Pictilisib-Induced Resistance Is Mediated through FOXO1-Dependent Activation of Receptor Tyrosine Kinases in Mucinous Colorectal Adenocarcinoma Cells.

The phosphatidylinositol (PI3K)/AKT/mTOR axis represents an important therapeutic target to treat human cancers. A well-described downstream target of the PI3K pathway is the forkhead box O (FOXO) transcription factor family. FOXOs have been implicated in many cellular responses, including drug-induced resistance in cancer cells. However, FOXO-dependent acute phase resistance mediated by pictilisib, a potent small molecule PI3K inhibitor (PI3Ki), has not been studied. Here, we report that pictilisib-induced adaptive resistance is regulated by the FOXO-dependent rebound activity of receptor tyrosine kinases (RTKs) in mucinous colorectal adenocarcinoma (MCA) cells. The resistance mediated by PI3K inhibition involves the nuclear localization of FOXO and the altered expression of RTKs, including ErbB2, ErbB3, EphA7, EphA10, IR, and IGF-R1 in MCA cells. Further, in the presence of FOXO siRNA, the pictilisib-induced feedback activation of RTK regulators (pERK and pAKT) was altered in MCA cells. Interestingly, the combinational treatment of pictilisib (Pi3Ki) and FOXO1i (AS1842856) synergistically reduced MCA cell viability and increased apoptosis. These results demonstrate that pictilisib used as a single agent induces acute resistance, partly through FOXO1 inhibition. Therefore, overcoming PI3Ki single-agent adaptive resistance by rational design of FOXO1 and PI3K inhibitor combinations could significantly enhance the therapeutic efficacy of PI3K-targeting drugs in MCA cells.

NF1 mutations in conjunctival melanoma.

BACKGROUND: Conjunctival melanoma is a potentially deadly eye tumour. Despite effective local therapies, tumour recurrence and metastasis remain frequent. The genetics of conjunctival melanomas remain incompletely understood. METHODS: A large cohort of 63 conjunctival melanomas was screened for gene mutations known to be important in other melanoma subtypes by targeted next-generation sequencing. Mutation status was correlated with patient prognosis. RESULTS: Frequent mutations in genes activating the MAP kinase pathway were identified. NF1 mutations were most frequent (n = 21, 33%). Recurrent activating mutations were also identified in BRAF (n = 16, 25%) and RAS genes (n = 12, 19%; 11 NRAS and 1 KRAS). CONCLUSIONS: Similar to cutaneous melanomas, conjunctival melanomas can be grouped genetically into four groups: BRAF-mutated, RAS-mutated, NF1-mutated and triple wild-type melanomas. This genetic classification may be useful for assessment of therapeutic options for patients with metastatic conjunctival melanoma.

The endocytic recycling regulatory protein EHD1 Is required for ocular lens development.

The C-terminal Eps15 homology domain-containing (EHD) proteins play a key role in endocytic recycling, a fundamental cellular process that ensures the return of endocytosed membrane components and receptors back to the cell surface. To define the in vivo biological functions of EHD1, we have generated Ehd1 knockout mice and previously reported a requirement of EHD1 for spermatogenesis. Here, we show that approximately 56% of the Ehd1-null mice displayed gross ocular abnormalities, including anophthalmia, aphakia, microphthalmia and congenital cataracts. Histological characterization of ocular abnormalities showed pleiotropic defects that include a smaller or absent lens, persistence of lens stalk and hyaloid vasculature, and deformed optic cups. To test whether these profound ocular defects resulted from the loss of EHD1 in the lens or in non-lenticular tissues, we deleted the Ehd1 gene selectively in the presumptive lens ectoderm using Le-Cre. Conditional Ehd1 deletion in the lens resulted in developmental defects that included thin epithelial layers, small lenses and absence of corneal endothelium. Ehd1 deletion in the lens also resulted in reduced lens epithelial proliferation, survival and expression of junctional proteins E-cadherin and ZO-1. Finally, Le-Cre-mediated deletion of Ehd1 in the lens led to defects in corneal endothelial differentiation. Taken together, these data reveal a unique role for EHD1 in early lens development and suggest a previously unknown link between the endocytic recycling pathway and regulation of key developmental processes including proliferation, differentiation and morphogenesis.

