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Introduction: Surgical management of level V in clinically node positive (cN+) oral squamous cell carcinomas (OSCCs) is controversial. The objectives of the study were to identify predictors of level V metastases in cN+ OSCC., Methods: This retrospective study is based on institutional data of operated cN+ OSCC between April 2018 and December 2022. Clinical and pathological parameters were subjected to univariate analysis. Significant parameters in univariate analysis were further subjected to multivariate analysis. A p value of less than 0.05 was considered statistically significant., Results: None of cN1 or pN1 patients had a level V metastasis. No skip metastasis to level-V was noticed. Total number of positive lymph nodes, lymph node ratio (LNR), extranodal extension, pN classification, and the presence of level II and III metastases were found to be significant predictors for level V metastases. The post hoc analysis suggested that ≥5 positive nodes, LNR >0.1, and pN3 status were independent risk factors for level V metastases., Conclusion: Selective neck dissection for N+ OSCC is feasible in the N1 neck, preferably where nodal metastases is limited to level-I only. Patients with a bulky nodal disease, particularly those with N3 neck, ≥5 positive nodes, ENE, and metastatic lymph nodes in levels II and III should be offered comprehensive neck dissection., (© 2024 The Author(s). Published by S. Karger AG, Basel.)

The semiconducting nature of CeFeO 3 /CeO 2 /Fe 2 O 3 nanocomposite has permitted the degradation of the organic toxic dye methylene blue under the irradiation of ultraviolet and visible light portions of solar radiation. Fullprof-assisted Rietveld refinement analysis, performed using the Match software, has revealed the orthorhombic nature of CeFeO 3 . In addition, in the synthesized material, the cubic phase byproduct CeO 2 was found due to the highly oxidizing nature of the cerium element. This occurred due to 6 min microwave irradiation because the microwave-assisted technique offered random distribution of heating during the supply of 800-W power for 30 s, followed by 12 cycles. Additionally, the presence of Fe 2 O 3 was also confirmed through Match software-assisted Rietveld refinement analysis through phase matching. During the synthesis, a certain portion of the synthesized CeFeO 3 experienced overheating, leading to phase transformation from CeFeO 3 into Fe 2 O 3 and CeO 2 . The unit cell compositions of CeFeO 3 , CeO 2 , and Fe 2 O 3 were found in the sample material with 35.54%, 52.43%, and 12.03%, respectively. The appearance of a fingerprint absorption region in the FTIR spectrum around 577.36 and 535.38 cm -1 further confirmed the similarity of these values (577.36 and 535.38 cm -1 ) to those obtained from the calculated values obtained by substituting Rietveld refined bond length parameters. The fourth step process in the thermal analysis curve (TGA) revealed the oxidation process, which led to the destruction of the CeFeO 3 phase, causing the transformation of CeFeO 3 into two byproducts (CeO 2 and Fe 2 O 3 ). This oxidation process permitted in an observable weight gain, which is observed in the thermal analysis curve (TGA). A cyclic voltammetry study (the experimentally measured current-voltage characteristic curve) revealed slightly distorted semirectangular CV curves, confirming the pseudo-capacitive behavior of the synthesized composite., (© 2024 The Author(s). Luminescence published by John Wiley & Sons Ltd.)

Glioblastoma (GBM) is a devastating brain cancer for which new effective therapies are urgently needed. GBM, after an initial response to current treatment regimens, develops therapeutic resistance, leading to rapid patient demise. Cancer cells exhibit an inherent elevation of endoplasmic reticulum (ER) stress due to uncontrolled growth and an unfavorable microenvironment, including hypoxia and nutrient deprivation. Cancer cells utilize the unfolded protein response (UPR) to maintain ER homeostasis, and failure of this response promotes cell death. In this study, as integrins are upregulated in cancer, we have evaluated the therapeutic potential of individually targeting all αβ1 integrin subunits using RNA interference. We found that GBM cells are uniquely susceptible to silencing of integrin α3. Knockdown of α3-induced proapoptotic markers such as PARP cleavage and caspase 3 and 8 activation. Remarkably, we discovered a non-canonical function for α3 in mediating the maturation of integrin β1. In its absence, generation of full length β1 was reduced, immature β1 accumulated, and the cells underwent elevated ER stress with upregulation of death receptor 5 (DR5) expression. Targeting α3 sensitized TRAIL-resistant GBM cancer cells to TRAIL-mediated apoptosis and led to growth inhibition. Our findings offer key new insights into integrin α3's role in GBM survival via the regulation of ER homeostasis and its value as a therapeutic target.

Unilateral Sensorineural Hearing Loss (USNHL) can present with varying symptoms apart from hearing loss, such as dizziness and tinnitus. Impaired sound localization, inability to identify speech in noise and decreased temporal summation can significantly impair the daily activities of an individual. This along with dizziness and tinnitus affects the physical, mental and social health as well. The purpose of this study is to assess and correlate the spectrum of handicap faced by USNHL patients. Validated questionnaires were used to evaluate handicap in 42 USNHL patients due to hearing loss (Hearing Handicap Inventory for Adults-HHIA), dizziness (Dizziness Handicap Inventory-DHI) and tinnitus (Tinnitus Handicap Inventory-THI). These handicaps were then correlated with each other using Spearman's correlation coefficient. Statistically significant strong positive correlation of HHIA was seen with DHI (rho = 0.60, p  ≤ 0.001) and THI (rho = 0.74, p  ≤ 0.001). Similarly, strong positive correlation was seen between emotional subscales of HHIA and DHI (rho = 0.73, p  ≤ 0.001). USNHL can lead to significant emotional and social handicap which is compounded in the presence of hearing loss, dizziness and tinnitus. These handicaps are strongly correlated to one another. Hence, a holistic approach is needed for USNHL rehabilitation. There exists a need to develop a comprehensive measure to assess and grade the handicap faced by these patients., Competing Interests: Conflicts of interestThere were no conflicts of interest., (© Association of Otolaryngologists of India 2021.)

