Exploring the Concordance Between High-Risk Human Papillomavirus Infections in Cervical and Oral Sites Among Females: A Cross-Sectional Study in Punjab, Pakistan.

PURPOSE: Human papillomavirus (HPV) is widely recognized as a key contributing factor in cervical and oropharyngeal cancers. However, there has been limited research on the prevalence of concurrent HPV infections in various anatomic regions. This study aimed to investigate the prevalence and specific types of high-risk HPV (HR-HPV) infections in the cervical and oral regions of females in Punjab, Pakistan. METHODS: We conducted a cross-sectional study involving women seeking care for general gynecologic issues at the gynecologic Outpatient Department of Lady Wallington Hospital in Lahore. After interviews and clinical examinations, we collected whole-saliva samples and high vaginal swabs from each participant. HR-HPV detection and genotyping were performed using real-time polymerase chain reaction at both the anatomic sites. RESULTS: In this study, 170 females, averaging 35.36 ± 8.305 years, participated. HR-HPV infection was more prevalent in the cervix (83/170 [48.8%]) than in the oral cavity (19/170 [11.2%]). Concordant HPV infections occurred in 10/170 participants (5.9%). HPV 16 was the most common genotype in cervical and oral locations, at rates of 21.8% and 5.3%, respectively, among concordant HR-HPV types. Socioeconomic status (P = .013), age at first sexual intercourse (P = .015), and history of oral sex (P = .01) were significantly associated with concurrent HR-HPV infection in both regions. CONCLUSION: This study suggests that HR-HPV cervical infections may increase the risk of oral transmission, especially during orogenital sexual practices. Thus, it is important to recognize that HPV infections may be linked in both areas. We emphasize the importance of comprehensive cervical and oral examinations and HPV vaccination in young women irrespective of their sexual practices.

Molecular characterization and risk factors of oral high-risk human papillomavirus among females in Punjab, Pakistan.

BACKGROUND: The development of oral and oropharyngeal cancers is associated with human papillomavirus (HPV) infection. However, debate exists about the prevalence of high-risk oral HPV (HR-HPV) and its associated risk factors. This study evaluated the distribution of genotypes and associated risk factors of oral HR-HPV infection in the normal oral mucosa of the Pakistani female population with general gynecological problems. METHODS: A total of 200 females from a regional gynecological hospital in Pakistan were included in this cross-sectional study. A self-administered questionnaire was carried out, followed by the study participants' oral and dental examinations. Whole saliva was collected by a drooling method. The viral DNA extraction and oral HR-HPV genotyping were detected via real-time polymerase chain reaction. RESULTS: Among the 200 females (mean age, 37.76 years), 22 females (11%) were positive for oral HR-HPV DNA. Single oral HR-HPV infection was seen in 8%, whereas co-infection was observed in 3% with HPV16 and HPV18 most common genotypes. Comparison of the risk factors like oral sex history odds ratio (OR 8.21, 95% confidence interval [CI] 1.85-28.16; p < 0.005), and open-mouth kissing (OR, 5.17; 95% CI 1.48-27.41; p = 0.005) with oral HR-HPV infection was found to be significantly associated. CONCLUSION: In Pakistan, oral HR-HPV-positive infection is not uncommon; HPV16 and HPV18 are the most frequent genotypes. Furthermore, this study also provides data that oral HR-HPV positive infection was significantly associated with sexual behaviors. Therefore, preventive health efforts, like health education and vaccination, should be practiced.

Whole-genome analysis and evolutionary characterization of cervical and oral human papillomavirus 16.

High-throughput genome-wide sequencing has revealed high genomic variability of HPV16 in different geographic regions which is the most predominant genotype in human papillomavirus (HPV)-associated malignancies. Analysis of the HPV16 by whole-genome sequence (WGS) is an advanced method for the identification of mutations in the genome. There is limited information about HPV16 diversity in Pakistan, especially at the genomic level. Till now, WGS for HPV16 has not been previously reported in Pakistan. The current study has sequenced three HPV16 viral genomes, from two cervical and one oral cavity positive sample of women presented with general gynecological problems without any evidence of precancerous or cancerous lesions using an ion ampliseq customized panel. Sequencing analysis detected 38 variations, including single-nucleotide polymorphisms (SNPs) and two Indels, across three samples with the highest number of SNPs present in E1, E2, and L2, respectively. A total of 20 non-synonymous and 11 synonymous mutations with amino acid substitutions (T1421C, G1515A, T2223C, T1389C, G1483A, and T2191C) were identified. The phylogenetic analysis revealed the genomes of HPV16 are closely associated with those reported from Thailand and the United States. These are the first HPV16 WGS from Pakistan. However, more research is needed with a large sample size from diversified areas to assess the carcinogenic consequences and impact of HPV vaccinations.

