However, 10- and 12--HODE alone cannot perfectly detect early impaired glucose tolerance (IGT) and/or insulin resistance, which ultimately lead to diabetes. In this study, we randomly recruited healthy volunteers (n = 57) who had no known history of any diseases, and who were evaluated using the OGTT, the HODE biomarkers, and several additional proposed biomarkers, including retinol binding protein 4 (RBP4), adiponectin, leptin, insulin, glycoalbumin, and high sensitivity-C-reactive protein. The OGTT revealed that our volunteers included normal individuals , “high-normal” individuals (fasting plasma glucose 100–109 mg/dL) with IGT , and diabetic individuals . We then used these groups to evaluate the potential biomarkers for the early detection of type 2 diabetes. Plasma levels of RBP4 and glycoalbumin were higher in Group HN+IGT, compared to those in Group N, and fasting levels of 10- and 12--HODE/linoleic acids were significantly correlated with levels of RBP4  and glycoalbumin . Furthermore, we developed a stepwise multiple linear regression models to predict the individuals’ insulin resistance index (the Matsuda Index 3). Fasting plasma levels of 10- and 12--HODE/linoleic acids, glucose, insulin, and leptin/adiponectin were selected as the explanatory variables for the models. The risks of type 2 diabetes, early IGT, and insulin resistance were perfectly predicted by comparing fasting glucose levels to the estimated Matsuda Index 3 -HODE/linoleic acids, insulin, and leptin/adiponectin).We have previously found that fasting plasma levels of totally assessed 10- and 12- Early detection and treatment of diabetes can postpone, or even prevent, the serious complications that are associated with diabetes . The criteria for diagnosing diabetes is a fasting plasma glucose (FPG) concentration of >126 mg/dL and a plasma concentration of >200 mg/dL at 120 min after the oral glucose tolerance test (OGTT). In comparison, healthy individuals have fasting or 120 min post-OGTT FPG concentrations of <110 and <140 mg/dL, respectively. The Japan Diabetic Society recommends that subjects with an FPG value of 100–109 mg/dL be classified as “high-normal” in the range of glucose metabolism disorders, and that subjects with a “high-normal” FPG value should undergo a 75-g OGTT to determine whether they are normal, borderline, or diabetic ; glucose, Cica liquid GLU J ; insulin, Lumipulse Presto Insulin [Fujirebio Inc. ]; leptin, Human leptin RIA kit ; adiponectin, CircuLex Human adiponectin ELISA Kit CY-8050 [MBL Co. Ltd. ]; RBP4, CircuLex Human RBP4 ELISA Kit [MBL Co. Ltd. ]; glycoalbumin, Lucia GA-L ; and hs-CRP, CircuLex Human HS-CRP ELISA Kit CY-8071 [MBL Co. Ltd. ]).p-values of <0.05 were considered statistically significant.Statistical analyses were performed on a Microsoft PC using SPSS software . One-factor repeated measures design analysis of variance (ANOVA) was used to examine the effect of elapsed time from the glucose injection on each index. Significant effects were followed by Tukey's honestly significant difference multiple comparisons, and correlations were analyzed with the Pearson test. Data were expressed as mean ± standard deviation, and -HODE/LA, HbA1c, RBP4, glycoalbumin, leptin/adiponectin, hs-CRP, and body mass index (BMI). These variables were used because they were significantly correlated with glucose levels at both 60 min and 120 min after the administration of glucose (p < 0.01) or with the Matsuda Index 3 (p < 0.05). Logarithmic transformation was applied to all variables to achieve normality before the analysis.We developed a multiple linear regression model to predict the Matsuda Index 3, as a function of 9 physiological variables Model 1, . The 9 vp-value of 0.05 -HODE/LA, RBP4, glycoalbumin, adiponectin, leptin, and hs-CRP. Glucose tolerance was estimated using HbA1c, FPG, and glucose levels at 60 min and 120 min after the glucose administration. Insulin resistance was estimated using fasting plasma insulin, insulin levels during the OGTT, HOMA-IR, and Matsuda Index 3. 