Search

Oral cavity hygiene has gained significance in recent years. Dental plaque is the community of microorganisms found on tooth surface as bio-film embedded in matrix of polymers of host and bacterial origin.Dental plaque is considered to be main causative factor for both dental caries as well periodontal disease. Although mechanical oral hygiene at home remains single most reliable means of cleaning teeth, however majority of people do not carry out regular activities like brushing daily due to unawareness of correct brushing techniques as well as duration hence use of mouth rinses has been widely advocated as an adjunctive treatment.Variety of antimicrobial mouthwashes are being used to treat plaque causing microorganisms however many times these mouthwashes are known give rise to adverse oral manifestations such as altered taste sensations soreness of oral mucosa/ tongue discoloration as well as staining.Herbal extracts have been used in traditional medicine for several thousand years as they are cheap and have little side effects.Herbal Medicine is still mainstay of about 75 to 80% world population mainly in developing countries for primary health care because of general belief that herbal drugs are without side effects besides cheap and locally available. According to World Health Organization, the use of herbal remedies throughout world exceeds that of the conventional drugs by two to three times.This article aims at brief description of common herbs such as Curcuma Longa, Acacia Arabica, Azadirachta indica Cymbopogon Citratus, Ocimum Sanctum, Salvadora Persica, Acacia Catechu, Aloe Barbadensis, Juglans Regia, Guaijaverin and their use. Herbs may appear to potential alternative to current treatment modalities however more research is required.

IKKepsilon expression in pancreatic tumor samples and PDAC cell lines (S1); Ectopic expression of IKKepsilon enhances growth of pancreatic cancer cells (S2); Transient knockdown of IKKepsilon in MiaPaCa and Colo357 cells by siRNA (S3); Forced expression of IKKepsilon in BxPC3 cells enhances glucose-uptake and consumption (S4); Overexpression of IKKepsilon in BxPC3 enhanced the expression and nuclear translocation of c-MYC (S5); IKKepsilon knockdown decreases half-life of c-MYC in MiaPaCa and Colo357 cells (S6); Ectopic expression of IKKepsilon in pancreatic cancer cell line BxPC3 activates Akt (S7).

List of primers used.

Description of additional methods and procedures used in the study.

Aberrant expression of the kinase IKKϵ in pancreatic ductal adenocarcinoma (PDAC) has been associated with poor prognosis. In this study, we define a pathobiologic function for IKKϵ in reprogramming glucose metabolism and driving progression in PDAC. Silencing IKKϵ in PDAC cells, which overexpressed it endogenously, was sufficient to reduce malignant cell growth, clonogenic potential, glucose consumption, lactate secretion, and expression of genes involved in glucose metabolism, without impacting the basal oxygen consumption rate. IKKϵ silencing also attenuated c-Myc in a manner associated with diminished signaling through an AKT/GSK3β/c-MYC phosphorylation cascade that promoted MYC nuclear accumulation. In an orthotopic mouse model, IKKϵ-silenced PDAC exhibited a relative reduction in glucose uptake, tumorigenicity, and metastasis. Overall, our findings offer a preclinical mechanistic rationale to target IKKϵ to improve the therapeutic management of PDAC in patients. Cancer Res; 76(24); 7254–64. ©2016 AACR.

PURPOSE: While pain is prevalent among survivors of head and neck cancer (HNC), there is a lack of data on pain management in the community oncology setting. We described sociodemographic correlates and disparities associated with patient-reported pain among patients with HNC. METHODS: We used the 2017-2021 nationwide community oncology data set from Navigating Cancer, which included electronic patient-reported outcomes. We identified a retrospective cohort of patients diagnosed with HNC (N = 25,572), with ≥ 1 patient-reported pain event. We adjusted for demographic (sex, age, smoking history, marital status) and clinical (cancer site) factors associated with pain reporting and pain resolution by new pain prescription on the basis of race (White v non-White patients), using multivariate logistic regression models. RESULTS: Our analytic cohort included 2,331 patients, 90.58% White, 58.62% married, with an average age of 66.47 years. Of these, 857 patients (36.76%) reported ≥ 1 pain event during study period. Mean resolution time (in minutes) for pain incidents was significantly longer for White patients than non-White patients (99.6 ± 3.2 v 74.9 ± 7.2, P < .05). After adjusting for covariates, smoking was associated with a 25% increased odds of reporting pain incidents (adjusted odds ratio [aOR], 1.25; 95% CI, 1.03 to 1.52). There was no statistically significant difference in odds of pain reporting between White versus non-White patients (aOR, 0.97; 95% CI, 0.73 to 1.30). However, White patients were significantly more likely to receive new prescription for pain than non-White patients (aOR, 2.52; 95% CI, 1.09 to 5.86). CONCLUSION: We found racial differences in patient-reported pain management, with White patients significantly more likely to receive new pain prescriptions. As pain management is a mainstay in cancer care, equity in pain management is critical to optimize quality of life for patients with HNC.

Chimney or Stacks is a long and flexible structure made of steel, concrete and stone construction. These are used to dispose of toxic water or gases at high altitudes that do not pollute the air and access to the floor is within the approved limits approved by the general pollution control officer. Various loads such as autonomy, fluid load, air load, earthquake load and heat exchanger operator. Most industrial steel chimneys are long structures with a circular section crossing; such small and medium-sized structures often have an air vibration in which the geometry of the supporting steel chimney plays an important role in its structural behavior under the corresponding strong loading. The basic dimensions of the industrial steel support chimney, such as height, width at the outlet etc. To validate the failure mode configuration code (IS-6533: 1989 Part 2) set several parameters in the geometry that range from top to base width width and height to the base width of the steel frame. The purpose of the current study is to obtain a comparative study of a supporting steel chimney for wind load and earthquake effect and also to justify code methods regarding the size of an industrial steel chimney foundation. In the present study of the static and flexible analysis of a steel chimney 45, 60, 75 and 90 m long, of the landscape II, III, IV, V with a base resting on hard and soft soil layers was performed. The wind speed operating in the chimney varied from 33 to 55 m / sec. The system analysis was developed using MATLAB 2013. The results obtained from the MATLAB system were compared with the data already available in the literature (Punmia et al. 2012]. , the system was developed using the MATLAB 2013 air analysis system with a strong air strength of 45m high-strength steel slabs of unstable cement with varying heights of the thickness of the scale and the maximum width to the base width. The results were compared with data already published in the literature (Rakshith et al, 2015). The results were consistent. It was evident from the results that the geometric limits specified by the code (IS-6533: 1989 Part 2) were not traded for dynamic analysis., {"references":["John, A. D., Gairola, A., Ganju, E., & Gupta, A. (2011). Design wind loads on reinforced concrete chimney–An experimental case study. Procedia engineering, 14, 1252-1257.","Gaczek, M., & Kawecki, J. (1989). A new method for prediction of steel chimney response to vortex shedding. In Int. Conf.: Dynamics of Structures- Preprints, Karlovy Vary (pp. 191- 194).","Gaczek, M., & Kawecki, J. (1996). Analysis of cross-wind response of steel chimneys with spoilers. Journal of wind engineering and industrial aerodynamics, 65(1-3), 87-96.","Galemann, T., & Ruscheweyh, H. (1992). Measurements of wind induced vibrations of a full-scale steel chimney. Journal of Wind Engineering and Industrial Aerodynamics, 41(1-3), 241-252.","IS 6533, part 1. Mechanical aspect, 1989, Code of practice for design and construction of steel chimney","IS 6533, part-2, Structural aspect. 1989. Code of practice for design and construction of steel chimney.","Jain, S.K. and Singh, B.P (1992). IS code provision for seismic design of tall chimneys. International Journal of Structure, 10 (2).103-111.","Jisha, S. V., Jayalekshmi, B. R., & Shivashankar, R. (2014). Analysis of foundation of tall RC chimney with 3D finite element method. In Computing in Civil and Building Engineering (2014) (pp. 1368-1375).","Jozwiak, R., Wysocki, Z., & Zuranski, J. A. (1991). Sensitivity of multi-flue chimneys to aero-elastic galloping instability. In Proceedings of Conference Krynica (Vol. 1, pp. 67- 73)."]}