Distribution of High-risk Human Papillomavirus Genotypes in Cervical Secretions in Punjab.

OBJECTIVE: To determine the high risk-human papillomavirus (HR-HPV) prevalence, genotype distribution, and associated risk factors in women with general gynaecological problems. STUDY DESIGN: Descriptive, cross-sectional study. PLACE AND DURATION OF STUDY: Lady Wallingdon Hospital, Lahore from August 2018 to May 2019. METHODOLOGY: One hundred females with general gynaecological problems constituted the sample. Real-time polymerase chain reaction and liquid base cytology were performed for HR-HPV detection, genotyping, and cytological changes in cervical samples. RESULTS: The overall prevalence of HR-HPV was found to be 57%, with the most frequent HR-HPV genotypes HPV 16 (18%), HPV 18 (6%), and HPV 45 (1%). The total prevalence of single HR-HPV infection was 25%; whereas, multiple co-infection of HR-HPV was seen in 32% of cases. In total, 81% of these women had normal cytology and 19% had cytologic abnormalities. A significant association was observed between HR-HPV infection and general gynaecological complaints and between HR-HPV infection and cytological abnormalities (p < 0.001). CONCLUSION: The HR-HPV prevalence was 57% with the most prevalent genotypes were HPV 16 and HPV 18. The results will also aid the identification of HR-HPV 51 and 31 genotypes, which should be added in the multi-type HPV vaccines. Key Words: Cancer of cervix, Human papillomavirus, Papanicolaou test, Pakistan.

A Cytological Study Enlightening the Unseen Effects of Concomitant Chemoradiotherapy in Contralateral Normal Buccal Mucosa of Oral Squamous Cell Carcinoma Patients.

Background/objectives In patients receiving concomitant chemoradiotherapy (CCRT) as a treatment for oral squamous cell carcinoma (OSCC), cytological changes were seen not only in neoplastic epithelial cells but the non-neoplastic epithelial cells are also affected, resulting in cytopathological atypical changes. The present study was designed to observe oral epithelial atypical cytopathologic changes induced in contralateral normal buccal mucosa in OSCC patients receiving CCRT. Methods The study included 150 patients with OSCC treated by CCRT whose details were collected from the Institute of Nuclear Medicine and Oncology Lahore (INMOL) Hospital Lahore. Cytological smears were obtained from the contralateral normal buccal mucosa of OSCC patients. The serial scrape smears were taken before, immediately after, on the 17th day (mid of treatment), and at the end of CCRT, whereas 20 patients were taken as normal healthy controls and were not exposed to CCRT. The smears were stained with hematoxylin and eosin and Papanicolaou stain. SPSS version 20 (Armonk, NY: IBM Corp.) was used for statistical analysis and p > 0.05 was considered to be significant. Results CCRT-induced oral epithelial atypical cytological changes were predominantly noted at end of therapy (19.7%) in the contralateral normal buccal mucosa. Nuclear atypia features were higher on the 17th day and end of treatment; whereas, epithelial atypia was mainly observed on the 17th day of CCRT (40%). A highly significant association was observed between epithelial atypia and radio-chemotherapy dose (p = 0.045), between CCRT-induced epithelial atypical cytological changes and days of treatment (p = 0.001), and between days of CCRT and nuclear atypia (0.000) accordingly. Atypia was not observed in any control group. Conclusion Varying degrees of oral epithelial atypical cytological changes may occur in otherwise normal contralateral mucosa of the patients receiving CCRT.

Assessment and prevalence of concomitant chemo-radiotherapy-induced oral mucositis in patients with oral squamous cell carcinoma

Background/aim: Quantification of oral mucositis that progresses during concomitant chemo-radiotherapy (CCRT) is essential for its management. It is important to determine the methods that are simple, reliable and beneficial in foreseeing mucositis at earlier stages of treatment. Materials and methods: A prospective study was conducted on 100 oral cancer patients receiving CCRT following the inclusion criteria. Patients were evaluated for mucositis i.e. erythema and ulcers by using the World Health Organization (WHO) scale and the oral mucositis assessment scale (OMAS), whereas mature and immature cells were identified by exfoliative cytology. Clinical examination and procedure of oral cavity were performed before, on days 5, 17, and at the end of treatment. Results: Oral mucositis was observed in all oral squamous cell carcinoma (OSCC) patients receiving CCRT on different days with noteworthy increase from day 5 of CCRT to the end of treatment. For OMAS grading related to ulceration and erythema, Grade 1 (7.2%; 34%) was most commonly seen on the 5th day of CCRT, Grade 2 (29%; 19%) and Grade 3 (19%) were most frequently seen at the 17th day and end of CCRT, accordingly. With respect to WHO scale grades 1 and 2 (18.3%; 21.5%) was most frequently observed at the 17th day of CCRT, whereas grades 3 and 4 (12.5%; 2%) was noted at the end of CCRT. There was statistically significant increase in the percentage of immature cells at the end of CCRT (99%). A significant association (P < 0.0000) was observed among the days of smear and maturation stages of epithelial cells as well as among WHO mucositis grading, OMAS and types of epithelial cells, respectively. Conclusion: According to the findings of the study, oral mucositis grade is directly proportional to the progressing days of CCRT. Oral mucositis is frequently related to adverse clinical outcomes, affecting the patient's quality of life. It is essential to develop methods that can be employed for the assessment of CCRT associated oral mucositis.

