Method for Inclusion Bodies Production via E. coli Host System: rGCSF as Model Biotherapeutic Protein.

Escherichia coli is an industrial-relevant microbial host system, which is highly preferred for the large-scale production of recombinant biotherapeutics. Overexpression of these recombinant biotherapeutics in the E. coli system often results in the formation of insoluble protein aggregates termed as inclusion bodies (IBs). The yield and quality of IBs are affected by a spectrum of parameters like temperature, optical density, medium composition, induction time, and amount of inducer. Here, we present a protocol for the formation and processing of IBs for production of recombinant human granulocytes colony-stimulating factor (rGCSF).

Labyrinthine Fistula-Our Experience at a Tertiary Hospital.

This study has aimed to determine the anatomical site of labyrinthine fistula in patients of chronic suppurative otitis media at our centre. Labyrinthine fistulae (LF) are caused by abnormal communications between the inner ear and surrounding structures resulting in perilymph leakage and hearing loss. Labyrinthine fistula represents as erosive loss of the enchondral bone overlying the semicircular canals without loss of perilymph. The manifestations of fistula like vertigo, hearing loss vary in severity and complexity, commonly ranging from very mild to incapacitating. Cholesteatoma induced fistula most commonly involves lateral semicircular canal probably because of its close proximity to the middle ear, but can involve other semicircular canals and rarely cochlea. This is a retrospective analysis of 36 patients of chronic suppurative otitis media with history of vertigo undergoing tympanomastoid surgery in whom there was an evidence of labyrinthine fistula on HRCT scan of temporal bone. The incidence of patients with labyrinthine fistula presenting with vertigo, nystagmus, sensorineural hearing loss, history of vertigo were analysed. The anatomical location of the fistula was supported by Radiological evidence. Patients underwent either canal wall down mastoidectomy or cortical mastoidectomy. The anatomical site and length of the labyrinthine fistula were analysed. Amongst the 36 patients of chronic suppurative otitis media with labyrinthine fistula 22 (61.1%) patients had atticoantral disease, 4 (11.1%) patients had chronic otitis media with extensive granulation, 2 (5.5%) patients had Tubotympanic disease with polyps, 4 (11.1%) patients had Tuberculous otitis media, 1 (2.77%) patient had Tubotympanic disease with extensive tympanosclerosis eroding the dome of lateral semicircular canal, 1 (2.77%) patient had extensive cholesteatoma with cerebellar abscess, 1 (2.77%) patient had fistula in the promontory following trauma, 1 (2.77%) patient had extensive tympanosclerosis with erosion of promontory. It was noticed that, in 14 (38.88%) patients the fistula was at the centre, in 17 (47.22%) patients the fistula is towards the ampullary end of horizontal semicircular canal and in 5 (13.88%) patients the fistula was towards the non ampullary end of lateral semicircular canal. The maximum length of fistula noticed was 6 mm and the minimum length of the fistula noticed was 2 mm. Labyrinthine fistula are most commonly noticed in the ampullary end of the lateral semicircular canal. The average length of the fistula was found to be 4 mm. Careful elevation of the cholesteatoma matrix over the endosteal membrane and immediate placement of temporal fascia over the exposed fistula is important to avoid injury to the inner ear. Maximum number of fistula were seen in the atticoantral type of Chronic suppurative otitis media. Prior knowledge of anatomical location of the fistulous tract in HRCT temporal bone is important to address the fistula.

Extensive Cholesteatomas: Presentation, Complications and Management Strategy.