Preflight performance characterization of spaceborne imaging systems offers high-quality images for scientific applications. A key challenge in the laboratory characterization process is to identify the candidate signal chains requiring performance optimization. For the Indian Remote Sensing (IRS) imaging systems, light transfer characterization has been identified as a standard process for performance characterization. Photon transfer curve (PTC) is another powerful and widely used tool to characterize the imaging systems in terms of camera gain ( $e^{-}$ /DN), read noise ( $e^{-}$ ), charge-to-voltage conversion factor (CVF), signal-to-noise ratio, mean–variance linearity, and so on. There have been no detailed investigations on the PTC characteristics of the IRS imaging systems. We present here PTC-based characterization studies on two high-resolution IRS imaging systems, namely, Cartosat-1 and Cartosat-2. For this, a quantitative analytical framework has been developed, which enables comparative studies among multiple signal chains by applying various statistical measures on the PTC derived parameters. This framework provides not only a quantitative assessment of performance deviations but also enables performance traceability up to detector level. Taken together, our analysis shows that all the signal chains have well behaved PTC characteristics, and performance deviations are less than 10%. In particular, performance traceability is established by the close match of the system-level CVF values within the detector manufacturer’s specified range. Studies on the adequacy of linear approximation of the PTC curve reveal large residual errors in the lower dynamic range due to an increase in read noise floor. The analytical framework developed here can significantly help optimize future IRS imaging systems.

In modern times reinforced concrete, composite concrete, is made. It has been used successfully to build its outstanding flexural strength, resistance, impact resistance and strength and frost resistance. It is an effective way to increase durability, shock resistance and cracking of plastic cement cracks. Fiber is added as a reinforcement material with some characteristic features. They can be round, triangular or flat in the cross section. Fiber is usually defined by simple parameters called-- factor ratio. Fiber feature measurement is its length and its back size. The main reason for inserting fibers in cement matrix is to increase the strength and durability and improve the resistance to cracking of the resulting cracks. For fiber Reinforced Concrete to be a viable construction material, it must be able to economically complete with the existing reinforcement system. The reinforced concrete slab is reinforced with concrete by adding briefly separated strands to the concrete. It shows many stable and advanced engineering structures such as compressive strength, strength, flexural strength etc. Fibers are able to prevent cracks in the ground by the act of cleaning which leads to better resistance to concrete. The combination of two or more different fibers is very common these days with the aim of improving the overall performance of the system. The goal is that the performance of these hybrid systems will outweigh the potential for each type of fiber. Hybrid is based on fiber-based reactions, in which one fiber is stronger and stronger and more powerful, while the other is ductile and provides difficulty in difficult areas (Banthiaand Nandakumar 2003 Hybrids are based on fiber size, where one fiber is too small and provides less control in the early stages of loading and the other fiber is larger, provides a way to close large cracks. Hybrid is also based on fiber performance, where one type of fiber provides strength or durability to a complex composite, while the second type offers new blending properties ready for processing., {"references":["Massoud, M. T. (2002). Interaction of silica fume and polypropylene fibers in high performance concrete.","AM Alkhaleefi, M MEI·Hawary, H Abdel·Fattah.(2002). On the behaviour of polymer portland cement concreteˮ in 27th Conference on Our world in concrete & structures: 29 - 30 August 2002, Singapore. Article Online Id: 100027015 pages 137 -144.","Neville, A., M.,& Brook., J.,J. Advance Concrete Technology.","F.Cambell.(2010). Structural Composite Materia.","Foud., F. Buildings and use of Fiber Reinforced Concrete. JKAU Eng. Sci.2(2):49-6.","Selina Ruby, G., Geethanjali, C., Varghese, J., & Muthu Priya, P. (2014). Influence of hybrid fiber on reinforced concrete. International Journal of Advanced Structures and Geotechnical Engineering, 3(01).","Patel, I., & Modhera, C. D. (2013). Experimental Investigation to Study Effect of Polyester Fibre on Durability. JERS.2(1):159-166","Patel, I., & Modhera, C. D. (2013). Experimental Investigation to Study Effect of Polyester Fibre on Durability of HVFA Concrete through RCPT Method. IOSR J. Eng, 3(6), 22-27.","Löfgren, I. (2005). Fibre-reinforced Concrete for Industrial Construction-a fracture mechanics approach to material testing and structural analysis. Chalmers University of Technology."]}

1575 Background: Digital health solutions (DHS) allow for enhanced remote communication between patients and clinical staff and the COVID-19 pandemic has brought these tools to the forefront of care delivery. Once adopted, barriers to adequate utilization still exist. Given the important need to decrease digital divides, and the diversity of patients and care settings across our clinic’s 220 sites of service, we sought to understand how utilization of oncology DHS may be limited among certain populations. Methods: We investigated utilization among cancer patients who enrolled and engaged with a portfolio of DHS between March 1, 2019 and January 15, 2022. This portfolio includes three tools: (1) an electronic patient-reported outcomes (ePRO) remote monitoring program for tracking symptoms and oral adherence, (2) a patient portal (PP) for securely accessing patient health records, and (3) digital education (DE) for patients regarding disease and treatments. ePRO completion rate, average number of PP logins, and DE read rate were used as measures of utilization for each tool, respectively, and compared among patients with different age (< 65 and ≥65 years), language preference [English (EL) or Spanish (SL)], and distance from clinic (non-rural: < 20 miles OR rural: ≥20 miles). Mann-Whitney U and Chi-Square tests were used to compare continuous and categorical variables, respectively. Results: This study included a total of 77,347 unique patients representing 651,004 digital encounters. 9,938 patients engaged in ePRO, 49,771 patients in PP, and 12,044 patients in DE. Engagement across all DHS was high in patients of age group < 65 (ePRO: 72.7%, PP: 79.67% and PE 54.7%) as compared to ≥65 years, but the ePRO completion rate is high in ≥65 age group (59.0% vs 55.6%), whereas no significant difference was observed in the PP login activity and DE read rate. EL patients were significantly (p-value < 0.01) more engaged (ePRO 68% vs. 54%, PP: 80% vs. 62%, DE: 57% vs. 37%) and had higher digital utilization (ePRO completion rate: 57.31% vs 53.23%, average PP logins: 7.48 vs 7.14 and DE read rate: 96.2% vs 90.8%) than SL patients across the DHS. Patients living in rural areas comprised roughly 25% of the population and participated across tools similarly as patients living in non-rural areas (ePRO 67% vs. 69%, PP: 79% vs. 79%, DE: 56.9% vs. 56.8%). Utilization of the portfolio was variable based on rural vs non-rural status (ePRO completion rate: 56.3% vs. 57.4%, average PP logins: 7.9 vs. 7.3, DE read rate: 96.02.7% vs 96.3%). Conclusions: Despite variable engagement based on age, language, and rural status across the portfolio, patients within these populations continue to utilize the DHS. How we understand and explore enhancements to DHS remain under investigation for tool optimization for patient-specific barriers to care.

1507 Background: There is a priority to accelerate the delivery of digital health solutions (DHS) to provide patients with enhanced means for accessing care, but lack of understanding of their utility in certain populations. There are concerns that equitable adoption translate into disparities. We sought to implement a portfolio of DHS across a large practice and characterize engagement across populations to enhance clinical informatics solutions that support care delivery. Methods: This is a retrospective evaluation of cancer patients who engaged with a portfolio of DHS between March 1, 2019 and January 15, 2022. We included four tools with opt-in and opt-out functionality: (1) a care management (CM) platform utilized by clinical staff to manage patient activities, (2) an electronic patient-reported outcomes (ePRO) remote monitoring program for tracking symptoms and oral adherence, (3) a patient portal (PP) for securely accessing patient health records, and (4) digital education (DE) for patients regarding disease and treatments. The engaged population was defined as the number of enrolled patients with at least one (1) record of triage activity, (2) completed ePRO assessment, (3) PP login, and (4) DE read activity, for each tool, respectively. The start of the index period was adjusted based on the first go-live date of each tool. We evaluated factors (age, gender, race/ethnicity, preferred-language, marital status, and distance from clinic) associated with patient engagement using Chi-Square test and multivariate logistic regression. Results: This analysis included a total of 267,375 unique patients. Of the enrolled population per tool, 172,840 (73.6%), 9,938 (67.7 %), 49,771 (79.2%), and 12,044 (56.9%) patients were engaged in CM, ePRO, PP and DE, respectively. The majority (>50%) of engaged patients were female, White and non-Hispanic/Latino, English-language, and aged 61-80 yrs. After adjusting for covariates, we observed that White and non-Hispanic/Latino [(CM: OR 1.15, ePRO OR 1.46, PP: OR 1.48, and DE: OR 1.36) and English-language (CM: OR 1.2, ePRO OR 1.67, PP: OR 1.8 and DE: OR 1.89) patients were significantly (p-value 80 years as compared to reference age of 0-20 years for any digital tools except CM. Conclusions: DHS can be used to support the cancer patient journey and we demonstrated high utilization in an array of sociodemographic variables in our population. However, tools designed and implemented with different populations in mind to reduce staff burden and lessen the digital divide should be further explored.

