Biochemical and immunomodulatory insights of extracellular matrix from decellularized human whole cervix: recellularization andin vivoECM remodeling interplay.

Extracellular matrix (ECM) rich whole organ bio-scaffolds, preserving structural integrity and essential growth factors, has potential towards regeneration and reconstruction. Women with cervical anomalies or trauma can benefit from clinical cervicovaginal repair using constructs rich in site specific ECM. In this study, complete human cervix decellularization was achieved using a modified perfusion-based stir bench top decellularization method. This was followed by physico-chemical processes including perfusion of ionic agents, enzymatic treatment and washing using detergent solutions for a duration of 10-12 d. Histopathological analysis, as well as DNA quantification confirmed the efficacy of the decellularization process. Tissue ultrastructure integrity was preserved and the same was validated via scanning electron microscopy and transmission electron microscopy studies. Biochemical analysis and structural characterizations like Fourier transform infrared, Raman spectroscopy of decellularized tissues demonstrated preservation of important proteins, crucial growth factors, collagen, and glycosaminoglycans.In vitrostudies, using THP-1 and human umbilical vein endothelial cell (HUVEC) cells, demonstrated macrophage polarization from M1 to M2 and vascular functional genes enhancement, respectively, when treated with decellularized human cervical matrix (DHCp). Crosslinked DHC scaffolds were recellularized with site specific human cervical epithelial cells and HUVEC, showing non-cytotoxic cell viability and enhanced proliferation. Furthermore, DHC scaffolds showed immunomodulatory effectsin vivoon small rodent model via upregulation of M2 macrophage genes as compared to decellularized rat cervix matrix scaffolds (DRC). DHC scaffolds underwent neo-vascularization followed by ECM remodeling with enhanced tissue integration.

Chromatin Dispersion Test to Asses DNA Damage in Cervical Epithelial Cells.

The chromatin dispersion test (CDT) is based on the removal of nuclear proteins under the assumption that cells with fragmented DNA produce a typical halo of circular DNA loops, which is absent in cells with non-fragmented DNA. This method represents a simple, rapid, accurate, highly reproducible, and inexpensive technique to assess nuclear DNA damage in somatic cells. The visualization of DNA damage and the capacity of the test to provide a threshold value to discriminate between high and low levels of cervical lesions would aid in determining the malignant transformation. All of these advantages associated with the CDT protocol could promote this technique as a tool for the quick and reliable diagnosis of cervical epithelial disorders, even at primary-care centers.

Intervención educativa basada en metodologías B-learning para mejorar las citologías cervicales: experiencias de enfermeras / Intervenção educacional através do B-learning para melhorar a citologia cervical: experiências de enfermeiras / Educational intervention through B-learning to improve cervical cytology: experiences of nurses

Resumen Objetivo analizar las experiencias de enfermeras en la toma de las citologías cervicales y otros factores organizacionales durante una intervención educativa asistida por metodologías B-learning. Método estudio cualitativo realizado en San Luis Potosí, México. Participaron 15 enfermeras. La recolección de datos se hizo a través de entrevistas semiestructuradas con base en una sistematización de experiencias. Para el análisis de la información se utilizó el programa Taguette y como referente teórico las metodologías B-learning. Resultados se identificaron debilidades en factores relacionados con la accesibilidad de las usuarias al servicio, insumos, infraestructura, bioseguridad, capacitación del personal de salud, entrega de resultados a las pacientes y conocimiento del programa por parte de las usuarias. Conclusiones e implicaciones para la práctica el cáncer cervical es un problema de salud pública. La citología cervical es la prueba de tamizaje más utilizada; sin embargo, existen limitantes en la calidad, por lo que se proponen acciones para mejorar los conocimientos y habilidades del personal de enfermería que tiene como función la toma. La intervención educativa fue efectiva para fomentar el aprendizaje integral sobre la toma de las citologías cervicales y permitió al personal de enfermería compartir sus experiencias.

