Reconstituting membrane fission using a high content and throughput assay.

Protein-mediated membrane fission has been analyzed both in bulk and at the single event resolution. Studies on membrane fission in vitro using tethers have provided fundamental insights into the process but are low in throughput. In recent years, supported membrane template (SMrT) have emerged as a facile and convenient assay system for membrane fission. SMrTs provide useful information on intermediates in the pathway to fission and are therefore high in content. They are also high in throughput because numerous fission events can be monitored in a single experiment. This review discusses the utility of SMrTs in providing insights into fission pathways and its adaptation to annotate membrane fission functions in proteins.

Polydactyly-Myopia Syndrome: Genetic and Ophthalmologic Perspectives.

Polydactyly-myopia syndrome is a rare genetic condition characterized by the co-occurrence of polydactyly and myopia. Herein, we present the case of a 28-year-old Muslim male, born of consanguineous parents, who presented with complaints of diminished vision since childhood. Ophthalmologic examination revealed severe myopia with characteristic fundus changes indicative of high myopia. Additionally, the patient exhibited polydactyly in all limbs, with a positive family history of both polydactyly and myopia. This case underscores the importance of recognizing and managing rare syndromes to provide appropriate genetic counseling and clinical care. Further research is warranted to elucidate the underlying genetic mechanisms and optimize therapeutic strategies for polydactyly-myopia syndrome. Awareness of this syndrome among healthcare providers is essential to facilitate early diagnosis and intervention for affected individuals and their families.

Comparative Analysis of Visual Outcomes and Complications in Intraocular Collamer Lens, Small-Incision Lenticule Extraction, and Laser-Assisted In Situ Keratomileusis Surgeries: A Comprehensive Review.

This review provides a comparative analysis of visual outcomes and complications associated with three prominent refractive surgical techniques: intraocular collamer lens (ICL) implantation, small-incision lenticule extraction (SMILE), and laser-assisted in situ keratomileusis (LASIK). Refractive surgeries aim to correct myopia, hyperopia, and astigmatism, offering patients an alternative to corrective lenses. The review highlights the importance of comparing these procedures to guide clinical decision-making effectively. Each technique is described, emphasizing its unique advantages and considerations. While LASIK remains widely favored for its rapid visual recovery and high patient satisfaction, ICL is suitable for patients with higher refractive errors or corneal irregularities. Although relatively newer, SMILE shows promise with potential benefits such as corneal biomechanical stability and a reduced risk of dry eye syndrome. However, each procedure carries its distinct complications, reinforcing the need for personalized patient care and informed decision-making. Understanding these techniques' relative efficacy and safety profiles is essential for optimizing outcomes and enhancing patient satisfaction. Continued advancements in technology and surgical techniques promise further improvements in refractive surgery outcomes, underscoring the importance of ongoing research and innovation.

Gender and sexual orientation of undergraduate medical students in India: A cross-sectional study.

Background: Although the potential for alternate conceptions of gender roles and sexual orientations are diverse, it is by-and-large not well tolerated. This study explores the self-reported gender-roles and sexual orientations of Indian undergraduate medical students. Aim: To study self-reported gender role and sexual orientation of undergraduate medical students in India. Method: One hundred twenty volunteers were included in the study consisting of 60 males and 60 females. A questionnaire comprising of a sociodemographic proforma, Bem Sex-Role Inventory (BSRI), and Epstein Sexual Orientation Inventory (ESOI) were given to each participant. The scales were scored, tabulated, and statistically analyzed. Results: The BSRI revealed that femininity was predominant in both female and male participants, at 68.33% and 55%, respectively. The ESOI revealed that females had significantly higher opposite-sex attraction than males. Though males had higher same-sex attraction than females, the difference was not statistically significant. Females also had a significantly higher sexual orientation range and a mean sexual orientation. Sexual drive was significantly higher in males. Significantly more females supported homosexuality and bisexuality as compared to males. Conclusion: This study helps establish that gender roles can be non-conforming. It helps ascertain that while heterosexual orientation predominates, alternate sexual orientations also exist. It paves the way for future studies and explorations to alleviate public misconceptions.

Insights into cargo sorting by SNX32 and its role in neurite outgrowth.