10- and 12--HODE/LA, RBP4, hs-CRP, and adiponectin were significantly correlated with both glucose tolerance and insulin resistance –HODE/LA, glycoalbumin, and hs-CRP were also well correlated with glucose tolerance and HbA1c. Interestingly, these markers were not correlated with insulin, nor with the insulin resistance indices (HOMA-IR and Matsuda Index 3). However, RBP4, adiponectin, and leptin/adiponectin were correlated with HOMA-IR and the Matsuda Index 3, and 10- and 12-–HODE/LA, RBP4, and, glycoalbumin were significantly correlated with glucose levels during the OGTT. –HODE/LA and RBP4 (p < 0.001), as well as glycoalbumin (p < 0.001), which are both prominent biomarkers for the early detection of diabetes [-HODE/LA alone can predict glucose tolerance, although not early-stage insulin resistance, we developed an algorithm to detect pre-diabetes using the above-mentioned markers.We analyzed the correlations of glucose tolerance and insulin resistance with the fasting levels of 10- and 12-sistance . 10- anddiabetes ,6. As 10First, we designed an algorithm that could predict IGT and early stage insulin resistance using the fasting plasma markers, although this study did not include any subjects with isolated IFG and only 3 subjects with isolated IGT. As shown in -HODE/LA, insulin, and leptin/adiponectin were selected as explanatory variables for the multiple linear regression model (Model 2). As shown in p < 10−20). Furthermore, FPG and the selected markers perfectly detected glucose tolerance and insulin resistance -HODE/LA, insulin, and leptin/adiponectin, perfectly predicted IGT and insulin resistance without using the OGTT. As it is generally accepted that IGT is largely due to insulin resistance, with IFG due to dysregulated gluconeogenesis, the OGTT is a useful tool for detecting the risk of both glucose tolerance and insulin resistance. However, as shown in Our multiple linear regression model -HODE/LA, insulin, and leptin/adiponectin) were able predict the risk of type 2 diabetes. However, 10- and 12--HODE/LA had a good correlation with RBP4 and glycoalbumin levels, which is unsurprising, given that they have been frequently studied as biomarkers for glucose tolerance [It is interesting that only 4 fasting plasma markers -HODEs, and created three models that did not include these markers: Model 3 (insulin and L/A), Model 4 , and Model 5 .The resulting AIC values were 32.6 (Model 3), 30.2 (Model 4), and 27.9 (Model 5). Therefore, it appears that 10- and 12--HODEs are prominent biomarkers for the detection of early stage diabetes.We investigated the usefulness of our biomarker models using Akaike’s information criterion (AIC). In this context, lower AIC values indicate that a model has a better predictive ability. The AIC values for the models using 10 biomarkers  and 4 bi-HPODE and 9- and 13--HPODE), while singlet oxygen oxidizes LA through non-radical oxidation to form 4 unique isomers -HPODE and 10- and 12--HPODE). Thus, the 10- and 12--HPODEs that we examined are specific oxidation products of singlet oxygen [in vivo. Furthermore, it has been reported that singlet oxygen is produced in vivo by the reaction of hydrogen peroxide with hypochlorite, which is produced by myeloperoxidase that is secreted by activated phagocytes [Hydroperoxyoctadecadienoic acids (HPODE) are formed by free-radical-mediated oxidation, and consist of 4 isomers -HODEs in a model of type 2 diabetes (Tsumura Suzuki Obese Diabetes mice) were significantly higher than those of control mice before IGT was observed. In addition, treating HaCaT cells with sub-lethal concentrations of 10- and 12--HODEs, although not 9- and 13--HODEs, caused resistance to hydrogen peroxide-induced oxidative damage, which indicates adaptive response [-HODE, although not 9- and 13-HODE, were highly correlated with clinical values for diabetes and had diagnostic potential. Nevertheless, if these biomarkers are affected by the individual’s diet, it is likely that the levels of all HETEs/HODEs isomers would increase. In addition, results from animal models indicate that plasma levels of HODE are not affected by the daily diet, if it does not contain fortified coenzyme Q 9 or vitamin Es as supplements [Z,E)-HODEs markers in the present study were not influenced by the subjects’ diet. Therefore, HODE levels after 10 h of fasting may be appropriate for clinical use, as subjects’ biomarker levels are typically stable in the morning after fasting.In an animal study , plasma response . Our preresponse  also demplements . Further-HODE/LA, insulin, and leptin/adiponectin, can be used for the early detection of diabetes. We have recently developed a second-generation prototype of a multi-marker analysis system, which is equipped with a CD-type microfluidic device [The present study clearly shows that multiple markers, including 10- and 12-c device  and an a