Corona pandemic: awareness of health care providers in Pakistan.

INTRODUCTION: Corona pandemic has resulted in a high mortality rate among health care professionals. The purpose of this study was to assess the knowledge and practices of health care providers during this pandemic in Punjab, Pakistan. METHODS: A web based cross sectional survey was conducted during 2nd of April to 20th April 2020 targeting health care professionals working in Punjab, Pakistan. A sample of 540 participants was collected using non probability, convenient sampling technique. Data was generated by using on line google forms after taking IRB approval from institution. RESULTS: Adequate knowledge was found among health care providers regarding diagnostic tests, modes of transmission, incubation period and preventive strategies. Significant association was seen in knowledge of post graduate and MBBS doctors (Bachelor of Medicine and Bachelor of Surgery) regarding viral etiology (p = 0.006), sign and symptoms (p = 0.000), risk factors (p = 0.000) and mortality rates (p = 0.001). Hand washing was considered as best preventive measure by 95% of the health care providers. CONCLUSION: Post graduate doctors have better knowledge regarding corona virus disease transmission, risk factors, incubation period and preventive strategies as compared to undergraduate doctors.

Efficiency of Crystal Violet Stain to Study Mitotic Figures in Oral Epithelial Dysplasia.

AIM: To evaluate mitotic activity in the different grades of oral epithelial dysplasia using 1% crystal violet stain. MATERIAL AND METHODS: A descriptive study was conducted in the Department of Histopathology of the Post Graduate Medical Institute, Lahore on a total of thirty-three cases of the Oral Epithelial Dysplasia (OED). Fresh, frozen paraffin-embedded archival tissue blocks were collected from Lahore General Hospital, Lahore & Oral & Maxillofacial Surgery Department of Nawaz Sharif Hospital, Yakki Gate, Lahore. The representative sections were taken and, after processing, mounted on glass slides and stained with H&E and crystal violet stains. The stained slides were then examined under an optical microscope. The efficacy of 1% crystal violet stain to identify mitotic figures in the different grades of oral epithelial dysplasia was assessed with the sample t-test. A difference of p < 0.05 was considered to be significant. RESULTS: A comparison of the mitotic figure count in two categories in sections stained with both stains showed a statistically significant difference. An increase in the mean mitotic count was noted in the sections of OED stained with crystal violet in comparison to the sections of OED stained with H&E which was statistically significant (p = 0.00). CONCLUSION: Counting of mitotic cell is the rapid and simplest way of evaluating the proliferative activity of cells. Crystal violet stain can be a rationalised step in the staining of mitotic figures compared to the usual H&E staining and can be employed as a selective stain during routine histopathological procedures.

Oral Ulcers Presentation in Systemic Diseases: An Update.

BACKGROUND: Diagnosis of oral ulceration is always challenging and has been the source of difficulty because of the remarkable overlap in their clinical presentations. AIM: The objective of this review article is to provide updated knowledge and systemic approach regarding oral ulcers diagnosis depending upon clinical picture while excluding the other causative causes. METHODS: For this, specialised databases and search engines involving Science Direct, Medline Plus, Scopus, PubMed and authentic textbooks were used to search topics related to the keywords such as oral ulcer, oral infections, vesiculobullous lesion, traumatic ulcer, systematic disease and stomatitis. Associated articles published from 1995 to 2019 in both dental and medical journals including the case reports, case series, original articles and reviews were considered. RESULTS: The compilation of the significant data reveals that ulcers can be classified according to (i) duration of onset, (ii) number of ulcers and (iii) etiological factors. Causation of oral ulcers varies from slight trauma to underlying systemic diseases and malignancies. CONCLUSION: Oral manifestations must be acknowledged for precise diagnosis and appropriate treatment.

Concomitant-chemoradiotherapy-associated oral lesions in patients with oral squamous-cell carcinoma.