The aim of this study is to analyze and characterize the antimicrobial effect of cerium oxide nanoparticles (NP) synthesized using neem and ginger. Finely grounded neem and ginger powder were taken and mixed with distilled water. This mixture was then heated and filtered. Ammonium cerium nitrate dissolved in distilled water. Both the mixtures were mixed and stirred magnetically. A double-beam ultraviolet-visible spectrophotometer was used to monitor color changes. The extract was centrifuged at 8000 rpm for 15 min. The final pellet was powdered using a hot air oven at 70°C for 24 h. Visualization was done by transmission electron microscopy and spherical morphology was noted, with an average diameter of 5 nm, in aggregated form. The sample containing 100 mg of cerium oxide shows the most significant effect on the zone of inhibition of 11 mm of Staphylococcus aureus . The results obtained in the current study confirmed that CeO-NP possessed antioxidant and cytotoxic properties., Competing Interests: There are no conflicts of interest., (Copyright: © 2022 Journal of Advanced Pharmaceutical Technology & Research.)

The present invention relates to dental compositions that include a phosphorescent material, more particularly, adding phosphorescent material to waxes that used in dentistry. The aim of the study is to develop a wax that has innate phosphorescent properties. Three groups of samples were taken in which the phosphorescent material was added. Three groups were later compared for their phosphorescent property in the material. All the samples in the initial molten state were poured into a mold made of silicone impression material and allowed to set. The material was placed in a dark room, and visual examination was done to compare the materials. From the samples obtained, paraffin wax showed more phosphorescent property than beeswax. The least phosphorescent property was seen in marginal wax. It can be concluded from this study that paraffin wax showed the most phosphorescent property., Competing Interests: There are no conflicts of interest., (Copyright: © 2022 Journal of Advanced Pharmaceutical Technology & Research.)

In countries with a high amount of geriatric patients, it is mandatory for a dental clinician to have working knowledge on complete denture (CD) procedures. The aim of this study was to troubleshoot the errors in denture fabrication by assessing the difficulties in clinical procedures. A questionnaire was distributed among dental undergraduates across the country in the present study. A total of 26 questions were included in the study. An online sharing medium was used for distribution and a total of 140 responses were received. SPSS for windows, version 20 was used for data tabulation. Descriptive statistics and Chi-square test were done. The results of this study revealed that jaw relation was the most difficult procedure followed by border molding. Multiple problems arise from the practitioner's end due to the lack of knowledge. One of the causes for these problems could be inability to learn the principles and procedures during the undergraduate program. This must be addressed at the base level so that better care can be given to patients., Competing Interests: There are no conflicts of interest., (Copyright: © 2022 Journal of Advanced Pharmaceutical Technology & Research.)

Introduction: Unilateral Sensorineural Hearing Loss (USNHL) is an age-old known entity. Patients with USNHL lack the merits of binaural hearing, i.e., temporal summation, sound localization, and speech recognition in a noisy environment. The lack of binaural hearing affects such individuals' quality of life (QOL). The present study is the most extensive Indian series related to QOL in USNHL patients., Objectives: This study aims to assess the impact of USNHL on the QOL of patients in the state of Uttarakhand., Methods: A cross-sectional study was carried out at a tertiary care center in Uttarakhand over a period of 18 months, from January 2018 to July 2019. A total of 115 patients with pure USNHL were evaluated using a validated questionnaire-Hearing Handicap Inventory for Adults-Hindi (HHIA-H)., Results: In our study, patients with higher HHIA-H scores were male, young adults (age group 18-30 years), students, and laborers. Most of our patients (64%) had significant handicaps, followed by moderate handicaps in 25%. However, no significant correlation of degree of handicap was seen with age, duration, and degree of hearing loss., Conclusion: USNHL can lead to a significant handicap that can severely affect the emotional and social aspects of life. Thus, early diagnosis and rehabilitation are essential to prevent handicap and uplift QOL in patients with USNHL. This trial is registered with Clinical Trials Registry of India (CTRI reg. no. CTRI/2018/06/014396)., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2022 Rachit Sood et al.)

Systemic sclerosis (SSc) is an autoimmune disorder leading to significant disability and loss of Quality of Life (QoL). Yoga has become popular in recent times for its potential therapeutic benefits. Since there are no scientific reports on the use of Yoga for SSc, we present two female cases (aged 49 and 29 respectively) of limited SSc (duration of illness 4 and 3 years respectively) who underwent Yoga therapy as an adjunct to conventional management in a residential setting for a period of five and four weeks, respectively. During their stay, they underwent a specifically designed Yoga module. After their discharge, they were followed-up for four weeks, during which they were asked to continue practicing Yoga for 1 h every day. Both of them reported a reduction in pain, stiffness, symptom scores, and improved QoL on discharge and at the follow-up compared to the values on admission. Erythrocyte Sedimentation Rate (ESR) and C-Reactive Protein (CRP), as biomarkers of inflammation, reduced on the discharge when compared to the baseline. No adverse events were noted during the stay and the follow-up. Thus, the present case series indicate a possible beneficial role of Yoga as an adjunct therapy to conventional management of SSc. Further studies in the area are warranted to ascertain the efficacy of Yoga for SSc., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