Sorting nexins (SNX) are a family of proteins containing the Phox homology domain, which shows a preferential endo-membrane association and regulates cargo sorting processes. Here, we established that SNX32, an SNX-BAR (Bin/Amphiphysin/Rvs) sub-family member associates with SNX4 via its BAR domain and the residues A226, Q259, E256, R366 of SNX32, and Y258, S448 of SNX4 that lie at the interface of these two SNX proteins mediate this association. SNX32, via its PX domain, interacts with the transferrin receptor (TfR) and Cation-Independent Mannose-6-Phosphate Receptor (CIMPR), and the conserved F131 in its PX domain is important in stabilizing these interactions. Silencing of SNX32 leads to a defect in intracellular trafficking of TfR and CIMPR. Further, using SILAC-based differential proteomics of the wild-type and the mutant SNX32, impaired in cargo binding, we identified Basigin (BSG), an immunoglobulin superfamily member, as a potential interactor of SNX32 in SHSY5Y cells. We then demonstrated that SNX32 binds to BSG through its PX domain and facilitates its trafficking to the cell surface. In neuroglial cell lines, silencing of SNX32 leads to defects in neuronal differentiation. Moreover, abrogation in lactate transport in the SNX32-depleted cells led us to propose that SNX32 may contribute to maintaining the neuroglial coordination via its role in BSG trafficking and the associated monocarboxylate transporter activity. Taken together, our study showed that SNX32 mediates the trafficking of specific cargo molecules along distinct pathways.

Expression of ß-catenin in oral leukoplakia and oral submucous fibrosis: An immunohistochemical study.

BACKGROUND: Oral potentially malignant disorders have increased propensity to turn malignant than its apparently normal counterparts. Histopathological examination, although gold standard, needs adjunct technique to give accurate diagnosis. Immunohistochemistry has proved to be a promising adjunct to aid in the diagnosis so far. The quest for a definitive marker is still on. Beta-catenin (ß-catenin), a structural protein has been evaluated to identify its likely role in malignant transformation of potentially malignant lesions and possibly designate it as one of the identifiable signature molecules in the transformation. AIM AND OBJECTIVE: To evaluate and estimate the expression of ß-catenin in different grades of dysplasia, oral submucous fibrosis (OSMF) and normal mucosa and compare the same. METHODOLOGY: A total number of 40 cases including different grades of dysplasia, OSMF and normal mucosa were immunohistochemically stained, location and intensity of its expression were evaluated for ß-catenin. The results were statistically analyzed using the one-way analysis of variance and Chi-square test. RESULTS: The expression of ß-Catenin in the cytoplasm as well as in the nucleus increased from mild-to-moderate dysplasia to OSMF and to severe epithelial dysplasia in an increasing order. The expression is seen to translocate from membranous to cytoplasm to nucleus indicating a proliferative potential in these group of lesions. CONCLUSION: ß-catenin is a promising marker which indicates the malignant transformation potential in the higher grades of dysplasia and OSMF.

Estimation of expression of beta-human chorionic gonadotropin levels through progression of disease from normal to epithelial dysplasia to malignancy.

BACKGROUND: Gonadotropins have been extensively studied in trophoblastic and nontrophoblastic tumors of breast, gastric, bladder, parathyroid, renal cell and cervical carcinomas, with a significant increase in tissue expressions. Serum levels of beta-human chorionic gonadotropin (ß-hCG) and its tissue expression were found more in oral squamous cell carcinoma (OSCC) patients with a significant diagnostic and prognostic value. No such study has been done on oral epithelial dysplasia (OED). AIMS AND OBJECTIVE: To evaluate the expression of ß-hCG in OED and the feasibility of using this marker for early diagnosis and to see its progression from normal to dysplasia to malignancy. MATERIALS AND METHODS: The study population consisted of thirty histologically confirmed cases of OED and thirty cases of OSCC. Fifteen normal tissues were also included in the study. All the tissue samples were subjected to immunohistochemical (IHC) staining using antimouse ß-hCG antibody. RESULTS: The IHC expression of ß-hCG was completely negative in normal cases (Group 1 [n = 15]), whereas 13 (43.3%) cases of OED (Group 2 [n = 30]) and 13 cases (43.3%) of OSCC (Group 3 [n = 30]) showed diffuse cytoplasmic staining in dysplastic surface epithelium and epithelial islands of OSCC. This difference was statistically significant with P = 0.007. CONCLUSION: We conclude that the expression of ß-hCG increased from normal mucosa to dysplasia to OSCC, suggesting that it is involved in the early stage of carcinogenesis and progression of the disease.