Histone posttranslational modifications and cell fate determination: lens induction requires the lysine acetyltransferases CBP and p300.

Lens induction is a classical embryologic model to study cell fate determination. It has been proposed earlier that specific changes in core histone modifications accompany the process of cell fate specification and determination. The lysine acetyltransferases CBP and p300 function as principal enzymes that modify core histones to facilitate specific gene expression. Herein, we performed conditional inactivation of both CBP and p300 in the ectodermal cells that give rise to the lens placode. Inactivation of both CBP and p300 resulted in the dramatic discontinuation of all aspects of lens specification and organogenesis, resulting in aphakia. The CBP/p300(-/-) ectodermal cells are viable and not prone to apoptosis. These cells showed reduced expression of Six3 and Sox2, while expression of Pax6 was not upregulated, indicating discontinuation of lens induction. Consequently, expression of αB- and αA-crystallins was not initiated. Mutant ectoderm exhibited markedly reduced levels of histone H3 K18 and K27 acetylation, subtly increased H3 K27me3 and unaltered overall levels of H3 K9ac and H3 K4me3. Our data demonstrate that CBP and p300 are required to establish lens cell-type identity during lens induction, and suggest that posttranslational histone modifications are integral to normal cell fate determination in the mammalian lens.

Implications of sea level rise scenarios on land use /land cover classes of the coastal zones of Cochin, India.

Physical responses of the coastal zones in the vicinity of Cochin, India due to sea level rise are investigated based on analysis of inundation scenarios. Quantification of potential habitat loss was made by merging the Land use/Land cover (LU/LC) prepared from the satellite imagery with the digital elevation model. Scenarios were generated for two different rates of sea level rise and responses of changes occurred were made to ascertain the vulnerability and loss in extent. LU/LC classes overlaid on 1 m and 2 m elevation showed that it was mostly covered by vegetation areas followed by water and urban zones. For the sea level rise scenarios of 1 m and 2 m, the total inundation zones were estimated to be 169.11 km(2) and 598.83 km(2) respectively using Geographic Information System (GIS). The losses of urban areas were estimated at 43 km(2) and 187 km(2) for the 1 m and 2 m sea level rise respectively which is alarming information for the most densely populated state of India. Quantitative comparison of other LU/LC classes showed significant changes under each of the inundation scenarios. The results obtained conclusively point that sea level rise scenarios will bring profound effects on the land use and land cover classes as well as on coastal landforms in the study region. Coastal inundation would leave ocean front and inland properties vulnerable. Increase in these water levels would alter the coastal drainage gradients. Reduction in these gradients would increase flooding attributable to rainstorms which could promote salt water intrusion into coastal aquifers and force water tables to rise. Changes in the coastal landforms associated with inundation generate concern in the background that the coastal region may continue to remain vulnerable in the coming decades due to population growth and development pressures.

Distribution and dynamics of mangrove forests of South Asia.

Mangrove forests in South Asia occur along the tidal sea edge of Bangladesh, India, Pakistan, and Sri Lanka. These forests provide important ecosystem goods and services to the region's dense coastal populations and support important functions of the biosphere. Mangroves are under threat from both natural and anthropogenic stressors; however the current status and dynamics of the region's mangroves are poorly understood. We mapped the current extent of mangrove forests in South Asia and identified mangrove forest cover change (gain and loss) from 2000 to 2012 using Landsat satellite data. We also conducted three case studies in Indus Delta (Pakistan), Goa (India), and Sundarbans (Bangladesh and India) to identify rates, patterns, and causes of change in greater spatial and thematic details compared to regional assessment of mangrove forests. Our findings revealed that the areal extent of mangrove forests in South Asia is approximately 1,187,476 ha representing ∼7% of the global total. Our results showed that from 2000 to 2012, 92,135 ha of mangroves were deforested and 80,461 ha were reforested with a net loss of 11,673 ha. In all three case studies, mangrove areas have remained the same or increased slightly, however, the turnover was greater than the net change. Both, natural and anthropogenic factors are responsible for the change and turnover. The major causes of forest cover change are similar throughout the region; however, specific factors may be dominant in specific areas. Major causes of deforestation in South Asia include (i) conversion to other land use (e.g. conversion to agriculture, shrimp farms, development, and human settlement), (ii) over-harvesting (e.g. grazing, browsing and lopping, and fishing), (iii) pollution, (iv) decline in freshwater availability, (v) floodings, (vi) reduction of silt deposition, (vii) coastal erosion, and (viii) disturbances from tropical cyclones and tsunamis. Our analysis in the region's diverse socio-economic and environmental conditions highlights complex patterns of mangrove distribution and change. Results from this study provide important insight to the conservation and management of the important and threatened South Asian mangrove ecosystem.

