Brachybacterium conglomeratum Is Associated with Cervicovaginal Infections and Human Papilloma Virus in Cervical Disease of Mexican Female Patients.

Brachybacterium conglomeratum, traditionally considered an environmental bacterium, has recently garnered attention for its potential involvement in human health. While prior research hinted at its pathogenic role in humans, our study aims to determine its prevalence and associations in diverse clinical contexts. We examined vaginal swabs from three distinct patient groups: patients with low-grade squamous intraepithelial lesions (LSIL), patients with cervicovaginal infections, and patients with a history of precancerous lesions undergoing follow-up. B. conglomeratum was present in all three patient groups, with the highest prevalence observed in the LSIL group. Statistically significant associations were primarily identified in the LSIL group, where B. conglomeratum was present in 60% of cases. Notably, the LSIL group exhibited coinfections with multiple high-risk oncogenotypes of human papillomavirus (HPV), suggesting potential synergistic effects, and understanding these microbial relationships and their influence on viral persistence, particularly with HPV, holds promise for mitigating HPV-related carcinogenesis. Furthermore, Gardnerella vaginalis and Atopobium vaginae were frequently detected in this group, along with Ureaplasma parvum as the predominant sexually transmitted bacterium. In all cases, B. conglomeratum was found in association with these microorganisms rather than as a sole pathogen. This coexistence underscores the intricate microbial interactions within cervicovaginal infections and precancerous lesions. This study marks the first report of B. conglomeratum prevalence in women with these clinical conditions.

An increase of microRNA-16-1 is associated with the high proliferation of squamous intraepithelial lesions in the presence of the integrated state of HR-HPV in liquid cytology samples.

Studies of cervical cancer (CC) have reported that microRNA-16-1 (miR-16-1), which is an oncomiR, is increased in the tissues and cell lines of CC. The aim of the present study was to investigate the association of miRNA-16-1 expression level with squamous cell carcinoma (SCC), the presence of squamous intraepithelial lesions (SIL) and the integration of high-risk human papillomavirus (HR-HPV) DNA. The current study analyzed 80 samples obtained from women by liquid-based cytology, which revealed that 20 were negative for SIL (NSIL) and without HPV, 20 were low-grade SIL (LSIL), 20 were high-grade SIL (HSIL), and 20 were diagnosed as SCC with HR-HPV. The genotyping of the viral DNA was conducted via an INNO-LiPA-HPV array, the expression of miR-16-1 was determined by reverse transcription-quantitative PCR, and the physical state of the HR-HPV was ascertained by in situ hybridization with amplification with tyramide. A total of eight HR-HPV genotypes were distinguished; the most frequent of these being HPV16, followed by multiple infection with HR-HPV (including HPV16). The mixed state of the HR-HPV was observed in 60 and 65% of LSIL and HSIL cases, respectively, while an integrated HR-HPV state was identified in 90% of cases with SCC. The expression level of miR-16-1 increased according to the grade of SIL, and cases with HSIL exhibited a significantly higher miR-16-1 expression level compared with women with NSIL (P<0.001; Table II). It can therefore be determined that the expression of miR-16-1 effects cellular proliferation, due to the viral integration of various HR-HPV genotypes in unique infection or in multiple infection. Thus, the overexpression of miR-16-1 could be monitored in women with LSIL, in order to discard a major lesion.

Evolución de las lesiones escamosas intraepiteliales de bajo grado del cérvix (2012-2013) / Evolution of Cervix Low-Grade Squamous Intraepithelial Lesions (2012-2013)

