Urinary microRNA-10a levels in diagnosis and prognosis of urinary bladder cancer.

Background: Urinary bladder cancer (UBC) is a disease quite common in developed countries; however, its incidence is increasing in developing countries as well. The diagnosis of UBC is generally based on a number of methods, of which urinary cytology is a very commonly used one. But it is not very reliable. Therefore many new markers and methods are being investigated to make non-invasive diagnosis of UBC easy and reliable. Objective: This study was carried out to find the usefulness of microRNA (miRNA)-10a as a diagnostic and prognostic marker in non-muscle-invasive urinary bladder carcinoma. Material and Method: Twenty patients with UBC were taken as cases with 20 controls. Urine cytological examination was done, as well as histopathological examination of tumor tissue of cases. Urinary miRNA-10a estimation of both the cases and controls were done. Result and Conclusion: It was found that miRNA-10a is significantly high in urine of patients with UBC. Its value also significantly correlated with the grade and stage of the tumor. Hence it can be concluded that urinary miRNA-10a is a potential candidate in the diagnosis and prognosis of UBC.

Structural interactions of phytoconstituent(s) from cinnamon, bay leaf, oregano, and parsley with SARS-CoV-2 nucleocapsid protein: A comparative assessment for development of potential antiviral nutraceuticals.

SARS-CoV-2 has been responsible for causing 6,218,308 deaths globally till date and has garnered worldwide attention. The lack of effective preventive and therapeutic drugs against SARS-CoV-2 has further worsened the scenario and has bolstered research in the area. The N-terminal and C-terminal RNA binding domains (NTD and CTD) of SARS-CoV-2 nucleocapsid protein represent attractive therapeutic drug targets. Naturally occurring compounds are an excellent source of novel drug candidates due to their structural diversity and safety. Ten major bioactive compounds were identified in ethanolic extract (s) of Cinnamomum zeylanicum, Cinnamomum tamala, Origanum vulgare, and Petroselinum crispum using HPLC and their cytotoxic potential was determined against cancer and normal cell lines by MTT assay to ascertain their biological activity in vitro. To evaluate their antiviral potential, the binding efficacy to NTD and CTD of SARS-CoV-2 nucleocapsid protein was determined using in silico biology tools. In silico assessment of the phytocomponents revealed that most of the phytoconstituents displayed a druglike character with no predicted toxicity. Binding affinities were in the order apigenin > catechin > apiin toward SARS-CoV-2 nucleocapsid NTD. Toward nucleocapsid CTD, the affinity decreased as apigenin > cinnamic acid > catechin. Remdesivir displayed lesser affinity with NTD and CTD of SARS-CoV-2 nucleocapsid proteins than any of the studied phytoconstituents. Molecular dynamics (MD) simulation results revealed that throughout the 100 ns simulation, SARS-CoV-2 nucleocapsid protein NTD-apigenin complex displayed greater stability than SARS-CoV-2 nucleocapsid protein NTD-cinnamic acid complex. Hence, apigenin, catechin, apiin and cinnamic acid might prove as effective prophylactic and therapeutic candidates against SARS-CoV-2, if examined further in vitro and in vivo. PRACTICAL APPLICATIONS: Ten major bioactive compounds were identified in the extract(s) of four medicinally important plants viz. Cinnamomum zeylanicum, Cinnamomum tamala, Origanum vulgare and Petroselinum crispum using HPLC and their biological activity was also evaluated against cancer and normal cell lines. Interestingly, while all extract(s) wielded significant cytotoxicity against cancer cells, no significant toxicity was found against normal cells. The outcome of the results prompted evaluation of the antiviral potential of the ten bioactive compounds using in silico biology tools. The present study emphasizes on the application of computational approaches to understand the binding interaction and efficacy of the ten bioactive compounds from the above plants with SARS-CoV-2 nucleocapsid protein N-terminal and C-terminal RNA binding domains in preventing and/or treating COVID-19 using in silico tools. Druglikeness and toxicity profiles of the compounds were carried out to check the therapeutic application of the components. Additionally, molecular dynamics (MD) simulation was performed to check the stability of ligand-protein complexes. The results provided useful insights into the structural binding interaction(s) that can be exploited for the further development of potential antiviral agents targeting SARS-CoV-2 especially since no specific therapy is still available to combat the rapidly evolving virus and the existing treatment is more or less symptomatic which makes search for novel antiviral agents all the more necessary and crucial.