Coalition of E-cadherin and vascular endothelial growth factor expression in predicting malignant transformation in common oral potentially malignant disorders.

BACKGROUND: Reduced E-cadherin expression and increased VEGF expression is known to be involved in tissue growth and transformation of Oral Potentially Malignant Disorders (OPMDs) and has been correlated with their differing histological grades in numerous studies. AIM: To evaluate Immunohistochemical (IHC) expression of both E-cadherin and VEGF in predicting the malignant transformation potential of common OPMDs. MATERIALS AND METHODS: Ten cases each of Normal Oral mucosa (NOM), Mild Oral Epithelial Dysplasia (OED), Moderate OED, Severe OED, Oral Submucous Fibrosis, (OSMF) and Oral Squamous Cell Carcinoma (OSCC) were stained and evaluated for the expression of Ecadherin and VEGF. Quick score (QS) for expression intensity in all epithelial layers was calculated for both markers and results statistically analysed using Kruskal -Wallis ANOVA and Mann-Whitney "U" test. RESULTS: E-cadherin expression was continuous and membranous in all the layers of NOM and reduced with progressing grades of OED to OSCC. In OSMF, expression was intermediate between moderate and severe OED. VEGF expression increased as the disease progressed from normal to increasing grades of OED to malignancy. In OSMF, expression was similar to that in mild OED. VEGF, E-cadherin expression for basal and parabasilar cells showed a strong statistically significant negative correlation in NOM. A very strong statistically significant positive correlation with perfect monotonic relation was noted in superficial cells in severe OED group and OSCC group. CONCLUSION: E-Cadherin and VEGF could be used as combination markers to predict the potential risk for malignant transformation in OEDs.

Beta-catenin in disease.

In continuation with the previous review on "ß-catenin in health", in this review we discuss the role of ß-catenin in the pathogenesis of common oral lesions in the oral and maxillofacial region- oral potentially malignant disorders, their progression to oral squamous cell carcinoma, salivary gland tumors and odontogenic tumours. This review is based on a pubmed search of all the lesions included in the review.

ß catenin in health: A review.

ß catenin belongs to the armadillo family of proteins. It plays a crucial role in developmental and homeostatic processes. Wnts are a family of 19 secreted glycoproteins that transduce multiple signaling cascades, including the canonical Wnt/ß catenin pathway, Wnt/Ca(2+) pathway and the Wnt/polarity pathway. This is a review on ß catenin, Wnt proteins and their secretion, the signaling pathway, the associated factors and the crucial role of ß catenin in odontogenesis.

Protozoan Parasite Growth Inhibitors Discovered by Cross-Screening Yield Potent Scaffolds for Lead Discovery.

Tropical protozoal infections are a significant cause of morbidity and mortality worldwide; four in particular (human African trypanosomiasis (HAT), Chagas disease, cutaneous leishmaniasis, and malaria) have an estimated combined burden of over 87 million disability-adjusted life years. New drugs are needed for each of these diseases. Building on the previous identification of NEU-617 (1) as a potent and nontoxic inhibitor of proliferation for the HAT pathogen (Trypanosoma brucei), we have now tested this class of analogs against other protozoal species: T. cruzi (Chagas disease), Leishmania major (cutaneous leishmaniasis), and Plasmodium falciparum (malaria). Based on hits identified in this screening campaign, we describe the preparation of several replacements for the quinazoline scaffold and report these inhibitors' biological activities against these parasites. In doing this, we have identified several potent proliferation inhibitors for each pathogen, such as 4-((3-chloro-4-((3-fluorobenzyl)oxy)phenyl)amino)-6-(4-((4-methyl-1,4-diazepan-1-yl)sulfonyl)phenyl)quinoline-3-carbonitrile (NEU-924, 83) for T. cruzi and N-(3-chloro-4-((3-fluorobenzyl)oxy)phenyl)-7-(4-((4-methyl-1,4-diazepan-1-yl)sulfonyl)phenyl)cinnolin-4-amine (NEU-1017, 68) for L. major and P. falciparum.