Resumo Objetivo analisar as experiências das enfermeiras na realização de esfregaços cervicais e outros fatores organizacionais durante uma intervenção educacional assistida por metodologias de b-learning. Método estudo qualitativo realizado em San Luis Potosí, México. Participaram 15 enfermeiras. A coleta de dados foi feita por meio de entrevistas semiestruturadas a partir de uma sistematização de experiências. Para a análise das informações, utilizou-se o programa Taguette e metodologias de b-learning como referencial teórico. Resultados foram identificadas fragilidades em fatores relacionados com a acessibilidade dos usuários ao serviço, insumos, infraestrutura, biossegurança, capacitação da equipe de saúde, entrega de resultados aos pacientes e conhecimento do programa pelos usuários. Conclusões e implicações para a prática o câncer do colo do útero é um problema de saúde pública. A citologia cervical é o teste de triagem mais utilizado; no entanto, existem limitações na qualidade, por isso são propostas ações para aprimorar os conhecimentos e habilidades das enfermeiras que estejam desempenhando essa função. A intervenção educacional foi eficaz para promover o aprendizado integral sobre a realização do esfregaço cervical e permitiu que as enfermeiras compartilhassem suas experiências.

Abstract Objective to analyze the nursing staff's experiences in taking cervical smears and other organizational factors during an educational intervention assisted by B-learning methodologies. Method a qualitative study was carried out in San Luis Potosí, Mexico, with 15 nurses. Data collection was done through semi-structured interviews based on a systematization of experiences. The Taguette program and B-learning methodologies as theoretical references were used to analyze the information. Results weaknesses were identified in factors related to the accessibility of users to the service, supplies, infrastructure, biosafety, training of health personnel, delivery of results to patients, and knowledge of the program by the users. Conclusions and implications for practice cervical cancer is a public health problem. Cervical cytology is the most widely used screening test; however, there are limitations in quality, so actions are proposed to improve the knowledge and skills of the nursing staff in their functions. The educational intervention effectively promoted comprehensive learning about taking cervical smears and allowed the nursing staff to share their experiences.

Immunometabolic and potential tumor-promoting changes in 3D cervical cell models infected with bacterial vaginosis-associated bacteria.

Specific bacteria of the human microbiome influence carcinogenesis at diverse anatomical sites. Bacterial vaginosis (BV) is the most common vaginal disorder in premenopausal women that is associated with gynecologic sequelae, including cervical cancer. BV-associated microorganisms, such as Fusobacterium, Lancefieldella, Peptoniphilus, and Porphyromonas have been associated with gynecologic and other cancers, though the pro-oncogenic mechanisms employed by these bacteria are poorly understood. Here, we integrated a multi-omics approach with our three-dimensional (3-D) cervical epithelial cell culture model to investigate how understudied BV-associated bacteria linked to gynecologic neoplasia influence hallmarks of cancer in vitro. Lancefieldella parvulum and Peptoniphilus lacrimalis elicited robust proinflammatory responses in 3-D cervical cells. Fusobacterium nucleatum and Fusobacterium gonidiaformans modulated metabolic hallmarks of cancer corresponding to accumulation of 2-hydroxyglutarate, pro-inflammatory lipids, and signs of oxidative stress and genotoxic hydrogen sulfide. This study provides mechanistic insights into how gynecologic cancer-associated bacteria might facilitate a tumor-promoting microenvironment in the human cervix.

The WID-BC-index identifies women with primary poor prognostic breast cancer based on DNA methylation in cervical samples.

Genetic and non-genetic factors contribute to breast cancer development. An epigenome-based signature capturing these components in easily accessible samples could identify women at risk. Here, we analyse the DNA methylome in 2,818 cervical, 357 and 227 matched buccal and blood samples respectively, and 42 breast tissue samples from women with and without breast cancer. Utilising cervical liquid-based cytology samples, we develop the DNA methylation-based Women's risk IDentification for Breast Cancer index (WID-BC-index) that identifies women with breast cancer with an AUROC (Area Under the Receiver Operator Characteristic) of 0.84 (95% CI: 0.80-0.88) and 0.81 (95% CI: 0.76-0.86) in internal and external validation sets, respectively. CpGs at progesterone receptor binding sites hypomethylated in normal breast tissue of women with breast cancer or in BRCA mutation carriers are also hypomethylated in cervical samples of women with poor prognostic breast cancer. Our data indicate that a systemic epigenetic programming defect is highly prevalent in women who develop breast cancer. Further studies validating the WID-BC-index may enable clinical implementation for monitoring breast cancer risk.