Uveal melanoma (UM) is the most prevalent primary intraocular malignancy in adults, and patients that develop metastases (~50%) survive <1 year, highlighting the urgent need for new therapies. TCGA has recently revealed that a hypoxia gene signature is associated with poor UM patient prognosis. Here we show that expression of hypoxia-regulated collagen prolyl-4-hydroxylase genes P4HA1 and P4HA2 is significantly upregulated in UM patients with metastatic disease and correlates with poor prognosis, suggesting these enzymes might be key tumor drivers. We targeted hypoxia-induced expression of P4HA1/2 in UM with KCN1, a hypoxia inducible factor-1 (HIF-1) pathway inhibitor and found potent inhibition of primary and metastatic disease and extension of animal survival, without overt side effects. At the molecular level, KCN1 antagonized hypoxia-induced expression of P4HA1 and P4HA2, which regulate collagen maturation and deposition in the extracellular matrix. The treatment decreased prolyl hydroxylation, induced proteolytic cleavage and rendered a disordered structure to collagen VI, the main collagen produced by UM, and reduced UM cell invasion. Together, these data demonstrate that extracellular collagen matrix formation can be targeted in UM by inhibiting hypoxia-induced P4HA1 and P4HA2 expression, warranting further development of this strategy in patients with uveal melanoma., (© 2021. The Author(s), under exclusive licence to Springer Nature Limited.)

Background: The relationship between the lack of quality care and adverse maternal outcomes is being highlighted globally. Respectful maternity care includes respect for women autonomy, dignity, feelings, privacy, choices and freedom from ill treatment, coercion, and consideration for personal preferences including option for companionship during the maternity care., Objectives: This study planned to determine the prevalence and types of disrespect and abuse among mothers during childbirth and its associated factors., Methods: This was a cross-sectional study conducted among 231 women having children <2 years, who had undergone institutional deliveries and accessing immunization clinics during October to December 2018 using a semi-structured questionnaire adapted from the Person-Centered Maternity Care Scale, a validated instrument which gives a holistic measure of women's experiences during childbirth wherein the questions were grouped under seven domains., Results: : The mean (standard deviation) age of the respondents was 28.9 ± 5.8 years. The prevalence of any form of abuse present was 96.5% with the highest found in "Facility and Environment" Domain (77.5%). Women delivering in public tertiary care facility were significantly more likely to face more abuse in the domains of "Dignity and Respect," "Support and Care," "Facility and Environment," "Predictability and Transparency," as compared to other types of health facilities (P < 0.05)., Conclusion: : Nearly almost all of the respondents (96.5%) faced some form of abuse in one domain or the other. There is a need for greater action, dialog, research, and advocacy on this important public health issue for promoting dignified maternal health-care services., Competing Interests: None

Purpose: Exploitation of altered glycosylation in cancer is a major goal for the design of new cancer therapy. Here, we designed a novel secreted chimeric signal peptide-Galectin-3 conjugate (sGal-3) and investigated its ability to induce cancer-specific cell death by targeting aberrantly N- glycosylated cell surface receptors on cancer cells., Experimental Design: sGal-3 was genetically engineered from Gal-3 by extending its N-terminus with a noncleavable signal peptide from tissue plasminogen activator. sGal-3 killing ability was tested on normal and tumor cells in vitro and its antitumor activity was evaluated in subcutaneous lung cancer and orthotopic malignant glioma models. The mechanism of killing was investigated through assays detecting sGal-3 interaction with specific glycans on the surface of tumor cells and the elicited downstream proapoptotic signaling., Results: We found sGal-3 preferentially binds to β1 integrin on the surface of tumor cells due to aberrant N -glycosylation resulting from cancer-associated upregulation of several glycosyltransferases. This interaction induces potent cancer-specific death by triggering an oncoglycan-β1/calpain/caspase-9 proapoptotic signaling cascade. sGal-3 could reduce the growth of subcutaneous lung cancers and malignant gliomas in brain, leading to increased animal survival., Conclusions: We demonstrate that sGal-3 kills aberrantly glycosylated tumor cells and antagonizes tumor growth through a novel integrin β1-dependent cell-extrinsic apoptotic pathway. These findings provide proof-of-principle that aberrant N -oncoglycans represent valid cancer targets and support further translation of the chimeric sGal-3 peptide conjugate for cancer therapy., (©2020 American Association for Cancer Research.)

Chiral sulfoxide based smart drug modafinil were studied experimentally and theoretically. Vibrational spectra were recorded in the mid IR region and electronic spectra were recorded in UV-Visible region. The molecular geometry, vibrational spectra, magnetic spectra and electronic spectra were simulated using Density Functional Theory (DFT) employed with B3LYP/6-311++G(d,p) basis set. The molecular geometry optimization, vibrational frequencies, chemical shifts and solvent effect on electronic properties were reported. The intermolecular interactions have been studied by Hirshfeld surface analysis. There is good agreement was found between calculated and observed values, thereby to confirm the molecular structure of modafinil., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Medulloblastoma (MB) is a malignant pediatric brain tumor for which new therapies are urgently needed. We demonstrate that treatment with EPZ-6438 (Tazemetostat), an enhancer of zeste homolog 2 (EZH2) inhibitor approved for clinical trials, blocks MB cell growth in vitro and in vivo, and prolongs survival in orthotopic xenograft models. We show that the therapeutic effect is dependent on epigenetic reactivation of adhesion G-protein-coupled receptor B1 (BAI1/ADGRB1), a tumor suppressor that controls p53 stability by blocking Mdm2. Histone 3 trimethylated on lysine 27 (H3K27me3), a marker of silent chromatin conformation is present at the ADGRB1 promoter, and inhibition of EZH2, the catalytic component of the Polycomb Repressive complex 2 (PRC2) that methylates H3K27, switches the gene into an active chromatin status and reactivates BAI1 expression. Mechanistically, targeting EZH2 promotes transition from H3K27me3 to H3K27ac at the promoter, recruits the C/EBPβ (CREB-binding protein) and CBP transcription factors and activates ADGRB1 gene transcription. Taken together, our results identify key molecular players that regulate ADGRB1 gene expression in MB, demonstrate that reactivation of BAI1 expression underlies EPZ-6438 antitumorigenic action, and provide preclinical proof-of-principle evidence for targeting EZH2 in patients with MB.

