Expression Profiling of EMT Transcriptional Regulators ZEB1 and ZEB2 in Different Histopathological Grades of Oral Squamous Cell Carcinoma Patients.

Background: Pakistan has a high burden of oral cancers, with a prevalence rate of around 9%. Oral Squamous Cell Carcinoma (OSCC) accounts for about 90% of oral cancer cases. Epithelial to Mesenchymal Transition (EMT) gets highly stimulated in tumor cells by adopting subsequent malignant features of highly invasive cancer populations. Zinc Finger E-Box binding factors, ZEB1 and ZEB2, are regulatory proteins that promote EMT by suppressing the adherent ability of cells transforming into highly motile cancerous cells. The present study aimed to analyze the expression of EMT regulators, ZEB1 and ZEB2, and their association with the clinicopathological features in different grades of OSCC patients. Methods: Tissue samples were collected for both case and control groups from the recruited study participants. Cancer tissues (cases) were collected from the confirmed OSCC patients, and healthy tissues (controls) were collected from third-molar dental extraction patients. The study participants were recruited with informed consent and brief demographic and clinical characteristics. The case group was further segregated with respect to the histological cancer grading system into well-differentiated (WD), moderately differentiated (MD), and poorly differentiated (PD) squamous cell carcinoma (SCC) groups. RNA was extracted from the tissue samples for expression profiling of ZEB1 and ZEB2 genes through quantitative real-time PCR (qRT-PCR). Results: All of the recruited participants had a mean age of 46.55 ± 11.7 (years), with most of them belonging to Urdu speaking ethnic group and were married. The BMI (kg/m2) of the healthy participants was in the normal range (18-22 kg/m2). However, BMI was found to be reduced with the proliferation in the pathological state of cancer. The oral hygiene of patients was better than the healthy participants, possibly due to the strict oral hygiene practice concerns of consultants. Every recruited OSCC patient had one or multiple addiction habits for more than a year. Patients reported health frailty (46.6%), unhealed mouth sores (40%), swallowing difficulties and white/reddish marks (80%), and restricted mouth opening (64.4%). Furthermore, 82.2% of the recruited patients observed symptoms within 1-12 months, and buccal mucosa was the most exposed tumor site among 55.6% of the patients. Expression profiling of EMT regulators showed gradual over-expressions of ZEB1 (8, 20, and 42 folds) and ZEB2 (4, 10, and 18 folds) in respective histological cancer grades. Conclusion: High expressions of ZEBs have been significantly associated with cancer progression and poor health. However, no association was found between OSCC with other clinicopathological features when compared to healthy controls.

Sublingual Epidermoid Cyst: Diagnosis, Surgical Treatment, and Follow-Up in a 22-Year-Old Male-A Case Report.

Epidermoid inclusion cysts are rare in the oral cavity, accounting for less than 0.01% of cysts found there. These cysts are defined as epidermoid if they contain only epithelial lining, dermoid if skin appendages, and teratoid if they contain other tissues such as muscle, cartilage, and bone. This case report describes a case of a plunging ranula that was later diagnosed as an epidermoid inclusion cyst. The patient is a 22-year-old male who presented with swelling in the floor of the mouth and difficulty eating solid foods for the past 4 months. Upon examination, a shiny, pink-colored mass measuring 1 cm × 5 cm was observed, which displaced the tongue upward and backward. MRI scan of the brain, face, and neck revealed a well-defined, non-enhancing cystic mass measuring 6.6 cm × 4.5 cm that extended beyond the sublingual area. Histopathological examination after surgical removal of the cyst revealed that the cyst wall was lined with stratified squamous keratinized epithelium with a retained granular layer, and the cavity was filled with lamellated keratin flakes. The patient recovered well after surgery; no recurrence was observed at the 6-month follow-up.

External Auditory Canal Carcinoma With An Unusually Extensive Invasion.

External auditory canal carcinoma, while starting out as a seemingly benign condition, if left untreated can have an aggressive course of disease and involve multiple lower cranial nerves. Squamous cell carcinoma remains the most frequent histological type of malignant neoplasm of the external auditory canal and temporal bone. Here we describe a patient with a history of chronic suppurative otitis media with an extensive spread, the tumour was reaching from the skull base to the oropharynx involving neurovasculature along with soft tissues and bones, as well as the cerebellum. The involvement of the recurrent laryngeal and hypoglossal nerves were the most unusual presentation here. The case differed from all previous reported cases as the spinal accessory nerve was to spared. Surgery and radiotherapy are the treatment options but for the inoperable cases presenting with an already poor prognosis, concomitant radiotherapy is the only choice.

Mammary analogue secretory carcinoma of the parotid gland: A rare tumour entity.

Mammary analogue secretory carcinoma (MASC) is a salivary gland tumour with low-grade potential and specific FTV6 derangement having translocation of chromosomes t (12;15) (p13;q25). It shares a similar morphological as well as an immunohistochemical profile with secretory carcinoma (SC) of the breast making it a diagnostic enigma. In this report, we discuss the case of a 65-year-old male patient, who presented with a complaint of right-sided facial swelling. To rule out the differential, he underwent various diagnostic modalities, including magnetic resonance imaging, fine-needle aspiration and it's the tumour's microscopic and immunohistochemical properties were also reviewed. Parotidectomy along with concurrent chemo-radiotherapy was performed to eradicate the growing mass.