Antiviral drug-resistant HBV mutants are complex and currently partly understood. This study was performed to analyze the profile of hepatitis B virus (HBV) resistance mutations against nucleos(t)ide analogues (NAs) in patients with chronic hepatitis B (CHB).This was a population-based cross-sectional study. Serum samples of 179 patients with virological breakthrough undergoing different NAs treatment were obtained between January 2008 and December 2012. The HBV reverse transcriptase region was sequenced and the following NAs-resistant changes including rtL80, rtI169, rtV173, rtL180, rtA181, rtT184, rtA194, rtS202, rtM204, rtN236 and rtM250 were analyzed.P=0.032]; while the HBV rtM204V mutations was significantly associated with HBV rtL180M mutations . Additionally, HBV rtA181 mutations were observed in 19.3% (22/114) of patients with LAM/LdT-based therapy and 23.9% (11/46) of patients with ADV-based therapy.In this cohort, 21.2% (38/179) were genotypes B and 78.8% (141/179) were genotypes C; and 89.4% (160/179) of them detected NAs-resistant mutations. The prevalence of HBV mutations at rtM204 was 93.0% (106/114) in patients with lamivudine (LAM) or telbivudine (LdT)-based therapies, and that of rtN236 mutations was 76.1% (35/46) in patients with adefovir dipivoxil (ADV)-based therapies. Among LAM/LdT based therapies, HBV rtM204I was significantly associated with HBV rtL80I/V mutations [rtM204I+rtL80I/V  vs. rtM204V+rtL80I/V , Majority of virological breakthrough is associated with NAs-resistant HBV, and the mutation patterns of NAs-resistant HBV are complicated in real clinical practice. For the ability to inhibit the activity of hepatitis B virus (HBV) reverse transcriptase (RT), nucleos(t)ide analogues (NAs) such as lamivudine (LAM), adefovir dipivoxil (ADV), entecavir (ETV) and telbivudine (LdT) are approved for the treatment of chronic hepatitis B (CHB) in China ,2. HowevAt present, many mutations in the RT gene of HBV have been reported to be associated with classical antiviral resistance, and mutations of rtA181, rtM204 and rtN236 have been reported to be associated with major primary drug resistance and are responsible for reduced treatment susceptibility resulting from antiviral agents . AccordiHowever, the profile data of classical antiviral resistance mutations belonging to different evolutionary pathways is limited in real clinical practice. Thus, this study is designed to characterize the molecular features of nucleos(t)ide analogues resistance (NAr) mutation positions belonging to different NAr pathways within HBV RT sequences.4 copies/mL. The exclusion criteria were patients with hepatitis C virus (HCV) or human immunodeficiency virus (HIV) coinfection, autoimmune liver disease and alcohol or drug abuse. This study was also approved by the Ethics Committee of Wuhan Neurologist Hospital and General Hospital of The Yangtze River Shipping in accordance with the Declaration of Helsinki.This was a population-based cross-sectional study. A total of 1308 CHB patients undergoing LAM, ADV, ETV or LdT treatment in single, sequential or combination between January 2008 and December 2012 were screened, and only 179 patients with virological breakthrough were finally included and analyzed in present study. The inclusion criteria were CHB patients received NAs treatment for more than 6 months and had virological breakthrough with HBV DNA ≥ 1.0×10Serum HBV DNA was quantified by fluorescence quantitative polymerase chain reaction (PCR)  with the lowest detection limit of 1,000 copies/mL. HBV gene fragment (nt54-1278) encompassing the complete RT gene was amplified by PCR. Sequencing was performed using the ABI 3730xl DNA Analyzer. Sequencing data were analyzed using the Vector NTI Suite software package . HBV genotypes assignment was based on the phylogenetic analysis of the 1225-bp-long S/P-gene sequence (nt 54–1278) of HBV genome. Mutations at 12 locations  in the RT gene belonging to different NAr pathways were analyzed using DNAStar version 5.0  software, and all above HBV NAs-resistant associated mutations were detected when virological breakthrough occurred.The clinical diagnosis of CHB was made according to guideline on prevention and treatment of chronic hepatitis B in