Introducción: las lesiones intraepiteliales cervicales son precursoras del cáncer de cuello uterino, enfermedad que causó 465 muertes en Cuba en el año 2015. Objetivo: describir la evolución de las lesiones intraepiteliales de bajo grado del cérvix durante un bienio según lo dispuesto en el Programa Nacional de Diagnóstico Precoz del Cáncer Cervicouterino. Métodos: se realizó un estudio descriptivo, longitudinal y prospectivo a 79 pacientes diagnosticadas citológicamente con una lesión escamosa intraepitelial de bajo grado en el período comprendido entre el 1ro de septiembre de 2012 y el 31 de agosto de 2013 en el Hospital Materno Ramón González Coro. Seguimiento evolutivo cada seis meses durante dos años. Resultados: 54,4 por ciento tenía entre 25 y 39 años de edad. Del total, 65,8 por ciento negativizó la citología entre los 6 y 18 meses. En los dos años de evolución, 25,3 por ciento del total progresó a una lesión de alto grado. Existió buena correlación citocolposcópica en 80 por ciento de las pacientes. El cálculo de la mediana de la zona transformación atípica en las lesiones de alto grado fue de 25 por ciento, muy superior a las de bajo grado. Hubo confirmación histológica en todos los casos cuya lesión progresó. Conclusiones: las pacientes que negativizaron la citología lo hizo durante los 18 meses de seguimiento. Existió buena relación citohistológica, no así colpohistológica. La evolución no guardó relación con las variables sociodemográficas excepto con el uso de DIU. Lesiones que ocupan más del 20 por ciento de la zona de transformación se relacionó con progresión hacia una lesión escamosa intraepitelial de alto grado(AU)

Introduction: Cervical intraepithelial lesions are signs of cervical cancer, a disease that caused 465 deaths in Cuba in 2015. Objective: To describe the evolution of low-grade intraepithelial lesions of the cervix during a biennium according to the provisions of the National Program of Early Diagnosis of Cervical Cancer. Methods: A descriptive, longitudinal and prospective study was carried out on 79 patients diagnosed by cytology with a low-grade squamous intraepithelial lesion from September 1, 2012 to August 31, 2013 at Ramón González Coro Gyneco-Obstetric Hospital. These subjects received an evolutionary follow-up every six months for two years. Results: 54.4 percent were between 25 and 39 years of age; 65.8 percent had negative cytology between 6 and 18 months. In the two years of evolution, 25.3 percent of the total progressed to a high-grade lesion. There was good cytocolposcopic correlation in 80 percent of the patients. The calculation of the median of the atypical transformation zone in the high-grade lesions was 25 percent, much higher than the low-grade ones. There was histological confirmation in all cases whose lesion progressed. Conclusions: Negative cytology in our patients was assessed during the 18 months of follow-up period. There was good cytohistological relationship, but not so for colpohistological relationship. The evolution was not related to the sociodemographic variables except with the use of IUD. Lesions that occupy more than 20 percent of the transformation zone were associated with progression towards a high-grade squamous intraepithelial lesion(AU)

The utility of pap cell block preparations with liqui-PREP™ cell pellets to clarify the cytological diagnosis of atypical squamous cells of undetermined significance and atypical glandular cells.

OBJECTIVE: Atypical squamous cells of undetermined significance (ASCUS) and atypical glandular cells (AGC) reflect cellular abnormalities insufficient for clear diagnosis. We used cell pellets obtained from liquid-based cytology (LBC) to prepare cell blocks (CB) and clarify the initial diagnosis of ASCUS and AGC. STUDY DESIGN: A total of 393 CBs with initial diagnosis of ASCUS or AGC were processed. Of those, 305 of the ASCUS and 20 of the AGC had adequate specimens. We compared results of CBs prepared from ASCUS and AGC to determine which had higher frequencies of higher-grade lesions. RESULTS: A majority of specimens (83%) were adequate for evaluation. Compared with the initial diagnosis, 14% (42/305) of ASCUS were diagnosed with low-grade squamous intraepithelial lesion (LSIL) in CB, while 10% (2/20) of AGC were diagnosed with LSIL or adenocarcinoma. No statistical relationship between the initial diagnosis of ASCUS and AGC and results of higher-grade lesions in CB is evident as determined by p value greater than 0.05 (p = 0.228). CONCLUSIONS: CBs prepared from Liqui-PREP cell pellets are, in most cases, assessable and can be useful as an adjunctive test to help clarify the initial diagnosis of ASCUS and AGC. Diagn. Cytopathol. 2017;45:520-525. © 2017 Wiley Periodicals, Inc.

HPV genotype distribution and anomalous association of HPV33 to cervical neoplastic lesions in San Luis Potosí, Mexico.