The DNA methylome of cervical cells can predict the presence of ovarian cancer.

The vast majority of epithelial ovarian cancer arises from tissues that are embryologically derived from the Müllerian Duct. Here, we demonstrate that a DNA methylation signature in easy-to-access Müllerian Duct-derived cervical cells from women with and without ovarian cancer (i.e. referred to as the Women's risk IDentification for Ovarian Cancer index or WID-OC-index) is capable of identifying women with an ovarian cancer in the absence of tumour DNA with an AUC of 0.76 and women with an endometrial cancer with an AUC of 0.81. This and the observation that the cervical cell WID-OC-index mimics the epigenetic program of those cells at risk of becoming cancerous in BRCA1/2 germline mutation carriers (i.e. mammary epithelium, fallopian tube fimbriae, prostate) further suggest that the epigenetic misprogramming of cervical cells is an indicator for cancer predisposition. This concept has the potential to advance the field of risk-stratified cancer screening and prevention.

Different expressions of aquaporin water channels and macrophages infiltration in human cervix remodeling during pregnancy.

Despite aquaporin water channels (AQPs) play a critical role in maintaining water homeostasis in female reproductive tract and prompt a gradual increase in water content in cervical edema as pregnancy progressed, their relationship with macrophage infiltration and collagen content in human cervical remodeling need to be further investigated. This is the first study to examine the expression and localization of AQP3, AQP4, AQP5, AQP8, and macrophages simultaneously in human cervical ripening. The immunoreactivity of these AQPs was 2.6 to 6-fold higher on gestational weeks 26 (GD26W) than that on GD6W and GD15W, but AQP4 expression on GD39W dropped a similar extent on GD15W, other AQPs continued to rise on GD39W. The AQP3, AQP4, and AQP5 intensity seemed more abundant in cervical stroma than in the perivascular area on GD26W; the distribution of AQP3, AQP5, and AQP8 in cervical stroma was equivalent to that in the perivascular area on GD39W. Macrophage numbers were 1.7-fold higher in subepithelium region and 3.0-fold higher in center area on GD26W than that on GD15W; such numbers remained elevated on GD39W. The electron micrographs showed that cervical extensibility increased significantly on GD26W and GD39W accompanied with increased macrophage infiltration, cervical water content, and much more space among collagen fibers. These findings suggest that the upregulation of AQPs expression in human cervix is closely related to enhanced macrophage infiltration during pregnancy; there may be a positive feedback mechanism between them to lead the increase of water content and the degradation of collagen.

Neoplasia intraepitelial escamosa cervical de alto grau: abordagem ambulatorial / High-grade cervical intraepithelial neoplasia: outpatient approach

As neoplasias intraepiteliais cervicais correspondem a alterações identificadas por rastreamento citológico cervical e estudo histológico, pós-biópsia incisional guiada por colposcopia ou procedimento diagnóstico excisional. Podem ser tratadas com abordagens conservadoras e procedimentos excisionais. A vacinação anti-HPV e o tratamento excisional oportuno constituem, respectivamente, prevenção primária e secundária contra o câncer do colo uterino.(AU)

Cervical intraephitelial neoplasms correspond to changes identified by cervical citological screening and histological study, post-incisional biopsy guided by colposcopy or excisional diagnostic procedure. They can be treated with conservative approaches and excision procedures. Anti-HPV vaccination and timely excional treatment are primary and secondary prevention against cervical cancer, respectively.(AU)

Acesso e acessibilidade ao rastreamento de câncer em mulheres brasileiras com lesão medular / Acceso y accesibilidad al rastreo de cáncer en mujeres brasileñas con lesión medular / Access and accessibility to cancer screening for Brazilian women with spinal cord injury