The concrete structures, after 1970 or after that made by use of high strength oriented steel bars with their maximum surface deformations, changed constituents and cement properties and different supplementary materials in cement and admixtures with their capacity of acceleration or deceleration. Now, in place of steel bars, fibers of steel, polypropylene, natural polymers etc. are in use [Anbuvelan, 2014]. The reasons for the demand are many, but as a building engineer, we have to think hard and architecturally using building materials. In anticipation of long-term sustainability, we need to be able to meet needs. Nowadays, the construction industry is turning to precast building materials and meeting pre-press and high-strength concrete delivery requirements. Also, engineers have to overcome these barriers, because in large concentrations, concrete sometimes mixes with different chemicals in the soil and makes changes in their properties. A variety of fiber materials other than steel, glass or other natural fibers have been developed and used in the industry to build reinforced fiber concrete. These fibers are classified as synthetic fibers, SNFRC editing. Synthetic fibers are man-made from R&D in the textile and petrochemical industries. Types of fiber that can be used in concrete matrices such as aramid, carbon, acrylic, nylon, polythene, polyester and polypropylene. Reinforced concrete concrete is a mixture that contains water, cement, composite and unfinished fibers of various shapes and sizes., {"references":["ACI Committee Report on Fiber Reinforced Concrete, ACI 506.1R-98","Rana, A. (2013). Some studies on steel fiber reinforced concrete. International journal of emerging technology and advanced engineering, 3(1), 120-127.","Ardeshana, A. L., & Desai, A. K. (2012). Durability of fiber reinforced concrete of marine structures. International J. of Eng. Research and Applications, 2(4), 215- 219.","Alani, A., &Aboutalebi, M. (2013). Mechanical properties of fibre reinforced concrete-a comparative experimental study. International Journal of Civil, Environmental, Structural, Construction and Architectural Engineering, 7(9), 646- 651.","Vasumathi A. M.,Rajkumar, K., GaneshPrabhu G., Compressive behavior of RC column with fiber reinforced concrete confined by CFRP sheets. 2014, Article ID- 601915","Rai, A., & Joshi, Y. P. (2014). Applications and properties of fibre reinforced concrete. Int. J. Eng. Res. Appl, 4(5), 123-131."]}

Precision cut Polyester fiber is a strong and durable man-made fiber that is widely used in many industries. Polyester is resistant to corrosion and shrinkage and perhaps the most important in industrial applications is resistant to corrosion damage. Polyester has a very high gravitational force, resulting in good water dispersion and a high melting point making it a good choice around a variety of applications., {"references":["Massoud, M. T. (2002). Interaction of Silica Fume and Polypropylene Fibers in High Performance Concrete (Doctoral dissertation, American University in Cairo).","Alkhaleefi, A.,M.,Hawary, M., Fattah, H., A.. On the behaviour of polymer portland cement concrete.27th Conference on Our world in Concrete &Structures: 29 - 30 August 2002, Singapore. Article Online Id: 100027015.137-144p.","Neville, A.,M., Brook, J.,J. Advance Concrete Technology.2010.","Campbell, F. C. (2010). Structural composite materials. ASM international.","Wafa, F. F. (1990). Properties & applications of fiber reinforced concrete. Engineering Sciences, 2(1).","Selina Ruby, G., Geethanjali, C., Varghese, J., &MuthuPriya, P. (2014). Influence of hybrid fiber on reinforced concrete. International Journal of Advanced Structures and Geotechnical Engineering, 3(01), 40- 43.","Patel, I., &Modhera, C. D. (2013). Experimental Investigation to Study Effect of Polyester Fibre on Durability of HVFA Concrete through RCPT Method. IOSR J. Eng, 3(6), 22-27.","Patel, I., &Modhera, C. D. (2013). Experimental Investigation to Study Effect of Polyester Fibre on Durability. IOSR J. Eng, 3(6), 22-27","Löfgren, I. (2005). Fracture Behaviour of Reinforced FRC Beams experiments and Analyses in Structural Concrete.Journal of the fib."]}

UNSTRUCTURED The world is facing an unprecedented situation, because of spread of coronavirus that causes the disease COVID-19 which is totally new, where no one can see light at the end of the tunnel. In last 3 months, this pandemic has spread to 204 countries, regardless of its being developed, developing or under developed. In comparison to developed countries and being a developing nation, India has to date curtailed the spread of virus to stage 2 of local transmissions, by limiting its progression. The government of India together with its states has taken stringent actions to prevent its spread, including a nationwide lockdown. The sudden lockdown in country of 1.4 billion have brought in many unseen challenges, which has created panic, confusion and inconvenience to the general public at large. Though the remedial steps are taken, but that may cause collateral damage in the long run, if not planned earlier. The main objective of the article is to apprise the world about certain hard realities, ignored situations and unseen challenges that are being faced by world’s 2nd most populous country due to sudden locking down.

Hydroxytyrosol (HT), a polyphenol from olives, is a potential anticancer agent. This study was designed to evaluate the anticancer activity of HT against prostate cancer cells, and the mechanism thereof.Treatment of LNCaP and C4-2 prostate cancer cells with HT resulted in a dose-dependent inhibition of proliferation. This was in contrast to HT's ineffectiveness against normal prostate epithelial cells RWPE1 and PWLE2, suggesting cancer-cell-specific effect. HT induced G1/S cell cycle arrest, with inhibition of cyclins D1/E and cdk2/4 and induction of inhibitory p21/p27. HT also induced apoptosis, as confirmed by flow cytometry, caspase activation, PARP cleavage, and BAX/Bcl-2 ratio. It inhibited the phosphorylation of Akt/STAT3, and induced cytoplasmic retention of NF-κB, which may explain its observed effects. Finally, HT inhibited androgen receptor (AR) expression and the secretion of AR-responsive prostate-specific antigen.Castration-resistant prostate cancers retain AR signaling and are often marked by activated Akt, NF-κB, and STAT3 signaling. Our results establish a pleiotropic activity of HT against these oncogenic signaling pathways. Combined with its nontoxic effects against normal cells, our results support further testing of HT for prostate cancer therapy.

Breast cancer (BC) continues to be the most frequently diagnosed cancer in American women, which disproportionately affects women of African-American (AA) descent. Previously, we reported greater serum levels of resistin in AA BC patients relative to Caucasian-American (CA) patients, and established its role in growth and aggressiveness of breast tumor cells. Here we have investigated the role of resistin in BC-chemoresistance. MDA-MB-231 and MDA-MB-468 BC cells of CA and AA origin, respectively, were incubated with resistin prior to doxorubicin treatment. Our data suggest that resistin conferred chemoresistance to both BC cell lines; however, the effect on AA cells was more profound. Furthermore, the resistin-induced doxorubicin-resistance was shown to occur due to suppression of apoptosis. Resistin treatment also affected the stemness of BC cells, as suggested by reduced cell surface expression of CD24, induced expression of CD44 and ALDH1, and increased capability of cells to form mammospheres. Mechanistic studies revealed that resistin-induced chemoresistance, apoptosis and stemness of BC cells were mediated through STAT3 activation. Taken together, our findings provide novel insight into the role of resistin in BC biology, and strengthen its role in racially disparate clinical outcomes.