China ,13. ViroQuantitative values expressed as mean plus standard deviation or range, and qualitative values were were presented as number and percent. Group comparisons between qualitative values were performed using Chi-square tests using SPSS 17.0 software . P value of less than 0.05 was considered statistically significant.In this cohort, there were 63 (35.2%) females and 116 (64.8%) males with an average age of 38 years (range 21–63), and the HBeAg positivity rate was 67.6% (121/179). Among those 179 HBV patients, 21.2% (38/179) and 78.8% (141/179) were infected with HBV genotypes B and C, respectively; 89.4% (160/179) of the patients detected NAs-resistant mutations including 114 patients with LAM/LdT-based therapies and 46 patients with ADV-based therapies. Among the 160 patients with NAs-resistant mutations, up to 7 antiviral strategies were involved which manifested as monotherapy , LAM plus ADV combination therapy (n=11), LAM switch-to ADV (n=18), or ETV sequential therapies (n=5).In present study, NAs associated mutations were detected at positions rtL80, rtV173, rtL180, rtA181, rtS202, rtM204, rtN236 and rtM250, but not at positions rtI169, rtT184, rtA194 or rtI233. Among LAM/LdT-based treatment (n=114), the prevalence rates of rtM204 mutations were 100% (85/85), 77.8% (14/18), 50.0% (4/8) and 100% (3/3) in patients receiving LAM or LdT monotherapy, ADV switch-to, ADV add-on, or ETV switch-to therapy respectively. And among those rtM204 mutations, rtM204I, rtM204V and rtM204I/V accounted for 60.4% (64/106), 31.1% (33/106) and 8.5% (9/106), respectively. Thus, our finding further confirms that rtM204 should be considered as an important primary LAM/LdT-resistance mutation position in real clinical practice, and rtM204I mutation is the most common type.vs. rtM204V+rtL80I/V , P=0.032]. By contrast, all rtM204V mutants were found to be coexistent with rtL180M mutations, which was significantly higher than that of rtM204I mutants .Among those rtM204 mutations, a total of 14 types of rtM204 containing mutant patterns were observed, and rtM204I/V+rtL180M  and rtM204 I/V+rtL80I/V  were the two most common mutation patterns (Table Among ADV-based treatment (n=46), the rtN236T mutations were detected in 78% (32/41), 66.7% (2/3) and 50% (1/2) of patients receiving ADV monotherapy, LAM add-on, or ETV switch-to therapies, respectively. Additionally, among LAM/LdT-based monotherapy, 50% of patients who (4/8) received ADV add-on also detected rtN236T mutations.Recently, HBV rtA181T/V mutations had been widely concerned and it was regarded to be a multidrug resistance which not only could result in resistance to ADV but also reduce sensitivity to LAM and LdT. In this cohort, HBV rtA181T/V mutation was observed in 19.3% (22/114) of patients with LAM/LdT-based therapies and 23.9% (11/46) of patients with ADV-based therapies. It was worth mentioning that this mutation was detected in all 11 patients who received LAM plus ADV combination therapy (8 from LAM/LdT-based combination therapy and 3 from ADV-based combination therapy). Our findings suggested that rtA181T/V mutations were common among patients with NAs resistance, and it should be regularly monitored in real clinical practice.In this study, classical antiviral resistance mutations within the HBV RT region, which belonged to diverse evolutionary pathways, were analyzed among 179 CHB patients with virological breakthrough. Among those patients, 89.4% 160/179) detected antiviral-resistant HBV mutations, which indicated that the emergency of resistance was the main reason for virological breakthrough and failure of treatment. In this cohort, three classic evolutionary pathways of L-nucleoside pathway against LAM, LdT or ETV (rtM204 I/V), the acyclic phosphonate pathway (rtN236T) against ADV and the shared pathway against both LAM and ADV (rtA181T T/V) were all involved, and our findings were also supported by recent studies ,16. Beca0/179 detRecently, rtI233 mutation has been reported to be associated with primary poor or failure response to ADV ; howeverIn this study, majority of patients had resistance that was associated with the L-nucleoside pathway [rtM204I (or V or I/V)], which could potentially lead to resistance to LAM, ETV and LdT treatment in LAM-experienced patients. The highest prevalence rate of genotypic resistance to the L-nucleoside pathway was in agreement