Identification & correlation of bacterial diversity in oral cancer and long-term tobacco chewers- A case-control pilot study.

Introduction. Squamous cell carcinoma is a highly aggressive type of oral cancer (OC). It is the most common cancer among men, and accounts for almost 90 % of all oral cancers in India. Consumption of tobacco is a leading factor contributing to maximum oral cancer incidences as per the WHO.Hypothesis/Gap statement. Researchers reported a direct association of microorganisms with dysbiosis in various oral lesions including oral cancer. However, there is a dearth of information related to compositional changes in the oral microbiome in long-term tobacco chewers and the Indian oral cancer population.Aim. The aim of this study was to identify and correlate the bacterial diversity in the oral cavity of tobacco chewers, patients with oral cancer and healthy subjects in the Indian population.Methods. Oral rinse samples were collected for ten subjects in each group followed by DNA extraction. The variable regions of the bacterial 16S rRNA gene (V6-V8) were amplified, sequenced, processed, and analysed using QIIME2 platform to assess alpha and beta diversity between the study groups.Results. This pilot study showed genus Streptococcus dominated the control group (18.54 %), and the abundance decreased in tobacco and OC group (9.63 and 5.45% respectively); whereas genus Prevotella dominated the tobacco and OC group (21.01 and 26.03% respectively). A shift in abundance of microbiome was observed from control population to oral cancer via the tobacco chewing population. Maximum alpha diversity of oral microbiome was found in Indian tobacco chewers. Beta diversity of tobacco chewers was similar to both the healthy population as well as oral cancer patients suggesting transitioning of the oral microbiome from healthy to oral cancer microbiome via the tobacco chewers microbiome.Conclusion. The data provides evidence of oral bacterial dysbiosis due to tobacco chewing habits that can further lead to progression towards cancer.

Three metachronous primary lung cancers in a chronic smoker: A case report and review of the literature.

RATIONALE: Lung cancer is a leading cause of cancer-related deaths. Smoking is major risk factor for initial and subsequent lung cancer especially in active smokers. Treatment of subsequent lung cancer depends on whether it is synchronous or metachronous. We report a rare case of triple metachronous lung cancer and review of literature of patients with triple metachronous cancers. This will be the second case reported of triple metachronous lung cancer. PATIENT CONCERNS: A 60-year-old male, active smoker with diabetes mellitus, chronic obstructive pulmonary disease (COPD) and peripheral arterial disease presented with cough and hemoptysis. Initial computed tomography (CT) scan showed right upper lobe spiculated mass. DIAGNOSIS: He underwent transthoracic needle biopsy for right upper lobe mass, showing primary lung adenocarcinoma (ADC)-Stage-IIIA. He continued to smoke and 9-years later had new left upper lobe spiculated nodule, which on surgical resection showed squamous cell carcinoma (SCC)-Stage-IA1. Despite counselling on smoking cessation, he was unable to quit. Six months later, he presented with shortness of breath and CT chest showing right hilar adenopathy in right upper and lower lobes. He underwent transbronchial biopsies of lesion which showed small cell lung carcinoma (SCLC). INTERVENTIONS: His initial lung ADC-Stage-IIIA, was treated with chemotherapy, weekly thoracic radiation and additional chemotherapy cycles. Nine years later, his left upper lobe mass showing SCC-Stage-IA1 was deemed curative after apical resection and he was kept on surveillance. Six months later, after diagnosis of SCLC in right upper and lower lobe, patient was not a candidate for systemic chemotherapy due to poor performance status and opted for hospice care. OUTCOMES: His initial lung ADC-Stage-IIIA showed complete radiological response with chemotherapy and radiation. Subsequent SCC-Stage-IA1 was deemed curative after resection. Due to his poor performance status, he was not a candidate for chemotherapy for SCLC and patient opted for hospice care. LESSONS: Smoking is a major risk factor for developing lung cancer and with continued smoking, patients are at higher risk for developing subsequent primary lung cancers. We recommend, patients with lung cancer must quit smoking, and those who do not, should remain on long-term surveillance.