Resumo Objetivo identificar e analisar a acessibilidade e o acesso de mulheres brasileiras com lesão medular para a realização de exames preventivos do câncer de mama e colo de útero. Método estudo quantitativo e transversal desenvolvido em plataforma virtual. Realizadas análises estatísticas descritivas e de associação entre as variáveis qualitativas por meio do teste exato de Fisher. Quando identificada a associação (p<0,05), foi realizada a regressão logística. Resultados participaram 120 mulheres brasileiras com lesão medular com idades entre 25 e 67 anos; 85,83% foram ao ginecologista após a lesão medular, 79,17% realizaram a citologia e 52,50%, a mamografia. Observou-se que as mulheres que utilizavam a saúde suplementar apresentaram maior probabilidade de terem ido ao ginecologista do que as usuárias do serviço público. Aquelas com companheiro e as de maior idade apresentaram maior probabilidade de terem realizado o exame de citologia. Para a mamografia, aquelas de maior idade e que utilizavam a saúde suplementar apresentaram maiores chances de terem realizado o exame de mamografia após a lesão medular. Conclusão mulheres com lesão medular buscam a realização de exames de rastreamento. Entretanto, encontram dificuldades relacionadas à estrutura física, aos equipamentos, transporte, profissionais da saúde, assim como dificuldades sociodemográficas e quanto ao serviço de saúde utilizado.

Resumen Objetivo este estudio tuvo como objetivo identificar y analizar la accesibilidad y el acceso de mujeres brasileñas con lesión medular para la realización de exámenes preventivos de cáncer de mama y de cuello uterino. Método se desarrolló un estudio cuantitativo y transversal, realizado en un entorno virtual. Los análisis estadísticos descriptivos y la asociación entre variables cualitativas se realizaron mediante la prueba exacta de Fisher, cuando se identificó una asociación se realizó una regresión logística. Resultados participaron 120 mujeres brasileñas con lesión medular, la edad de las participantes varió de 25 a 67 años. Con relación al rastreo, el 85,83% de las mujeres acudió al ginecólogo tras la LM, el 79,17% se sometió a citología y el 52,50% a mamografía. Se observó que las mujeres que utilizaban un seguro médico privado tenían más probabilidades de haber visto a un ginecólogo que las usuarias del servicio público. Las que tenían pareja y mayores tenían más probabilidades de someterse a citología oncótica. Para la mamografía, las que eran mayores y que usaban un seguro médico privado tenían más probabilidades de someterse al examen después de la LM. Conclusión las mujeres con LM buscan pruebas de detección. Sin embargo, enfrentan dificultades relacionadas con la estructura física, equipamientos, transporte, profesionales de la salud, así como dificultades sociodemográficas relacionadas con el tipo de servicio de salud utilizado.

Abstract Objective to identify and analyze the accessibility and accessibility of Brazilian women with spinal cord injury to preventive examinations for breast and cervical cancer. Method quantitative and cross-sectional study developed in a virtual platform. Descriptive statistical analysis was performed, as well as association analysis between qualitative variables using Fisher's exact test. When identified the association (p<0.05), logistic regression was performed. Results a total of 120 Brazilian women with spinal cord injury, aged between 25 and 67 years participated in the study; 85.83% visited a gynecologist after the spinal cord injury, 79.17% underwent cytology and 52.50% underwent mammography. It was observed that women who used the supplementary health plan were more likely to have visited a gynecologist than those who used the public service. Those who had a partner and were older were more likely to have undergone the cytology exam. For mammography, those who were older and who used supplementary health care were more likely to have had mammography exams after the spinal cord injury. Conclusion women with spinal cord injury seek screening tests. However, they encounter difficulties related to the physical structure, equipment, transportation, health professionals, as well as socio-demographic difficulties and difficulties regarding the health service used.

Factores que afectan la suficiencia e interpretación de la citología de cuello uterino / Factors affecting cervical cytological sample adequacy and interpretation

