples applying thermal shock.

The extraction of DNA in taeniid eggs shows complications attached to the composition of stool samples and the high resistance of eggs to degradation. The objective of this study was to test a method of DNA extraction in taeniid eggs by applying a thermal shock to facilitate the chemical-enzymatic degradation of these elements. A group of six tubes containing 1 ml of dog stool sample was spiked with eggs of Echinococcus granulosus and another group of six with Taenia pisiformis. Samples were floated with supersaturated sugar solution and centrifuged. The upper portion of each tube (500 µl) was aspirated and deposited in 1.5 ml tubes. Three tubes from each group were incubated at -20 °C and then at 90 °C, the remaining three from each group, incubated at room temperature. Proteinase K and lysis buffer were added to each tube and incubated for 12 h at 58 °C. The lysis effect was evaluated by microscopy at 3, 6 and 12 h and integrity by electrophoresis in 1% agarose gels. With the same experimental scheme, the thermal shock effect was evaluated in extractions of 1, 2, 3 and 4 eggs of each species and the DNA was quantified. Additionally, the protocol was applied in samples of 4 dogs diagnosed with natural infection by Taeniidae worms. Finally, all the extractions were tested by PCR amplification. Both E. granulosus and T. pisiformis eggs showed a similar response in the tests. In samples without treatment, the lysis effect was poor and showed no differences over time, but in those subjected to thermal shock, eggs degradation increased with time. In both treatments, there was no DNA loss integrity. The protocol applied to limited amounts of eggs yielded PCR products in 100% of the samples exposed to thermal shock, allowing PCR amplifications up to 1 egg. In non-exposed samples, the results were not replicable. However, DNA quantification showed low values in both treatments. In turn, DNA extractions with thermal shock in infected dog samples finally yielded PCR amplifications in 100%. It was concluded that thermal shock facilitates the DNA extraction for molecular analysis in taeniid eggs. The technique is effective extracting DNA even from a single egg and also to analyze natural infections samples with a relatively simple implementation.

Frequency of Human papillomavirus in women attending cervical cancer screening program in Chile.

BACKGROUND: Human papillomavirus (HPV) is the etiological factor for cervical cancer and its precursor lesions. The characterization of HPV genotypes in preneoplastic lesions and cervical cancer could establishes the effectiveness of vaccination plan in Chilean population. The aim of this study was to determine HPV frequency in a group of women including in a cervical screening program in the public health care system in Chile. METHODS: We analyzed 985 cervical smears samples from women with different histological diagnosis, attending to public health care in Temuco-Chile between 2004 and 2012, to detect HPV genotypes, through PCR followed by reverse line blotting assay. RESULTS: HPV was found present in 80.8% (n = 796) of samples. Only a 5.6% of 985 samples were infected with a low-risk HPV, considering multiple infections. 10.5% (n = 8/76) of normal cervical epithelia, 83.5% (n = 208/249) and 87.6% (n = 557/636) of low and high grade squamous intraepithelial lesions, respectively, and 95.8% (n = 23/24) of squamous cervical carcinomas tested positive for HPV. HPV 16 was the most frequent genotype found (Overall 44.9%, n = 442/985; SCC: 62.5%, n = 15/24). A high variability of HPV types was also found in preneoplastic lesions, whereas there was a selection of genotypes in neoplasia. Also, there was a higher risk of infection with HPV 16 in women ≤26 years and 34-41 years old (p < 0.05), meanwhile infections with HPV 16 or HPV 18 have related with cancer development (p < 0.01). CONCLUSIONS: These data provide further information about the frequency of HPV genotypes in women with cervical lesions in Chile, and the introduction of new targeted vaccines against a wider spectrum of HPV is suggested.

[Human papilloma virus and Chlamydia trachomatis by number of sexual partners and time of sexual activity on university students in the Region of La Araucanía, Chile]. / Virus papiloma humano y Chlamydia trachomatis según número de parejas sexuales y tiempo de actividad sexual en estudiantes universitarias en la Región de La Araucanía, Chile.