Purpose: Uveal melanoma (UM) is the most prevalent and lethal intraocular malignancy in adults. Here, we examined the importance of hypoxia in UM growth and tested the antitumor effects of arylsulfonamide 64B, an inhibitor of the hypoxia-induced factor (HIF) pathway in animal models of UM and investigated the related mechanisms., Experimental Design: UM cells were implanted in the uvea of mice eyes and mice systemically treated with 64B. Drug effect on primary eye tumor growth, circulating tumor cells, metastasis formation in liver, and survival were examined. 64B effects on UM cell growth, invasion and hypoxia-induced expression of C-X-C chemokine receptor type 4 (CXCR4) and mesenchymal-epithelial transition factor (c-Met) were measured. Luciferase reporter assays, chromatin immunoprecipitation, co-immunoprecipitation, and cellular thermal shift assays were used to determine how 64B interferes with the HIF transcriptional complex., Results: Systemic administration of 64B had potent antitumor effects against UM in several orthotopic mouse models, suppressing UM growth in the eye (∼70% reduction) and spontaneous liver metastasis (∼50% reduction), and extending mice survival ( P < 0.001) while being well tolerated. 64B inhibited hypoxia-induced expression of CXCR4 and c-Met, 2 key drivers of tumor invasion and metastasis. 64B disrupted the HIF-1 complex by interfering with HIF-1α binding to p300/CBP co-factors, thus reducing p300 recruitment to the MET and CXCR4 gene promoters. 64B could thermostabilize p300, supporting direct 64B binding to p300., Conclusions: Our preclinical efficacy studies support the further optimization of the 64B chemical scaffold toward a clinical candidate for the treatment of UM., (©2018 American Association for Cancer Research.)

Adhesion G protein-coupled receptors (ADGRs) encompass 33 human transmembrane proteins with long N termini involved in cell-cell and cell-matrix interactions. We show the ADGRB1 gene, which encodes Brain-specific angiogenesis inhibitor 1 (BAI1), is epigenetically silenced in medulloblastomas (MBs) through a methyl-CpG binding protein MBD2-dependent mechanism. Knockout of Adgrb1 in mice augments proliferation of cerebellar granule neuron precursors, and leads to accelerated tumor growth in the Ptch1 +/- transgenic MB mouse model. BAI1 prevents Mdm2-mediated p53 polyubiquitination, and its loss substantially reduces p53 levels. Reactivation of BAI1/p53 signaling axis by a brain-permeable MBD2 pathway inhibitor suppresses MB growth in vivo. Altogether, our data define BAI1's physiological role in tumorigenesis and directly couple an ADGR to cancer formation., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Prostaglandins and leukotrienes are produced in the COX and 5-LOX pathways of the inflammatory process. The current drugs target the upstream enzymes of either of the two pathways, leading to side effects. We have attempted to target the downstream enzymes simultaneously. Two compounds 2 and 3 (10 μM), identified by virtual screening, inhibited mPGES-1 activity by 53.4 ± 4.0 and 53.9 ± 8.1%, respectively. Structural and pharmacophore studies revealed a set of common residues between LTC4S and mPGES-1 as well as four-point pharmacophore mapping onto the inhibitors of both these enzymes as well as 2 and 3. These structural and pharmacophoric features may be exploited for ligand- and structure-based screening of inhibitors and designing of dual inhibitors.

The arachidonic acid pathway consists of several enzymes and targeting them is favored for developing anti-inflammatory drugs. However, till date the current drugs are generally active against a single target, leading to undesirable side-effects. Phytochemicals are known to inhibit multiple targets simultaneously and hence, an attempt is made here to investigate their suitability. A pharmacophore based study is performed with three sets of reported phytochemicals namely, dual 5-LOX/mPGES1, alkaloids and FLAP inhibitors. The analysis indicated that phenylpropanoids (including ferulic acid) and benzoic acids derivatives, and berberine mapped onto these pharmacophores with three hydrophobic centroids and an acceptor feature. 2,4,5-trimethoxy (7) and 3,4-dimethoxy cinnamic acids (8) mapped onto all the three pharmacophores. Experimental studies indicated that berberine inhibited 5-LOX (100 μM) and PGE 2 (50 μM) production by 72.2 and 72.0% and ferulic acid by 74.3 and 54.4% respectively. This approach offers a promising theoretical combined with experimental strategy for designing novel molecules against inflammatory enzymes., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Purpose: Elevation in D-2-Hydroxyglutarate (D-2HG) has recently emerged as a mandatory byproduct of mutated Isocitrate Dehydrogenase (IDH) genes 1 and 2 in glioma patients. The goal of the present study was to demonstrate the feasibility of detection of elevated levels of D-2HG in the cerebrospinal fluid (CSF) of glioma patients that carry point substitutions in the IDH gene., Experimental Design: We developed a mass spectrometry (MS)-based platform to detect and quantify the D- and L-forms of 2HG in the CSF of glioma patients. Three independent cohorts of patients were analyzed, comprising a total of 176 samples derived from 84 patients. The levels of D- and L-2HG were used to stratify patients into IDH wild-type or IDH-mutated groups using an empirically obtained threshold of 0.69 μmol/L., Results: Using this platform, a greater than 17-fold mean increase in D-2HG was observed in the CSF of patients with IDH mutant versus wild-type gliomas. The means for the D-2HG levels in CSF were 0.427 μmol/L in wild-type and 7.439 μmol/L in mutant groups. The C statistic for the receiver operator curve was 0.938, with 84% sensitivity, 90% specificity, and 89% accuracy to detect D-2HG. The levels of D- and L-2HG in CSF from wild-type patients varied by location of CSF draw (cisternal > ventricular > lumbar)., Conclusions: Our findings demonstrate that the CSF of patients harboring IDH mutant gliomas contain increased levels of D-2HG, which can be reliably detected with a MS-based platform. Clin Cancer Res; 22(24); 6256-65. ©2016 AACR., (©2016 American Association for Cancer Research.)