ASCUS (Atypical Squamous Cells of Undetermined Significance) in the Cervical Smears of Women from Rural Population of Lucknow West.

BACKGROUND: Though Atypical squamous cells of undetermined significance (ASCUS) are not always precursors of cervical pre-malignancy but they need timely diagnosis and careful periodical follow-up. AIM: The present study was carried out to investigate the incidence of ASCUS and SIL (squamous intraepithelial lesion of cervix) in women under rural setting. MATERIALS AND METHODS: Cervical cancer screening is in progress in rural population of West Lucknow, India, since May 2013 under auspices of Era's Lucknow Medical College and Hospital, Lucknow, and a total of 2478 women have been cytologically examined till October 2017. RESULTS: The incidence of ASCUS was found to be 8.8%, which was approximately less than half of the squamous intraepithelial lesions of cervix (SIL) rate (17.9%). The ASCUS rate was higher in the symptomatic women (9.4%) than (7.4%) seen in women without symptoms. The ASCUS incidence was higher in younger sexually active group up to 40 years after which it declined. The ASCUS rate was higher in all parity groups and was commonly associated with pain in lower abdomen and vaginal discharge. The ASCUS rate was higher with erosion cervix, while the commonly associated sexually transmitted diseases (STD) were found to be Candida albicans (2.4%). Follow-up after 18-24 months was available in 40 cases of ASCUS, and progression to low-grade SIL (LSIL) was seen in 7 (17.5%) and to high-grade SIL (HSIL) in one case (2.5%). CONCLUSION: The findings suggest that the ASCUS are not always precursors of SIL, but they need periodical follow-up after every 6 months. This will be ideal management of ASCUS under low-resource settings.

Oct-4: a prognostic biomarker of urinary bladder cancer in North India.

BACKGROUND: The objective of this study was to evaluate Octamer-binding transcription factor 4 (Oct-4), neutrophil to lymphocyte ratio (NLR) and body mass index (BMI) as independent prognostic biomarkers for prediction of urinary bladder cancer (UBC) outcomes. With the advancement in prognostic biomarker discovery, tumor recurrence is difficult to accurately predict in UBC. UBC is costly to treat due to the requirement of frequent invasive follow-up sessions. Therefore, it is of utmost importance to evaluate good prognostic biomarkers for UBC surveillance. METHODS: We studied 39 UBC tissue samples. Oct-4 protein expression was evaluated semiquantitatively by immunohistochemistry (IHC). Complete blood count data and body weight as well as the height of the patients were retrieved and recorded before the date of the first transurethral resection of bladder tumor (TURBT). The follow-up period was 48 months for recurrence-free survival (RFS), progression-free survival (PFS), and overall survival (OS). RESULTS: Oct-4 expression profile was found to be significantly associated with gender (p = 0.028), tumor grade (p = 0.038), tumor stage (p = 0.003), lymph node status (p = 0.029), recurrence (p = 0.004), progression (p = 0.011), and treatment modality (p = 0.016). Tumor grade and progression were found significant with NLR values (tumor grade, p = 0.006; progression, p = 0.038) and BMI (tumor grade, p = 0.036; progression, p = 0.014). Moreover, BMI was also significantly associated with UBC recurrence (p = 0.014). Kaplan-Meier survival analysis showed poor prognosis with both high Oct-4 expression (RFS, p = 0.001; PFS, p = 0.004; OS, p = 0.014) and high NLR values (RFS, p = 0.049; PFS, p = 0.004; OS, p = 0.005). Patients with high BMI too had poor RFS (p = 0.025) and poor PFS (p = 0.032). Furthermore, multivariate Cox regression analysis, indicated Oct-4 as an independent prognostic biomarker for RFS (HR = 0.240, 95% CI, 0.072-0.804, p = 0.021). CONCLUSIONS: We conclude that the expression profile of Oct-4 will be beneficial in prediction of UBC recurrence, and could have profound implications on the development of new therapeutic targets for UBC treatment.

