Study of morphology with assessment of expression of proliferation marker Ki67 antigen and P53 protein in lesions of gall bladder.

OBJECTIVE: To study the spectrum and distribution of histopathological changes and evaluate immunohistochemistry markers p53 protein and Ki67 antigen in various lesions of gall bladder. MATERIALS AND METHODS: A total of 804 consecutive gall bladder specimens were evaluated. Forty cases were selected for immunohistochemical analysis to evaluate expression of p53 and ki67 proliferation index, including 20 carcinoma gall bladder cases and 20 cases of inflammatory pathology associated with metaplasia, atypia, hyperplasia, dysplasia, and adenoma. p53 immunostaining was categorized as wild type and mutant type. ki67 of >20% was considered high expression. RESULTS: The majority of the gall bladder lesions were inflammatory in origin, most common being chronic cholecystitis. In the group of 20 gall bladder carcinoma cases, 65% were p53 mutant and the remaining 35% cases had a p53 wild-type immunophenotype. 55% cases showed high expression for ki67 labeling. However, significant correlation ( P < 0.05) was seen with lympho-vascular invasion. Among non-malignant lesions, normal/wild-type p53 expression was seen with increasing intensity and positivity in lesions with atypia and intra-epithelial neoplasms. Ki67 index also showed the same trend in all cases. CONCLUSIONS: p53 and ki-67 expression increases in inflammation, and further increment occurs in premalignant and malignant lesions of the gall bladder epithelium and can be used as a marker of aggression of histopathological lesions. The results emphasize the potential of Ki-67 and p53 as biomarkers of carcinogenesis in gall bladder carcinoma.

Widespread amyloid aggregates formation by Zika virus proteins and peptides.

Viral pathogenesis typically involves numerous molecular mechanisms. Protein aggregation is a relatively unknown characteristic of viruses, despite the fact that viral proteins have been shown to form terminally misfolded forms. Zika virus (ZIKV) is a neurotropic one with the potential to cause neurodegeneration. Its protein amyloid aggregation may link the neurodegenerative component to the pathogenicity associated with the viral infection. Therefore, we investigated protein aggregation in the ZIKV proteome as a putative pathogenic route and one of the alternate pathways. We discovered that it contains numerous anticipated aggregation-prone regions in this investigation. To validate our prediction, we used a combination of supporting experimental techniques routinely used for morphological characterization and study of amyloid aggregates. Several ZIKV proteins and peptides, including the full-length envelope protein, its domain III (EDIII) and fusion peptide, Pr N-terminal peptide, NS1 ß-roll peptide, membrane-embedded signal peptide 2K, and cytosolic region of NS4B protein, were shown to be highly aggregating in our study. Because our findings show that viral proteins can form amyloids in vitro, we need to do a thorough functional study of these anticipated APRs to understand better the role of amyloids in the pathophysiology of ZIKV infection.

Coronaviruses spike glycoprotein endodomains: The sequence and structure-based comprehensive study.

Any protein's flexibility or region makes it available to interact with many biomolecules in the cell. Specifically, such interactions in viruses help them to perform more functions despite having a smaller genome. Therefore, these flexible regions can be exciting and essential targets to be explored for their role in pathogenicity and therapeutic developments as they achieve essential interactions. In the continuation with our previous study on disordered analysis of SARS-CoV-2 spike protein's cytoplasmic tail (CTR), or endodomain, here we have explored the endodomain's disordered potential of six other coronaviruses using multiple bioinformatics approaches and molecular dynamics simulations. Based on the comprehensive analysis of its sequence and structural composition, we report the varying disorder propensity in endodomains of spike proteins of coronaviruses. The observations of this study may help to understand the importance of spike glycoprotein endodomain and creating therapeutic interventions against them.

An Exposition on Surgical Experiences in Identification, Exposure, and Injuries of Recurrent Laryngeal Nerve (RLN) During Thyroid Operations: Gleanings, Narrative, and the Reflections.