Objetivo: evaluar los factores que afectan la suficiencia e interpretación de la citología de cuello uterino. Materiales y métodos: estudio transversal y retrospectivo en el Hospital Central "Dr. Urquinaona", Maracaibo, Venezuela con la revisión de las historias clínicas e informes de las citologías de cuello uterino de la consulta de ginecología y obstetricia de enero a diciembre 2019. Se analizan las características generales y la clasificación de los informes (satisfactorios para la evaluación, satisfactorios pero limitados por y no satisfactorios). Resultados: se seleccionaron 581 informes de los cuales 329 (56,6%) eran muestras satisfactorias, 233 (40,1%) satisfactorias pero limitadas y 19 (3,3%) insatisfactorios. El análisis univariante demostró que la muestra insatisfactoria, la presencia síntomas al momento de la toma y el tipo de método anticonceptivo fueron factores que se asociaron significativamente (p < 0,0001). Los que influyeron para un resultado anormal de la citología cervical fueron frotis satisfactorio (razón de probabilidad, 4,78; intervalo de confianza del 95%, 3,127-8,136) y presencia de síntomas (razón de probabilidad, 11,652; intervalo de confianza del 95%, 2,992-38,55). Esta asociación continuó siendo significativa luego de ajustarlos a los factores de edad, paridad, edad al momento de la toma de la primera citología y método de anticoncepción (p < 0,0001). Conclusión: la suficiencia de la muestra de citología es un factor importante para la detección de anomalías celulares de cuello uterino y evitar resultados falsos negativos, retrasando la detección del cáncer.

Objective: to evaluate factors that affect cervical cytological sample adequacy and interpretation. Materials and methods: a retrospective cross-sectional study conducted at Hospital Central "Dr. Urquinaona", Maracaibo, Venezuela by a review of the gynecology and obstetrics outpatient clinic medical records and cervical smear reports from January to December 2019. The general characteristics and classification of the reports (as, satisfactory, satisfactory but limited by and not satisfactory for cytological evaluation), were analyzed. Results: out of 581 reports selected, 329 (56.6%) were satisfactory, 233 (40.1%) satisfactory but limited and 19 (3.3%) not satisfactory. A univariate analysis showed that not satisfactory samples, presence of symptoms at the time of collection and type of contraceptive method were significantly associated factors (p < 0.0001). Those influencing an abnormal result were satisfactory smears (odds ratio, 4.78; confidence interval 95%, 3.127-8.136) and the presence of symptoms (odds ratio, 11.652; confidence interval 95%, 2.992-38.55). This association remained significant after considering other variables such as age, parity, age at first Pap smear and contraceptive method (p < 0.0001). Conclusion: cytological sample adequacy is an important factor for identifying cell abnormalities and avoiding false negative results which delay cancer detection.

Inhibition of miR-155 Promotes TGF-ß Mediated Suppression of HIV Release in the Cervical Epithelial Cells.

TGF-ß has been shown to play a differential role in either restricting or aiding HIV infection in different cell types, however its role in the cervical cells is hitherto undefined. Among females, more than 80% of infections occur through heterosexual contact where cervicovaginal mucosa plays a critical role, however the early events during the establishment of infection at female genital mucosa are poorly understood. We earlier showed that increased TGF-ß level has been associated with cervical viral shedding in the HIV infected women, however a causal relationship could not be examined. Therefore, here we first established an in vitro cell-associated model of HIV infection in the cervical epithelial cells (ME-180) and demonstrated that TGF-ß plays an important role as a negative regulator of HIV release in the infected cervical epithelial cells. Inhibition of miR-155 upregulated TGF-ß signaling and mRNA expression of host restriction factors such as APOBEC-3G, IFI-16 and IFITM-3, while decreased the HIV release in ME-180 cells. To conclude, this is the first study to decipher the complex interplay between TGF-ß, miR-155 and HIV release in the cervical epithelial cells. Collectively, our data suggest the plausible role of TGF-ß in promoting HIV latency in cervical epithelial cells which needs further investigations.

Human genital antibody-mediated inhibition of Chlamydia trachomatis infection and evidence for ompA genotype-specific neutralization.