AgNOR Pleomorphic Count as a Tumor Marker in Cervical Carcinogenesis and Feasibility of Its Introduction in Cervical Cancer Screening Programs to Discriminate High-Risk Cases of Squamous Intraepithelial Lesions of the Cervix.

BACKGROUND: AgNOR pleomorphism has been widely used for its diagnostic importance in differentiating premalignant and malignant lesions of different human neoplasms. However, an evaluation of its potential for discriminating cases of high-risk squamous intraepithelial lesions of the cervix (SIL) has been rarely attempted. AIM: The tumor marker potential of AgNOR pleomorphism counts was assessed by correlating high and low mean counts in low-grade SIL (LSIL) cases with persistence or regression of the lesion and HPV positivity. MATERIALS AND METHODS: The 115 LSIL cases selected for the study were registered from the ongoing cervical cancer screening of the rural population of Lucknow West. Silver nitrate staining for AgNOR counts and HPV DNA testing were done in all 115 cases. RESULTS: The AgNOR counts in the 115 LSIL cases revealed low counts in 92 and high counts in 23 cases. Follow-up, available in 107 cases, revealed persistence of the lesion in 21 of the 23 cases with high counts and in 4 of the 84 cases with low counts. HPV positivity showed a strong correlation with high counts. Persistence of LSIL was also more frequent with high AgNOR counts and in HPV-positive cases. CONCLUSIONS: The study showed a correlation of high mean AgNOR counts with HPV positivity and persistence of LSIL.

Immunohistochemical expression of human telomerase reverse transcriptase in oral cancer and precancer: A case-control study.

INTRODUCTION: Telomere Length is critically important in normal cells and telomere shortening in combination with other oncogenic changes- promotes genome instability, potentially stimulating initiation of the early stages of cancer. AIM: The present study was carried out to detect human telomerase reverse transcriptase expression in oral cancer and pre-cancerous lesions by immunohistochemistry. MATERIALS AND METHODS: An observational study was planned in which a total of 45 biopsy specimen of oral mucosa was obtained. Of these, 15 (33.3%) belonged to normal subjects, 15 (33.3%) to subjects found to have Oral submucousal fibrosis and 15 (33.3%) subjects with Oral squamous cell carcinoma. RESULTS: Among cases of OSCC, majority was of well differentiated grade (80.0%), only 1 (6.7%) case was poorly differentiated and rest was of moderately differentiated (13.3%) Labelling intensity of OSCC (78.07 ± 22.31) was maximum followed by that of Normal (44.47 ± 6.32) and minimum of OSMF (26.67 ± 15.05) and intergroup difference and between group differences were also found to be significant. Labelling score of OSCC (154.47 ± 94.74) was maximum followed by that of Normal (84.73 ± 51.51) and minimum of OSMF (46.73 ± 44.25) and intergroup difference and between groups differences (Normal vs OSCC, and OSMF and OSCC) were found to be statistically significant. CONCLUSION: The present study highlights only the discriminating ability of hTERT for differentiating the malignant condition from premalignant and normal mucosa. Hence, further studies on a larger sample size, with inclusion of other premalignant conditions too are recommended in order to understand the pattern of hTERT expression changes.

A prospective study of association of cancer stem cell marker aldehyde dehydrogenase 1 with clinicopathological profile in lung carcinoma patients.