with the largest percentage  of LAM-based patients in our cohort. Indeed, this high prevalence of rtM204 mutation was caused by the long-term use of LAM in China. So we speculated the presence of rtM204 mutation carriers might constitute an unavoidable challenge for the effective control of antiviral-resistant HBV in the future because it is difficult to clear such resistant viral strains .Although preliminary data using ADV belonging to acyclic phosphonates on LAM-resistance patients seem promising, a shared pathway (rtA181T/V) of both L-nucleosides and acyclic phosphonates could be selected, which would affect both LAM and ADV sensitivity . In presThe most obvious limitation of the study is its limited sample size, thus clinical trials with large sample sizes were required to reveal the comprehensive and accurate profile of HBV resistance mutations against NAs.Our findings indicate that there is a complex pool of resistant HBV in real life of CHB patients with virological breakthrough. Thus, monitoring HBV genotypic resistance and shared resistant pathway pertaining to antiviral agents would help to optimize or rescue current antiviral therapies and avoid the outbreak of clinical deterioration.The contents are solely the responsibility of the authors.XLY was involved in the study concept and design, statistical analysis and interpretation of the data, drafting and critical revision of the manuscript. LJ was involved in the study concept and design, acquisition of the data, statistical analysis and interpretation of the data, drafting and critical revision of the manuscript and study supervision. WLL was involved in statistical analysis and interpretation of the data and critical revision of the manuscript. ZSJ, CW and BZJ were involved in the study concept and design, and in the critical revision of the manuscript. All authors have read and approved the final manuscript.


Craniosynostosis results from premature ossification of one or more cranial sutures and leads to alterations in the shape of the skull and/or premature closure of cranial fontanels, causing impairment of brain perfusion, vision and hearing, airway obstruction, learning difficulties, severe cosmetic deformities and high intracranial pressure . To dateA newborn female born at term (39 weeks) with birth weight 3.405 kg and length 45 cm, an occipitofrontal head circumference (OFC) of 36.5 cm, 2nd pregnancy of a mother at 39 years old and a father at 41 years old, not consanguineous and without family history of congenital anomalies was examined by neonatologists using the Mercks protocol (2003) for early identification of major and minor anomalies. GTG Banding analyzes were performed on metaphase obtained by stimulating peripheral blood lymphocytes from patient and their parents in accordance with standard procedures. The technique Multiplex ligation-dependent probe amplification (MLPA) was performed using kits for the subtelomeric regions of all chromosomes (P036 and P070-E1-B2 - MRC-Holland). Array Comparative Genome Hybridization (aCGH) (OGT CytoSure ISCA 8x60k) was performed according to the manufacturer's recommendations.de novo deletion in region 10q26.2-q2, confirmed by aCGH that showed a 12.9 Mb deletion (122085501-135053489) (The UCSC Human Genome Project Working Draft Build 36/hg 18). Mutations in the FGFR2 gene cause several syndromes that result in craniosynostosis  and familial scaphocephaly [The newborn had sagittal craniosynostosis , microcephaly, facial asymmetry, short forehead and bitemporal narrowing, microtia, hypoplastic and low-set ears, thin lips, strabismus, retromicrognathia, anteriorized anus, right single palmar crease, short neck, syndactyly of 2-3rd fingers and congenital heart disease. The karyotype was normal and MLPA detected a