BACKGROUND: Human papilloma virus (HPV) and Chlamydia trachomatis are the most prevalent sexually transmitted infections (STIs), among teenagers and young people, with risk factors: active sex life and multiple partners. Chlamydia trachomatis infection may favor HPV infection and this, the development of cervical cancer. Both infections can lead to consequences on sexual and reproductive health. OBJECTIVE: To determine frequency of HPV and C. trachomatis in asymptomatic university women less than 25 years, associating them with number of sexual partners (n°SxP) and time of sexual activity (TSxA). Material andMethods: 151 cervical samples for HPV and C. trachomatis, were processed by conventional and in real time reaction polymerase chain. RESULTS: HPV 21, 8%, C. trachomatis 11, 2% and co-infection (HPV/C.trachomatis), 4.6%. Aimong HPV +, 80, 6% showed high risk HPV. The n°SxP was strongly associated with HPV. Aimong young coinfected HPV/C. trachomatis, 71.4% had 3 or more PSx. Chlamydia trachomatis was more frequent (64,7%) that HPV within range of 3-5 years according to the TSxA, Discussion: A high prevalence of HPV and C. trachomatis was observed. Young women with coinfection HPV/C. trachomatis could be a high-risk group need to monitor their infections. It suggests the implementation of university programs in education, counseling and prevention in sexual health.

Virus papiloma humano y Chlamydia trachomatis según número de parejas sexuales y tiempo de actividad sexual en estudiantes universitarias en la Región de La Araucanía, Chile / Human papilloma virus and Chlamydia trachomatis by number of sexual partners and time of sexual activity on university students in the Region of La Araucanía, Chile

Introducción: El virus papiloma humano (VPH) y Chlamydia trachomatis son las infecciones de transmisión sexual (ITS) más frecuentes, en adolescentes y jóvenes, con factores de riesgo: vida sexual activa y múltiples parejas. Chlamydia trachomatis puede favorecer el ingreso de VPH y éste el desarrollo del cáncer cérvico uterino. Ambas infecciones pueden dejar secuelas en la salud sexual y reproductiva. Objetivo: Determinar frecuencias VPH y C trachomatis en estudiantes universitarias asinto-máticas bajo 25 años, asociándolas con número de parejas sexuales (n°PSx) y tiempo de actividad sexual (TASx). Material y Método: Se procesaron 151 muestras/exo y endo cervicales para VPH y C. trachomatis, mediante reacción de polimerasa en cadena convencional y en tiempo real. Resultados: La frecuencia fue: VPH 21,8%, C. trachomatis 11,2% y co-infección 4,6%. De las jóvenes con infección por VPH, 80,6% presentó VPH alto riesgo oncogénico. El n°PSx se asoció fuertemente a VPH. Entre las jóvenes con co-infección VPH/C. trachomatis, 71,4% tenían tres o más PSx. Según TASx, C. trachomatis fue más frecuente (64,7%) entre 3-5 años que VPH. Conclusión: Se observó alta prevalencia de VPH y C trachomatis. Mujeres jóvenes con co-infección VPH/C. trachomatis podrían ser un grupo de alto riesgo con necesidad de mo-nitorear sus infecciones. Es sugerida la implementación de programas universitarios en educación, orientación y prevención en ITS.

Background: Human papilloma virus (HPV) and Chlamydia trachomatis are the most prevalent sexually transmitted infections (STIs), among teenagers and young people, with risk factors: active sex life and multiple partners. Chlamydia trachomatis infection may favor HPV infection and this, the development of cervical cancer. Both infections can lead to consequences on sexual and reproductive health. Objective: To determine frequency of HPV and C. trachomatis in asymptomatic university women less than 25 years, associating them with number of sexual partners (n°SxP) and time of sexual activity (TSxA). Material andMethods: 151 cervical samples for HPV and C. trachomatis, were processed by conventional and in real time reaction polymerase chain. Results: HPV 21, 8%, C. trachomatis 11, 2% and co-infection (HPV/C.trachomatis), 4.6%. Aimong HPV +, 80, 6% showed high risk HPV. The n°SxP was strongly associated with HPV. Aimong young coinfected HPV/C. trachomatis, 71.4% had 3 or more PSx. Chlamydia trachomatis was more frequent (64,7%) that HPV within range of 3-5 years according to the TSxA, Discussion: A high prevalence of HPV and C. trachomatis was observed. Young women with coinfection HPV/C. trachomatis could be a high-risk group need to monitor their infections. It suggests the implementation of university programs in education, counseling and prevention in sexual health.