BACKGROUND: The association of human papillomavirus (HPV) types to neoplastic lesions increase as a function of their oncogenicity and the duration of the infection since lesion severity progresses from low-grade to high-grade and cancer. In an outbreak, the prevalence of the HPV type involved would increase and the proportion of the associated low-grade lesions would predominate over severe lesions. In this study, the prevalence of HPV types and their association to neoplastic lesions was determined in women subjected to colposcopy in San Luis Potosí, Mexico. METHODS: DNA from high-risk (HR) and low-risk (LR) HPV types was identified by E6 nested multiplex PCR in cervical scrapes from 700 women with normal cytology, atypical squamous cells of undetermined significance (ASCUS), low-grade squamous intraepithelial lesions (LSIL), high-grade squamous intraepithelial lesions (HSIL) or invasive cervical cancer (CC). RESULTS: Overall HPV-DNA prevalence was 67.7 %, that of HR-HPV was 63.1 %, and that of LR-HPV was 21.3 %. The highest prevalence (78.2 %) occurred in the 15-24 year group, whereas that of single infections was 52 % and that of multiple infections (i.e., by 2-6 HPV types) was 48 %. The most prevalent HR types were HPV33 (33.1 %), HPV16 (16.6 %), HPV18 and HPV51 (6.7 % each). HR-HPV prevalence was 29.6 % in normal cytology, 26.7 % in ASCUS, 63.3 % in LSIL, 68.2 % in HSIL, and 90.5 % in CC. Three prevalence trends for HR-HPV types were found in neoplastic lesions of increasing severity: increasing (LSIL < HSIL < CC) for HPV16, HPV39, HPV18, HPV58, HPV31 and HPV35; asymptotic (LSIL < HSIL ≈ CC) for HPV51 and HPV68; U-shaped (LSIL < HSIL > CC) for HPV33. CONCLUSIONS: Two-thirds of the women subjected to colposcopy from 2007 to 2010 in San Luis Potosí have HPV infections which predominate in the 15-24 years group. Around half of the infections are by one viral type and the rest by 2-6 types. HPV33 is the most prevalent type, followed by HPV16. Overall HR-HPV prevalence increases with the severity of neoplastic lesions. HPV33 prevalence is highest in LSIL and its U-shaped trend with progressing neoplastic lesions differs from the growing/asymptotic trends of other HR-HPV types. An ongoing or recent HPV33 outbreak is consistent with its high prevalence and anomalous association to LSIL.

Follow-up outcomes in a large cohort of patients with HPV-negative LSIL cervical screening test results.

OBJECTIVES: Cervical screening guidelines now recommend repeat cotesting of patients aged 30 years and older having a human papillomavirus (HPV)-negative low-grade squamous intraepithelial lesion (LSIL) in 1 year as preferred management. Only limited follow-up data on patients with HPV-negative LSILs are available from routine US clinical practice settings. METHODS: In total, 680 patients with Hybrid Capture 2 (Qiagen, Hinden, Germany) high-risk HPV-negative LSIL ThinPrep (Hologic, Marlborough, MA) results were identified. Patients' ages and histopathologic, cytologic, and HPV follow-up results were identified. RESULTS: Among 680 patients with HPV-negative LSILs, 468 had follow-up within 1 year. During the study period, 14 (3.0%) of 468 had follow-up high-grade squamous intraepithelial lesion (HSIL) and 184 (39.3%) LSIL findings. No diagnoses of cervical carcinoma were documented. There were no significant follow-up differences between age groups. Of the 321 patients who had follow-up HPV testing, 271 (84.4%) had negative and 50 (15.6%) had positive HPV results. CONCLUSIONS: This is the largest study documenting follow-up results for patients with HPV-negative LSIL results based on prevalent US FDA-approved co-testing methods from one collection vial. These data document that risk for follow-up HSILs in these patients is low and also that no cervical cancers were diagnosed. These findings support recent recommendations for repeat co-testing after 1 year as an appropriate option for patients with HPV-negative LSIL results.

The integration of HR-HPV increases the expression of cyclins A and E in cytologies with and without low-grade lesions.