Spry1 and Spry2 are necessary for eyelid closure.

Sproutys (Sprys) are downstream targets and negative feedback regulators of the FGF-Ras-ERK signaling pathway. Our previous studies have shown that Spry1 and Spry2, through negative modulation of FGF-ERK signaling, allow lens vesicle separation from the overlying ectoderm and regulate corneal epithelial proliferation. Here we show that Spry1 and Spry2 are necessary for eyelid closure. Murine palpebral conjunctival epithelial cells that differentiate as inner eyelids and adjacent mesenchymal cells express Spry1 and Spry2 prior to eyelid closure. Conditional deletion of both Spry1 and Spry2, but not either one alone, in the ocular surface epithelial cells result in the "EOB" (eyes open at birth) phenotype suggesting redundant roles for these proteins during eyelid closure. Spry mutant eyelids show increased proliferation of conjunctival epithelial cells with concomitant induction of FGF targets, Erm, Pea3 and Dusp6 and elevated ERK phosphorylation. Peridermal cells at the leading edge of Spry-mutant eyelids showed reduced c-Jun, but not ERK, phosphorylation, reduced F-actin polymerization and reduced motility in vitro. Spry mutant eyelids also showed disruptions in epithelial mesenchymal interactions reflected in the enhanced mesenchymal Spry1 and Spry4 expression, disaggregation of BMP4-positive mesenchymal cells and loss of Shh in the eyelid epithelium. Spry mutant eyelids also showed increased Wnt signaling and reduced expression of Foxc1 and Foxc2, two transcription factors previously shown to be necessary for eyelid closure. Collectively, our results show that conjunctival epithelial Spry1 and Spry2 redundantly promote eyelid closure by (a) stimulating ERK-independent, c-Jun-mediated peridermal migration, (b) suppressing conjunctival epithelial proliferation through FGF-ERK signaling, (c) mediating conjunctival epithelial-mesenchymal interactions and (d) maintaining expression of Foxc1 and Foxc2.

Bacterial bioluminescence response to long-term exposure to reverse osmosis treated effluents from dye industries.

The bacterial bioluminescence assay is one of the novel means for toxicity detection. The bioluminescence response of 2 marine bioluminescent bacteria was tested upon their long-term exposure to 9 different reverse osmosis (RO) rejects with varying chemical composition sampled from various dye industries. Bioluminescent bacteria were cultured in the RO reject samples, at different concentrations, and their growth rate and luminescence was measured for 24 h. The RO reject samples caused sublethal effects upon exposure and retarded the growth of bacteria, confirming their toxic nature. Further, continuation of the exposure showed that the initial luminescence, though reduced, recovered and increased beyond the control cultures irrespective of cell density, and finally decreased once again. The present study emphasizes the need of evolving a long-term exposure assay and shows that the method followed in this study is suitable to evaluate the toxicants that exert delayed toxicity, using lower concentrations of toxicants as well as coloured samples.

TERT promoter mutations in ocular melanoma distinguish between conjunctival and uveal tumours.

BACKGROUND: Recently, activating mutations in the TERT promoter were identified in cutaneous melanoma. We tested a cohort of ocular melanoma samples for similar mutations. METHODS: The TERT promoter region was analysed by Sanger sequencing in 47 uveal (ciliary body or choroidal) melanomas and 38 conjunctival melanomas. RESULTS: Mutations of the TERT promoter were not identified in uveal melanomas, but were detected in 12 (32%) conjunctival melanomas. Mutations had a UV signature and were identical to those found in cutaneous melanoma. CONCLUSION: Mutations of TERT promoter with UV signatures are frequent in conjunctival melanomas and favour a pathogenetic kinship with cutaneous melanomas. Absence of these mutations in uveal melanomas emphasises their genetic distinction from cutaneous and conjunctival melanomas.