CONTEXT: In India, lung carcinoma is the fifth-most common tumor and second-most common tumor in the males as per the Indian Council of Medical Research registry of 2002. It has been seen that ALDH1 expression in non-small cell lung cancer (NSCLC) and the presence of marker was linked to a more tumorigenic potential in the in vivo assessment and shorter disease-free survival in NSCLC patients with platinum treatment. AIMS: Hence, our objective was to detect association of cancer stem cell (CSC) marker aldehyde dehydrogenase 1 (ALDH1) with clinicopathological profile in lung carcinoma patients. SETTINGS AND DESIGN: This is a Pilot study. SUBJECTS AND METHODS: It was a Pilot study where biopsies from 55 fresh previously untreated lung cancer patients visiting the Pulmonary Medicine Department of Era's Lucknow Medical College and Hospital Lucknow and King George's Medical University were taken for 18 months November 2014-April 2016, after taking proper informed consent from them. Paraffin blocks were taken and stained by hematoxylin and eosin (Sigma) to make the histopathological diagnosis and immunohistochemistry was done for detection of CSC marker ALDH1 (Daco). STATISTICAL ANALYSIS USED: The statistical analysis was done using Statistical Package for Social Sciences Version 15.0 Statistical Analysis Software. The values were represented in number (%) and mean ± standard deviation. RESULTS: Expression of stem cell marker ALDH1 with the staging of the tumor was observed in 62.5% of Stage I, 80% of Stage II, 94.1% of Stage III, and 100% of Stage IV cases. Statistically, there was a significant association between ALDH1expression and stage of disease (P < 0.001). Diagnostic efficacy of ALDH1 expression in the detection of any positive clinical stage, it was found to be 88.6% sensitive and 90.9% specific. CONCLUSIONS: Strong ALDH1 expression correlates with higher stage of lung carcinoma making it a prognostic marker needing in-depth study.

Results of Cervical Cancer Screening in the Rural Population of Lucknow West, India, through a Camp Approach.

OBJECTIVE: Cytological screening was carried out in rural women of Lucknow West, India, through a camp approach to detect cervical cancer in its preinvasive phase and to see whether the primitive living conditions in the villages have any effect on the incidence of precancer and carcinoma of the cervix and the associated predisposing factors of cervical carcinogenesis. STUDY DESIGN: During a span of 4 years (May 2013 to March 2017), a total of 135 camps were organized under the auspice of Era's Lucknow Medical College and Hospital (Lucknow, India). Only 4,269 (31.2%) out of 13,500 women who were motivated and counseled attended the camp and 2,369 (55.1%) of them underwent a Pap smear examination. RESULTS: The incidence of squamous intraepithelial lesions of the cervix (SIL) was found to be very high (18.2%) in the 2,369 women who were screened, but the majority of them were of a low grade. The SIL incidence was very high for all predisposing factors like gynaecological symptoms, age, and clinical lesions of the cervix. However, the SIL rate showed a rise with increasing parity. Candida albicans was more common in rural women (4.7%) than Trichomonas vaginalis (1.1%), and a low incidence of viral sexually transmitted diseases (STD) was seen. CONCLUSIONS: Organizing camps in the villages through proper counseling and motivation may help in the screening of larger numbers of women for early detection of cervical cancer, the adequate treatment of which will check the progression of the disease and thus minimize the incidence of carcinoma of the cervix and its associated mortality in the rural population of India.

Effect of ethyl acetate aroma on viability of human breast cancer and normal kidney epithelial cells in vitro.

BACKGROUND: Aromatherapy is used in clinical settings for patients suffering from several chronic and critical diseases such as cancer. Ethyl acetate (EA) is a colorless liquid with a characteristic fruity smell and is naturally present in fruits and wines. METHODS: In the present study, the effect of the aroma of EA was evaluated on human breast cancer cell line MDA-MB-231 and normal cell line, Vero. Cell line viability and mechanism of EA cytotoxicity were determined by Trypan blue dye exclusion assay and phase contrast microscopy. RESULTS: It was found that EA at a concentration of 0.026 M was effective in causing considerable cytotoxicity in breast cancer cells (without even coming in contact with the culture medium and cells), while showing no effect on normal cells. Mechanism of action of EA on cancer and Vero cells was investigated by DNA fragmentation and dye binding assays using agarose gel electrophoresis (AGE) and fluorescence microscopy/cytometry, respectively. It was found that EA aroma induced predominantly necrosis in the cancer cells exposed to it. CONCLUSION: A study such as this has not been attempted before and results need further investigation before EA aroma can be used as a complementary therapy.