The endocervix, the primary site of Chlamydia trachomatis (Ct) infection in women, has a unique repertoire of locally synthesized IgG and secretory IgA (SIgA) with contributions from serum IgG. Here, we assessed the ability of genital and serum-derived IgG and IgA from women with a recent positive Ct test to neutralize Ct elementary bodies (EBs) and inhibit inclusion formation in vitro in human endocervical epithelial cells. We also determined if neutralization was influenced by the major outer membrane protein (MOMP) of the infecting strain, as indicated by ompA gene sequencing and genotyping. At equivalent low concentrations of Ct EB (D/UW-3/Cx + E/UW-5/Cx)-specific antibody, genital-derived IgG and IgA and serum IgA, but not serum IgG, significantly inhibited inclusion formation, with genital IgA being most effective, followed by genital IgG, then serum IgA. The well-characterized Ct genotype D strain, D/UW-3/Cx, was neutralized by serum-derived IgG from patients infected with genotype D strains, genital IgG from patients infected with genotype D or E strains, and by genital IgA from patients infected with genotype D, E, or F strains. Additionally, inhibition of D/UW-3/Cx infection by whole serum, rather than purified immunoglobulin, was associated with levels of serum EB-specific IgG rather than the genotype of infecting strain. In contrast, a Ct genotype Ia clinical isolate, Ia/LSU-56/Cx, was neutralized by whole serum in a genotype and genogroup-specific manner, and inhibition also correlated with EB-specific IgG concentrations in serum. Taken together, these data suggest that (i) genital IgA most effectively inhibits Ct infection in vitro, (ii) human antibody-mediated inhibition of Ct infection is significantly influenced by the ompA genotype of the infecting strain, (iii) the genital antibody repertoire develops or matures differently compared to systemic antibody, and (iv) ompA genotype-specificity of inhibition of infection by whole serum can be overcome by high concentrations of Ct-specific IgG.

Segmentation of Overlapping Cervical Cells with Mask Region Convolutional Neural Network.

The task of segmenting cytoplasm in cytology images is one of the most challenging tasks in cervix cytological analysis due to the presence of fuzzy and highly overlapping cells. Deep learning-based diagnostic technology has proven to be effective in segmenting complex medical images. We present a two-stage framework based on Mask RCNN to automatically segment overlapping cells. In stage one, candidate cytoplasm bounding boxes are proposed. In stage two, pixel-to-pixel alignment is used to refine the boundary and category classification is also presented. The performance of the proposed method is evaluated on publicly available datasets from ISBI 2014 and 2015. The experimental results demonstrate that our method outperforms other state-of-the-art approaches with DSC 0.92 and FPRp 0.0008 at the DSC threshold of 0.8. Those results indicate that our Mask RCNN-based segmentation method could be effective in cytological analysis.

Inhibition of HMGB1/RAGE axis suppressed the lipopolysaccharide (LPS)-induced vicious transformation of cervical epithelial cells.

The chronic inflammation operates as one of the critical causes of cervical cancer. Activation of HMGB1/RAGE axis could induce the inflammation and lead to multiple types of cancer. However, whether the HMGB1/RAGE axis could affect the development of cervical cancer by regulating the inflammation is unclear. Here, we stimulated normal cervical epithelial cells with lipopolysaccharide (LPS). Next, the expression of RAGE in these cells was suppressed by the RAGE inhibitor. CCK-8 and wound healing assays were performed to detect the proliferation and invasion. To determine how inflammatory factors (IL-1ß, IL-6 and TNF-α) expressed in supernatant of these cells, ELISA was conducted. Western blotting was used for the detection of the expression of pyroptosis-related proteins (NLRP3 and caspase4). It was found that stimulation of LPS enhanced the proliferation and invasion of normal cervical epithelial cells. The expression of inflammatory factors (IL-1ß, IL-6 and TNF-α) in these cells was promoted as well. Application of RAGE inhibitor abolished the efficacy of LPS on these cells. Furthermore, LPS promoted the expression of NLRP3 and caspase4 in these cells while RAGE inhibitor exerted suppressive effects on the expression of these proteins. In summary, LPS-induced inflammation of normal cervical epithelial cells resulted in the malignant transformation of these cells by activating HMGB1/RAGE axis.

High-Resolution Quantitative Mapping of Macaque Cervicovaginal Epithelial Thickness: Implications for Mucosal Vaccine Delivery.