BACKGROUND: Identifying and preserving the recurrent laryngeal nerve (RLN) is of paramount importance during thyroid surgeries. Iatrogenic injuries to RLN (RLNI) are considered one of the most serious and feared complications of thyroidectomies. Surgically, there are four routes/approaches (lateral, inferior, superior, and medial) for localizing and identifying the RLN. This study aims to estimate the incidence of RLNI in the context of various approaches taken intra-operatively for nerve localization and identification. MATERIALS AND METHODS: This retrospective analytical study included 54 cases of thyroidectomies operated for various benign and malignant thyroid disorders in a tertiary care center from January 2018 to December 2020. Intraoperative search, identification, and dissection of the nerve were done with superior, inferior, medial, and lateral approaches. The chi-square test and exact test were used to analyze the data and p-value < 0.05 was considered significant. Pre- and post-operative recurrent laryngeal nerve evaluation was done with 90 degrees Hopkins laryngoscope. RESULTS: Overall in this series, the incidence of post-thyroidectomy RLNI was 3.7% and 3.7% for permanent and temporary nerve insults, respectively. Non-recurrent RLN on the right side was identified in one case and extra-laryngeal branching of RLN was identified in two cases. There was no statistically significant difference (p = 0.929) between the different approaches taken and the incidence of RLNI. The type of surgery and pathology also expressed no statistically significant relevance with the incidence of RLNI (p = 0.463 and p = 0.277, respectively). CONCLUSION: Adoption of a particular surgical approach to localize and identify RLN during thyroid surgery carries no statistically significant difference between RLNI and approaches taken. Meticulous handling and dissection of the tissue in the correct surgical plane are crucial determinants in preventing RLNIs.

Amyloidogenic proteins in the SARS-CoV and SARS-CoV-2 proteomes.

The phenomenon of protein aggregation is associated with a wide range of human diseases. Our knowledge of the aggregation behaviour of viral proteins, however, is still rather limited. Here, we investigated this behaviour in the SARS-CoV and SARS-CoV-2 proteomes. An initial analysis using a panel of sequence-based predictors suggested the presence of multiple aggregation-prone regions (APRs) in these proteomes and revealed a strong aggregation propensity in some SARS-CoV-2 proteins. We then studied the in vitro aggregation of predicted aggregation-prone SARS-CoV and SARS-CoV-2 proteins and protein regions, including the signal sequence peptide and fusion peptides 1 and 2 of the spike protein, a peptide from the NSP6 protein, and the ORF10 and NSP11 proteins. Our results show that these peptides and proteins can form amyloid aggregates. We used circular dichroism spectroscopy to reveal the presence of ß-sheet rich cores in aggregates and X-ray diffraction and Raman spectroscopy to confirm the formation of amyloid structures. Furthermore, we demonstrated that SARS-CoV-2 NSP11 aggregates are toxic to mammalian cell cultures. These results motivate further studies about the possible role of aggregation of SARS proteins in protein misfolding diseases and other human conditions.

Evaluation of variable p16 immunostaining patterns, Ki-67 indices and HPV status in cervical SILs and squamous cell carcinomas: An institutional experience.

Objective: The present study evaluates the immunoexpression of p16 and Ki-67 in cervical squamous intraepithelial lesion (SIL) and carcinomas and correlates their expression with clinicopathological features and HPV-DNA status. Material and Methods: A total 36 included cases of SIL and squamous cell carcinoma (SCC) were subjected to p16 and Ki-67 immunostaining. p16 staining was evaluated depending on grading, distribution, localization pattern, intensity and IHC score. Ki-67 expression was graded based on percentage of positive cells. Results: Incidence of HSIL and SCC cases was found to be significantly increased with parity > 5. p16 grade III diffuse nucleocytoplasmic immunostaining was observed in 62.5% LSIL, 80% HSIL and 87% SCC cases. Significant association of p16 staining intensity, IHC score and Ki-67 indices was noted with increasing grades of SILs and carcinomas. Conclusion: Our experience indicates that a combination of p16 and Ki-67 immunostaining may be useful to determine the severity of dysplastic change.

Extra-Rhino Cerebral Manifestations of Mucormycosis in Head and Neck Region: An Insight.

Mucormycosis is a potentially life-threatening fungal infection with a high mortality rate. The difficulty and delay in diagnosis due to its rarity usually results in a poor prognosis. Most common site in head and neck region is the nose and paranasal sinuses. However there are other very unusual areas in head and neck region where mucormycosis is encountered. Knowledge of these unusual areas is must and can save a patient's life. To elaborate and highlight the unusual areas in head and neck region where mucormycosis can mimic other common diseases. This retrospective study was done from May 2010 to May 2019 over a period of 9 years. All histopathologically confirmed cases of Head and Neck mucormycosis were evaluated and data analyzed. Total 35 cases of head neck mucormycosis were encountered from May 2010 to May 2019 over a period of 9 years. Out of which 30 cases (85.72%) were of rhino-orbito-cerebral mucormycosis and 5 cases (14.28%) were EXTRA rhino-orbito-cerebral mucormycosis in head neck region. Mucormycosis at such unusual sites can cause diagnostic dilemma for the treating doctor. Clinical knowledge with anticipation is a must for success in treatment of mucormycosis in unusual places in head neck region especially when normal looking diseases does not respond to usual treatment.