Lumbar drains may reduce the need for permanent CSF diversion in spontaneous subarachnoid haemorrhage.

Subarachnoid haemorrhage (SAH) is well known to induce hydrocephalus. This is often, initially, treated with external ventricular drainage (EVD). We recently started, also, using lumbar drains (LD) in patients refractory to removal of their EVD as a bridge to permanent CSF diversion. LD were placed in 25 patients with spontaneous SAH. LD remained in place a mean of 6.7 days (range 4-16) prior to removal. Patients had a median Fisher Grade of 4, Hunt-Hess score of 4, and WFNS score of 4. Only 4 of 25 patients (16%) progressed to the need of permanent shunting, one of which occurred after delayed recurrent aneurysm rupture. Only 7 of 25 patients developed symptomatic vasospasm despite their high median Fisher Grade. Both the shunt rate and the symptomatic vasospasm rate in this series are much less than the historical series predict. This suggests that lumbar drains may reduce the need for shunting and decrease the rate of symptomatic vasospasm.

Fracture reattachment in an immature permanent incisor with talon's cusp. A rare case report.

AIM: To report the successful and conservative management of a fractured immature permanent maxillary incisor tooth with talon cusp by fracture reattachment. SUMMARY: Coronal fractures of the maxillary anterior teeth are common dental injuries. Among these, the complicated fractures especially in immature teeth require an unambiguous treatment without any delay. When the tooth fragment is available and there is no (or minimal) violation of the biological width, reattachment is the preferable choice. It is a conservative procedure in which the original anatomic form, color, tooth contour, surface texture and aesthetics are preserved. The prognosis of this procedure depends on the patient's cooperation, good understanding about the treatment limitations and periodic follow up. This report emphasises the management of coronal fracture in immature teeth by reattachment.

Centric relation definition: a historical and contemporary prosthodontic perspective.

Centric relation (CR) is a core topic of dentistry in general and prosthodontics in particular. The term CR has become thoroughly confusing because of many conflicting definitions. Unfortunately definition of CR changed repeatedly over past ten decades. All the existing definitions in the dental literature, for the past 81 years, are segregated into definitions from 1929 to 1970, 1970-1980, and 1980-2010 and are critically analyzed. Both PubMed (key words: centric relation/centric jaw relation) and hand searches were employed, from citation in other publications, to identify relevant articles in English language peer reviewed PubMed journals from 1956 to 2010; although the review is from 1929. Numerous definitions for CR have been given, however, no consensus exists and the definition given by a current glossary of prosthodontic terms is confusing. It relates CR to many clinically invisible parts and cannot guide a dental surgeon to record the CR following its description. The purpose of this article is not only to review all the definitions critically but to propose a self explanatory definition to minimize the confusion in the minds of dental practitioners and students for better understanding of the concept of CR. Centric relation is clinically significant since it is the only clinically repeatable jaw relation and the logical position to fabricate prosthesis.

Repurposing of clinically proven bioactive compounds for targeted treatment of Alzheimer's disease using molecular docking approach.

Neurodegenerative diseases like Alzheimer's have become a growing concern as it is difficult to cure. Tau protein is found to be playing a major role in Alzheimer's disease, and the majority of drugs that are currently on the market are not only prohibitively expensive but also come packaged with side effects that the body cannot tolerate. Repurposing existing compounds is a successful and optimistic strategy that offers reduced risk and increased possibility. We aim to retrieve the existing drugs and analyze them using in-silico techniques. We have retrieved the compounds from the Selleckchem natural product library, and the ability of the drug to cross Blood Brain Barrier (BBB) and ADMET (Absorption, Distribution, Metabolism, Excretion, and Toxicity) properties were examined using SwissADME. The structure of Tau protein (2MZ7) was then retrieved from PDB, and molecular docking of the compounds was performed using the PyRx-Virtual Screening Tool. Initially, 92 compounds passed the ADMET screening criteria, out of which the compound Ligustroflavone was found to have the most favourable binding affinity without violating Lipinski's rule of 5 of the compounds in the library.