[Genotyping of human papillomavirus in women under 25 years old treated in the screening program for cervical cancer]. / Genotipificación del virus papiloma humano en mujeres bajo 25 años de edad participantes del Programa Nacional del Cáncer Cérvico-uterino en la Región de la Araucanía, Chile.

BACKGROUND: In Chile, cervical cancer (CC) is the second leading cause of death from malignancy in women. The main causal agent of cervical cancer is the human papillomavirus (HPV). This virus is the most common sexually transmitted infection among sexually active youth. An early onset of sexual life increases the chances of HPV infection; this may involve a possible early development of cervical intraepithelial neoplasia and CC, creating a major public health problem. OBJECTIVE: To present HPV frequency in women under the age of 25, treated in the CC screening program and their follow-up after histopathological diagnosis. METHODS: 173 cervical samples were genotyped by polymerase chain reaction and non-radioactive reverse hybridization (line blot). RESULTS: The overall frequency of HPV was 84.8%. HPV16 was the most prevalent. In 12.1% of women the cervical lesion persisted or progressed. 28.9% of women had irregular follow-up; in this group, 88% were HPV(+) and 52% had no record of Pap smear in the past 3 years. DISCUSSION: The results reaffirm the usefulness of complementing the Pap and HPV detection as a primary screening tool in sexually active women. They also suggest the possibility of extending the age coverage of the national screening program.

Genotipificación del virus papiloma humano en mujeres bajo 25 años de edad participantes del Programa Nacional del Cáncer Cérvico-uterino en la Región de la Araucanía, Chile / Genotyping of human papillomavirus in women under 25 years old treated in the screening program for cervical cancer

Background: In Chile, cervical cancer (CC) is the second leading cause of death from malignancy in women. The main causal agent of cervical cancer is the human papillomavirus (HPV). This virus is the most common sexually transmitted infection among sexually active youth. An early onset of sexual life increases the chances of HPV infection; this may involve a possible early development of cervical intraepithelial neoplasia and CC, creating a major public health problem. Objective: To present HPV frequency in women under the age of 25, treated in the CC screening program and their follow-up after histopathological diagnosis. Methods: 173 cervical samples were genotyped by polymerase chain reaction and non-radioactive reverse hybridization (line blot). Results: The overall frequency of HPV was 84.8%. HPV16 was the most prevalent. In 12.1% of women the cervical lesion persisted or progressed. 28.9% of women had irregular follow-up; in this group, 88% were HPV(+) and 52% had no record of Pap smear in the past 3 years. Discussion: The results reaffirm the usefulness of complementing the Pap and HPV detection as a primary screening tool in sexually active women. They also suggest the possibility of extending the age coverage of the national screening program.

Introducción: En Chile, el cáncer cérvico-uterino (CCU) es la segunda causa de muerte por neoplasias malignas en la mujer. El principal agente causal es el virus papiloma humano (VPH), descrito como la infección de transmisión sexual más frecuente entre jóvenes sexualmente activas. El comienzo precoz de la vida sexual incrementa las posibilidades de infección con VPH; esto puede implicar un eventual desarrollo prematuro de neoplasia intraepitelial cervical y CCU, creando un importante problema de salud pública. Objetivo: Presentar la frecuencia del VPH en mujeres bajo 25 años de edad, participantes del programa de CCU y su seguimiento post-lesión. Material y Métodos: Se genotipificaron 173 muestras cervicales, mediante reacción de polimerasa en cadena e hibridación no radioactiva (reverse line blot). Resultados: La frecuencia global del VPH fue 84,8%. El genotipo más frecuente fue VPH16. En 12,3% la lesión cervical persistió o evolucionó a una mayor. Se encontró 28,9% de mujeres con seguimiento post-lesión irregular; en este grupo, 88% fue VPH (+) y 52% no tuvo registro de Papanicolaou en los últimos tres años. Discusión: Los resultados obtenidos reafirman la utilidad de complementar el Papanicolaou con detección del VPH como herramienta de tamizaje primario en mujeres sexualmente activas. Además sugieren la posibilidad de ampliar la edad de cobertura del programa de tamizaje.