BACKGROUND: Cyclin-A and cyclin-E are regulators of G1-S phase of normal cell cycle. Integration of human papilloma virus high-risk (HR-HPV) could alter this mechanism, and its overexpression has been associated with poor prognosis in cervical cancer. AIM: To determine the expression of cyclin-A and cyclin-E, types of HR-HPV and physical state of DNA in cytologies with the diagnosis of low-grade squamous intraepithelial lesion (LSIL). MATERIALS AND METHODS: 115 cytological specimens in liquid base (liquid-PREP(™)) were analyzed. 25 specimens were with no signs of SIL (NSIL) and without HPV; 30 with NSIL with low-risk HPV (LR-HPV); 30 with NSIL with HR-HPV; and 30 with both LSIL and HR-HPV. The expression of cyclins was evaluated by immunocytochemistry; and the detection of viral DNA was done by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLPs) for genotyping or sequencing of HPV. The physical state of HPV was evaluated by in situ hybridization with amplification with tyramide. RESULTS: In the cytologies NSIL with LR-HPV, the expression of cyclin-A and cyclin-E was found respectively in 23.3% and 33.3% of the specimens. Among the specimens of NSIL with HR-HPV, 33.3% expressed cyclin-A and 40% cyclin-E, while 100% of the LSILs expressed the 2 cyclins. On the other hand, 100% of the samples NSIL with LR-HPV presented an episomal pattern. Of the specimens of NSIL with HR-HPV, 56.6% exhibited an episomal pattern, 23.3% integrated and 20%, mixed. Among the LSILs, 90% were mixed and 10% integrated. CONCLUSIONS: The cyclins A and E are present in the LSILs that occur predominantly in mixed state in the presence of HR-HPV.

Urothelial cells in smears from cervix uteri.

OBJECTIVES: To establish the cytological criteria to identify the urothelial cells in cervical smears in order to avoid mistakes in the cytological diagnosis. MATERIALS AND METHODS: Cervical smears from 34 post menopausal women with vesicovaginal fistulas, advanced bladder prolapse and genital erosive lichen planes (vulvar kraurosis) (Group 1) and transitional cell metaplasia of the cervix (TCM, Group 2) were stained with Papanicolaou technique. The cervical samples were taken during the routine annual examination for prevention of the uterine cancer. RESULTS: The smears of cervix from Group 1 showed urothelial cells from the three layers of the transitional epithelium. The umbrella cells are the bigger ones with relatively large nuclei. Frequently, they are multinucleated with single or multiple nucleoli and a typical "frothy" cytoplasm (cytoplasmic vacuoles). The cells of the Group 2 showed nuclei with oval to spindled shapes, some tapered ends, less cytoplasm than squamous metaplastic cells, powdery chromatin, small nucleoli and nuclear grooves. CONCLUSIONS: The umbrella cells may be mistaken for dysplastic cells originating in low grade squamous intraepithelial lesions lesions (LSILs) due to their nuclear and cytoplasm sizes. Therefore, it is important to know the possibility of their appearance in the cervical smears, especially in post menopausal patients in order to avoid a false diagnosis of an intraepithelial lesion. It is unlikely that deeper cells of urothelium would be confused with high grade squamous intraepithelial lesion (HSIL) cells. However, their presence might be a reason of mistake in the diagnosis. TCM is an under-recognized metaplastic phenomenon of the cervix and vagina, which is a mimicker of high-grade squamous intraepithelial lesion. The differential characteristic between umbrella cells, cells from TCM and the deeper urothelial cells, and LSIL and HSIL are detailed in the present paper.

Lesión escamosa intraepitelial de bajo grado y su correlación citocolpohistológica / Low-Grade Squamous Intraepithelial Lesion and its cyto-colpo-histologic correlation