Atypical Endovascular Cells in SARS-CoV-2 Pneumonia.

The autopsy findings for 3 cases of SARS-(CoV-2) pneumonia-related deaths are reported with pulmonary histology and immunohistochemistry findings. In 2 cases (cases 1 and 2), the time interval from presentation to death was approximately 1 week, whereas for case 3, the time interval from presentation to death was hours. Case 1 and case 2 presented with shortness of breath, cough, and flu-like symptoms. The decedent from case 3 died shortly after presenting to a local emergency room with high fever, chest and abdominal pain, and shortness of breath. All 3 cases had 1 or more comorbidities. The postmortem interval for cases 1 and 2 was 2 weeks as they died at sea and were stored on board within the respective cruise ships' refrigeration units, whereas case 3 was examined within 24 hours of death. The autopsies were conducted at the Miami-Dade County Medical Examiners Department under routine infectious precautions. Salient clinical history and autopsy findings are summarized. Microscopic examination revealed pneumonia with associated atypical endovascular cells.

Usefulness of salivary sialic acid as a tumor marker in tobacco chewers with oral cancer.

AIM: This study aims to assess the usefulness of salivary sialic acid (SA) as a tumor marker in the detection of oral squamous cell carcinoma (OSCC) among tobacco chewers. MATERIALS AND METHODS: After the approval of study protocol by the Institutional Ethics Committee and informed voluntary consent, salivary samples were collected from 96 participants in each group of tobacco chewers with OSCC, tobacco chewers without precancerous or cancerous lesion, and healthy controls. Salivary protein-bound SA (PBSA) and salivary-free SA (FSA) were measured by Yao et al.'s method of acid ninhydrin reaction, and the data were subjected to appropriate statistical analysis. RESULTS: The salivary PBSA and FSA levels in the Groups 1, 2, and 3 participants were 31.17 ± 7.6 mg/dL and 63.45 ± 9.8 mg/dL, 25.45 ± 16.61 mg/dL and 33.18 ± 11.38 mg/dL, and 22.73 ± 3.01 mg/dL and 21.62 ± 8.86 mg/dL, respectively. Salivary FSA levels were significantly increased among the tobacco chewers with OSCC patients (Group 1) and tobacco chewers with no premalignant lesions of the oral cavity (Group 2) compared to the healthy controls (Group 3) with P < 0.05 being statistically significant. Salivary FSA levels were significantly increased in Group 1 as compared with Group 2. The salivary PBSA was high among Group 1 as compared to the control Group 3; there was however no significant difference in the levels of salivary PBSA between Group 1 and Group 2. There was no significant difference in the PBSA levels between OSCC patients of Group 1 and the tobacco chewers without precancerous or cancerous lesion in the oral cavity of Group 2. CONCLUSION: Salivary PBSA and FSA are significantly raised in both tobacco chewers with OSCC and in tobacco chewers with no precancerous or cancerous lesions in the oral cavity. SA should therefore be used cautiously while considering it as a marker for the early detection of oral cancer. Tobacco can be a crucial confounding factor when SA is used as a biomarker in OSCC since their levels are elevated to some extent even in tobacco chewers without any clinically obvious precancerous or cancerous lesions in the oral cavity.

Recent tobacco use has widespread associations with adolescent white matter microstructure.