Vaginal mucosal surfaces naturally offer some protection against sexually transmitted infections (STIs) including Human Immunodeficiency Virus-1, however topical preventative medications or vaccine designed to boost local immune responses can further enhance this protection. We previously developed a novel mucosal vaccine strategy using viral vectors integrated into mouse dermal epithelium to induce virus-specific humoral and cellular immune responses at the site of exposure. Since vaccine integration occurs at the site of cell replication (basal layer 100-400 micrometers below the surface), temporal epithelial thinning during vaccine application, confirmed with high resolution imaging, is desirable. In this study, strategies for vaginal mucosal thinning were evaluated noninvasively using optical coherence tomography (OCT) to map reproductive tract epithelial thickness (ET) in macaques to optimize basal layer access in preparation for future effective intravaginal mucosal vaccination studies. Twelve adolescent female rhesus macaques (5-7kg) were randomly assigned to interventions to induce vaginal mucosal thinning, including cytobrush mechanical abrasion, the chemical surfactant spermicide nonoxynol-9 (N9), the hormonal contraceptive depomedroxyprogesterone acetate (DMPA), or no intervention. Macaques were evaluated at baseline and after interventions using colposcopy, vaginal biopsies, and OCT imaging, which allowed for real-time in vivo visualization and measurement of ET of the mid-vagina, fornices, and cervix. P value ≤0.05 was considered significant. Colposcopy findings included pink, rugated tissue with variable degrees of white-tipped, thickened epithelium. Baseline ET of the fornices was thinner than the cervix and vagina (p<0.05), and mensing macaques had thinner ET at all sites (p<0.001). ET was decreased 1 month after DMPA (p<0.05) in all sites, immediately after mechanical abrasion (p<0.05) in the fornix and cervix, and after two doses of 4% N9 (1.25ml) applied over 14 hrs in the fornix only (p<0.001). Histological assessment of biopsied samples confirmed OCT findings. In summary, OCT imaging allowed for real time assessment of macaque vaginal ET. While varying degrees of thinning were observed after the interventions, limitations with each were noted. ET decreased naturally during menses, which may provide an ideal opportunity for accessing the targeted vaginal mucosal basal layers to achieve the optimum epithelial thickness for intravaginal mucosal vaccination.

Fractal geometry applied to the analysis of cervix biopsy.

BACKGROUND: the measurement of the spaces of occupation of irregular objects in the context of fractal geometry has had some applications at a cellular morphometric level, where characterizations of normality and disease have been established. The objective of the present study is to apply a fractal methodology to characterize images from cervical colposcopy. MATERIALS AND METHODS: a mathematical and geometrical characterization of 67 cell samples was performed by measuring cellular fractal characteristics through the Box-Counting method, being nine normal, eight low-intraepithelial lesions, 16 high-intraepithelial lesions, eight carcinomas in situ, 20 squamous cell carcinomas and six endocervical carcinomas. RESULTS: the values of fractal dimension of the nuclear and cytoplasmic borders with respect to the totality varied between 0.719 to 1128 and 0.81 to 1024 while the occupation spaces in the 2 pixels grid were between 293 to 1606 and 64 to 693 respectively and in the 4 pixels grid oscillated between 153 to 894 and 36 to 361, respectively. Exocervical cells values had sensitivities between 78.3% to 100% in order to differentiate them from different types of cervical lesions. CONCLUSIONS: according to the results obtained, the mathematical values found are suggestive of being able to differentiate between normality and some colposcopy-guided cervical biopsy lesions.

Human Herpesviruses 6A and 6B in Reproductive Diseases.

Human herpesviruses 6A (HHV-6A) and human herpesvirus 6B (HHV-6B)-collectively, HHV-6A/B-are recently-discovered but ancient human viruses. The vast majority of people acquire one or both viruses, typically very early in life, producing an ineradicable lifelong infection. The viruses have been linked to several neurological, pulmonary and hematological diseases. In early human history, the viruses on multiple occasions infected a germ cell, and integrated their DNA into a human chromosome. As a result, about 1% of humans are born with the full viral genome present in every cell, with uncertain consequences for health. HHV-6A may play a role in 43% of cases of primary unexplained infertility. Both the inherited and acquired viruses may occasionally trigger several of the factors that are important in the pathogenesis of preeclampsia. Transplacental infection occurs in 1-2% of pregnancies, with some evidence suggesting adverse health consequences for the child. While emerging knowledge about these viruses in reproductive diseases is not sufficient to suggest any changes in current practice, we write this review to indicate the need for further research that could prove practice-changing.