Oxidative stress in non-small cell lung cancer patients after chemotherapy: association with treatment response.

UNLABELLED: Levels of lipid peroxidation, nitric oxide and glutathione, as well as superoxide dismutase activity were evaluated in non-small cell lung cancer patients before and after chemotherapy. Oxidative stress was shown to influence treatment efficacy and survival of these patients. BACKGROUND AND OBJECTIVE: The aim of this study was to assess the level of oxidative stress after chemotherapy in non-small cell lung cancer patients, and its association with treatment response and survival. METHODS: Two hundred and three previously untreated non-small cell lung cancer patients and 150 healthy subjects were selected for the study. Patients received cisplatin+etoposide, and were followed for up to six cycles, for evaluation of oxidative stress. Blood levels of lipid peroxidation products (LPO), glutathione (GSH) and nitric oxide (NO), and superoxide dismutase (SOD) activity were measured at day 0 and after the third and sixth cycles of chemotherapy. Response and survival were measured at the end of follow up. Survival was estimated by the Kaplan-Meier method using the log-rank test. RESULTS: In the patients, pretreatment levels of LPO and NO were low, while GSH and SOD levels were high compared with those after the third and sixth cycles of chemotherapy. Among the 203 patients, there were 51 deaths, 82 non-responders and 70 responders at the end of the sixth cycle. Overall mean survival was higher among responders than non-responders (24.6 vs 21.2 weeks, P<0.01). The hazard ratio was 2.4 (95% CI: 1.3-3.77). Pretreatment levels of oxidative stress were similar among responders and non-responders (P>0.05). After the third and sixth cycles of chemotherapy, LPO and NO levels were low (P<0.05 or P<0.01) and GSH levels and SOD activity were high (P<0.05 or P<0.01) in responders as compared with non-responders. CONCLUSIONS: In lung cancer patients, oxidative stress increased and anti-oxidant enzymes decreased as the disease progressed. Chemotherapy may suppress oxidative stress and decreased anti-oxidant enzyme activity in responders as compared with non-responders. These effects may contribute to improved survival among responders.

Tuberculous otitis media with postaural abscess and submandibular lymphadenopathy.

We are reporting a case of right-sided tuberculous otitis media with postaural abscess and multiple submandibular lymphadenopathy which has been reported very infrequently. A high level of suspicion by the treating physician is mandatory to avoid long delay in diagnosis and increased complications in the modern chemotherapy era.

Correlation of serum vascular endothelial growth factor with clinicopathological parameters in cervical cancer.

Angiogenesis plays an important role in cervical cancer progression. Currently among several factors known to promote angiogenesis, vascular endothelial growth factor (VEGF) is most important. To evaluate the effect of treatment on VEGF levels and their correlation with other predictive factors, pre-and post treatment levels of VEGF were estimated in cervical cancer patients. 110 cases of frank cancer and 50 controls were enrolled for the present study: 18 in Stage I, 32 in Stage II, 48 in Stage III, and 12 in Stage IV. Serum VEGF levels were estimated by ELISA in patients on the day of recruitment and post treatment follow-up at a fixed time interval of 6-8 weeks. VEGF levels were highly significant among patients as compared to controls (p = 0.001). The pre-treatment VEGF levels among different stages of the disease were marginally insignificant (p = 0.07). However, they were significantly different for (i) various grades (p < 0.001), (ii) tumor size (p = 0.026), and (iii) smoking habits (p = 0.018). Post treatment levels were highly significant, as compared to pre-treatment values (p = 0.001). The pre-treatment and post-treatment VEGF levels were associated with (i) disease stage (p = 0.002), (ii) grade (p = 0.001), and (iii) tumor size (p = 0.001). In conclusion, VEGF is a potent angiogenic factor and can be considered as an effective prognostic marker in cervical cancer.