IMPORTANCE: Given the prevalence of alcohol, cannabis, and tobacco use during adolescence, it is important to explore the relative relationship of these three substances with brain structure. OBJECTIVE: To determine associations between recent alcohol, cannabis, and tobacco use and white and gray matter in a large sample of adolescents. DESIGN, SETTING, AND PARTICIPANTS: MRI data were collected in N = 200 adolescents ages 14-18 (M = 15.82 years; 67% male; 61% Hispanic/Latino). On average, during the past month, participants reported consuming 2.05 drinks per 1.01 drinking day, 0.64 g per 6.98 cannabis use days, and 2.49 cigarettes per 12.32 smoking days. MAIN OUTCOMES AND MEASURES: General linear models were utilized to examine past 30-day average quantities of alcohol, cannabis, and tobacco use, age, sex, and sex by substance interactions in skeletonized white matter (fractional anisotropy and axial, radial, and mean diffusivity) and voxel-based morphometry of gray matter (volume/density). RESULTS: Tobacco use was negatively associated with white matter integrity (radial and mean diffusivity) with peak effects in inferior and superior longitudinal fasciculi. Cannabis use was negatively associated with white matter integrity (axial diffusivity) in a small cluster in the left superior longitudinal fasciculus. No associations were observed between recent alcohol use and white or gray matter overall, but interactions showed significant negative associations between alcohol use and white matter in females. CONCLUSIONS AND RELEVANCE: It is important to note that recent tobacco use, particularly given the popularity of e-tobacco/vaping in this age group, had widespread associations with brain structure in this sample of adolescents.

Pathway based prognostic gene expression profile of buccal and gingivo-buccal oral squamous cell carcinoma in smokeless tobacco chewers.

BACKGROUND: The objective was to study comprehensive mRNA expression profiles of buccal mucosa oral squamous cell carcinoma (OSCC-BM) and gingivo-buccal OSCC (OSCC-GB) in smokeless tobacco chewers to understand the biological behavior of OSCC at these specific sites and identify diagnostic and prognostic markers. METHODS: High throughput RNA sequencing transcriptome of fresh buccal mucosa (4 samples) and gingivo-buccal (4 samples) OSCC with normal oral mucosa (3 samples) was performed on Illumina NextSeq500 paired end sequencing with 75x2bp. RESULTS: In the comparison between OSCC and normal, there were 402 differentially expressed genes (DEGs); between OSCC-BM and normal, there were 467 DEGs; and between OSCC-GB and normal oral tissue, there were 608 DEGs. Pathway-based analysis of gene expression was done. The inflammation mediated by chemokine and cytokine signaling pathway had the maximum gene hits. CONCLUSIONS: FZD2 and its interactions with the cadherins have a role in invasion and metastasis. immunosurveillance is evident in OSCC-GB with the downregulation of CADM1.

Expression analysis of cyclooxygenase-2 in patients suffering from esophageal squamous cell carcinoma.

Esophageal squamous cell carcinoma (ESCC) is one of the aggressive malignancies and mechanisms underlying its pathogenesis remain unclear. Cyclooxygenase-2 (COX-2) enzyme system plays a crucial role in many gastrointestinal malignancies and is an important regulator of cell growth, proliferation, apoptosis, differentiation and transformation. More precise outcome of COX-2 in ESCC is less investigated. In this study we investigated the risk factors of ESCC and expression of COX-2 in Carcinoma in situ (CIS) and ESCC compared to normal esophageal mucosa. ESCC relationship to clinico-pathological parameters using immunohistochemistry was also part of this investigation. Current study was conducted in the Institute of Basic Medical Sciences, Khyber Medical University, Peshawar, Pakistan. A total of 69 diagnosed patients of ESCC, both Pakistanis and Afghans were enrolled. Various risk factors associated with ESCC were recorded. Mean age at the time of diagnosis was 55 years. Out of 69 patients of ESCC 46 (67%) were users of dipping tobacco (Naswar). Expression of COX-2 was determined in normal esophageal mucosa, CIS and invasive ESCC using Immunohistochemistry (IHC). Differences of mean were computed using ANOVA followed by applying Post Hoc test. Patients were categorized as positive with high expression or negative with low to nil expression. ANOVA showed large differences in expression of COX-2 in normal healthy mucosa compared with CIS and ESCC with the mean difference of -9.529 and -7.370 respectively, p-value being <.05 at 95% confidence interval (CI). No significant difference was noticed in the expression of COX-2 in CIS compared with ESCC with p-value >.05 at 95% CI. Our complete cohort (23-85 years) showed statistically significant difference in the expression of COX-2 gene in ESCC and CIS tissue samples compared with normal healthy mucosa. Results of this study indicate that over-expression of COX-2 is positively associated with ESCC.