Background: Chemoresistance is a significant clinical problem in pancreatic cancer (PC) and underlying molecular mechanisms still remain to be completely understood. Here we report a novel exosome-mediated mechanism of drug-induced acquired chemoresistance in PC cells. Methods: Differential ultracentrifugation was performed to isolate extracellular vesicles (EVs) based on their size from vehicle- or gemcitabine-treated PC cells. Extracellular vesicles size and subtypes were determined by dynamic light scattering and marker profiling, respectively. Gene expression was examined by qRT-PCR and/or immunoblot analyses, and direct targeting of DCK by miR-155 was confirmed by dual-luciferase 3′-UTR reporter assay. Flow cytometry was performed to examine the apoptosis indices and reactive oxygen species (ROS) levels in PC cells using specific dyes. Cell viability was determined using the WST-1 assay. Results: Conditioned media (CM) from gemcitabine-treated PC cells (Gem-CM) provided significant chemoprotection to subsequent gemcitabine toxicity and most of the chemoresistance conferred by Gem-CM resulted from its EVs fraction. Sub-fractionation grouped EVs into distinct subtypes based on size distribution and marker profiles, and exosome (Gem-Exo) was the only sub-fraction that imparted chemoresistance. Gene expression analyses demonstrated upregulation of SOD2 and CAT (ROS-detoxifying genes), and downregulation of DCK (gemcitabine-metabolising gene) in Gem-Exo-treated cells. SOD/CAT upregulation resulted, at least in part, from exosome-mediated transfer of their transcripts and they suppressed basal and gemcitabine-induced ROS production, and partly promoted chemoresistance. DCK downregulation occurred through exosome-delivered miR-155 and either the functional suppression of miR-155 or restoration of DCK led to marked abrogation of Gem-Exo-mediated chemoresistance. Conclusions: Together, these findings establish a novel role of exosomes in mediating the acquired chemoresistance of PC.

Background: Exosomes are important mediators of intercellular communications and play pivotal roles in cancer progression, metastasis and chemoresistance. CD63 and CD9 are widely accepted exosomal markers. In patients with pancreatic ductal adenocarcinoma (PDAC), positive correlation between CD9 expression and overall survival (OS) was reported. CD63 expression was conserved in all patients with no reported prognostic significance. This study explored the prognostic significance of CD63 and CD9 expression using immunohistochemistry (IHC) in patients with PDAC of mixed racial background. Methods: Between 2012 and 2016, 49 patients with PDAC had available tissues for CD63 and CD9 staining using IHC. Two pathologists independently scored the CD63 and CD9 expression. Staining intensity was graded from 1–3 and staining percentage was estimated in 10% increments. Mean Quick-score (Q-score) (Intensity X Percentage of staining) was calculated. Results: The mean Q-score for CD63 and CD9 are higher in primary tumor from the pancreas compared to pancreatic tumor from metastatic sites (185 vs . 102, P=0.0002) and (48 vs . 20, P=0.0418) respectively. We fitted Cox proportion hazard regression models to investigate the impact of the covariates CD63 and CD9 on progression free survival (PFS) and OS. CD63 has significant impact on PFS (P=0.0135) and OS (P=0.003). The higher the CD63 Q-score, the longer the PFS and OS. CD9 doesn’t have significant impact on PFS (P=0.5734) or OS (P=0.2682). The mean CD63 and CD9 Q-scores are slightly higher in African American (AA) compared to Caucasians (157 vs . 149, P=0.76) and (45 vs . 29, P=0.43) respectively. Conclusions: CD63 and CD9 expression is higher in primary tumor from the pancreas compared to pancreatic tumor from metastatic sites. There is correlation between CD63 expression (but not CD9 in this cohort) and PFS and OS. To our knowledge, this is the first study to show prognostic significance of CD63 expression in patients with PDAC using IHC. A trend of higher expression of CD63 and CD9 among AA compared to Caucasians was also noticed.

Introduction: Amalgam, the most commonly used restorative material, is composed of nearly 50% mercury and 69% silver. It cannot be disposed along with biomedical waste (BMW) because mercury-contaminated waste cannot be incinerated or autoclaved. Objectives: To assess the knowledge and observance of proper mercury hygiene and amalgam waste management among general dental practitioners (GDPs). Materials and Methods: A confidential questionnaire containing 14 questions regarding handling and disposal of amalgam was randomly distributed to 175 GDPs in Chandigarh, Panchkula, and Mohali. A response rate of 78% was obtained, and results were statistically analyzed. Results: Out of total dentists surveyed, 71% were found to be using amalgam as restorative material, 63% were doing

Prostate cancer incidence and mortality rates are remarkably higher in African-American men as compared to their European-Americans counterparts. Despite these recognitions, precise causes underlying such prevalent racial disparities remain poorly understood. Although socioeconomic factors could account for such differences up to a certain extent, it is now being increasingly realized that such disparity has a molecular basis. Indeed, several differences, including genetic polymorphism, gene mutations, epigenetic modifications, miRNAs alterations, etc., have been reported in malignant prostate tissues from patients of diverse racial backgrounds. Here, we attempt to provide a molecular perspective on prostate cancer racial disparities by gathering available information on these associated factors and discussing their potential significance in disproportionate incidence and clinical outcomes.

Background: MYB encodes for a transcription factor regulating the expression of a wide array of genes involved in cellular functions. It is reported to be amplified in a sub-set of pancreatic cancer (PC) cases; however, its pathobiological association has remained unclear thus far. Methods: Expression of MYB and other cellular proteins was analysed by immunoblot or qRT-PCR analyses. MYB was stably overexpressed in non-expressing (BxPC3) and silenced in highly expressing (MiaPaCa and Panc1) PC cells. Effect on growth was analysed by automated cell counting at 24-h interval. Cell-cycle progression and apoptotic indices of PC cells with altered MYB expression were measured through flow cytometry upon staining with respective biomarkers. Cell motility/invasion was examined in a Boyden's chamber assay using non-coated or Matrigel-coated membranes. Effect on tumorigenicity and metastatic potential was examined by non-invasive imaging and through end-point measurements of luciferase-tagged MYB-altered PC implanted in the pancreas of nude mice. Results: MYB was aberrantly expressed in all malignant cases of pancreas, whereas remained undetectable in normal pancreas. All the tested established PC cell lines except BxPC3 also exhibited MYB expression. Forced expression of MYB in BxPC3 cells promoted their growth, cell-cycle progression, survival and malignant behaviour, whereas its silencing in MiaPaCa and Panc1 cells produced converse effects. More importantly, ectopic MYB expression was sufficient to confer tumorigenic and metastatic capabilities to non-tumorigenic BxPC3 cells, while its silencing resulted in significant loss of the same in MYB-overexpressing cells as demonstrated in orthotopic mouse model. We also identified several MYB-regulated genes in PC cells that might potentially mediate its effect on tumour growth and metastasis. Conclusions: MYB is aberrantly overexpressed in PC cells and acts as a key determinant of pancreatic tumour growth and metastasis.

Background: Previously, miR-345 was identified as one of the most significantly downregulated microRNAs in pancreatic cancer (PC); however, its functional significance remained unexplored. Methods: miR-345 was overexpressed in PC cells by stable transfection, and its effect on growth, apoptosis and mitochondrial-membrane potential was examined by WST-1, Hoechst-33342/Annexin-V, and JC-1 staining, respectively. Gene expression was examined by quantitative reverse-transcription-PCR and/or immunoblotting, and subcellular fractions prepared and caspase-3/7 activity determined by commercially available kits. miR-345 target validation was performed by mutational analysis and luciferase-reporter assay. Results: miR-345 is significantly downregulated in PC tissues and cell lines relative to normal pancreatic cells, and its expression decreases gradually in PC progression model cell lines. Forced expression of miR-345 results in reduced growth of PC cells because of the induction of apoptosis, accompanied by a loss in mitochondrial membrane potential, cytochrome-c release, caspases-3/7 activation, and PARP-1 cleavage, as well as mitochondrial-to-nuclear translocation of apoptosis-inducing factor. These effects could be reversed by the treatment of miR-345-overexpressing PC cells with anti-miR-345 oligonucleotides. BCL2 was characterised as a novel target of miR-345 and its forced-expression abrogated the effects of miR-345 in PC cells. Conclusions: miR-345 downregulation confers apoptosis resistance to PC cells, and its restoration could be exploited for therapeutic benefit.

