Differential Gene Expression Patterns in HPV-Positive and HPV-Negative Oropharyngeal Carcinomas

Head and neck (HN) cancer is the fifth most prevalent malignancy worldwide with 90% of them categorized as squamous cell carcinomas (SCC). Carcinomas of the oropharynx, pharynx and the oral cavity, comprise a subset of HNSCC and are referred to as oropharyngeal squamous cell carcinomas (OPSCC). Up to 60% of OPSCC and 25% of HNSCC are positive for high-risk human papillomavirus (HR-HPV), primarily HPV16. HPV positive and HPV negative OPSCC’s are molecularly and biologically distinct with differences in risk factors, age of presentation and clinical behavior. The precise molecular signatures of each have been well studied with respect to gene expression, genetic-epigenetic modifications and mutational analysis. However, recent studies have identified HPV as a racially linked marker for OPSCC, where African American (AA) patients present with a higher percentage of HPV-negative and aggressive tumors as compared to European American (EA) patients. Thus, we aimed to study the HPV status and differential gene expression (DEG) profile of OPSCC in AA and EA patients from South Carolina, using Agilent 8x60k arrays and a dual marker system for HPV status. We characterized all the tumors into HPV-active (DNA+/E7RNA+), HPV-inactive (DNA+/E7RNA-) and HPV-negative (DNA-/RNA-) based on INNO-LiPA and RTqPCR assays. Overall, 59% of OPSCC tissue samples tested positive for HPV DNA, while only 48% of those harbored an active HPV infection and significant differences were observed when compared by race. We observed a higher prevalence of HPV in EA patients, both by DNA (69%) as well as by E7 RNA (39%), when compared to that of AA patients (40% and 10% respectively). Microarray analysis over a set of 36 oropharyngeal tumors and 4 normals revealed that HPV-inactive tumors have gene expression profiles distinct from those of both HPVactive and HPV-negative SCC, suggesting that HPV-inactive tumors may constitute a group of their own. The expression of a selected panel of genes was confirmed by RTqPCR which was in concordance with our microarray data. Our RT-qPCR assays conformed to the latest MIQE guidelines and data were normalized using three reference genes through Biogazelle qbase+ software and the results were scaled to the control group (normal samples). Normalized relative quantities for each gene of interest were then calculated using qbase+. Selection of the most stably expressed candidate reference genes was done through a comprehensive study of 8 well known housekeeping/reference genes through Normfinder, BestKeeper and GeNorm. Our microarray studies and RTqPCR assays categorized HPV-inactive tumors as a distinct entity in comparison to the HPV-negative and –active tumors by race. Also, HPV-inactive tumors clustered closer to the HPV-negative tumors with one exception, suggesting the loss of function of HPV in the former. Overall, our results confirmed that AA patients more often present with HPVnegative tumors in comparison to EA patients, and indicated that HPV-negative tumors exhibit gene expression profiles indicative of activation of epithelial mesenchymal transition, while HPV active tumors are characterized by alterations of cell cycle and growth control

Dengue: epidemiology, prevention and pressing need for vaccine development

AbstractDengue fever is a mosquito born viral infection, and the complicated form of dengue is dengue hemorrhagic fever (DHF). In the recent decades incidence and distribution of dengue has increased dramatically. Dengue viruses belong to family flaviviridae with four serotypes and are transmitted mainly by mosquito Aedes aegypti. Today almost two-fifth of world's population (2.5 million) is at risk of dengue and no specific antiviral drug or vaccine is available against it. Uncontrolled population growth in Africa and South East Asia has increased number of susceptible hosts in urban and semi urban areas. About 40% of world population resides in the high risk area for dengue transmission. According to latest estimates by WHO, yearly 50 to 100 million infections occur globally, this includes around 500 000 DHF and 22 000 deaths, mostly among children. Only symptomatic treatment in the form of analgesic, antipyretics and body fluid management is provided to the patient. Prevention strategies mainly focus on two approaches, firstly on activities to control vector and secondly on activities to protect human from mosquito bite but there is always concerns regarding their sustainably and effectiveness. Theoretically development of an effective dengue vaccine is feasible and production of an effective and affordable vaccine could be a viable option to save humans from this dreadful disease. Conceptually vaccine production is possible, but it has to be tetravalent, providing immunity against all serotypes. Few candidate vaccines are in advance stage of their development; however international cooperation is needed to make these vaccines available on cheaper rates to the poor and vulnerable countries. Objective of this review is to discuss various aspects related to dengue, its epidemiology, available preventive methods, need for vaccine and challenges in its development

Effects of Seed Culture and Attached Growth System on the Performance of Anammox Hybrid Reactor Treating Nitrogenous Wastewater