Andersen health care utilization model: A survey on factors affecting the utilization of dental health services among school children.

BACKGROUND: Children's quality of life, academic performance, and future achievement can all be negatively affected by poor dental health. The present study aimed to assess the need for dental health services and the factors influencing their utilization using the Andersen health care utilization model among school children. METHODS: The current cross-sectional study was conducted among schoolchildren aged 13 to 15 in Bangalore, India (n = 1100). A questionnaire was developed using the concepts of the Andersen healthcare usage model. The parents of the children filled out the questionnaire. The factors were investigated using bivariate analysis and multivariate logistic regression analysis. RESULTS: About 78.1% of the children did not utilize dental health services. Regarding the reasons for not visiting a dentist, 65.8% said they did not have a dental problem, and 22.2% said they could not afford it. Bivariate analysis showed that age, gender, education level, occupation of the family's head of household, monthly family income, socioeconomic status, perceived oral health problems, accessibility of dental health facilities, and parental attitudes toward their children's oral health were significantly associated with using dental health services (p<0.05). Multiple regression analysis showed dental health service utilization was directly related to age (OR = 2.206), education, family size (OR = 1.33), and brushing frequency twice a day (OR = 1.575) with no significant relationship between distance to reach the dental facility, the number of dental visits, and socioeconomic status. CONCLUSION: Dental health service utilization was low in the past year. The age, number of family members, parent's education level, travel time to the dental facility, the child's oral health behaviors, and positive parental attitude all play a role in a children's utilization of dental health service.

Aberrant hypermethylation in primary tumours and sentinel lymph node metastases in paediatric patients with cutaneous melanoma.

BACKGROUND: Debate on how to manage paediatric patients with cutaneous melanoma continues, particularly in those with sentinel lymph node (SLN) metastases who are at higher risk of poor outcomes. Management is often based on adult algorithms, although differences in clinical outcomes between paediatric and adult patients suggest that melanoma in paediatric patients differs biologically. Yet, there are no molecular prognostic studies identifying these differences. OBJECTIVES: We investigated the epigenetic (methylation) regulation of several tumour-related genes (TRGs) known to be significant in adult melanoma progression in histopathology(+) SLN metastases (n = 17) and primary tumours (n = 20) of paediatric patients with melanoma to determine their clinical relevance. METHODS: Paediatric patients (n = 37; ≤ 21 years at diagnosis) with American Joint Committee on Cancer stage I-III cutaneous melanoma were analysed. Gene promoter methylation of the TRGs RASSF1A, RARß2, WIF1 and APC was evaluated. RESULTS: Hypermethylation of RASSF1A, RARß2, WIF1 and APC was found in 29% (5/17), 25% (4/16), 25% (4/16) and 19% (3/16) of histopathology(+) SLNs, respectively. When matched to adult cutaneous melanomas by Breslow thickness and ulceration, hypermethylation of all four TRGs in SLN(+) paediatric patients with melanoma was equivalent to or less than in adults. With a median follow-up of 55 months, SLN(+) paediatric patients with melanoma with hypermethylation of > 1 TRG vs. ≤ 1 TRG had worse disease-free (P = 0·02) and overall survival (P = 0·02). CONCLUSIONS: Differences in the methylation status of these TRGs in SLN(+) paediatric and adult patients with melanoma may account for why SLN(+) paediatric patients have different clinical outcomes. SLN biopsy should continue to be performed; within SLN(+) paediatric patients with melanoma, hypermethylation of TRGs can be used to identify a subpopulation at highest risk for poor outcomes who warrant vigilant clinical follow-up.

Co-Variation of Serum Osteoprotegerin and Pigment-Epithelial Derived Factor as Biomarker of Pancreatic Cancer.