The Intersection of Sex Differences, Tobacco Use, and Inflammation: Implications for Psychiatric Disorders.

PURPOSE OF REVIEW: Tobacco use, sex differences, and psychiatric disorders are associated with altered immune function. There are also sex differences in tobacco use and psychiatric disorders. This review summarizes findings from the small, but growing literature examining sex differences in the effects of tobacco use on inflammation and the implications for psychiatric disorders. RECENT FINDINGS: We identified four studies that tested the interaction between sex and tobacco/nicotine on inflammation. Although males and females generally exhibited differential tobacco-induced immune responses, the pattern varied depending on the sample (rodents vs. humans) and the method to evaluate inflammation. Evidence suggests that sex modulates the effects of tobacco smoke on inflammation. Many inflammation markers associated with sex differences and tobacco use are related to psychiatric disorders. We propose a model in which sex, tobacco use, and inflammation interact to increase risk for psychiatric disorders. Future studies are needed to examine the mechanisms that explain this relationship.

Types of tobacco consumption and the oral microbiome in the United Arab Emirates Healthy Future (UAEHFS) Pilot Study.

Cigarette smoking alters the oral microbiome; however, the effect of alternative tobacco products remains unclear. Middle Eastern tobacco products like dokha and shisha, are becoming globally widespread. We tested for the first time in a Middle Eastern population the hypothesis that different tobacco products impact the oral microbiome. The oral microbiome of 330 subjects from the United Arab Emirates Healthy Future Study was assessed by amplifying the bacterial 16S rRNA gene from mouthwash samples. Tobacco consumption was assessed using a structured questionnaire and further validated by urine cotinine levels. Oral microbiome overall structure and specific taxon abundances were compared, using PERMANOVA and DESeq analyses respectively. Our results show that overall microbial composition differs between smokers and nonsmokers (p = 0.0001). Use of cigarettes (p = 0.001) and dokha (p = 0.042) were associated with overall microbiome structure, while shisha use was not (p = 0.62). The abundance of multiple genera were significantly altered (enriched/depleted) in cigarette smokers; however, only Actinobacillus, Porphyromonas, Lautropia and Bifidobacterium abundances were significantly changed in dokha users whereas no genera were significantly altered in shisha smokers. For the first time, we show that smoking dokha is associated to oral microbiome dysbiosis, suggesting that it could have similar effects as smoking cigarettes on oral health.

Complementary clinical effects of red complex bacteria on generalized periodontitis in a caucasian population.

OBJECTIVES: To relate five periodontopathogenic bacteria, including the red complex, to the severity, extent, and inflammation of the periodontal lesion in Caucasian patients with generalized aggressive and chronic periodontitis and to explore whether tobacco use is associated with a specific bacterial profile. MATERIALS AND METHODS: A cross-sectional and analytic study was conducted in patients with aggressive and chronic periodontitis. Data were gathered on socio-demographic and periodontal variables, and RH-PCR was used to determine subgingival bacterial profile. Linear and logistic regression analyses were performed. RESULTS: The study included 60 patients with aggressive and 123 with chronic periodontitis. Total red complex bacteria count was higher in aggressive periodontitis, mainly due to T. denticola (P = 0.015). In both periodontitis types, models showed an association between T. forsythia count and probing depth (B = 0.157, P = 0.030) and between T. denticola count and higher bleeding scores (B = 2.371, P = 0.027). Smoking did not affect the red complex bacteria count in either disease. CONCLUSIONS: The prevalence of red complex bacteria was similar between aggressive and chronic periodontitis, but their count was higher in the former. In both diseases, T. forsythia was associated with greater severity and T. denticola with more severe bleeding. Tobacco smoking was not associated with the presence of red complex bacteria in either disease.

Alcohol and Tobacco Increases Risk of High Risk HPV Infection in Head and Neck Cancer Patients: Study from North-East Region of India.