Case report of a female child with right nasal chondromyxoid fibroma.

Chondromyxoid fibroma is a rare tumour, representing <1% of all primary bone neoplasm. We report the case of a four-year-old female child with a one-year history of nasal obstruction and facial swelling. A large enhanced lesion with amorphous densities spreading into the right cribriform plate and floor of sphenoid sinus, laterally into the right lamina papyracea, inferolaterally into the medial wall of maxillary sinus, posteriorly into the nasopharynx and superior aspect of oropharynx was observed on CT scan. The mass was excised by Caldwell Luc's endoscopic medial maxillectomy via sublabial approach. CMF was confirmed histopathologically.

Long-Term Prospective Outcomes of Patients with Non-Muscle Invasive Bladder Cancer After Holmium Laser Ablation.

Objectives: To evaluate the long-term tumor recurrence, progression, and cancer-specific mortality (CSM) of patients after holmium laser ablation of non-muscle invasive bladder cancer (NMIBC) under topical anesthesia. To identify variables that help to predict risk of tumor recurrence, progression, and CSM. Materials and Methods: A prospective study of 251 consecutive patients with 399 recurrent NMIBC who underwent outpatient Holmium yttrium aluminium garnet laser ablation under topical anesthesia was carried out between September 2006 and September 2018. All patients underwent primary transurethral resection of bladder tumor to obtain histology. All patients completed a patient satisfaction questionnaire and visual analogue scale (VAS) for procedure-related pain. Results: All 251 patients were analyzed with an average age of 75.2 years and follow-up of 69.8 months. The majority of patients had multiple comorbidities with an American Society of Anesthesiologists of 3 or 4. The proportion of tumor grade 1, 2, and 3 disease and tumor stage Ta and T1 was 43.4%, 41.5%, 15.1%, 78.5%, and 21.5%, respectively. Overall, 57.8% of patients had at least one tumor recurrence with an average time of 16.3 months. Risk factors were tumor grade, number of tumors at time of first laser ablation, and tumor stage. Overall, 19.1% of patients demonstrated tumor progression with an average time of 51 months. Only 5.6% of patients had progression to muscle invasive bladder cancer, and the CSM rate was 5.2% with an average time to death of 71.2 months. Risk factors for CSM were tumor grade, stage, and the patient's age. The average VAS for pain was 0.6 out of 10, with all patients willing to have the procedure repeated if required. Conclusion: Laser ablation of NMIBC is a safe, effective, and well-tolerated long-term option for patients, particular in those who have moderate-to-severe co-morbidities. The long-term cancer outcomes are best for younger patients with primary low-grade and low-stage disease.

The Oncogene Metadherin Interacts with the Known Splicing Proteins YTHDC1, Sam68 and T-STAR and Plays a Novel Role in Alternative mRNA Splicing.

Oncogenic metadherin is a key contributor to tumourigenesis with metadherin expression and cytoplasmic localisation previously linked to poor survival. A number of reports have shown metadherin localises specifically to nuclear speckles known to be rich in RNA-binding proteins including the splicing proteins YTHDC1, Sam68 and T-STAR, that have been shown to select alternative splice sites in mRNA of tumour-associated proteins including BRCA, MDM2 and VEGF. Here we investigate the interaction and relationship between metadherin and the splice factors YTHDC1, T-STAR and Sam68. Using a yeast two-hybrid assay and immunoprecipitation we show that metadherin interacts with YTHDC1, Sam68 and T-STAR and demonstrate that T-STAR is significantly overexpressed in prostate cancer tissue compared to benign prostate tissue. We also demonstrate that metadherin influences splice site selection in a dose-dependent manner in CD44v5-luc minigene reporter assays. Finally, we demonstrate that prostate cancer patients with higher metadherin expression have greater expression of the CD44v5 exon. CD44v5 expression could be used to discriminate patients with poor outcomes following radical prostatectomy. In this work we show for the first time that metadherin interacts with, and modulates, the function of key components of splicing associated with cancer development and progression.

Immunohistochemical biomarker validation in highly selective needle biopsy microarrays derived from mpMRI-characterized prostates.

