A FACS-based novel isolation technique identifies heterogeneous CTCs in oral squamous cell carcinoma.

Purpose: Isolating circulating tumour cells (CTCs) from the blood is challenging due to their low abundance and heterogeneity. Limitations of conventional CTC detection methods highlight the need for improved strategies to detect and isolate CTCs. Currently, the Food and Drug Administration (FDA)-approved CellSearch™ and other RUO techniques are not available in India. Therefore, we wanted to develop a flexible CTC detection/isolation technique that addresses the limitation(s) of currently available techniques and is suitable for various downstream applications. Methods: We developed a novel, efficient, user-friendly CTC isolation strategy combining density gradient centrifugation and immuno-magnetic hematogenous cell depletion with fluorescence-activated cell sorting (FACS)-based positive selection using multiple CTC-specific cell-surface markers. For FACS, a stringent gating strategy was optimised to exclude debris and doublets by side scatter/forward scatter (SSC/FSC) discriminator, remove dead cells by 4',6-diamidino-2-phenylindole (DAPI) staining, and eliminate non-specific fluorescence using a "dump" channel. APC-labelled anti-CD45mAB was used to gate remaining hematogenous cells, while multiple epithelial markers (EpCAM, EGFR, and Pan-Cytokeratin) and an epithelial-mesenchymal transition (EMT) marker (Vimentin) labelled with fluorescein isothiocyanate (FITC) were used to sort cancer cells. The technique was initially developed by spiking Cal 27 cancer cells into the blood of healthy donors and then validated in 95 biopsy-proven oral squamous cell carcinoma (OSCC) patients. CTCs isolated from patients were reconfirmed by Giemsa staining, immuno-staining, and whole transcriptome amplification (WTA), followed by qRT-PCR. In vitro culture and RNA sequencing (RNA-Seq) were also performed to confirm their suitability for various downstream applications. Results: The mean detection efficiency for the Cal 27 tongue cancer cells spiked in the whole blood of healthy donors was 32.82% ± 12.71%. While ~75% of our patients (71/95) had detectable CTCs, the CTC positivity was independent of the TNM staging. The isolated potential cancer cells from OSCC patients were heterogeneous in size. They expressed different CTC-specific markers in various combinations as identified by qRT-PCR after WTA in different patients. Isolated CTCs were also found to be suitable for downstream applications like short-term CTC culture and RNA-Seq. Conclusion: We developed a sensitive, specific, flexible, and affordable CTC detection/isolation technique, which is scalable to larger patient cohorts, provides a snapshot of CTC heterogeneity, isolates live CTCs ready for downstream molecular analysis, and, most importantly, is suitable for developing countries.

Feasibility of Establishing an Artificial Intelligence Based Head and Neck Cancer Registry: Experience from a Tertiary Care Hospital.

Introduction Establishing and maintaining a cancer registry in a tertiary high volume centre is fraught with difficulty, inaccuracy and missed data entry. Further, the raw unstructured data must be converted into a structured digital data, so that scientists can identify trends in cancer diagnoses and treatment responses. Objective We test the feasibility of establishing a cancer registry of Head and Neck malignancy patients through a research oriented artificial intelligence (AI) enabled data collection platform, using its smartphone application version. Materials and Methods This prospective observational study was conducted in the Department of Otolaryngology & Head and Neck Surgery, Post Graduate Institute of Medical Education And Research, Chandigarh in collaboration with Departments of Radiotherapy and Community Medicine. After taking due clearance from the Institute ethical committee, HNC patients, who were biopsy proven, were enrolled from October 2019 up to March 2021. The obtained data was entered, followed up and analysed through Jiyyo Research application which is a commercially available dedicated research oriented AI enabled data collection platform. Results The Jiyyo Research site was browsed and after proper registration, the patient data was entered into a proforma/questionnaire. The entered patient details were browsed for review, follow up and addition of new information. The whole process of data capture for each patient, took approximately 5-8 min, while any updates or review for the same patient required less than a minute. Search and data retrieval was very quick, and can be done in 1-2 min. Through this platform, a total of 1214 HNC patients were collected, followed and analysed during the study period. Conclusion It was feasible to establish a Head and Neck Cancer Registry using an AI based smartphone app. This AI based tumor registry could benefit in further studies with longer follow up of 5 and 10 years and in future AI studies. Supplementary Information: The online version contains supplementary material available at 10.1007/s12070-022-03173-3.