BACKGROUND: Human papilloma virus (HPV) associated Head and Neck Cancers (HNCs) have generated significant amount of research interest in recent times. Due to high incidence of HNCs and lack of sufficient data on high-risk HPV (hr-HPV) infection from North -East region of India, this study was conceived to investigate hr-HPV infection, its types and its association with life style habits such as tobacco, alcohol consumption etc. METHODS: A total of one hundred and six primary HNC tumor biopsy specimens were collected. These samples were analyzed for hr-HPV DNA (13 HPV types) using hybrid capture 2 (HC2) assay and genotyping was done by E6 nested multiplex PCR (NMPCR). RESULTS: The presence of hr-HPV was confirmed in 31.13% (n = 33) and 24.52% (n = 26) of the HNC patients by nested multiplex PCR (NMPCR) and HC2 assay respectively. Among hr-HPV positive cases, out of thirteen hr- HPV types analyzed, only two prevalent genotypes, HPV-16 (81.81%) followed by HPV-18 (18.18%) were found. Significant association was observed between hr-HPV infection with alcohol consumption (p <0.001) and tobacco chewing (p = 0.02) in HNC cases. Compared to HPV-18 infection the HPV-16 was found to be significantly associated with tobacco chewing (p = 0.02) habit. CONCLUSIONS: Our study demonstrated that tobacco chewing and alcohol consumption may act as risk factors for hr-HPV infection in HNCs from the North-East region of India. This was the first study from North-East India which also assessed the clinical applicability of HC2 assay in HNC patient specimens. We suggest that alcohol, tobacco and hr- HPV infection act synergistically or complement each other in the process of HNC development and progression in the present study population.

Chronic exposure to chewing tobacco selects for overexpression of stearoyl-CoA desaturase in normal oral keratinocytes.

Chewing tobacco is a common practice in certain socio-economic sections of southern Asia, particularly in the Indian subcontinent and has been well associated with head and neck squamous cell carcinoma. The molecular mechanisms of chewing tobacco which leads to malignancy remains unclear. In large majority of studies, short-term exposure to tobacco has been evaluated. From a biological perspective, however, long-term (chronic) exposure to tobacco mimics the pathogenesis of oral cancer more closely. We developed a cell line model to investigate the chronic effects of chewing tobacco. Chronic exposure to tobacco resulted in higher cellular proliferation and invasive ability of the normal oral keratinocytes (OKF6/TERT1). We carried out quantitative proteomic analysis of OKF6/TERT1 cells chronically treated with chewing tobacco compared to the untreated cells. We identified a total of 3,636 proteins among which expression of 408 proteins were found to be significantly altered. Among the overexpressed proteins, stearoyl-CoA desaturase (SCD) was found to be 2.6-fold overexpressed in the tobacco treated cells. Silencing/inhibition of SCD using its specific siRNA or inhibitor led to a decrease in cellular proliferation, invasion and colony forming ability of not only the tobacco treated cells but also in a panel of head and neck cancer cell lines. These findings suggest that chronic exposure to chewing tobacco induced carcinogenesis in non-malignant oral epithelial cells and SCD plays an essential role in this process. The current study provides evidence that SCD can act as a potential therapeutic target in head and neck squamous cell carcinoma, especially in patients who are users of tobacco.

Tobacco carcinogen-metabolizing genes CYP1A1, GSTM1, and GSTT1 polymorphisms and their interaction with tobacco exposure influence the risk of head and neck cancer in Northeast Indian population.