Background: FKBP51 is overexpressed in melanoma and impacts tumour cell properties. However, its comprehensive role in melanoma pathogenesis and underlying mechanism(s) remain elusive. Methods: FKBP51 was stably silenced in aggressive melanoma cell lines and its effect examined in vitro and in mouse model. Histological/immunohistochemical analyses were performed to confirm metastasis, angiogenesis and neutrophil infiltration. Gene expression was analyzed by qRT–PCR, immunoblot and/or ELISA. NF-κB transcriptional activity and promoter binding were monitored by luciferase-based promoter-reporter and ChIP assays, respectively. Interleukin (IL)-8 inhibition was achieved by gene silencing or neutralising-antibody treatment. Results: FKBP51 silencing reduced melanoma growth, metastasis, angiogenesis and neutrophil infiltration and led to IL-8 downregulation through NF-κB suppression in cell lines and tumour xenografts. IL-8 inhibition drastically decreased growth, migration and invasiveness of FKPB51-overexpressing cells; whereas its treatment partially restored the suppressed phenotypes of FKBP51-silenced melanoma cells. Interleukin-8 depletion in conditioned medium (CM) of FKBP51-overexpressing melanoma cells inhibited endothelial cell proliferation and capillary-like structure formation, whereas its treatment promoted these effects in endothelial cells cultured in CM of FKBP51-silenced melanoma cells. Conclusions: FKBP51 promotes melanoma growth, metastasis and angiogenesis, and IL-8 plays a key role in these processes. Thus, targeting of FKBP51 or its upstream or downstream regulatory pathways could lead to effective therapeutic strategies against melanoma.

// Mohammad Aslam Khan 1, * , Sanjeev K. Srivastava 1, * , Arun Bhardwaj 1 , Seema Singh 1 , Sumit Arora 1 , Haseeb Zubair 1 , James E. Carter 2 , Ajay P. Singh 1, 3 1 Department of Oncologic Sciences, Mitchell Cancer Institute, University of South Alabama, Mobile, Alabama, USA 2 Department of Pathology, College of Medicine, University of South Alabama, Mobile, Alabama, USA 3 Department of Biochemistry and Molecular Biology, College of Medicine, University of South Alabama, Mobile, Alabama, USA * These authors have contributed equally to this work Correspondence to: Ajay P. Singh, e-mail: asingh@health.southalabama.edu Keywords: pancreatic cancer, gemcitabine, angiogenesis, IL-8 Received: March 11, 2015 Accepted: April 10, 2015 Published: April 23, 2015 ABSTRACT Pancreatic tumor microenvironment (TME) is characterized by poor tumor-vasculature and extensive desmoplasia that together contribute to poor response to chemotherapy. It was recently shown that targeting of TME to inhibit desmoplasiatic reaction in a preclinical model resulted in increased microvessel-density and intratumoral drug concentration, leading to improved therapeutic response. This approach; however, failed to generate a favorable response in clinical trial. In that regard, we have previously demonstrated a role of gemcitabine-induced CXCR4 signaling as a counter-defense mechanism, which also promoted invasiveness of pancreatic cancer (PC) cells. Here, we investigated the effect of gemcitabine on endothelial cell phenotype. Gemcitabine-treatment of human-umbilical-vein-endothelial-cells (HUVECs) did not promote the growth of HUVECs; however, it was induced when treated with conditioned media from gemcitabine-treated (Gem-CM) PC cells due to increased cell-cycle progression and apoptotic-resistance. Moreover, treatment of HUVECs with Gem-CM resulted in capillary-like structure (CLS) formation and promoted their ability to migrate and invade through extracellular-matrix. Gemcitabine-treatment of PC cells induced expression of various growth factors/cytokines, including IL-8, which exhibited greatest upregulation. Further, IL-8 depletion in Gem-CM diminished its potency to promote angiogenic phenotypes. Together, these findings suggest an indirect effect of gemcitabine on angiogenesis, which, in light of our previous observations, may hold important clinical significance.

African-American (AA) women with breast cancer (BC) are diagnosed with more aggressive disease, have higher risk of recurrence and poorer prognosis as compared to Caucasian American (CA) women. Therefore, it is imperative to define the factors associated with such disparities to reduce the unequal burden of cancer. Emerging data suggest that inherent differences exist in the tumor microenvironment of AA and CA BC patients, however, its molecular bases and functional impact have remained poorly understood. Here, we conducted cytokine profiling in serum samples from AA and CA BC patients and identified resistin and IL-6 to be the most differentially-expressed cytokines with relative greater expression in AA patients. Resistin and IL-6 exhibited positive correlation in serum levels and treatment of BC cells with resistin led to enhanced production of IL-6. Moreover, resistin also enhanced the expression and phosphorylation of STAT3, and treatment of BC cells with IL-6-neutralizing antibody prior to resistin stimulation abolished STAT3 phosphorylation. In addition, resistin promoted growth and aggressiveness of BC cells, and these effects were mediated through STAT3 activation. Together, these findings suggest a crucial role of resistin, IL-6 and STAT3 in BC racial disparity.

OBJECTIVES Exosomes are important mediators in intercellular communications and play a role in cancer progression and metastasis. Exosomal membranes are enriched in endosome-specific tetraspanins (CD9 and CD63). Here, we explored the expression of CD63 and CD9 utilizing immunohistochemistry in malignant and nonmalignant cells in 29 resected pancreatic specimens (RPSs) of mixed racial background. METHODS The pathologic tissues (PTs) and adjacent normal tissues (ANTs) in each RPS were stained for CD63 and CD9. Two pathologists independently scored the expression of CD63 and CD9. Staining intensity was graded from 1 to 3. Staining percentage was estimated in 10% increments. An average Quick score (Q score) (intensity × percentage of staining) was calculated. Unpaired t test was used for statistical analysis. RESULTS The mean multiplicative Q score for CD63 and CD9 expression is higher in PTs (209 and 72) compared with ANTs (154 and 24) (P = 0.0041, P = 0.0018), respectively. The Mean Q score for CD63 and CD9 expression is higher in the malignant PTs (231 and 85) compared with ANTs (129 and 25) (P < 0.0001 and P < 0.0124). CONCLUSIONS Exosomal markers (CD63 and CD9) expression assessment using immunohistochemistry is feasible in RPS. The expression of CD63 and CD9 is higher in PTs and malignant PTs compared with their ANTs.

Pancreatic cancer (PC) continues to rank among the most lethal cancers. The consistent increase in incidence and mortality has made it the seventh leading cause of cancer-associated deaths globally and the third in the United States. The biggest challenge in combating PC is our insufficient understanding of the molecular mechanism(s) underlying its complex biology. Studies during the last several years have helped identify several putative factors and events, both genetic and epigenetic, as well as some deregulated signaling pathways, with implications in PC onset and progression. In this review article, we make an effort to summarize our current understanding of molecular and cellular events involved in the pathogenesis of pancreatic malignancy. Specifically, we provide up-to-date information on the genetic and epigenetic changes that occur during the initiation and progression of PC and their functional involvement in the pathogenic processes. We also discuss the impact of the tumor microenvironment on the molecular landscape of PC and its role in aggressive disease progression. It is envisioned that a better understanding of these molecular factors and the mechanisms of their actions can help unravel novel diagnostic and prognostic biomarkers and can also be exploited for future targeted therapies.