Introducción: La detección de las lesiones tempranas y precursoras al carcinoma del cérvix uterino se realiza con la interpretación del papanicolaou. Objetivo: Determinar la correlación entre la citología cervical comparada con el diagnóstico por colposcopia y de histología en la lesión escamosa intraepitelial de bajo grado (LEIBG). Material y métodos: A 100 pacientes con citología compatible con LEIBG se les realiza colposcopia y se toma biopsia dirigida para estudio histopatológico. Resultados: La edad promedio fue de 37 años, y el inicio de vida sexual activa 19.25 años, con un número de parejas sexuales de 2.57, uso de condón en 0.3%, y eran fumadoras 45% de las pacientes. La citología diagnosticó: LEIBG en 77 de las pacientes (77%); proceso inflamatorio 20% y lesiones escamosas intraepiteliales de alto grado (LEIAG) en 3%; sensibilidad de 88.23%; especificidad de 42.85%; valor predictivo positivo (VPP) de 94.93%, y valor predictivo negativo (VPN) de 23.07%. La citología de la unidad diagnosticó: LEIBG en 85% de las pacientes; proceso inflamatorio en 13%; LEIAG en 2%; sensibilidad en 79.12%; especificidad de 28.57%; VPP de 93.50%, y VPN de 9.52%. La colposcopia con el índice de Reid diagnosticó: LEIBG en 92%; proceso inflamatorio de 6%; LEIAG en 2%; sensibilidad en 93.47%; especificidad de 33.33%; VPP de 95.55%, y VPN de 25%. El estándar de oro fue por histopatología: en un 100% con diagnóstico a LEIBG. Conclusión: Ante una citología anormal debe realizarse colposcopia con toma de biopsia dirigida para un diagnóstico más exacto. La citología seguirá siendo un buen estudio de escrutinio ya que logra detectar lesiones tempranas.

Introduction: The detection of the early lesions precursors of uterine cervix carcinoma is carried out based on the interpretation of Papanicolaou smear. Objective: To determine the correlation that exists betweeen cervical cytology compared with the diagnosis by either colposcopy or histology in the Low-Grade Squamous Intraepithelial Lesion (LSIL). Material and methods: 100 patients with cytology compatible with LSIL are practiced colposcopy taking directed biopsy for histopathologic study. Results: Average age: 37; Average onset of sexually active life: 19.25 years old; number of sexual partners: 2.57; condom use in 0.3%. 45% of the patients were smokers. Diagnosis by cytology: LSIL in 77 (77%) of the patients, inflammatory process 20 (20%), and HSIL 3 (3%). Sensibility: 88.23%; specificity 42.85%; positive predictive value (PPV): 94.93%, and negative predictive value (NPV): 23.07%. LSIL diagnosis by cytology: 85 patients (85%); inflammatory process: 13 (13%); HSIL: 2 (2%); Sensibility: 79.12%; specificity: 28.57%; PPV: 93.50%; NPV: 9.52%. The colposcopy with Reid index and diagnosis of LSIL: 92 (92%) patients; inflammatory process: 6 (6%); HSIL: 2 (2%); Sensibility: 93.47%; Specificity: 33.33%; PPV: 95.55%; NPV: 25%. The gold standard was histopathology: 100% with a diagnosis of LSIL. Conclusion: Before an abnormal cytology, colposcopy with directed biopsy should be performed for a more accurate diagnosis. Cytology will remain a good screening test due to its usefulness to detect early cervical lesions.

Clinical response to glycyrrhizinic acid in genital infection due to human papillomavirus and low-grade squamous intraepithelial lesion.

Human papilloma virus (HPV) can infect any of the mucosal areas of the body and cause cervical cancer. Until recently, no specific treatments were available for this condition; therefore, any damaged tissue had to be removed or destroyed, which may have presented obstetrical repercussions for some women. Recently, new drugs have been developed that have shown to be effective for the cure of HPV infection. Glycyrrhizinic acid (GA) has shown fewer side effects and its systemic use makes it possible to reach difficult-to-treat lesions. The purpose of this study was to evaluate the clinical outcome of GA to eliminate the epithelial lesion and HPV. We carried out a longitudinal, descriptive study that included women of reproductive age who were diagnosed with HPV associated with low-grade squamous intraepithelial lesion (LSIL). Subjects began treatment based on GA using two routes of administration - systemic (oral) and topical (spray) - with assessments every month to determine the clinical changes of the lesions through colposcopy and Papanicolaou (Pap) smear. Simple statistics were used along with two-tailed Student's t-test; P<0.05 was considered statistically significant before and after treatment. There were 70 eligible patients, of whom 62 fulfilled the inclusion criteria. Age of subjects was 27.8±9.5 years. At the time of the study, 100% of the patients had HPV infection, 40% were associated with LSIL, and only 16% used a barrier contraceptive (condom) method. Resolution was achieved in all patients from 4 weeks of treatment initiation and improvement was achieved in the majority of patients at 12 weeks (74%) (P<0.001). However, there was persistence of LSIL in 27.7% of patients and only one patient progressed to cervical intraepithelial neoplasia (CIN) II. The use of GA proved to be effective in resolving clinical HPV lesions. For cervical lesions with epithelial changes (LSIL), treatment may be required for a longer period as with other drugs used for this infection, as well as monitoring for at least 1 year according to the natural evolution of the disease.