An Easy to use Periosteal Flap for Cochlear Implantation: Experience from a Tertiary Care Centre.

This report describes a novel and easy periosteal flap design for cochlear implantation. This technique has been used in 37 patients between June 2019 and August 2020. The patients have been followed up for a period of 2 months to 15 months. There were no flap related complications attributed to this flap. There was no wound hematoma, wound breakdown or implant migration. The flap design is safe, easy, less time consuming and results in better coverage of the receiver stimulator unit without any tension.

As the virus sowed, the fungus reaped! A comparative analysis of the clinico-epidemiological characteristics of rhino-orbital mucormycosis before and during COVID-19 pandemic.

BACKGROUND: The sudden surge of mucormycosis cases which happened during the second wave of COVID-19 pandemic was a significant public health problem in India. OBJECTIVES: The aim of this study was to analyse the clinico-epidemicological characteristics of the mucormycosis cases to determine the changes that had occurred due to COVID-19 pandemic. METHODOLOGY: A retrospective cross-sectional study was conducted at the Department of Otolaryngology Head and Neck Surgery, PGIMER, Chandigarh, India. Patients diagnosed with rhino-orbital mucormycosis were categorised into the following groups: Pre-pandemic(May 2019 to April 2020), Pandemic Pre-epidemic (May 2020 to April 2021) and Epidemic (1 May 2021 to 12 July 2021). The epidemiological, clinical and surgical data of all the patients were retrieved from the hospital records and analysed. RESULTS: The epidemic period had 370 cases, compared with 65 during pandemic period and 42 in the pre-pandemic period. Diabetes mellitus was seen in 87% of cases during epidemic period, 92.9% in the pre-pandemic period and 90.8% in the pre-pandemic pre-epidemic period. The proportion of patients suffering from vision loss, restricted extra-ocular movements, palatal ulcer and nasal obstruction was higher in the pre-epidemic groups, and the difference was significant (p, <.01). There was no history of oxygen use in 85.9% of patients and no steroid use in 76.5%. The death rates were the lowest during epidemic (10%). CONCLUSION: COVID-19 has caused a statistically significant increase in the number of mucormycosis infections. The mortality and morbidity which showed an increase during the first wave of COVID-19 decreased significantly during the epidemic period.

Prognostic Value of HPV Infection Assessed by p16 Immunohistochemistry and the Influence of Tobacco Usage in Oropharyngeal Cancers: Real World Scenario.

To find if an association could be established between Human Papilloma Virus (HPV) infection and oropharyngeal cancers (OPCs) in a group of patients known to be regular users of tobacco, and to determine the impact of HPV status on clinical outcomes.Case records of 212 patients with AJCC-7 (The American Joint Committee on Cancer 7th edition) stages II-IVB non metastatic squamous cell carcinoma of the oropharynx treated using radical radiotherapy with or without chemotherapy during the years 2015-2018 were retrieved. Formalin-fixed, paraffin-embedded blocks from oropharyngeal biopsies were available for 177 patients and were evaluated for p16 expression by immunohistochemical (IHC) staining. More than 50% nuclear staining with or without cytoplasmic staining was considered HPV+ . The association between tobacco use and HPV, as well as the influence of HPV status on survival outcomes were assessed. p16 expression was found to be positive in 23(13%) patients. Significant association was found between chewable tobacco usage and HPV positivity (p = 0.051). The median follow up was 20.5 months (range: 3-80). 5-year Overall Survival was 43.4% and 29.8% (p = 0.044) in HPV+ and HPV- patients, respectively. Local control was significantly better in HPV+ patients (38.6% vs. 25.3%, p = 0.049). There was also a trend towards improved Disease-free Survival in HPV+ patients (31 months vs. 15 months, p = 0.078). Though less in prevalence among the Indian population, improved outcomes in HPV+ OPC patients and widely available IHC HPV assays signifies the routine implementation of p16 testing in day-to-day clinical practice.