To document the clinical presentation, complications, management strategy and post-operative outcomes of extensive cholesteatomas. Cholesteatoma is a well demarcated cystic lesion derived from an abnormal growth of keratinizing squamous epithelium in the temporal bone. Cholesteatomas commonly involve the middle ear, epitympanum, mastoid antrum and air cells and can remain within these confines for a considerable period. Bony erosion is present confined to ossicular chain and scutum initially, but as the cholesteatoma expands, erosion of the otic capsule, fallopian canal and tegmen can occur. Erosion of the tegmen tymapani or tegmen mastoideum may lead to development of a brain hernia or cerebrospinal fluid leakage. Invasion of jugular bulb, sigmoid sinus, internal carotid artery are noticed in extensive cholesteatoma and are quite challenging and requires expertise. Neurosurgical intervention should be considered along with the otological management in the same sitting in all possible cases. A retrospective review of 12 patients were carried out to assess the clinical presentation, complications, surgical management and postoperative outcomes of extensive cholesteatomas presenting at our centre between January 2017 and December 2019. CT or MRI findings, extent of cholesteatoma intra-operatively along with the status of major neurovascular structures and disease clearance, and the post-operative outcomes including morbidity and mortality were noted. All patients underwent canal wall down mastoidectomy with or without ossiculoplasty. Post operatively all patients were treated with intravenous antibiotics and if required intravenous steroids. Amongst the 12 patients of extensive cholesteatoma (EC), all of them (100%) presented with foul smelling, purulent ear discharge. 9 (75%) patients presented with otalgia. 4 (33.33%) patients had temporal headache. 10 (83.33%) patients complained of hard of hearing. 7 (58.33%) patients gives history of vertigo at the time of presentation. In 8 (66.66%) patients there was tegmen plate erosion noticed in CT scan. In 3 (25%) patients, the disease was invading the sigmoid sinus and in 1 (8.33%) patient jugular bulb was involved. In 3 (25%) cases of EC, blind sac closure was performed. In two patients who developed cerebellar abscess, drainage procedure was performed. 2 (16.66%) patients developed sigmoid sinus thrombosis, 1 (8.33%) patient had petrositis.

Outcome of Submucosal Inferior Turbinoplasty in Perennial Allergic Rhinitis.

Pharmacotherapy forms mainstay of treatment for allergic rhinitis, and has adverse effects associated with it. Topical steroid therapy is the preferred medication and considered best for long term prophylaxis but with limited compliance. Submucosal turbinoplasty reduces the duration of treatment in comparison to topical steroid which has to be taken daily for a long time. The aim was to evaluate the outcome of submucosal inferior turbinoplasty in patients with perennial allergic rhinitis. A prospective interventional study was performed on 35 patients diagnosed with perennial allergic rhinitis, diagnosed as per ARIA criteria from July 2016 to July 2018. The severity of the disease was assessed using mini RQLQ scoring system. The patients were then subjected to bilateral submucosal inferior turbinoplasty under endoscopic guidance under local anesthesia. 50% significant improvement (p value < 0.05) seen in symptoms were need to blow nose, sneezing, nasal obstruction, nasal discharge, watery eyes, need to rub eye, regular house work, recreational activities, sore eyes, tiredness, irritability and thirst. 100% improvement (p value < 0.05) seen in symptoms were sleep, need to blow nose, sneezing, nasal discharge, watery eyes, need to rub eye, recreational activities and irritability. Nasal obstruction was not severe in 17 (48.5%) patients giving a very good symptom relief and improving quality of life. This is due to reduction in the erectile tissue and roominess in the nasal cavity. All patients with allergic rhinitis with associated hypertrophied turbinates should invariably be given option of inferior turbinoplasty along with proper counselling regarding its advantages and disadvantages.

Outcome of Treatment in Verrucous Carcinoma of Oral Cavity: A Tertiary Rural Hospital Experience.

Verrucous carcinoma of oral cavity is a highly well differentiated variant of squamous cell carcinoma with a low potential for invasion and metastasis. It is prevalent in the tobacco quid chewing population in our region. In this observational study, we reviewed the medical case records of 58 patients treated for oral verrucous carcinoma staged T2 to T4a. All patients underwent wide excision of tumour which included marginal mandibulectomy in 22 and hemimandibulectomy in 23 patients along with neck dissection saving the accessory nerve and internal jugular vein. 5 patients were found to have bone involvement along the alveolar sockets. 11 patients had other associated premalignant lesions in oral cavity. Only 2 patients had lymph node metastasis without extra nodal spread in submandibular region. With a mean follow up of 6 years and minimum follow up of 1 year, 3 patients had local recurrence. All these 3 patients had bone involvement and 2 of them had lymph node metastasis on histopathological examination. 3 patients who had associated premalignant lesions developed second primary in oral cavity after 3 years. In our experience, verrucous carcinoma has good prognosis when treated by surgery. Bone involvement along alveolar sockets and associated oral premalignant lesions adversely affect the outcome. There was no difference in the outcome between selective and modified radical neck dissection. Therefore selective neck dissection (supraomohyoid) would be adequate in treating these patients. Adjuvant radiotherapy can be reserved for T4a lesions or for positive margins.