OBJECTIVE: : Oral squamous-cell carcinoma (OSCC) accounts for >90% of oral cancers affecting adults mostly between the fourth to seventh decades of life. The most common OSCC treatment is concomitant chemoradiotherapy (CCRT) having both loco-regional and distant control, but CCRT has acute and chronic toxic effects on adjacent normal tissue. This study aimed to determine the side effects of CCRT on the oral mucosa and to characterize the clinicopathology of oral lesions in patients with OSCC. METHODS: This descriptive, cross-sectional study was certified by the Ethical Review Committee (UHS/Education/126-12/2728) of the University of Health Sciences, Lahore, Pakistan. OSSC patients (n=81) with various histological subtypes, grades, and stages were recruited, and findings on their oral examination were recorded. These patients received 70, 90, and 119 Gy of radiotherapy dosages in combination with the chemotherapy drugs cisplatin and 5-fluorouracil. Data were analyzed using SPSS 20.0. RESULTS: : The most common presentation of OSCC was a nonhealing ulcer (63%) involving tongue (55.6%). Clinical findings included mucositis (92.6%) and xerostomia of mild, moderate, and severe degrees in 11.1%, 46.9%, and 35.8% cases, respectively. Ulcers (87.7%), palpable lymph nodes (64.2%), limited mouth opening (64.2%) and fistula (40.7%) were also observed. In females, the association of radiotherapy dosage with limited mouth opening, xerostomia, and histological grading was statistically significant (P<0.05). The association of chemotherapy drugs with xerostomia (P=0.003) was also statistically significant. CONCLUSIONS: : CCRT induced mucositis, xerostomia, and trismus in patients with OSCC.

Stereotactic radiosurgery versus whole-brain radiotherapy after intracranial metastasis resection: a systematic review and meta-analysis.

BACKGROUND: In patients with one to three brain metastases who undergo resection, options for post-operative treatments include whole-brain radiotherapy (WBRT) or stereotactic radiosurgery (SRS) of the resection cavity. In this meta-analysis, we sought to compare the efficacy of each post-operative radiation modality with respect to tumor recurrence and survival. METHODS: Pubmed, Embase and Cochrane databases were searched through June 2016 for cohort studies reporting outcomes of SRS or WBRT after metastasis resection. Pooled effect estimates were calculated using fixed-effect and random-effect models for local recurrence, distant recurrence, and overall survival. RESULTS: Eight retrospective cohort studies with 646 patients (238 with SRS versus 408 with WBRT) were included in the analysis. Comparing SRS to WBRT, the overall crude risk ratio using the fixed-effect model was 0.59 for local recurrence (95%-CI: 0.32-1.09, I2: 3.35%, P-heterogeneity = 0.36, 3 studies), 1.09 for distant recurrence (95%-CI: 0.74-1.60, I2: 50.5%, P-heterogeneity = 0.13; 3 studies), and 2.99 for leptomeningeal disease (95% CI 1.55-5.76; I2: 14.4% p-heterogeneity: 0.28; 2 studies). For the same comparison, the risk ratio for median overall survival was 0.47 (95% CI: 0.41-0.54; I2: 79.1%, P-heterogeneity < 0.01; 4 studies) in a fixed-effect model, but was no longer significant (0.63; 95%-CI: 0.40-1.00) in a random-effect model. SRS was associated with a lower risk of leukoencephalopathy (RR: 0.15, 95% CI: 0.07-0.33, 1 study), yet with a higher risk of radiation-necrosis (RR: 19.4, 95% CI: 1.21-310, 1 study). CONCLUSION: Based on retrospective cohort studies, the results of this study suggest that SRS of the resection cavity may offer comparable survival and similar local and distant control as adjuvant WBRT, yet may be associated with a higher risk for developing leptomeningeal disease. Future research on SRS should focus on achieving a better understanding of the various factors that may favor SRS over WBRT.

Evaluation of Various Nuclear Cytological Changes in Normal Buccal Mucosa and Peritumoural Area in Patients with Oral Squamous Cell Carcinoma Receiving Concomitant Chemoradiotherapy.

Objectives. To evaluate the role of serial cytological assay in calculating the nuclear response of contralateral normal buccal mucosa and peritumoural area of squamous cell carcinoma of oral cavity in patients receiving fractionated radiotherapy (RT) and chemotherapy. Materials and Methods. This prospective, nonrandomized study was comprised of 76 histologically confirmed cases of oral squamous cell carcinoma on cyclical chemoradiation treatment. Chemoradiosensitivity was evaluated using serial scrape smears taken before and after immediate exposure to CCRT, at 17th day of CCRT (mid of treatment), and at the end of treatment. The nuclear changes, such as multinucleation, micronucleation, karyorrhexis, karyolysis, nuclear budding, prominent nucleoli, and binucleation occurring in both irradiated cancer cells and contralateral normal buccal mucosa, had a statistically significant dose related increase with concomitant chemoradiotherapy (p < 0.05). Conclusion. We recommend regular use of serial cytological assay during CCRT as it may prove to be a valuable tool for assessment of chemoradiosensitivity and persistence of tumour/dysplastic cells after radiotherapy.