Desmoplastic fibroma of the mandible: a rare gnathic bone tumor with a review of the literature

Desmoplastic fibroma (DF) is a rare bone tumor, which is known to involve mostly the gnathic bones. In this setting, the clinical presentation is usually represented by a bulging tumor of the face. Radiologically, the tumor is usually characterized by an expansile lytic bone lesion. The histopathology of the tumor shows a poorly circumscribed hypocellular lesion rich in collagen fibers with interspersed spindle cells having bland nuclear chromatin. Despite the lack of mitoses and nuclear pleomorphism, DF has an aggressive nature, presenting as a destructive growth causing entrapment of neuro-vascular bundles, sinusitis, or trismus. Some cases of DF show mutations in the adenomatous polyposis coli pathway shown by nuclear localization of the ß-catenin protein. Few reports showed an association with tuberous sclerosis, though most of these cases were sporadic. We discuss a rare case of desmoplastic fibroma involving the mandible, and a review of the literature of the DF cases involving the gnathic bones.

Desmoplastic fibroma of the mandible: a rare gnathic bone tumor with a review of the literature.

Desmoplastic fibroma (DF) is a rare bone tumor, which is known to involve mostly the gnathic bones. In this setting, the clinical presentation is usually represented by a bulging tumor of the face. Radiologically, the tumor is usually characterized by an expansile lytic bone lesion. The histopathology of the tumor shows a poorly circumscribed hypocellular lesion rich in collagen fibers with interspersed spindle cells having bland nuclear chromatin. Despite the lack of mitoses and nuclear pleomorphism, DF has an aggressive nature, presenting as a destructive growth causing entrapment of neuro-vascular bundles, sinusitis, or trismus. Some cases of DF show mutations in the adenomatous polyposis coli pathway shown by nuclear localization of the ß-catenin protein. Few reports showed an association with tuberous sclerosis, though most of these cases were sporadic. We discuss a rare case of desmoplastic fibroma involving the mandible, and a review of the literature of the DF cases involving the gnathic bones.

☆Comparison of transcutaneous laryngeal ultrasound with video laryngoscope for assessing the vocal cord mobility in patients undergoing thyroid surgery.

OBJECTIVE: We evaluated the accuracy and feasibility of transcutaneous laryngeal ultrasonography as an alternative to videolaryngoscopy for assessing vocal cord mobility to rule out recurrent laryngeal nerve injury following thyroidectomy. METHODS: Forty-five adult patients scheduled to undergo elective thyroidectomy under general anesthesia were included. Preoperatively, indirect laryngoscopy and transcutaneous laryngeal ultrasonography was done for assessing vocal cord mobility. Intraoperatively, following induction, patients were intubated using videolaryngoscope. On completion of the surgical procedure, one anesthetist performed videolaryngoscopy so as to record vocal cord mobility while the patients were being extubated in deep plane of anesthesia. Simultaneously another anesthesiologist performed transcutaneous laryngeal ultrasonography.Vocal cord mobility, changes in hemodynamics and total time duration for the two procedures was recorded. Indirect laryngoscopic assessment and flexible fiberoptic laryngoscopy was done on postoperative day 1 and 7 respectively. RESULTS: Postoperative videolaryngoscopy picked up bilaterally mobile vocal cords in 88.8% cases. Transcutaneous laryngeal ultrasonography could correctly identify 39(86.6%) of these patients, with 1(2.5%) patient being misdiagnosed as having bilaterally immobile vocal cords. Further, videolaryngoscopy identified 5 patients of vocal cord palsy, of which transcutaneous laryngeal ultrasonography correctly identified 3 (60%) patients. Hence, in comparison to videolaryngoscopy, the sensitivity, specificity, positive predictive value, and negative predictive value of transcutaneous laryngeal ultrasonography for assessment of vocal cords was 75%, 95.1%, 60%, and 97.5% respectively. CONCLUSION: In patients undergoing thyroidectomy, transcutaneous laryngeal ultrasonography can serve as a non-invasive, bedside screening tool for assessing vocal cord palsy postoperatively.

A rare case of asymptomatic radioiodine-avid renal and brain metastases 20 years after hemi-thyroidectomy for adenomatous goiter.