Origin of ameloblastoma from Basal cells of the oral epithelium- establishing the relation using neuroectodermal markers.

BACKGROUND AND OBJECTIVES: Basal cell layer of the oral epithelium has been rightfully regarded as a potential source of odontogenic tumours and cysts, but, without substantial evidence. Also, whether the basal cell layer retains within it, some properties of ectomesenchyme, which was imbibed during the early embryogenesis and hence its neuroectodermal relation, is not known. Here, an attempt is made to establish the hidden neuroectodermal potential of the oral epithelium, especially the basal layer, by observing the expression of known neuroectodermal markers, NSE (Neuron Specific Enolase), Synaptophysin and CD99. The expression of the same markers has also been studied in Ameloblastoma, connecting it with oral epithelium, in turn establishing basal cell layer as a potential source of Ameloblastoma. MATERIALS AND METHODS: Sections of formalin fixed, paraffin embedded tissue samples of 20 cases of Ameloblastoma and 10 cases of Normal Retromolar mucosa, were stained immunohistochemically with NSE, Synaptophysin, CD99 and also with CK-19 and evaluated for positive expression. RESULTS: Positive reaction was obtained in all the cases of Ameloblastoma and NRM (Normal Retromolar mucosa) with NSE, all the cases of Ameloblastoma and eight cases of NRM with Synaptophysin and in six cases of Ameloblastoma and NRM with CD99. The staining was diffuse and more marked in case of NSE than Synaptophysin and CD99. CK19 staining done to assure that the tissue antigenicity was maintained was positive in all the samples. INTERPRETATION AND CONCLUSION: A strong relationship between the neuroectoderm, Ameloblastoma and the basal layer of the oral epithelium is established by the study. It favours the hypothesis that the basal cell layer of oral mucosa may be the sought out culprit in most cases of the Ameloblastomas, especially those occurring in the non-tooth bearing area. This would call for the need to incorporate additional therapy in the form of mucosal striping along with the conventional treatment.

Immunohistochemical analysis of laminin expression in adenoid cystic carcinoma.

BACKGROUND AND OBJECTIVES: This study aims at the observation of the immunohistochemical expression of laminin in adenoid cystic carcinoma (ACC) of salivary gland origin and to analyze the distribution of laminin in various components of the tumor and correlate the expression of laminin with the growth and differentiation of the tumor. MATERIALS AND METHODS: THIRTY CASES OF ACC WERE SUBJECTED TO IMMUNOHISTOCHEMICAL STUDY USING POLYCLONAL ANTIHUMAN LAMININ PRIMARY ANTIBODY, DISTRIBUTION OF LAMININ IN EACH CASE OF ACC WAS OBSERVED IN THE FOLLOWING AREAS: Intracellularly, inner borders of the pseudocystic spaces, within the lumen of the pseudocysts, around the tumor islands and in the intervening stroma. RESULTS: Laminin positivity was observed in the inner aspect of the pseudocystic spaces in 15 cases, within the lumen of pseudocystic spaces in 22 cases, in the intervening stroma in 20 cases, bordering the tumor islands in 16 cases and intracellularly in 4 cases. INTERPRETATION AND CONCLUSION: Based on these observations, it can be assumed that laminin plays a major role in proliferation of the tumor cells and in pseudocyst formation. Thus, laminin might play a significant role in the growth and differentiation of ACC and also help in assessing the prognosis of the tumor.

Evaluation and comparison of expression of p63 in odontogenic keratocyst, solid ameloblastoma and unicystic ameloblastoma.