Human papillomavirus false positive cytological diagnosis in low grade squamous intraepithelial lesion

The purpose of this study was to investigate the number of Human Papillomavirus false positive cytological diagnosis in low grade squamous intraepithelial lesions (LSIL). Three hundred and two women who assisted to an Out-Patient Gynecologic Clinic in Maracaibo, Venezuela, were recruited for this study. Each patient had the Pap smear and a cervical swab for Hybrid Capture 2 (HC2). Three cytotechnologists reviewed the Pap smears and two pathologists rescreened all of them. The cytotechnologists reported 161 (53.3 percent) Pap smears negatives for intraepithelial lesion (IL) or malignancy, and 141 cases (46.7 percent) with epithelial abnormalities. They reported 46 percent of 302 patients with HPV infection in Pap smear slides. The pathologists found that 241 (79.8 percent) Pap smears were negatives for IL or malignancy and 61 (20.2 percent), with abnormal Pap smears. They found 14.6 percent HPV infection in all Pap smears (p<0.0001; 46 percent vs 14.6 percent). The HC2 study showed that 47 samples (15.6 percent) were positive for HPV. The study found that 114 Pap smears (False Positive: 85 percent) of 134 reported by the cytotechnologists and 24 (False Positive: 43 percent) of 56 cytologies reported by the pathologists as LSIL, were negative for HPV infection determined by HC2 (p<0.00003). The present study suggests that the cytotechnologists overdiagnosed cellular changes associated with HPV infection in the Pap smear, increasing the FP cytological diagnosis of LSIL.

El presente trabajo tuvo por objeto el investigar el número de falsos positivos reportados en la citología cervicovaginal (CCV) de la presencia del Virus del Papiloma Humano (VPH) con diagnóstico de Lesión Intraepitelial Escamosa de bajo grado (LIE-BG). Se estudiaron 302 mujeres que asistieron a la Consulta de Patología de Cuello Uterino del Hospital Manuel Noriega Trigo, en Maracaibo, Venezuela. A cada paciente se le practicaron una CCV y muestra para la captura de híbridos 2 (CH2). Tres citotecnólogos y 2 patólogos estudiaron las CCV. Los citotecnólogos reportaron 161(53,3 por ciento) de CCV negativas para lesión intraepitelial o malignidad y 141 casos (46,7 por ciento) con anomalías epiteliales. Éstos encontraron 46 por ciento de presencia de VPH en las 302 CCV. Los patólogos reportaron 241 CCV (79,8 por ciento) negativas y 61 CCV (20,2 por ciento) anormales. Estos encontraron en 14,6 por ciento de las CCV, la presencia de VPH (p < 0, 0001; 46 por ciento vs 14,6 por ciento). La CH2 mostró que 47 muestras (15, 6 por ciento) fueron positivas a VPH. Esta investigación mostró que 112 CCV de 134 (Falso Positivo: 85 por ciento) reportados por los citotecnólogos y 24 de 56 CCV (Falso Positivo: 43 por ciento) reportados por los patólogos como LIE-BG, fueron negativos a la infección del VPH determinados por la CH2 (p < 0,00003). La investigación sugiere un sobrediagnóstico de la presencia de cambios celulares debidos al VPH en la CCV, por parte de los citotecnólogos, incrementando los falsos positivos de la presencia del VPH en CCV con diagnóstico de LIE-BG.