Emerging data highlight the significance of chemokine (C-X-C motif) ligand 12/chemokine (C-X-C motif) receptor 4 (CXCL12/CXCR4) signaling axis in the chemoresistance of several malignancies, including prostate cancer (PCa); however, underlying mechanisms remain largely elusive. Here, we demonstrate that CXCL12 treatment rescues the PCa cells from docetaxel (DTX)-induced toxicity by overriding its effect on cell cycle (G2/M phase arrest). We further demonstrate that the chemoprotective effect of CXCL12 is abolished upon pharmacological inhibition or RNA interference-mediated silencing of CXCR4. Moreover, microtubule stabilization caused by DTX is suppressed in CXCL12-stimulated PCa cells as revealed by immunofluorescence and immunoblot analyses. The effect of CXCL12 on microtubule stabilization is abrogated when PCa cells are pre-treated with a CXCR4 antagonist. In additional studies, we show that the chemoprotective action of CXCL12/CXCR4 signaling is mediated by p21-activated kinase 4 (PAK4)-dependent activation of Lim domain kinase 1 (LIMK1), and inhibition of either PAK4 or LIMK1 leads to re-sensitization of PCa cells to DTX-induced tubulin polymerization and cellular toxicity even in the presence of CXCL12. Altogether, our findings uncover a novel mechanism underlying CXCL12/CXCR4 signaling-induced PCa chemoresistance and suggest that targeting of this signaling axis or its downstream effector pathway could lead to therapeutic enhancement of DTX.

Identification of novel molecular targets and understanding the mechanisms underlying the aggressive nature of pancreatic cancer (PC) remain prime focus areas of research. Here, we investigated the expression and pathobiological significance of p21-activated kinase 4 (PAK4), a gene that was earlier shown to be amplified in a sub-set of PC. Our data demonstrate PAK4 overexpression in PC tissues and cell lines with little or no expression in the normal pancreas. PAK4 silencing in two PC cell lines, MiaPaCa and T3M4, by RNA interference causes suppression of growth and clonogenic ability due to decreased cell cycle progression and apoptosis-resistance. PAK4-silenced PC cells exhibit altered expression of proliferation- and survival-associated proteins. Moreover, we observe decreased nuclear accumulation and transcriptional activity of NF-κB in PAK4-silenced PC cells associated with stabilization of its inhibitory protein, IκBα. Transfection of PAK4-silenced PC cells with constitutively-active mutant of IKKβ, an upstream kinase of IκBα, leads to restoration of NF-κB activity and PC cell growth. Furthermore, we show that PAK4-induced NF-κB activity is mediated through activation and concerted action of ERK and Akt kinases. Together, these findings suggest that PAK4 is a regulator of NF-κB pathway in PC cells and can serve as a novel target for therapy.

Recently, we have shown that CXCL12/CXCR4 signaling plays an important role in gemcitabine resistance of pancreatic cancer (PC) cells. Here, we explored the effect of gemcitabine on this resistance mechanism. Our data demonstrate that gemcitabine induces CXCR4 expression in two PC cell lines (MiaPaCa and Colo357) in a dose- and time-dependent manner. Gemcitabine-induced CXCR4 expression is dependent on reactive oxygen species (ROS) generation because it is abrogated by pretreatment of PC cells with the free radical scavenger N-acetyl-L-cysteine. CXCR4 up-regulation by gemcitabine correlates with time-dependent accumulation of NF-κB and HIF-1α in the nucleus. Enhanced binding of NF-κB and HIF-1α to the CXCR4 promoter is observed in gemcitabine-treated PC cells, whereas their silencing by RNA interference causes suppression of gemcitabine-induced CXCR4 expression. ROS induction upon gemcitabine treatment precedes the nuclear accumulation of NF-κB and HIF-1α, and suppression of ROS diminishes these effects. The effect of ROS on NF-κB and HIF-1α is mediated through activation of ERK1/2 and Akt, and their pharmacological inhibition also suppresses gemcitabine-induced CXCR4 up-regulation. Interestingly, our data demonstrate that nuclear accumulation of NF-κB results from phosphorylation-induced degradation of IκBα, whereas HIF-1α up-regulation is NF-κB-dependent. Lastly, our data demonstrate that gemcitabine-treated PC cells are more motile and exhibit significantly greater invasiveness against a CXCL12 gradient. Together, these findings reinforce the role of CXCL12/CXCR4 signaling in gemcitabine resistance and point toward an unintended and undesired effect of chemotherapy.

Failure of conventional as well as target-based therapy against the advanced metastatic cancers is a significant clinical problem. While some cancers, such as pancreatic cancer, respond poorly to any kind of therapies, tumor relapse is often observed in many other cancer types after initial therapeutic response. Hence, significant research is being conducted to understand the mechanisms underlying therapeutic refractoriness of cancer. During the past decade, cancer stem cell (CSC) hypothesis has gained significant experimental and clinical support, and CSCs have emerged as potentially useful pharmacologic targets. MicroRNAs (miRNAs) are a group of small (~18-25 nucleotides) non-protein encoding RNAs that are now established as important regulators of gene expression. They can function as tumor promoters (oncomirs) or suppressors (anti-oncomirs) and thus hold profound implications for cancer therapy. Recent studies have identified several miRNAs to be differentially expressed in CSCs and established their role in targeting genes and pathways supporting cancer stemness properties. Here, we discuss these findings and review recent advances in miRNA-based strategies to exploit therapeutic potential of miRNAs in cancer treatment.

MicroRNAs (miRNAs) have emerged as novel post-transcriptional regulators of gene expression. These short non-coding RNAs are involved in diverse biological processes and their dysregulation is often observed under diseased conditions. Therefore, miRNAs hold great potential as clinical biomarkers of physiological and pathological states and extensive efforts are underway to develop efficient approaches for their detection. We review recent advances and discuss the promises and pitfalls of emerging methods of miRNA profiling and detection.

Aberrant expression of the kinase IKKϵ in pancreatic ductal adenocarcinoma (PDAC) has been associated with poor prognosis. In this study, we define a pathobiologic function for IKKϵ in reprogramming glucose metabolism and driving progression in PDAC. Silencing IKKϵ in PDAC cells, which overexpressed it endogenously, was sufficient to reduce malignant cell growth, clonogenic potential, glucose consumption, lactate secretion, and expression of genes involved in glucose metabolism, without impacting the basal oxygen consumption rate. IKKϵ silencing also attenuated c-Myc in a manner associated with diminished signaling through an AKT/GSK3β/c-MYC phosphorylation cascade that promoted MYC nuclear accumulation. In an orthotopic mouse model, IKKϵ-silenced PDAC exhibited a relative reduction in glucose uptake, tumorigenicity, and metastasis. Overall, our findings offer a preclinical mechanistic rationale to target IKKϵ to improve the therapeutic management of PDAC in patients. Cancer Res; 76(24); 7254–64. ©2016 AACR.

We have recently demonstrated that the transcription factor MYB can modulate several cancer-associated phenotypes in pancreatic cancer. In order to understand the molecular basis of these MYB-associated changes, we conducted deep-sequencing of transcriptome of MYB-overexpressing and -silenced pancreatic cancer cells, followed by in silico pathway analysis. We identified significant modulation of 774 genes upon MYB-silencing (p in silico analysis. Further analyses placed genes in our RNA sequencing-generated dataset to several canonical signalling pathways, such as cell-cycle control, DNA-damage and -repair responses, p53 and HIF1α. Importantly, we observed downregulation of the pancreatic adenocarcinoma signaling pathway in MYB-silenced pancreatic cancer cells exhibiting suppression of EGFR and NF-κB. Decreased expression of EGFR and RELA was validated by both qPCR and immunoblotting and they were both shown to be under direct transcriptional control of MYB. These observations were further confirmed in a converse approach wherein MYB was overexpressed ectopically in a MYB-null pancreatic cancer cell line. Our findings thus suggest that MYB potentially regulates growth and genomic stability of pancreatic cancer cells via targeting complex gene networks and signaling pathways. Further in-depth functional studies are warranted to fully understand MYB signaling in pancreatic cancer.