Pancreatic cancer is one the most lethal cancers. Currently, there are reliable predictive markers to assess cancer development. Widely used CA19-9 molecular marker has been less effective in the diagnosis of early stages of cancer. Objective: To study if the soluble Osteoprotegerin (OPG) and pigment-epithelial derived factor (PEDF) levels in serum will be an indicator of cancer progression. Methods: Soluble OPG and PEDF were measured from human pancreatic cancer patients by ELISA. Results: We show that while OPG has been less predictive features, PEDF is more sensitive than CA19-9 in cancer detection. More importantly, PEDF and CA19-9 as combined markers showed higher sensitivity in stratifying early stages of pancreatic cancer. Conclusion: Results from the pilot studies suggest that PEDF is useful biomarker for pancreatic cancer.

Evaluating the failure to bloom in dark-cutting and lactate-enhanced beef longissimus steaks.

Previous studies have noted lower L* (lightness) values for both dark-cutting beef and normal-pH beef enhanced with lactate. In the current study, absorption-coefficient, scattering-coefficient, CIE L*a*b* values, refractive index of sarcoplasm, and inter-muscle bundle space were evaluated for dark-cutting beef, normal-pH beef enhanced with lactate, normal-pH beef enhanced with water, and normal-pH beef not enhanced with either water or lactate. Compared with non-enhanced loins, lactate-enhancement had lower a*, chroma, oxymyoglobin, reflectance, scattering, and inter-muscle bundle space as well as greater absorption and refractive index. Dark-cutting steaks had lower a*, chroma, oxymyoglobin values, reflectance, and scattering as well as less inter-muscle bundle space compared with lactate-enhanced steaks. Sarcoplasm refractive index values were greater in lactate-enhanced steaks than dark-cutting steaks. The results suggest that changes in muscle structure and optical properties due to either pH or lactate addition can alter muscle darkening and blooming properties.

Microwave based nanogenerator using the ratchet effect in Si/SiGe heterostructures.

Ratchet based microwave current generators and detectors were developed in Si/SiGe heterostructures for wireless communication with the possibility of extending the detection limit to the terahertz range. A microwave induced ratchet current was generated in the two-dimensional electron gas by patterning an array of semicircular antidots in hexagonal geometry. The spatial asymmetry created by the semicircular antidots forces the electrons under the influence of the microwave electric field to move preferentially towards the direction of the semidisc axis. A photovoltage of the order of few millivolts was observed. Such a photovoltage was completely absent in a symmetric system consisting of circular antidots. The induced photovoltage increased monotonically with microwave power and was found to be independent of the microwave polarization. This device opens the possibility of employing silicon based heterostructures for nanogenerators and other wireless communication devices using microwaves.

Role of cell-free network communication in alcohol-associated disorders and liver metastasis.

The aberrant use of alcohol is a major factor in cancer progression and metastasis. Contributing mechanisms include the systemic effects of alcohol and the exchange of bioactive molecules between cancerous and non-cancerous cells along the brain-gut-liver axis. Such interplay leads to changes in molecular, cellular, and biological functions resulting in cancer progression. Recent investigations have examined the role of extracellular vesicles (EVs) in cancer mechanisms in addition to their contribution as diagnostic biomarkers. Also, EVs are emerging as novel cell-free mediators in pathophysiological scenarios including alcohol-mediated gut microbiome dysbiosis and the release of nanosized EVs into the circulatory system. Interestingly, EVs in cancer patients are enriched with oncogenes, miRNA, lipids, and glycoproteins whose delivery into the hepatic microenvironment may be enhanced by the detrimental effects of alcohol. Proof-of-concept studies indicate that alcohol-associated liver disease is impacted by the effects of exosomes, including altered immune responses, reprogramming of stromal cells, and remodeling of the extracellular matrix. Moreover, the culmination of alcohol-related changes in the liver likely contributes to enhanced hepatic metastases and poor outcomes for cancer patients. This review summarizes the numerous aspects of exosome communications between organs with emphasis on the relationship of EVs in alcohol-associated diseases and cancer metastasis. The potential impact of EV cargo and release along a multi-organ axis is highly relevant to the promotion of tumorigenic mechanisms and metastatic disease. It is hypothesized that EVs target recipient tissues to initiate the formation of prometastatic niches and cancer progression. The study of alcohol-associated mechanisms in metastatic cancers is expected to reveal a better understanding of factors involved in the growth of secondary malignancies as well as novel approaches for therapeutic interventions.