The start-up of anammox (anaerobic ammonium oxidation) process in hybrid reactor delineated four distinct phases i.e. cell lysis, lag phase, activity elevation and stationary phase. Cell lysis phase was marked by death and decay of heterotrophic denitrifiers resulting in breakdown of organic nitrogen into ammonium. Lag phase showed initiation of anammox activity with turnover of heterotrophic denitrifiers, which is evident from appearance of NO3-N in the effluent. In activity elevation phase, anammox became the dominant reaction, which can be attributed to consequent reduction of NH4-N into N2 with increased NO3-N in the effluent. Proper selection of mixed seed culture at influent NO2-/NH4+ ratio (1:1) and hydraulic retention time (HRT) of 1 day led to early start up of anammox within 70 days. Pseudo steady state removal efficiencies of NH4+ and NO2- were found as 94.3% and 96.4% respectively, at nitrogen loading rate (NLR) of 0.35 kg N/m3d at an HRT of 1 day. Analysis of the data indicated that attached growth system contributes an additional 11% increase in the ammonium removal and results an average of 29% reduction in sludge washout rate. Mass balance study of nitrogen indicated that 74.1% of total input nitrogen is converted into N2 gas followed by 11.2% being utilized in biomass development. Scanning electron microscope (SEM) observation of the granular sludge clearly showed the presence of cocci and rod shaped microorganisms intermingled on the external surface of the granules

Utjecaj fluorida proizvedenih aluminijskim industrijskim postrojenjem na biljke i područja nastanjena ljudima

Fluorine is a highly reactive common element that does not occur in nature in the elemental state. It exists in the form of fluorides and accounts for about 0.3 g/kg of the Earth’s crust. Generally, it is found in the form of a number of minerals like fluorspar, cryolite and fluor-apatite. Fluoride has both positive and negative effects on individual health. Fluoride, in the form of fluorspar and cryo- lite is distributed extensively in the lithosphere, and is renowned as the thir- teenth most common among elements in the earth’s crust. Hydrogen fluorides in gaseous form accumulate in the leaves of generally sensitive plants against a concentration gradient and therefore, considered as a most phytotoxic air pollut- ant and affects plants at extremely low concentration. As per our study, it is found that the fluoride impacts on fauna are in normal condition, but in coming times it may have adverse impact on fauna and flora of surroundings of Hindalco Industries Limited.Fluor je vrlo reaktivan element koji se u elementarnom obliku ne pojavljuje u prirodi. Postoji u obliku fluorida u zemljinoj kori, gdje se nalazi u omjeru od oko 0,3 g/kg. Općenito, nalazi se u obliku velikog broja minerala poput fluorita, kriolita i apatita. Fluorid ima i pozitivne i negativne učinke na ljudsko zdravlje. Fluorid, u obliku fluorita i kriolita, široko je rasprostranjen u litosferi, a poznat je i kao trinaesta najčešća tvar u Zemljinoj kori. Fluorovodici u plinovitom obliku nagomilavaju se u listovima uglavnom osjetljivih biljaka prema gradijentu koncentracije. Stoga se smatraju najfitotoksičnijim onečišćujućim tvarima u zraku, a djeluju na biljke i pri ekstremno niskim koncentracijama. Prema našoj studiji utvrđeno je da su učinci fluorida na faunu u uobičajenim okvirima, ali u budućnosti može imati štetan utjecaj na faunu i floru u okruženju tvornice Hindalco Industries Limited

Utjecaj fluorida proizvedenih aluminijskim industrijskim postrojenjem na biljke i područja nastanjena ljudima

Fluorine is a highly reactive common element that does not occur in nature in the elemental state. It exists in the form of fluorides and accounts for about 0.3 g/kg of the Earth’s crust. Generally, it is found in the form of a number of minerals like fluorspar, cryolite and fluor-apatite. Fluoride has both positive and negative effects on individual health. Fluoride, in the form of fluorspar and cryo- lite is distributed extensively in the lithosphere, and is renowned as the thir- teenth most common among elements in the earth’s crust. Hydrogen fluorides in gaseous form accumulate in the leaves of generally sensitive plants against a concentration gradient and therefore, considered as a most phytotoxic air pollut- ant and affects plants at extremely low concentration. As per our study, it is found that the fluoride impacts on fauna are in normal condition, but in coming times it may have adverse impact on fauna and flora of surroundings of Hindalco Industries Limited.Fluor je vrlo reaktivan element koji se u elementarnom obliku ne pojavljuje u prirodi. Postoji u obliku fluorida u zemljinoj kori, gdje se nalazi u omjeru od oko 0,3 g/kg. Općenito, nalazi se u obliku velikog broja minerala poput fluorita, kriolita i apatita. Fluorid ima i pozitivne i negativne učinke na ljudsko zdravlje. Fluorid, u obliku fluorita i kriolita, široko je rasprostranjen u litosferi, a poznat je i kao trinaesta najčešća tvar u Zemljinoj kori. Fluorovodici u plinovitom obliku nagomilavaju se u listovima uglavnom osjetljivih biljaka prema gradijentu koncentracije. Stoga se smatraju najfitotoksičnijim onečišćujućim tvarima u zraku, a djeluju na biljke i pri ekstremno niskim koncentracijama. Prema našoj studiji utvrđeno je da su učinci fluorida na faunu u uobičajenim okvirima, ali u budućnosti može imati štetan utjecaj na faunu i floru u okruženju tvornice Hindalco Industries Limited