Recent evidence suggests a major role of tumor-stromal interactions in pancreatic cancer pathobiology. The chemokine CXCL12 (stromal cell-derived factor 1 (SDF-1)), abundantly produced by stromal cells, promotes progression, metastasis, and chemoresistance of pancreatic cancer cells. On the other hand, pancreatic tumor cell-derived sonic hedgehog (SHH) acts predominantly on stromal cells to induce desmoplasia and, thus, has a paracrine effect on tumorigenesis and therapeutic outcome. In this study, we examined the association between these two proteins of pathological significance in pancreatic cancer. Our data demonstrate that CXCL12 leads to a dose- and time-dependent up-regulation of SHH in pancreatic cancer cells. CXCL12-induced SHH up-regulation is specifically mediated through the receptor CXCR4 and is dependent on the activation of downstream Akt and ERK signaling pathways. Both Akt and ERK cooperatively promote nuclear accumulation of NF-κB by inducing the phosphorylation and destabilization of its inhibitory protein, IκB-α. Using dominant negative IκB-α, a SHH promoter (deletion mutant) reporter, and chromatin immunoprecipitation assays, we demonstrate that CXCL12 exposure enhances direct binding of NF-κB to the SHH promoter and that suppression of NF-κB activation abrogates CXCL12-induced SHH expression. Finally, our data demonstrate a strong correlative expression of CXCR4 and SHH in human pancreatic cancer tissues, whereas their expression is not observed in the normal pancreas. Altogether, our data reveal a novel mechanism underlying aberrant SHH expression in pancreatic cancer and identify a molecular link facilitating bidirectional tumor-stromal interactions.

Myb, a cellular progenitor of v-Myb oncogenes, is amplified in prostate cancer and exhibits greater amplification frequency in hormone-refractory disease. Here, we have investigated the functional significance of Myb in prostate cancer. Our studies demonstrate Myb expression in all prostate cancer cell lines (LNCaP, C4-2, PC3 and DU145) examined, whereas it is negligibly expressed in normal/benign prostate epithelial cells (RWPE1 and RWPE2). Notably, Myb is significantly upregulated, both at transcript (>60-fold) and protein (>15-fold) levels, in castration-resistant (C4-2) cells as compared with androgen-dependent (LNCaP) prostate cancer cells of the same genotypic lineage. Using loss and gain of function approaches, we demonstrate that Myb promotes and sustains cell cycle progression and survival under androgen-supplemented and -deprived conditions, respectively, through induction of cyclins (A1, D1 and E1), Bcl-xL and Bcl2 and downregulation of p27 and Bax. Interestingly, Myb overexpression is also associated with enhanced prostate-specific antigen expression. Furthermore, our data show a role of Myb in enhanced motility and invasion and decreased homotypic interactions of prostate cancer cells. Myb overexpression is also associated with actin reorganization leading to the formation of filopodia-like cellular protrusions. Immunoblot analyses demonstrate gain of mesenchymal and loss of epithelial markers and vice versa, in Myb-overexpressing LNCaP and -silenced C4-2 cells, respectively, indicating a role of Myb in epithelial to mesenchymal transition. Altogether, our studies provide first experimental evidence for a functional role of Myb in growth and malignant behavior of prostate cancer cells and suggest a novel mechanism for castration resistance.

Nisin-, pediocin 34-, and enterocin FH99-resistant variants of Listeria monocytogenes ATCC 53135 were developed. In an attempt to clarify the possible mechanisms underlying bacteriocin resistance in L. monocytogenes ATCC 53135, sensitivity of the resistant strains of L. monocytogenes ATCC 53135 to nisin, pediocin 34, and enterocin FH99 in the absence and presence of different divalent cations was assessed, and the results showed that the addition of divalent cations significantly reduced the inhibitory activity of nisin, pediocin 34, and enterocin FH99 against resistant variants of L. monocytogenes ATCC 53135. The addition of EDTA, however, restored this activity suggesting that the divalent cations seem to affect the initial electrostatic interaction between the positively charged bacteriocin and the negatively charged phospholipids of the membrane. Nisin-, pediocin 34-, and enterocin-resistant variants of L. monocytogenes ATCC 53135 were more resistant to lysozyme as compared to the wild-type strain both in the presence as well as absence of nisin, pediocin 34, and enterocin FH99. Ultra structural profiles of bacteriocin-sensitive L. monocytogenes and its bacteriocin-resistant counterparts revealed that the cells of wild-type strain of L. monocytogenes were maximally in pairs or short chains, whereas, its nisin-, pediocin 34-, and enterocin FH99-resistant variants tend to form aggregates. Results indicated that without a cell wall, the acquired nisin, pediocin 34, and enterocin FH99 resistance of the variants was lost. Although the bacteriocin-resistant variants appeared to lose their acquired resistance toward nisin, pediocin 34, and enterocin FH99, the protoplasts of the resistant variants appeared to be more resistant to bacteriocins than the protoplasts of their wild-type counterparts.

The bacteriocin susceptibility of Listeria monocytogenes MTCC 657, Enterococcus faecium DSMZ 20477, E. faecium VRE, and E. faecalis ATCC 29212 and their corresponding bacteriocin resistant variants was assessed. The single and combined effect of nisin and pediocin 34 and enterocin FH99 bacteriocins produced by Pediococcus pentosaceus 34, and E. faecium FH99, respectively, was determined. Pediocin34 proved to be more effective in inhibiting L. monocytogenes MTCC 657. A greater antibacterial effect was observed against E. faecium DSMZ 20477 and E. faecium (VRE) when the a combination of nisin, pediocin 34 and enterocin FH99 were used whereas in case of L. monocytogenes MTCC 657 a combination of pediocin 34 and enterocin FH99 was more effective in reducing the survival of pathogen. Bacteriocin cross-resistance and the antibiotic susceptibility of wild type and their corresponding resistant variants were assessed and results showed that resistance to a bacteriocin may extend to other bacteriocins within the same class and also the acquired resistance to bacteriocins can modify the antibiotic susceptibility/resistance profile of the bacterial species used in the study. According to the hydrophobicity nisin resistant variant of L. monocytogenes was more hydrophobic (p

Earlier we identified PPP2CA, which encodes for the α-isoform of protein phosphatase 2A (PP2A) catalytic subunit, as one of the downregulated genes in androgen-independent prostate cancer. PP2A is a serine/threonine phosphatase and a potent tumor suppressor involved in broad cellular functions; however, its role in prostate cancer has not yet been determined. Here, we have investigated the effect of PP2A activity modulation on the androgen-independent growth of prostate cancer cells. Our data show that the PPP2CA expression and PP2A activity is downregulated in androgen-independent (C4-2) prostate cancer cells as compared with androgen-dependent (LNCaP) cells. Downregulation of PP2A activity by pharmacologic inhibition or short interfering RNA-mediated PPP2CA silencing sustains the growth of LNCaP cells under an androgen-deprived condition by relieving the androgen deprivation–induced cell-cycle arrest and preventing apoptosis. Immunoblot analyses reveal enhanced phosphorylation of Akt, extracellular signal–regulated kinase (ERK), BAD, increased expression of cyclins (A1/D1), and decreased expression of cyclin inhibitor (p27) on PP2A downregulation. Furthermore, our data show that androgen receptor (AR) signaling is partially maintained in PP2A-inhibited cells through increased AR expression and ligand-independent phosphorylation. Pharmacologic inhibition of Akt, ERK, and AR suggest a role of these signaling pathways in facilitating the androgen-independent growth of LNCaP cells. These observations are supported by the effect of ceramide, a PP2A activator, on androgen-independent C4-2 cells. Ceramide inhibited the growth of C4-2 cells on androgen deprivation, an effect that could be abrogated by PP2A downregulation. Altogether, our findings suggest that modulation of PP2A activity may represent an alternative therapeutic approach for the treatment of advanced androgen-independent prostate cancer. Mol Cancer Ther; 10(5); 720–31. ©2011 AACR.