BACKGROUND AND OBJECTIVES: The behavior of odontogenic lesions varies with some tumors behaving like a cyst and some cysts behaving like tumors. p63, a member of the p53 family of tumor suppressor genes has recently come into light in view of its role as an oncogene. The aim of the present study was to investigate the expression of p63 protein in OKC, Solid ameloblastoma, Unicystic Ameloblastoma and Follicular tissue. MATERIALS AND METHODS: p63 expression was compared in 12 cases of OKC, 12 Solid Ameloblastoma, 14 cases of Unicystic ameloblastoma and 10 cases of Follicular tissue using immunohistochemical technique. All 48 cases were subjected to heat-induced antigen retrieval method using citrate buffer in a pressure cooker. Then the sections were stained with anti-p63 polyclonal antibody and visualized using super sensitive polymer HRP detection system. In each case, number of cells showing p63 positivity were assessed in two compartments - basal and suprabasal and compared. RESULTS: Statistical analysis showed that p63 expression in the suprabasal compartment in Odontogenic keratocysts was equivalent to that of central neoplastic cells of Solid Ameloblastoma and Unicystic Ameloblastoma type 3. Statistically significant difference in the expression of p63 was observed between OKC and Unicystic Ameloblastoma Type 1 and Solid Ameloblastoma and Unicystic Ameloblastoma Type 1. CONCLUSION: We conclude that the higher expression of p63 in these odontogenic lesions correlates well with their aggressive behavior and thereby suggesting alterations in treatment modalities.

Immunohistochemical expression of basement membrane laminin in histological grades of oral squamous cell carcinoma: A semiquantitative analysis.

BACKGROUND AND OBJECTIVES: To determine the immunohistochemical (IHC) localization of basement membrane component laminin in histological grades of oral squamous cell carcinoma (OSCC). The purpose of this study was to demonstrate the loss of continuity of the basement membrane in OSCC using an antibody directed against laminin using advanced polymer staining system. MATERIALS AND METHODS: A total of 30 cases of OSCC: 10 cases of well differentiated squamous cell carcinom (WDSCC), 10 cases of moderately differentiated squamous cell carcinoma (MDSCC), and 10 cases of poorly differentiated squamous cell carcinoma (PDSCC) were subjected to heat-induced antigen retrieval method using ethylene-di-amine-tetraacetic acid buffer in a microwave oven. Then the sections were stained with anti-laminin polyclonal antibody and visualized using super sensitive polymer horseradish peroxidase detection system. In each case, the integrity of the basement membrane laminin was assessed by using statistical analysis. RESULTS: Statistical analysis showed a decreased distribution of laminin from WDSCC to MDSCC to PDSCC (P value 0.0573). The intracytoplasmic staining of laminin gradually increased from WDSCC to MDSCC to PDSCC (P value 0.0198). INTERPRETATION AND CONCLUSION: WDSCC cases showed more laminin expression in basement membrane around the tumor islands and less loss of continuity compared to MDSCC and PDSCC cases suggesting a greater enzymatic degradation of basement membrane components in MDSCC and PDSCC than WDSCC. The loss of structural basement membrane laminin and the presence of laminin in the tumor cells of PDSCC cases suggest that laminin helps in tumor invasion. The expression of laminin in the basement membrane may be a useful parameter to evaluate tumor histologic differentiation and aggressiveness.

Expression of p53 and Cyclin D1 in oral squamous cell carcinoma and normal mucosa: An Immunohistochemical study.

AIM: To assess p53 and Cyclin D1 expression using Immunohistochemistry in normal mucosa and oral squamous cell carcinoma. MATERIALS AND METHODS: Twenty cases of Oral squamous cell carcinoma (OSCC) and 10 normal mucosa were used and the primary antibodies used were p53 (DAKO-DO7) and Cyclin D1 Mouse Anti human Cyclin D1 (DCS-6) 1: 100 dilution. STATISTICAL ANALYSIS: Labelling index was calculated and mean LI and SD were calculated, using Descriptive Statistics and t-test was used to compare mean LI between antibodies used in OSCC. Percentage positivity was done by Chi-Square test. Comparison of LI between p53 and Cyclin D1 was studied using t test. RESULTS: p53 was positive in 30% and Cyclin D1 in40% of normal cases and 65% and 95% of OSCC were positive for p53 and Cyclin D1 respectively. Mean LI of p53 and Cyclin D1 were found to be statistically significant between the normal mucosa and OSCC. The correlation of mean LI of p53 and Cyclin D1 in OSCC was found to be statistically significant. LI of p53 was found to be higher than Cyclin D1 in OSCC. CONCLUSION: In the present study, increased p53 and Cyclin D1 expression were seen in OSCC when compared to the normal mucosa and a positive correlation was seen between increased p53 and Cyclin D1 expression in OSCC.