Purpose: The hypoxia-inducible factor-1 (HIF-1) plays a critical role in tumor adaptation to hypoxia, and its elevated expression correlates with poor prognosis and treatment failure in patients with cancer. In this study, we determined whether 3,4-dimethoxy-N-[(2,2-dimethyl-2H-chromen-6-yl)methyl]-N-phenylbenzenesulfonamide, KCN1, the lead inhibitor in a novel class of arylsulfonamide inhibitors of the HIF-1 pathway, had antitumorigenic properties in vivo and further defined its mechanism of action., Experimental Design: We studied the inhibitory effect of systemic KCN1 delivery on the growth of human brain tumors in mice. To define mechanisms of KCN1 anti-HIF activities, we examined its influence on the assembly of a functional HIF-1α/HIF-1β/p300 transcription complex., Results: KCN1 specifically inhibited HIF reporter gene activity in several glioma cell lines at the nanomolar level. KCN1 also downregulated transcription of endogenous HIF-1 target genes, such as VEGF, Glut-1, and carbonic anhydrase 9, in a hypoxia-responsive element (HRE)-dependent manner. KCN1 potently inhibited the growth of subcutaneous malignant glioma tumor xenografts with minimal adverse effects on the host. It also induced a temporary survival benefit in an intracranial model of glioma but had no effect in a model of melanoma metastasis to the brain. Mechanistically, KCN1 did not downregulate the levels of HIF-1α or other components of the HIF transcriptional complex; rather, it antagonized hypoxia-inducible transcription by disrupting the interaction of HIF-1α with transcriptional coactivators p300/CBP., Conclusions: Our results suggest that the new HIF pathway inhibitor KCN1 has antitumor activity in mouse models, supporting its further translation for the treatment of human tumors displaying hypoxia or HIF overexpression., (©2012 AACR.)

Brain-specific angiogenesis inhibitor 1 (BAI1), an orphan G protein-coupled receptor-type seven transmembrane protein, was recently found mutated or silenced in multiple human cancers and can interfere with tumor growth when overexpressed. Yet, little is known about its regulation and the molecular mechanisms through which this novel tumor suppressor exerts its anti-cancer effects. Here, we demonstrate that the N terminus of BAI1 is cleaved extracellularly to generate a truncated receptor and a 40-kDa fragment (Vasculostatin-40) that inhibits angiogenesis. We demonstrate that this novel proteolytic processing event depends on a two-step cascade of protease activation: proprotein convertases, primarily furin, activate latent matrix metalloproteinase-14, which then directly cleaves BAI1 to release the bioactive fragment. These findings significantly augment our knowledge of BAI1 by showing a novel post-translational mechanism regulating BAI1 activity through cancer-associated proteases, have important implications for BAI1 function and regulation, and present novel opportunities for therapy of cancer and other vascular diseases.

The hypoxia inducible factor (HIF) pathway is an attractive target for cancer, as it controls tumor adaptation to growth under hypoxia and mediates chemotherapy and radiation resistance. We previously discovered 3,4-dimethoxy-N-[(2,2-dimethyl-2H-chromen-6-yl)methyl]-N-phenylbenzenesulfonamide as a novel, small-molecule HIF-1 pathway inhibitor in a high-throughput cell-based assay, but its in vivo delivery is hampered by poor aqueous solubility (0.009 μM in water; log P(7.4) = 3.7). Here we describe the synthesis of 12 N-alkyl-N-[(8-R-2,2-dimethyl-2H-chromen-6-yl)methyl]heteroarylsulfonamides, which were designed to possess optimal lipophilicities and aqueous solubilities by in silico calculations. Experimental log P(7.4) values of 8 of the 12 new analogs ranged from 1.2-3.1. Aqueous solubilities of three analogs were measured, among which the most soluble N-[(8-methoxy-2,2-dimethyl-2H-chromen-6-yl)methyl]-N-(propan-2-yl)pyridine-2-sulfonamide had an aqueous solubility of 80 μM, e.g., a solubility improvement of ∼9000-fold. The pharmacological optimization had limited impact on drug efficacy as the compounds retained IC(50) values at or below 5 μM in our HIF-dependent reporter assay.