A 65-year-old patient, with a history of left hemi-thyroidectomy for adenomatous goiter 20 years previously, was found to have pulmonary lesions on chest X-ray, a brain lesion on computerized tomography (CT), and elevated serum thyroglobulin (Tg). While completion thyroidectomy revealed that no pathological evidence of thyroid malignancy, radioiodine-avid pulmonary, brain, and renal and bone lesions were identified on diagnostic as well as posttherapy whole body planar scintigraphy and single photon emission computed tomography-CT. Subsequent ultrasonography-guided biopsy of a renal nodule showed thyroid follicular cells. This case suggests that metastatic differentiated thyroid carcinoma should be suspected in asymptomatic patients with incidentally detected lesions, raised serum Tg, and history of thyroid lesions.

Giant cystic primary mucoepidermoid carcinoma of mandible: a rare case and literature review.

Primary intra-osseous mucoepidermoid carcinoma arising from jaw is an extremely rare condition accounting to less than 2% of all mucoepidermoid carcinomas. In the jaw, it occurs more commonly in mandible than maxilla. They are low-grade cancers and affect jaw as uni- or multi-locular radiographic lesions. Here we discuss a rare case of giant cystic primary intra-cystic mucoepidermoid carcinoma of the mandible which was excised in toto. Here we discuss the clinical features, radiological and histological characteristics of this rare lesion, and review the literature.

Contrasting radiological presentation of cervical sympathetic chain schwannoma: case series of two cases.

Splaying of carotid bifurcation on imaging, known as Lyre sign, is seen characteristically in carotid body tumors. Is positive Lyre sign always confirmatory of carotid body tumor? Here we discuss two cases of cervical sympathetic chain schwannoma. The first case presented radiologically as an enhancing lesion with splaying of external carotid artery and internal carotid artery (positive Lyre sign) and misled us to the diagnosis of carotid body tumor. The second case presented as an enhancing lesion causing anterior displacement of external carotid artery and internal carotid artery (negative Lyre sign). Both lesions were confirmed as cervical sympathetic schwannoma. Post-operatively both patients developed Horner's syndrome. Lyre sign is not always confirmatory of carotid body tumor.

Aneurysmal bone cyst of maxilla with ectopic molar tooth - a case report.

Aneurysmal cysts of bone are rare non-neoplastic, locally aggressive lesion of bone with propensity for rapid growth, affecting mainly the long bones and spine. It rarely occurs in the head and neck region and within the head and neck mandible (especially the molar areas) is common. Aneurysmal cyst of bone involving the maxilla in the first decade of life is even rarer. We report a case of giant aneurysmal bone cyst of maxilla in eight-year-old male with ectopic molar tooth within the cyst. Is endoscopic excision possible? Endoscopic excision of the cyst was done successfully and no recurrence was noted even after 1 year of follow up.

Primary giant cell rich osteosarcoma of maxilla: an unusual case report.

Giant cell rich osteosarcomas, histological variant of conventional osteosarcomas account for 3% of all cases of osteosarcomas and most of them are arise from the appendicular skeleton. Giant cell rich osteosarcomas have been rarely reported in the head and neck region. This is the first case report of giant cell rich variant of osteosarcoma in the maxilla. It is important to recognize this variant, as its prognosis is worse as compared to conventional osteosarcomas of the head and neck region. We here present a case of giant cell rich variant of osteosarcoma of the maxilla and along with its characteristic radiological and histopathological picture.

Autotymapanomastoidectomy in a case of cholesteatoma with foreign body.

A unique case of foreign body of the external auditory canal (EAC) in a 16 year old boy with normal intelligence is presented. The foreign body caused the cholesteatoma to flourish behind the obstruction resulting in complete destruction of middle ear cleft and external auditory canal to form a single large cavity. The diagnosis of foreign body remained elusive till the last and could be proved only after biochemical analysis. We also discuss the Keratosis obturans, cholesteatoma of external auditory canal and the process of calcification in cholesteatoma ear, which was thought to represent the foreign body in this case.

Isolated intracranial metastasis from an early glottic cancer: how rare a presentation?

The incidence of distant metastasis with laryngeal tumors varies from 1-4% [1]. The lungs are the most frequent site, followed by bone and liver. Intracranial metastases are rarely clinically diagnosed in head neck cancer patients and if present are mainly located in the cerebrum and cerebellum. Early stage glottic squamous carcinoma with subsequent distant metastasis is rare [2]. We report a patient with T1N0 glottic tumor who underwent a curative radiotherapy and on 3 months follow up presented with isolated intracranial metastasis.