Role of structural disorder in the multi-functionality of flavivirus proteins.

INTRODUCTION: The life cycle of a virus involves interacting with the host cell, entry, hijacking host machinery for viral replication, evading the host's immune system, and releasing mature virions. However, viruses, being small in size, can only harbor a genome large enough to code for the minimal number of proteins required for the replication and maturation of the virions. As a result, many viral proteins are multifunctional machines that do not directly obey the classic structure-function paradigm. Often, such multifunctionality is rooted in intrinsic disorder that allows viral proteins to interact with various cellular factors and remain functional in the hostile environment of different cellular compartments. AREAS COVERED: This report covers the classification of flaviviruses, their proteome organization, and the prevalence of intrinsic disorder in the proteomes of different flaviviruses. Further, we have summarized the speculations made about the apparent roles of intrinsic disorder in the observed multifunctionality of flaviviral proteins. EXPERT OPINION: Small sizes of viral genomes impose multifunctionality on their proteins, which is dependent on the excessive usage of intrinsic disorder. In fact, intrinsic disorder serves as a universal functional tool, weapon, and armor of viruses and clearly plays an important role in their functionality and evolution.

The Emerging Surgical Concepts and Principles of Attic and Postero-Superior Wall Reconstruction by Auricular Cartilage and Free Fibro-Periosteal Tissue in Middle Ear Surgery. An Exposition on Surgical Experiences, Observations and Mistakes.

In middle ear surgeries, addressing the pathologies (cholesteatoma, retractions or tympanosclerosis) in attic or postero-superior area, are associated with difficulties like reconstruction of attic and postero-superior canal wall after clearing the disease. There are various graft materials available for such attic reconstruction in intact canal -wall surgeries. We describe our experience of attic and postero-superior canal wall reconstruction in combined approach tympanoplasty (CAT) using free auricular cartilage and free fibro-periosteal tissue and to present our results in anatomical and functional perspectives. This retrospective observational study was conducted in a tertiary care center which comprised 25 cases of combined approach tympanoplasty (with attic wall reconstruction) which were operated for attic cholesteatoma, tympanosclerosis or adhesive otitis media from January 2017 to December 2019 in our otolaryngology department. Outcomes were evaluated in audiological and morphological dimensions and paired "t" test (significant p value < 0.05) was used to analyse the audiological data. A total of 25 cases (22 primary and 3 revision cases) were included in the study. Defect in attic and posterosuperior canal wall was caused by pathology (disease itself) in 2 cases, surgically created (intraoperatively to clear the disease) in 11 cases and both by pathology and surgically in 12 cases. 22 cases had successful graft uptake, with a morphological success rate of 88%, whereas graft failure was encountered in 3 cases. Complications observed were post-operative otorrhea (2 cases), recurrence (3 cases), no hearing improvement despite intact graft (1 case) and worsening of hearing post operatively in 1 case. Pre-operative ACT (Air conduction threshold) was 45.96 ± 12.47 dB and the post-operative ACT was 38.61 ± 9.76 dB (p value = 0.0246). Pre-operative ABG (Air-bone gap) was reduced from 30.58 ± 11.06 dB to the post-operative ABG of 21.89 ± 6.24 dB (p value = 0.0013). Reconstruction of postero-superior canal wall and attic defects in CAT using auricular cartilage and fibro-periosteal tissue (over mastoid cortex) is a novel method and associated with satisfactory morphological and audiological results. It is also effective in preventing postoperative retractions.

Diagnostic Utility of Epithelial Membrane Antigen (EMA) and Calretinin (CAL) in Effusion Cytology.

INTRODUCTION: Fluid cytology and subsequent utilization of immunocytology on cell block is being used commonly for increasing the diagnostic accuracy in effusion cytology. AIM: To do cytological analysis of fluids and evaluate the role of Epithelial Membrane Antigen (EMA) and Calretinin (CAL) to differentiate between reactive and malignant cell in pleural and peritoneal fluids. MATERIALS AND METHODS: This was a prospective study carried out in Department of Pathology, Shri Guru Ram Rai Institue of Medical and Health Science, for a period of 18 months from January 2014 to June 2015. A total of 253 samples of pleural and peritoneal fluid were studied by May-Gruenwald-Giemsa (MGG) stain and Papanicolaou (Pap) staining. In 73 cases which were suspicious for malignancy, cell blocks were prepared and IHC was done using two immunomarkers- EMA and CAL. RESULTS: A total of 253 cases of effusion cytology were studied out of which 73 were found positive for malignant cells. Maximum cases of malignant cells or atypical cells were seen in peritoneal fluid. A total of 34 cases were histologically positive for malignancy. All these cases showed strong membranous and cytoplasmic positivity for EMA. Most of cases also showed 2%-5% positivity for CAL. There were 38 cases categorized as atypical or reactive mesothelial cell hyperplasia. These cases showed nuclear and cytoplasmic staining for calretinin and none of these were positive for EMA, although 5 cases showed score 2-3, which was considered as negative. CONCLUSION: The distinction between reactive and malignant mesothelial cells in cytological specimens can be problematic. A combination of CAL and EMA may help in accurate diagnosis.