We have discovered that 3,4-dimethoxy-N-[(2,2-dimethyl-2H-chromen-6-yl)methyl]-N-phenylbenzenesulfonamide, a novel small molecule HIF-1 pathway inhibitor, can antagonize tumor growth in animal models of cancer, but the treatment necessitates its delivery in a formulation, due to poor water solubility (<15 μg/mL; pH 7.4), evidencing that the chemotype needs further exploration of its amenability to additional chemical modifications for ultimate optimization of function and pharmacology. As a first step towards this goal we investigated the structure-activity relationships of 15 lipophilic 2,2-dimethyl-2H-chromene based arylsulfonamide analogs of 3,4-dimethoxy-N-[(2,2-dimethyl-2H-chromen-6-yl)methyl]-N-phenylbenzenesulfonamide to find out strategies of modification. A 3,4-dimethoxybenzenesulfonyl group in region 1 showed the strongest inhibition among five arylsulfonyl groups tested. The presence of propan-2-amine in region 2 conferred the strongest inhibitory effect of the compound on HIF-1 activated transcription in a reporter assay. These findings are important as they help define the structural motifs where the 3,4-dimethoxy-N-[(2,2-dimethyl-2H-chromen-6-yl)methyl]-N-phenylbenzenesulfonamide can be chemically modified to improve its pharmacological properties towards development as a cancer therapeutic., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Leukotrienes are important mediators of pain and inflammation and they are produced in the arachidonic acid pathway via 5-lipoxygenase. They have been shown to have important roles in pyresis following antigen attack and in aspirin- intolerant asthma. They promote inflammation processes including eosinophil migration, increase in vascular permeability and bronchoconstriction. Hence, targeting the enzymes involved in the synthesis of these mediators can lead to the development of novel anti-inflammatory drugs. However, no drugs have yet been developed targeting leukotriene C4 synthase, a key enzyme leading to the synthesis of cysteinyl leukotrienes. The recent elucidation of its crystal structure now opens up the possibility of drugs against it. The inhibitors developed for this enzyme until now and the structural features responsible for their activity are discussed in this review. This understanding could lead to the design of new chemical entities.

Hypoxia inducible factors (HIFs) are transcription factors that activate expression of multiple gene products and promote tumor adaptation to a hypoxic environment. To become transcriptionally active, HIFs associate with cofactors p300 or CBP. Previously, we found that arylsulfonamides can antagonize HIF transcription in a bioassay, block the p300/HIF-1α interaction, and exert potent anticancer activity in several animal models. In the present work, KCN1-bead affinity pull down, (14)C-labeled KCN1 binding, and KCN1-surface plasmon resonance measurements provide initial support for a mechanism in which KCN1 can bind to the CH1 domain of p300 and likely prevent the p300/HIF-1α assembly. Using a previously reported NMR structure of the p300/HIF-1α complex, we have identified potential binding sites in the p300-CH1 domain. A two-site binding model coupled with IC50 values has allowed establishment of a modest ROC-based enrichment and creation of a guide for future analogue synthesis.

Hypoxia, a reduction in partial oxygen pressure, is a salient property of solid tumors. Hypoxia drives malignant progression and metastasis in tumors and participates in tumor resistance to radio- and chemotherapies. Hypoxia activates the hypoxia-inducible factor (HIF) family of transcription factors, which induce target genes that regulate adaptive biological processes such as anaerobic metabolism, cell motility, and angiogenesis. Clinical evidence has demonstrated that expression of HIF-1 is strongly associated with poor patient prognosis and activation of HIF-1 contributes to malignant behavior and therapeutic resistance. Consequently, HIF-1 has become an important therapeutic target for inhibition by small molecules. Herein, we describe the design and synthesis of small molecules that inhibit the HIF-1 signaling pathway. Many of these compounds exhibit inhibitory activity in the nanomolar range. Separate mechanistic studies indicate that these inhibitors do not alter HIF-1 levels but interfere with the ability of HIF-1α/HIF-1β to interact with cofactors p300/CBP to form an active transcriptional complex.

Solid tumors generally grow under hypoxic conditions, a pathophysiological change, which activates the expression of genes responsible for malignant, aggressive, and treatment-refractory properties. Hypoxia inducible factor (HIF) is the chief transcription factor regulating hypoxia-driven gene expression. Therefore, the HIF pathway has become a critical target for cancer therapeutics development. We screened a privileged library of about 10,000 natural-product-like compounds using a cell-based assay for HIF-dependent transcriptional activity and identified several arylsulfonamide HIF pathway inhibitors. Among these compounds, the most potent ones showed an IC(50) of ∼0.5 μM in the hypoxia-responsive element (HRE)-luciferase reporter system. Further studies are needed to fully elucidate the mechanism of action of this class of compounds and their structure-activity relationship., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The purpose of the present study was to investigate the effects of prenatal exposure to octylphenol (OP) at the dose of 50mg/kg body weight on days 1, 7 and 14 of pregnancy on reproductive health of male rats at adulthood. F1 male rats from control and OP exposed animals were weaned and maintained up to postnatal day (PND) 100. The indices of testis, epididymis and seminal vesicles were significantly decreased in male rats exposed to OP during embryonic development when compared with controls. Significant reduction in the epididymal sperm count, viable sperms and motile sperms and number of tail coiled sperms (HOS-test) were observed in experimental rats when compared to control rats. The levels of serum testosterone and also activity levels of testicular hydroxysteroid dehydrogenases were significantly decreased with a significant increase in the serum follicle stimulating and leutinizing hormones in experimental rats. Furthermore, embryonic exposure to OP caused significant down regulation of StAR, 3ß hydroxysteroid dehydrogenase and 17ß hydroxysteroid dehydrogenase mRNAs in testis of adult rats as compared to control rats. The results of fertility studies revealed that there was an increase in the mating index in experimental rats with an increase in the pre- and post-implantation losses in rats cohabited with treated animals indicating poor male reproductive performance., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

A facile, convenient and high yielding synthesis of a combinatorial library of 3-alkanoyl/aroyl/heteroaroyl-2H-chromene-2-thiones has been developed by the condensation of easily accessible beta-oxodithioesters and salicylaldehyde/substituted 2-hydroxybenzaldehydes under solvent-free conditions. The assessment of radical scavenging capacity of the compounds towards the stable free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) was measured and these compounds were found to scavenge DPPH free radical efficiently. Five selected compounds were able to protect curcumin from the attack of sulfur free radical generated by radiolysis of glutathione (GSH). The newly synthesized compounds exhibited profound antioxidant activities. Five of them rendered comparatively high antioxidant capacity., (Copyright (c) 2010 Elsevier Masson SAS. All rights reserved.)