Smoking intensity, oxidative stress and chemotherapy in nonsmall cell lung cancer: a correlated prognostic study.

Cigarette smoking is a well known environmental risk factor for lung cancer; furthermore it can also enhance lung carcinogenesis by free radical mediated reactions. In addition smoking affects the rates of metabolism of several drugs and may contribute to poor cancer survival. The purpose of the present work, therefore, was to see the relationship of different smoking intensities with oxidative stress and survival after platinum based chemotherapy in non-small cell lung cancer (NSCLC). The oxidative stress levels (LPO, NO, SOD, and GSH) of 144 control subjects and 203 advanced stage NSCLC patients were assessed at day '0', after the 3rd and 6th cycle of chemotherapy. Pack year (PY) was stratified in groups (1-20, 21-50, > 50) for further analysis. Groups were compared using repeated measured ANOVA, while survival curves were compared by Kaplan-Meier methods. Oxidative stress levels of smokers were significantly high (p < 0.01 or p < 0.05) as compared to non-smoker at pretreatment, after the 3rd cycle and 6th cycle of chemotherapy but not well correlated with the PY exposures. Overall mean survival of smoker patients were significantly low when compared to non-smokers. The survival of > 50 PY group was significantly lowered (p < 0.01) as compared to others PY groups, indicating that survival after chemotherapy in smoker NSCLC patients may be dependent on their PY exposures. In conclusion, smoking is a bad prognostic factor in lung cancer therapy, besides its role in oxidative stress, and poor survival. Therefore, this factor can be used in patient selection for chemoprevention.

AgNOR counts in cervical smears under normal and other cytopathologic conditions.

OBJECTIVE: To investigate the diagnostic value of AgNOR counts in cervical smears in the process of cervical carcinogenesis and in discriminating the different grades of squamous intraepithelial lesion (SIL). STUDY DESIGN: Silver nitrate staining for AgNOR counts was performed in 50 cervical smears of cytologically diagnosed normal, inflammatory, low grade SIL (LSIL) (mild dysplasia), high grade SIL (HSIL) (moderate and severe dysplasia) and squamous cell carcinoma. The smears were derived from the ongoing routine outpatient cytology screening at Queen Mary's Hospital, Lucknow, India. RESULTS: In normal and inflammatory smears, the number of AgNOR dots varied from 1 to 2, in mild dysplasia from 2 to 4, in moderate dysplasia from 4 to 6 and in severe dysplasia from 6 to 8. Frank cervical carcinoma cases revealed 8-10 dots. Thus, a progressive increase in AgNOR counts was observed when the severity of pathologic lesions increased. Statistical analysis revealed a significant difference in AgNOR counts between normal and inflammatory smears, but it was highly significant between inflammatory and LSIL cases, between LSIL and HSIL, and between severe dysplasia and frank malignancy. CONCLUSION: This study underscored the diagnostic importance of AgNOR counts, especially in discriminating between LSIL and HSIL of the cervix. Another study is under way to assess the potentiality of AgNOR counts as tumor markers in cervical carcinogenesis.

Salivary duct carcinoma of the larynx: report of a rare case.

Salivary duct carcinomas are primarily high-grade, aggressive malignancies that affect men in the fifth and sixth decades of life. These tumors are usually found in the major salivary glands; rarely do they originate in the minor salivary glands. The distinctive feature of these neoplasms is their remarkable histologic resemblance to infiltrating ductal carcinomas of the mammary gland; both types of tumor feature epithelial and myoepithelial cells arrayed in solid, papillary, and cribriform patterns. To the best of our knowledge, only one case of a primary salivary duct carcinoma of the larynx has been previously reported. In this article, we describe a new case, and we review the literature on salivary duct carcinomas.