Asymmetric effect of oxygen adsorption on electron and hole mobilities in bilayer graphene: long- and short-range scattering mechanisms.

We probe electron and hole mobilities in bilayer graphene under exposure to molecular oxygen. We find that the adsorbed oxygen reduces electron mobilities and increases hole mobilities in a reversible and activated process. Our experimental results indicate that hole mobilities increase due to the screening of long-range scatterers by oxygen molecules trapped between the graphene and the substrate. First principle calculations show that oxygen molecules induce resonant states close to the charge neutrality point. Electron coupling with such resonant states reduces the electron mobilities, causing a strong asymmetry between electron and hole transport. Our work demonstrates the importance of short-range scattering due to adsorbed species in the electronic transport in bilayer graphene on SiO2 substrates.

[Human papillomavirus genotyping in cervical adenocarcinoma in the Region of La Araucania-Chile]. / Genotipificación del virus papiloma humano en mujeres con adenocarcinoma cervical de la Región de La Araucania-Chile.

UNLABELLED: Human papillomavirus (HPV) is the main cause of cervical cancer. Thus, HPV detection and typing becomes important in order to know the frequency of genotypes present in the region. In this paper we studied 44 biopsies of cervical adenocarcinoma. For HPV detection nested polymerase chain reaction (PCR) was used to amplify the L1 gene. For viral typing restriction enzymes (Rsa I, Dde I, Pst I) and DNA sequencing were used. Viral DNA was detected by nested L1 PCR in 100% of biopsies; 38/44 cases could be typed: 81.6% HPV16; 13.2% HPV 18; 2.6% VPH 33 and 2.6% HPV 18/33. CONCLUSIONS: The technique was successful in identifying the virus type in 86% of biopsies. There was a strong association ACC-HPV, especially with the viral type 16, detected in 81.6% of established cases.

Description of Echinococcus granulosus genotypes in human hydatidosis in a region of southern Chile.

INTRODUCTION: Echinococcus granulosus species has a wide variety in both geography and hosts; indeed, 10 genotypes have been reported in studies on material of animal origin. The aim of this study was to genotype E. granulosus obtained from human hydatid cysts. MATERIALS AND METHODS: The hydatid fluid and sand was collected from patients who underwent surgery for hepatic and pulmonary hydatidosis at Hospital Regional in Temuco, Chile, between 2004 and 2005. Two PCR systems were used: PCR Eg 9 and PCR Eg 16. The RsaI enzyme was used for RFLP. The genotype was confirmed using the sequence of one fragment of 366 bp from a mitochondrial gene (cox1). RESULTS: The DNA of protoscolices from 24 samples was analyzed, 4 of them from pulmonary cysts and 20 from hepatic cysts. The 366 bp fragment was amplified in 20 out of 24 samples (83.3%). Enzymatic digestion revealed the presence of 3 possible genotypes: in 20 out of 21 samples (95,2%), a restriction was observed corresponding to the G1 or G7 genotypes; in the remaining sample genotype G4 or G7 was observed. Sequencing confirmed the presence of G1 genotype for 19 samples and G6 genotype for the remaining sample (G4 or G7 according to PCR-RFLP). CONCLUSION: The PCR-RFLP technique enabled three possible genotypes present (G1 or G7, G4 or G7) to be established. Sequencing allowed us to decisively identify the G1 and G6 genotypes in our study group. Previous studies agree with the identification of the G1 genotype in our country. We consider it significant that the G6 genotype is present in Chile for its epidemiological implications.