A facile route to hitherto unknown 5-methylmercaptothiocarbonyl-4-aryl-3,4-dihydropyrimidin-2(1H)-ones and substituted 2H-chromene-2-thiones has been developed. SnCl(2)-catalyzed cyclocondensation of beta-oxodithioesters with a variety of readily accessible aldehydes and urea affords the dihydropyrimidinones. The methodology involves the three-component Biginelli reaction. On the other hand, substituted salicylaldehyde and beta-oxodithioesters reacted under the same condition to afford the substituted 2H-chromene-2-thiones in high yields.

Angiogenesis is a critical physiologic process that is appropriated during tumorigenesis. Little is known about how this process is specifically regulated in the brain. Brain angiogenesis inhibitor-1 (BAI1) is a brain-predominant seven-transmembrane protein that contains five antiangiogenic thrombospondin type-1 repeats (TSR). We recently showed that BAI1 is cleaved at a conserved proteolytic cleavage site releasing a soluble, 120 kDa antiangiogenic factor called vasculostatin (Vstat120). Vstat120 has been shown to inhibit in vitro angiogenesis and suppress subcutaneous tumor growth. Here, we examine its effect on the intracranial growth of malignant gliomas and further study its antitumor mechanism. First, we show that expression of Vstat120 strongly suppresses the intracranial growth of malignant gliomas, even in the presence of the strong proangiogenic stimulus mediated by the oncoprotein epidermal growth factor receptor variant III (EGFRvIII). This tumor-suppressive effect is accompanied by a decrease in tumor vascular density, suggesting a potent antiangiogenic effect in the brain. Second, and consistent with this interpretation, we find that treatment with Vstat120 reduces the migration of cultured microvascular endothelial cells in vitro and inhibits corneal angiogenesis in vivo. Third, we show that these antivascular effects critically depend on the presence of the cell surface receptor CD36 on endothelial cells in vitro and in vivo, supporting the role of Vstat120 TSRs in mediating these effects. These results advance the understanding of brain-specific angiogenic regulation, and suggest that Vstat120 has therapeutic potential in the treatment of brain tumors and other intracerebral vasculopathies.

Genes carry the instructions for making proteins that are found in a cell as a specific sequence of nucleotides that are found in DNA molecules. But, the regions of these genes that code for proteins may occupy only a small region of the sequence. Identification of the coding regions plays a vital role in understanding these genes. In this paper we have explored an Artificial Immune System (AIS) that can be used to strengthen and identify the protein coding regions in a genomic DNA system in changing environments and the CA technique for protein structure prediction of small alpha/beta proteins using Rosetta. From an initial round of Rosetta sampling, we learn properties of the energy landscape that guide a subsequent round of sampling toward lower-energy structures. Three different approaches to improve tertiary fold prediction using the genetic algorithm are discussed: refinement of the search strategy; combination of prediction and experiment; inclusion of experimental data as selection criteria into the genetic algorithm. It has been developed using a slight variant of genetic algorithm. Good classifiers can be produced, especially when the number of the antigens is increased. However, an increase in the range of the antigens somehow affects the fitness of the immune system. Experimental results confirm the scalability of the proposed AIS FMACA based classifier to handle large volume of datasets irrespective of the number of classes, tuples and attributes. We note an increase in accuracy of more than 5.2%, over any existing standard algorithms that address this problem. This was the first algorithm to identify protein coding regions in mixed and also non-overlapping exon-intron boundary DNA sequences. The accuracy of prediction of the structure of proteins was also found comparable.

BRCA1-associated protein-1 (BAP1), a deubiquitinating enzyme of unknown cellular function, is mutated in breast and lung cancers. In this study, we have shown for the first time that BAP1 has tumor suppressor activity in vivo by showing that BAP1 can suppress tumorigenicity of lung cancer cells in athymic nude mice. We show that BAP1 fulfills another criterion of a genuine tumor suppressor because cancer-associated BAP1 mutants are deficient in deubiquitinating activity. We show for the first time that one of the two predicted nuclear targeting motifs is required for nuclear localization of BAP1 and that a truncation mutant found in a lung cancer cell line results in BAP1 that fails to localize to the nucleus. Furthermore, we show that deubiquitinating activity and nuclear localization are both required for BAP1-mediated tumor suppression in nude mice. We show that BAP1 exerts its tumor suppressor functions by affecting the cell cycle, speeding the progression through the G(1)-S checkpoint, and inducing cell death via a process that has characteristics of both apoptosis and necrosis. Surprisingly, BAP1-mediated growth suppression is independent of wild-type BRCA1. Because deubiquitinating enzymes are components of the ubiquitin proteasome system, this pathway has emerged as an important target for anticancer drugs. The identification of the deubiquitinating enzyme BAP1 as a tumor suppressor may lead to further understanding of how the ubiquitin proteasome system contributes to cancer and aid in the identification of new targets for cancer therapy.