Investigation of the acetic acid stress response in Saccharomyces cerevisiae with mutated H3 residues

Enhanced levels of acetic acid reduce the activity of yeast strains employed for industrial fermentation-based applications. Therefore, unraveling the genetic factors underlying the regulation of the tolerance and sensitivity of yeast towards acetic acid is imperative for optimising various industrial processes. In this communication, we have attempted to decipher the acetic acid stress response of the previously reported acetic acid-sensitive histone mutants. Revalidation using spot-test assays and growth curves revealed that five of these mutants, viz., H3K18Q, H3S28A, H3K42Q, H3Q68A, and H3F104A, are most sensitive towards the tested acetic acid concentrations. These mutants demonstrated enhanced acetic acid stress response as evidenced by the increased expression levels of AIF1, reactive oxygen species (ROS) generation, chromatin fragmentation, and aggregated actin cytoskeleton. Additionally, the mutants exhibited active cell wall damage response upon acetic acid treatment, as demonstrated by increased Slt2-phosphorylation and expression of cell wall integrity genes. Interestingly, the mutants demonstrated increased sensitivity to cell wall stress-causing agents. Finally, screening of histone H3 N-terminal tail truncation mutants revealed that the tail truncations exhibit general sensitivity to acetic acid stress. Some of these N-terminal tail truncation mutants viz., H3 [del 1-24], H3 [del 1-28], H3 [del 9-24], and H3 [del 25-36] are also sensitive to cell wall stress agents such as Congo red and caffeine suggesting that their enhanced acetic acid sensitivity may be due to cell wall stress induced by acetic acid

Hormonal and non-hormonal oral contraceptives given long-term to pubertal rats differently affect bone mass, quality and metabolism

IntroductionWe investigated the effects of hormonal and non-hormonal oral contraceptives (OCs) on bone mass, mineralization, composition, mechanical properties, and metabolites in pubertal female SD rats.MethodsOCs were given for 3-, and 7 months at human equivalent doses. The combined hormonal contraceptive (CHC) was ethinyl estradiol and progestin, whereas the non-hormonal contraceptive (NHC) was ormeloxifene. MicroCT was used to assess bone microarchitecture and BMD. Bone formation and mineralization were assessed by static and dynamic histomorphometry. The 3-point bending test, nanoindentation, FTIR, and cyclic reference point indentation (cRPI) measured the changes in bone strength and material composition. Bone and serum metabolomes were studied to identify potential biomarkers of drug efficacy and safety and gain insight into the underlying mechanisms of action of the OCs.ResultsNHC increased bone mass in the femur metaphysis after 3 months, but the gain was lost after 7 months. After 7 months, both OCs decreased bone mass and deteriorated trabecular microarchitecture in the femur metaphysis and lumbar spine. Also, both OCs decreased the mineral: matrix ratio and increased the unmineralized matrix after 7 months. After 3 months, the OCs increased carbonate: phosphate and carbonate: amide I ratios, indicating a disordered hydroxyapatite crystal structure susceptible to resorption, but these changes mostly reversed after 7 months, indicating that the early changes contributed to demineralization at the later time. In the femur 3-point bending test, CHC reduced energy storage, resilience, and ultimate stress, indicating increased susceptibility to micro-damage and fracture, while NHC only decreased energy storage. In the cyclic loading test, both OCs decreased creep indentation distance, but CHC increased the average unloading slope, implying decreased microdamage risk and improved deformation resistance by the OCs. Thus, reduced bone mineralization by the OCs appears to affect bone mechanical properties under static loading, but not its cyclic loading ability. When compared to an age-matched control, after 7 months, CHC affected 24 metabolic pathways in bone and 9 in serum, whereas NHC altered 17 in bone and none in serum. 6 metabolites were common between the serum and bone of CHC rats, suggesting their potential as biomarkers of bone health in women taking CHC.ConclusionBoth OCs have adverse effects on various skeletal parameters, with CHC having a greater negative impact on bone strength

Racial disparities in oropharyngeal cancer

10.1007/978-3-319-21100-8_3Human Papillomavirus (HPV)-Associated Oropharyngeal Cancer43-6