INTRODUCTION: Diagnosing prostate cancer routinely involves tissue biopsy and increasingly image guided biopsy using multiparametric MRI (mpMRI). Excess tissue after diagnosis can be used for research to improve the diagnostic pathway and the vertical assembly of prostate needle biopsy cores into tissue microarrays (TMAs) allows the parallel immunohistochemical (IHC) validation of cancer biomarkers in routine diagnostic specimens. However, tissue within a biopsy core is often heterogeneous and cancer is not uniformly present, resulting in needle biopsy TMAs that suffer from highly variable cancer detection rates that complicate parallel biomarker validation. MATERIALS AND METHODS: The prostate cores with the highest tumor burden (in terms of Gleason score and/or maximum cancer core length) were obtained from 249 patients in the PICTURE trial who underwent transperineal template prostate mapping (TPM) biopsy at 5 mm intervals preceded by mpMRI. From each core, 2 mm segments containing tumor or benign tissue (as assessed on H&E pathology) were selected, excised and embedded vertically into a new TMA block. TMA sections were then IHC-stained for the routinely used prostate cancer biomarkers PSA, PSMA, AMACR, p63, and MSMB and assessed using the h-score method. H-scores in patient matched malignant and benign tissue were correlated with the Gleason grade of the original core and the MRI Likert score for the sampled prostate area. RESULTS: A total of 2240 TMA cores were stained and IHC h-scores were assigned to 1790. There was a statistically significant difference in h-scores between patient matched malignant and adjacent benign tissue that is independent of Likert score. There was no association between the h-scores and Gleason grade or Likert score within each of the benign or malignant groups. CONCLUSION: The construction of highly selective TMAs from prostate needle biopsy cores is possible. IHC data obtained through this method are highly reliable and can be correlated with imaging. IHC expression patterns for PSA, PSMA, AMACR, p63, and MSMB are distinct in malignant and adjacent benign tissue but did not correlate with mpMRI Likert score.

Transport inhibition of digoxin using several common P-gp expressing cell lines is not necessarily reporting only on inhibitor binding to P-gp.

We have reported that the P-gp substrate digoxin required basolateral and apical uptake transport in excess of that allowed by digoxin passive permeability (as measured in the presence of GF120918) to achieve the observed efflux kinetics across MDCK-MDR1-NKI (The Netherlands Cancer Institute) confluent cell monolayers. That is, GF120918 inhibitable uptake transport was kinetically required. Therefore, IC50 measurements using digoxin as a probe substrate in this cell line could be due to inhibition of P-gp, of digoxin uptake transport, or both. This kinetic analysis is now extended to include three additional cell lines: MDCK-MDR1-NIH (National Institute of Health), Caco-2 and CPT-B2 (Caco-2 cells with BCRP knockdown). These cells similarly exhibit GF120918 inhibitable uptake transport of digoxin. We demonstrate that inhibition of digoxin transport across these cell lines by GF120918, cyclosporine, ketoconazole and verapamil is greater than can be explained by inhibition of P-gp alone. We examined three hypotheses for this non-P-gp inhibition. The inhibitors can: (1) bind to a basolateral digoxin uptake transporter, thereby inhibiting digoxin's cellular uptake; (2) partition into the basolateral membrane and directly reduce membrane permeability; (3) aggregate with digoxin in the donor chamber, thereby reducing the free concentration of digoxin, with concomitant reduction in digoxin uptake. Data and simulations show that hypothesis 1 was found to be uniformly acceptable. Hypothesis 2 was found to be uniformly unlikely. Hypothesis 3 was unlikely for GF120918 and cyclosporine, but further studies are needed to completely adjudicate whether hetero-dimerization contributes to the non-P-gp inhibition for ketoconazole and verapamil. We also find that P-gp substrates with relatively low passive permeability such as digoxin, loperamide and vinblastine kinetically require basolateral uptake transport over that allowed by +GF120918 passive permeability, while highly permeable P-gp substrates such as amprenavir, quinidine, ketoconazole and verapamil do not, regardless of whether they actually use the basolateral transporter.

Significance of ultrasound features in predicting malignant solid thyroid nodules: need for fine-needle aspiration.

OBJECTIVE: The purpose of this study was to provide sonographic and colour flow criteria helpful for differentiation between benign and malignant solid thyroid nodules. METHODS: This prospective study was carried out at Sindh Institute of Urology and Transplantation (SIUT), Karachi Pakistan from 01.05.07 to 31.12.08. Sonographic scans of 78 thyroid nodules in 66 patients were performed and characteristics of thyroid nodules that were studied included microcalcifications, an irregular or microlobulated margins, marked hypoechogenicity, a shape that was taller than it was wide and color flow pattern in Color Doppler ultrasound. The presence and absence of characteristics of nodules were classified as having positive or negative findings. If even one of these sonographic features was present, the nodule was classified as positive (malignant). If a nodule had none of the features described, it was classified as negative (benign). The final diagnosis of a lesion as benign (n = 53) or malignant (n = 25) was confirmed by fine needle aspiration biopsy, and patients who were proved to have benign lesions were followed-up for 6 months and malignant lesions which were proved on histopathology after FNA were subjected to surgery. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were then calculated on the basis of our proposed classification method. RESULTS: Among 78 solid thyroid nodules 35 lesions were classified as positive considering the sonographic characteristics and 23 of them were proved to be malignant on histopatology. Out of 43 lesions which were classified as negative, 2 were proved to be malignant. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy based on our sonographic classification method were 93.8%, 66%, 56.1%, 95.9%, and 74.8%, respectively. CONCLUSION: Ultrasound is valuable for identifying many malignant or potentially malignant thyroid nodules. No single ultrasound criterion is reliable in differentiating all benign from malignant thyroid nodules, but many US features aid in predicting the benign or malignant nature of a given nodule. Fine-needle aspiration biopsy should be performed on thyroid nodules classified as positive, regardless of palpability.