Background: Pancreatic cancer cells are highly resistant to drug therapy; however, underlying causes remain largely unknown. We hypothesised that the activation of CXCL12–CXCR4 signalling confers drug resistance to pancreatic cancer cells by potentiating survival. CXCR4 is overexpressed in precancerous/malignant pancreatic lesions and cancer stem cells, and implicated in its pathogenesis. Methods: Effect of CXCR4 activation by CXCL12 on restricting the gemcitabine-induced cytotoxicity and stimulating the survival signalling was examined in pancreatic cancer cells by MTT, DNA laddering, caspase activity, immunoblot, and promoter-reporter assays. Subsequently, we examined the effect of CXCR4 antagonist, AMD3100, in abrogating the rescue effect of activated CXCL12–CXCR4 signalling. Results: The pancreatic cancer cells treated with gemcitabine exhibited reduced cytotoxicity in the presence of CXCL12 as compared with the cells treated with drug alone. CXCL12 induced the activation of FAK, ERK, and Akt signalling pathways, enhanced transcriptional activities of β-catenin and NF-κB, and expression of survival proteins. AMD3100 arrested the CXCL12-induced pancreatic cancer cell growth and drug resistance. Conclusion: Our findings demonstrate, for the first time, a role of CXCL12–CXCR4 signalling axis in conferring drug resistance to pancreatic cancer cells and suggest that it could serve as a novel therapeutic target for pancreatic cancer therapy, alone and in combination with the cytotoxic drug.

Resistance to chemotherapeutic drugs remains a major cause of therapeutic failure in pancreatic cancer (PC) patients. Efficacy of chemotherapy is further limited by its debilitating effects on immune system. Apart from genetic abnormalities in tumor cells, the tumor immune- microenvironment plays a crucial role in PC chemoresistance. However, it is unclear how chemotherapy affects the cancer immunity or how it affects the tumor microenvironment (TME). In the present study, we demonstrate gemcitabine induces an increased infiltration of CD68+ M2 macrophages in pancreatic tumors isolated from an orthotopic xenograft murine model. Moreover, the conditioned media from PC cell lines cultured with gemcitabine (Gem-CM) promotes migration, invasion, and growth of RAW264.7 macrophage. Interestingly, Gem-CM induces an upregulation of tumor-associated or M2-polarized macrophage markers, arginase-1 and TGF-β1. Additionally, gemcitabine treatment of PC cell lines induces the expression of M2-polarization associated cytokines, growth factors and chemokines, including IL-8. IL-8 exhibits the greatest upregulation after culture with Gemcitabine. Further, IL-8 neutralization in Gem-CM diminished its ability to induce growth, migration, and invasion of RAW264.7 macrophages. Together, these findings suggest an indirect effect of gemcitabine on increasing macrophage trafficking and M2-polarization that potentially supports a pro-tumor microenvironment. These actions may contribute to the chemoresistance of PC. Prevention of gemcitabine-induced macrophage infiltration, growth and M2-polarization will offer a better strategy to counter chemotherapy-induced immunosuppression in PC. Citation Format: Sachin Kumar Deshmukh, Arun Bhardwaj, Nikhil Tyagi, Mohammad Aslam Khan, Sanjeev K. Srivastava, Ahmed AL-Ghadhban, Kari Dugger, James E. Carter, Ajay P. Singh, Seema Singh. Gemcitabine treatment induces immunosuppressive microenvironment in pancreatic cancer by promoting the infiltration, growth, and polarization of macrophages [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 113.

In the present study 14 bacteriocinogenic strains of Enterococcus faecium isolated from dairy foods and faecal sample were evaluated for the presence of virulence determinants, production of biogenic amines and their susceptibility to various antibiotics. Genetic diversity among them was evaluated by RAPD-PCR method. Further, they were evaluated for their probiotic potential under in vitro trials. The efaAfm was the only virulence trait detected in all E. faecium and tyramine was the only biogenic amine produced by 9 tested strains. No strain was resistant to all antibiotics and for some strains, multiple resistances were observed. E. faecium FH 99 showed highest good ability to tolerate acid and bile, while good bile salt hydrolase activity and were able to assimilate cholesterol from growth media. These results suggest that the tested E. faecium are generally free from virulence traits and having good probiotic potential and may be exploit in dairy industry and probiotic preparations.

342 Background: Exosomes are important mediators of intercellular communications and play pivotal roles in cancer progression, metastasis and chemoresistance. CD63 and CD9 are exosomal markers. In patients with pancreatic ductal adenocarcinoma (PDAC), positive correlation between CD9 expression and overall survival (OS) was reported. CD63 expression was conserved in all patients with no reported prognostic significance. This study explored the prognostic significance of CD63 and CD9 expression using immunohistochemistry (IHC) in patients with PDAC of mixed racial background. Methods: Between 2012 and 2016, 49 patients with PDAC had available tissues for CD63 and CD9 staining using IHC. Two pathologists independently scored CD63 and CD9 expression. Staining intensity was graded from 1-3. Staining percentage was estimated in 10% increments. Mean Quick-score (Intensity X Percentage of staining) was calculated. Results: Median age was 64 (range 42-85). 53% are males. 67% Caucasians, 27% African Americans (AA) and 6% are other ethnicities. 41% had stage IV while 59% had stage I-III. The mean CD63 and CD9 Q scores are higher in primary tumor from the pancreas compared to pancreatic tumor from metastatic sites (185 vs 102, p = 0.0002) and (48 vs 11, p = 0.0418) respectively. We fitted accelerated failure-time models to investigate the impacts of the covariates CD63 and CD9 on progression free survival (PFS) and OS. CD63 has significant impact on PFS (p = 0.0058) and OS (p = 0.0012). The higher the CD63 Q score, the longer the PFS and OS. CD9 doesn’t have significant impact on PFS (p = 0.8950) or OS (p = 0.7182). The mean CD63 and CD9 Q scores are slightly higher in AA compared to Caucasians (157 vs 149, p = 0.76) and (45 vs 29, p = 0.43) respectively. Conclusions: CD63 and CD9 expression is higher in primary tumor from the pancreas compared to pancreatic tumor from metastatic sites. There is correlation between CD63 expression (but not CD9 in this cohort) and PFS and OS. To our knowledge, this is the first study to show prognostic significance of CD63 expression in patients with PDAC using IHC. A trend of higher expression of CD63 and CD9 among AA compared to Caucasians was noticed.

The presence of biogenic amines (BAs) in foods is not acceptable as the consumption of food and beverages containing high levels of BAs could result in toxicological effects for human health. BAs are mainly produced by the microbial decarboxylation of certain amino acids. Among the various BAs, tyramine is the most commonly cited and abundant BA produced by several bacterial genera. Various biochemical, chromatographic and molecular methods are used for the detection of BAs. Among different lactic acid bacteria, enterococci have been found to be the most abundant tyramine producers. In the present study, 28 previously isolated bacteriocinogenic strains of Enterococcus sp. were tested for their ability to produce tyramine by qualitative, quantitative and molecular methods. Correlations between these methods were also investigated. A total of 19 enterococcal strains were found to produce tyramine by all the methods used. A low level of correlation was found between the results of two different decarboxylating media used. “Improved medium” detected more tyrosine-positive colonies than “tyrosine production medium”. However, a 100% correlation was observed between the results observed with “tyrosine production medium”, tdc gene-targeted polymerase chain reaction and tyramine quantification by high performance liquid chromatography. Therefore, these methods may be used to complement each other in the detection of tyramine-producing enterococci in foods.

The genus Enterococcus like other LAB has also been featured in dairy industry for decades due to its specific biochemical traits such as lipolysis, proteolysis, and citrate breakdown, hence contributing typical taste and flavor to the dairy foods. Furthermore, the production of bacteriocins by enterococci (enterocins) is well documented. These technological applications have led to propose enterococci as adjunct starters or protective cultures in fermented foods. Moreover, enterococci are nowadays promoted as probiotics, which are claimed for the maintenance of normal intestinal microflora, stimulation of the immune system and improvement of nutritional value of foods. At the same time, enterococci present an emerging pool of opportunistic pathogens for humans as they cause disease, possess agents for antibiotic resistance, and are frequently armed with potential virulence factors. Because of this “dualistic” nature, the use of enterococci remains a debatable issue. However, based on a long history of safe association of particular enterococci with some traditional food fermentations, the use of such strains appears to bear no particular risk for human health. Abundance of knowledge as well as progress in molecular techniques has, however, enabled exact characterization and safety assessment of strains. Therefore, a balanced evaluation of both, beneficial and undesirable nature of enterococci is required. A clear understanding of their status may, therefore, allow their safe use as a starter, or a probiotic strain. The present review describes the broader insight of the benefits and risks of enterococci in dairy foods and their safety assessment.