Genetic polymorphisms in tobacco-metabolizing genes may modulate the risk of head and neck cancer (HNC). In Northeast India, head and neck cancers and tobacco consumption remains most prevalent. The aim of the study was to investigate the combined effect of cytochrome P450 1A1 (CYP1A1) T3801C, glutathione S-transferases (GSTs) genes polymorphisms and smoking and tobacco-betel quid chewing in the risk of HNC. The study included 420 subjects (180 cases and 240 controls) from Northeast Indian population. Polymorphisms of CYP1A1 T3801C and GST (M1 & T1) were studied by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) and multiplex PCR, respectively. Logistic regression (LR) and multifactor dimensionality reduction (MDR) approach were applied for statistical analysis. LR analysis revealed that subjects carrying CYP1A1 TC/CC + GSTM1 null genotypes had 3.52-fold (P < 0.001) increase the risk of head and neck squamous cell carcinoma (HNSCC). Smokers carrying CYP1A1 TC/CC + GSTM1 null and CYP1A1 TC/CC + GSTT1 null genotypes showed significant association with HNC risk (odds ratio [OR] = 6.42; P < 0.001 and 3.86; P = 0.005, respectively). Similarly, tobacco-betel quid chewers carrying CYP1A1 TC/CC + GSTM1 null genotypes also had several fold increased risk of HNC (P < 0.001). In MDR analysis, the best model for HNSCC risk was the four-factor model of tobacco-betel quid chewing, smoking, CYP1A1 TC/CC, and GSTM1 null genotypes (testing balance accuracy [TBA] = 0.6292; cross-validation consistency [CVC] = 9/10 and P < 0.0001). These findings suggest that interaction of combined genotypes of carcinogen-metabolizing genes with environmental factors might modulate susceptibility of HNC in Northeast Indian population.

Consumo de tabaco y alcohol en pacientes con cáncer esofágico en el Hospital Vargas de Caracas durante el período 2004-2009 / Tobacco and alcohol consumption in patients with esophageal cancer at

El Cáncer de Esófago ocupa el noveno lugar entre las neoplasias malignas a nivel mundial y está asociado al hábito tabáquico y alcohólico. En el Hospital Vargas de Caracas ocupa el lugar número 11 entre las primeras 15 (2.5%) causas de egresos por cáncer al año. Determinar el consumo de tabaco y alcohol en pacientes ingresados por Cáncer de esófago en el Hospital Vargas de Caracas durante el período 2004 - 2009. Se realizó estudio retrospectivo y descriptivo, luego de la revisión de historias de 24 pacientes ingresados por cáncer de esófago con diagnóstico de cáncer de esófago. Se utilizaron medidas de tendencia central para interpretación de resultados. 21(87.5%) de los pacientes fueron masculinos y 3(12.5%), femeninos. La edad promedio fue de 61 años. El 83.3%(20) refirieron hábito tabáquico de más de 24 paquetes/año y hábito alcohólico mayor a 120 gr/día. Histológicamente 87.5% (21) correspondió a Carcinoma Epidermoide localizados en 1/3 medio de esófago en 58.3% (14). Adenocarcinoma fue diagnosticado en 3 pacientes (12.5%) El hallazgo histopatológico más frecuente fue el carcinoma epidermoide en el 87.5% de las historias revisadas. El 83% de los pacientes tenían una asociación importante al hábito tabáquico y alcohólico, por lo que recomendamos implementación de Programa conjunto de promoción y prevención en salud entre la Sociedad Venezolana de Gastroenterología y la Sociedad Anticancerosa de Venezuela, de lucha contra los factores de riesgo de esta enfermedad...

Esophageal cancer ranks ninth among malignancies worldwide and is associated with smoking and alcoholism. In Hospital Vargas de Caracas is ranked number 11 among the top 15 (2.5%) causes of cancer discharges per year. To determine the consumption of tobacco and alcohol in patients admitted due to esophageal cancer in Hospital Vargas de Caracas during 2004-2009. A retrospective descriptive study was performed after reviewing the charts of 24 patients admitted with esophageal cancer. We used central tendency measures for interpretation of results. 21 (87.5%) patients were male and three (12.5%) female. The average age was 61 years-old. 83.3% (20) reported smoking over 24 pack/years and become an alcoholic greater than 120 g/day. Histologically, 87.5% (21) corresponded to epidermoid carcinoma located in 1/3 of the esophagus in 58.3% (14). Adenocarcinoma was diagnosed in 3 patients (12.5%). The most common histopathological fi nding was squamous cell carcinoma in 87.5% of the charts reviewed. 83% of patients had a signifi cant association with smoking and alcoholism, that’s why, we recommend the implementation of a joint program in health promotion and disease prevention among the Venezuelan Society of Gastroenterology and Cancer Society of Venezuela, to fight against the risk factors of this disease...