The CXCL12/CXCR7 signalling axis promotes proliferation and metastasis in cervical cancer.

C-X-C chemokine receptor 7 (CXCR7), a novel receptor of C-X-C motif chemokine ligand 12 (CXCL12), is associated with the occurrence and metastasis of various malignant tumours. However, the role, function and underlying mechanisms of CXCR7 expression in cervical cancer remain undefined. The expression level of CXCR7 was evaluated in cervical cancer samples by immunohistochemistry and real-time PCR analyses. Western blot analysis was used to examine the expression level of CXCR7 in cervical cancer cell lines. HeLa cells were genetically silenced or pharmacologically inhibited for CXCR7 or CXCR4. Transwell and CCK-8 assays were used to examine cell migration and proliferation. The expression levels of MMP2, MMP9, TIMP-1 and TIMP-2 in HeLa cells were assessed by western blot or real-time PCR. HeLa cells silenced for CXCR7 were subcutaneously injected into nude mice to form tumours. The expression of CXCR7 in nude mice was investigated by immunohistochemical staining. Tumour volumes and weights were measured. The in vivo expression levels of MMP2, MMP9, TIMP-1 and TIMP-2 were determined by western blot analysis and real-time PCR. CXCR7 was overexpressed in cervical cancer tissues and cell lines. CXCL12 was highly expressed in cervical cancer lines. CXCR7 silencing or CCX733 treatment rather than CXCR4 silencing or AMD3100 treatment suppressed the proliferation, migration and invasion of cervical cancer cells stimulated by CXCL12. In a xenograft tumour model, CXCR7 silencing or CCX733 treatment inhibited the volumes and weights of xenograft tumours. In addition, downregulation of CXCR7 decreased the expression levels of MMP2 and MMP9 but increased the expression levels of TIMP-1 and TIMP-2 in vivo. These data support the finding that the downregulation of CXCR7 suppresses the proliferation and metastasis of cervical cancer cells. Inhibition of CXCR7 may be a potential targeted therapy for cervical cancer.

Progesterone alters human cervical epithelial and stromal cell transition and migration: Implications in cervical remodeling during pregnancy and parturition.

The cervix undergoes extensive remodeling throughout pregnancy and parturition. This process involves both ECM collagen degradation and cellular remodeling, which includes cell proliferation, transition and migration. Progesterone (P4) has been used clinically to delay cervical ripening and prevent preterm birth (PTB). However, the mechanisms by which progesterone affects cell transition and the migration of cervical epithelial and stromal cells are not yet fully known. In this study, we documented the role of a gestational level of P4 in the cellular transition (epithelial-mesenchymal transition [EMT] and mesenchymal-epithelial transition [MET]), cell migration, and inflammatory responses of endocervical epithelial cells (EEC) and cervical stromal cells (CSC). EEC and CSC were treated with LPS and P4 for 6 days. The epithelial:mesenchymal ratio (regular microscopy and cell shape index analysis), shift in intermediate filaments (immunofluorescence microscopy and western blot analyses for cytokeratin [CK]-18 and vimentin), adhesion molecules and transcription factors (western blot analyses for E-cadherin, N-cadherin and SNAIL), were used to determine growth characteristics and EMT and MET changes in EEC and CSC under the indicated conditions. To test cell remodeling, scratch assays followed by cellular analyses as mentioned above were performed. Inflammatory cytokines (interleukin-6 [IL-6], tumor necrosis factor α [TNFα]) and matrix metallopeptidase 9 (MMP9) were measured by ELISA. LPS promoted EMT (decreased cell shape index, decreased CK-18 and E-cadherin, increased vimentin, N-cadherin, and SNAIL), and increased IL-6 and MMP9 production by EEC. A gestational level of P4 prevented LPS-induced EMT in EEC and exhibited anti-inflammatory effect in both EEC and CSC. LPS slowed down wound healing in CSC but P4 treatment prevented the negative impact of LPS in CSC wound healing. These results may explain the cellular mechanisms by which P4 helps to stabilize the cervical epithelial barrier and preserve the mechanical and tensile strength of the cervical stromal layer, which are important in normal cervical remodeling processes during pregnancy.