Role of p53 and Ki-67 immunomarkers in carcinoma of urinary bladder.

BACKGROUND: Urothelial carcinoma is common urinary malignancy responsible for a significant proportion of cancer morbidity and mortality. We carried out the present study to demonstrate the clinicohistopathological features and to correlate the p53 and Ki-67 immunoexpression with grade and stage of bladder carcinomas. MATERIALS AND METHODS: We investigated 110 cases of bladder tumor. Grading and staging were done according to the WHO-2004 and American Joint Committee on Cancer-TNM staging recommendations. Immunohistochemical staining for p53 and Ki-67 was performed in all the cases, categorized as high and low expression taking 20% positivity as cutoff value. Statistical analysis was done using McNemar's Chi-square test and Fisher's exact test. RESULTS: There were 61 cases of high grade and 49 cases of low grade exhibiting urothelial carcinoma as the most common variant (97.3%). Muscle invasive carcinomas (pT2) noted in 29 cases whereas 23 and 58 cases revealed stage pTa and pT1, respectively. Evaluation of p53 and Ki-67 immunoexpression showed a significant association with histological grade and stage individually and also in combination (P < 0.05). CONCLUSION: Our results corroborate with the opinion that combined use of p53 and Ki-67 immunomarkers may provide additional prognostic information along with histological grading and staging in bladder carcinomas.

Diffuse intestinal submucosal lipomatosis with incidental epidermal inclusion cyst of caecum clinically masquerading as carcinoma caecum.

Symptomatic diffuse submucosal intestinal lipomatosis is a rare entity. Also few cases of epidermal inclusion cyst of caecum have been reported in literature. Here, we are presenting a rare case of intestinal submucosal lipomatosis with coincidence of epidermal inclusion cyst of caecum and presumptively diagnosed as carcinoma of ileocaecal region during surgery in a 55 years old male. Both are rare entity considering the location even they should be kept as a differential diagnosis in unusual cases of intestinal perforations with inconclusive radiological findings or clinical uncertainity.

First branchial cleft anomaly: clinical insight into its relevance in otolaryngology with pediatric considerations.

First branchial cleft anomalies (FBCA) represent a small subset of congenital malformations in neck. Prime objective of this study is to share our experience with FBCA, emphasize its relevance in otolaryngology and deal with its pediatric perspective. Embryology, pathologic anatomy and varied spectra of clinical presentations of FBCA are discussed. Along with this we have illustrated three different cases; all of them were of pediatric age group and were misdiagnosed by their treating specialists elsewhere. In this article we have also laid special emphasis on its pediatric considerations. FBCA are mostly misdiagnosed due to their unfamiliar clinical signs and symptoms. Swellings may masquerade as other neck masses. Majority of patients give a history of previous incision and drainage. While dealing with pediatric patients the important factors to be kept in mind are the age of child, superficial course of facial nerve, any associated agenesis of parotid gland. Alteration in surgical technique may be required in children. A thorough medical examination with high index of clinical suspicion should be kept in mind while dealing with such anomalies. Owing to their complex presentation and close relation with facial nerve they are challenging lesions for surgeons.

Primary MPNST in Childhood- A Rare Case Report.

Malignant peripheral nerve sheath tumour usually occurs between 20-50 years of age, comprising about 5-10% of soft tissue sarcomas. Only 1.7% of them have been reported to occur in children < 5 months of age according to the literature. Here, we are describing 18 mnth old male child presented with a swelling in the lower back. MRI showed a sacrcoccygeal swelling extending to and communicating with CSF at lower sacral level. Birth history of the child was normal with normal apgar score. The histological diagnosis was malignant peripheral nerve sheath tumour. IHC showed focal positivity of GFAP and S100. Primary spinal MPNST in children are rarer. A careful neurological examination is warranted in children. Early diagnosis and referral to multidisciplinary team are important in ensuring the best diagnosis and optimal therapy in this young age.