There is a need for novel therapies targeting hypoxic cells in tumors. These cells are associated with tumor resistance to therapy and express hypoxia inducible factor-1 (HIF-1), a transcription factor that mediates metabolic adaptation to hypoxia and activates tumor angiogenesis. We previously developed an oncolytic adenovirus (HYPR-Ad) for the specific killing of hypoxic/HIF-active tumor cells, which we now armed with an interleukin-4 gene (HYPR-Ad-IL4). We designed HYPR-Ad-IL4 by cloning the Ad E1A viral replication and IL-4 genes under the regulation of a bidirectional hypoxia/HIF-responsive promoter. The IL-4 cytokine was chosen for its ability to induce a strong host antitumor immune response and its potential antiangiogenic activity. HYPR-Ad-IL4 induced hypoxia-dependent IL-4 expression, viral replication, and conditional cytolysis of hypoxic, but not normoxic cells. The treatment of established human tumor xenografts with HYPR-Ad-IL4 resulted in rapid and maintained tumor regression with the same potency as that of wild-type dl309-Ad. HYPR-Ad-IL4-treated tumors displayed extensive necrosis, fibrosis, and widespread viral replication. Additionally, these tumors contained a distinctive leukocyte infiltrate and prominent hypoxia. The use of an oncolytic Ad that locally delivers IL-4 to tumors is novel, and we expect that HYPR-Ad-IL4 will have broad therapeutic use for all solid tumors that have hypoxia or active HIF, regardless of tissue origin or genetic alterations.

Brain angiogenesis inhibitor 1 (BAI1) is a transmembrane protein with unknown function expressed primarily in normal but not tumoral brain. The finding of thrombospondin type 1 repeats in its extracellular domain suggested an antiangiogenic function, but the mechanisms by which a transmembrane receptor could inhibit angiogenesis remained unexplained. Here we demonstrate that BAI1 is proteolytically cleaved at a conserved G-protein-coupled receptor proteolytic cleavage site (GPS), releasing its 120 kDa extracellular domain. We named this secreted fragment Vasculostatin as it inhibited migration of endothelial cells in vitro and dramatically reduced in vivo angiogenesis. Both constitutive and doxycycline-induced expression of Vasculostatin elicited dose-dependent suppression of tumor growth and vascular density in mice, implicating Vasculostatin in the regulation of vascular homeostasis and tumor prevention. Generation of a soluble antiangiogenic factor by cleavage of a pre-existing transmembrane protein represents a novel mechanism for regulating vascular homeostasis and preventing tumorigenesis. Modulation of this cleavage or delivery of Vasculostatin may constitute novel treatment modalities for cancer and other diseases of aberrant angiogenesis, especially in the brain.

Hypoxia plays a critical role in driving tumor malignancy and is associated with poor patient survival in many human cancers. Novel therapies targeting hypoxic tumor cells are urgently needed, because these cells hinder tumor eradication. Here we demonstrate than an anticancer strategy based on intratumoral delivery of a novel type of oncolytic adenovirus targeting tumor hypoxia is therapeutically efficient and can augment standard chemotherapy. We used a conditionally replicative adenovirus (HYPR-Ad) to specifically kill hypoxic tumor cells. Viral infection and conditional replication occurred efficiently in hypoxic/hypoxia-inducible factor-active cells in culture and in vivo, prevented tumor formation, and reduced the growth of established tumors. Combining HYPR-Ad with chemotherapy effective against normoxic cells resulted in strongly enhanced antitumor efficacy. These studies demonstrate that targeting the hypoxic microenvironment of tumors rather than an intrinsic gene expression defect is a viable and novel antitumor therapeutic strategy that can be used in combination with existing treatment regimens. The replication and oncolytic potential of this virus was made dependent on hypoxic/hypoxia-inducible factor, a transcription factor activated in the tumor hypoxic microenvironment, broadening its therapeutic use to solid tumors of any genetic make-up or tissue of origin.

TMS1/ASC is an intracellular signaling molecule with proposed roles in the regulation of apoptosis, nuclear factor-kappaB activation, and cytokine maturation. Previous studies have shown that TMS1/ASC is silenced by epigenetic means in human breast tumors. In this study, we examined methylation and expression of TMS1/ASC in glioblastoma multiforme (GBM). Whereas normal brain tissue was unmethylated at the TMS1 locus and expressed TMS1 message, 11 of 23 human GBM cell lines exhibited reduced or absent expression of TMS1 that was associated with aberrant methylation of a CpG island in the promoter of the TMS1 gene. Quantitative analysis showed that there was an inverse correlation between the degree of methylation and level of TMS1 expression. Treatment of GBM cell lines lacking TMS1 expression with the methyltransferase inhibitor 5-aza-2'deoxycytidine resulted in partial demethylation and re-expression of TMS1. Analysis of primary tissues indicated that the TMS1 gene is unmethylated and expressed in normal brain, where its expression is restricted to astrocytes. In contrast, TMS1 was aberrantly methylated in 43% (10 of 23) primary GBM specimens. Tumors that exhibited aberrant methylation of TMS1 generally expressed reduced or absent expression of TMS1 as compared to unmethylated cases. Methylation of TMS1 was not associated with patient age, gender, or treatment status. Although the relationship did not reach statistical significance, there was a trend toward increased overall survival for patients with unmethylated tumors. For one patient, disease progression from astrocytic astrocytoma (World Health Organization grade III) to GBM (World Health Organization grade IV) was associated with selective expansion of TMS1-negative cells. The data suggest a role for the epigenetic silencing of TMS1 in the pathogenesis of human GBM. Methylation of TMS1 may prove to be a useful prognostic marker and/or predictor of patient survival and tumor malignancy.