Immune cell infiltration in (white) adipose tissue (AT) during obesity is associated with the development of insulin resistance. In AT, the main population of leukocytes are macrophages. Macrophages can be classified into two major populations: M1, classically activated macrophages, and M2, alternatively activated macrophages, although recent studies have identified a broad range of macrophage subsets. During obesity, AT M1 macrophage numbers increase and correlate with AT inflammation and insulin resistance. Upon activation, pro-inflammatory M1 macrophages induce aerobic glycolysis. By contrast, in lean humans and mice, the number of M2 macrophages predominates. M2 macrophages secrete anti-inflammatory cytokines and utilize oxidative metabolism to maintain AT homeostasis. Here, we review the immunologic and metabolic functions of AT macrophages and their different facets in obesity and the metabolic syndrome. Obesity is a prevalent metabolic disease characterized by excess accumulation of white adipose tissue (AT) due to increased food intake and changes in lifestyle , 2. ObesA major player in systemic low-grade chronic inflammation in obesity is the increased numbers of AT pro-inflammatory macrophages and deregulated production and function of AT hormones and cytokines , 4. BesiInsulin resistance is defined as a reduced response to insulin in liver, muscle, and AT. This impairment is due to the inhibition of the insulin-signaling pathway, leading to hyperglycemia. Insulin resistance is commonly associated with obesity and may precede the onset of T2D –13. One The finding that infiltration of monocytes, which differentiate into macrophages, is augmented in obesity is fundamental , 16. ThiIn this review, we highlight the different functions of AT macrophages (ATMs) in the maintenance AT tissue homeostasis during lean, obese, and insulin resistant states.in vitro. In AT, the secretion of TNF-α is primarily derived from macrophages  was the first cytokine identified to be capable of inducing insulin resistance in adipocytes rophages –22, and rophages . This suMacrophages make up to 40% of all AT cells in obese mice compared to 10% in lean mice . These cIn addition to the activation and inflammatory profile of macrophages in the obese state, ATMs are highly adaptive to its lipid-rich environment. To maintain AT homeostasis in this lipid-rich microenvironment, macrophages increase their adiposity by activating lysosomal lipid metabolism . This maSeveral immune cells regulate AT inflammation, insulin resistance , and macIn vivo, this phenotype is due to continuous and excessive exposure of ATMs to FFAs, such as palmitate, in a microenvironment that is saturated with glucose and insulin. In obesity, this differentiated macrophage subtype indicates the importance and the necessity to identify differentiated profiles of immune cells. Since there is a large spectrum of ATMs that have different immune profiles, we choose to focus on M1 and M2 subtypes of ATMs to better understand how metabolic alterations in ATMs impact obesity and insulin resistance.However, Kratz et al. recently described a different subtype of macrophage . They obM1 macrophages are associated with a pro-inflammatory profile. These macrophages are generally stimulated by T-helper 1 (Th1) type of cytokines, such as interferon-γ (IFN-γ), or by pathogen-associated molecular patterns (PAMPs), such as LPS . In turnIn addition to IFN-γ and LPS, there are several other molecules involved in M1 macrophage polarization, such as interferon regulatory factor (IRF), signal transducers and activators of transcription (STAT), and suppressor of cytokine signaling (SOCS). IRF5 is involved in M1 polarization by inducing the transcription of interleukin-12 subunit p40 (IL-12p40), IL-12p35, and IL-23p19, and by repressing the transcription of IL-10 . M1 macrin vitro, which do not express CD11c, M1 ATMs express CD11c concomitant with F4/80 and CD11b   , inhibit1 and 2) , extrace1 and 2) , and mit1 and 2) . These a1 and 2) , 68–72. 1 and 2) . Hematop1 and 2) . Corrobo1 and 2) . Similar1 and 2) .Toll-like receptors and inflammasomes are activated in obesity by damage-associated molecular pattern molecules (DAMPs), such as high-mobility group box 1 (HMGB1) and oxidized low-density lipoprotein (Ox-LDL), RBP4 or PAMPs, such as LPS , 76–80. Toll-like receptor 4 deficiency in HFD-fed mice ameliorates AT inflammation, insulin resistance, and adiposity , 85, 86.Nod-like receptor protein 3 inflammasome also plays a key role in the development of AT inflammation and insulin resistance , 89. ExpFunctional deletion of NLRP3 and caspase-1 ameliorate HFD-induced insulin resistance and AT inflammation , 90, 92.In addition to these important signaling pathways, the mammalian target of rapamycin (TOR) has an important function in insulin resistance. It is able to sense nutrients and respond by altering the cellular metabolism in different kind of cells, including ATMs . Insulinin vitro and in vivo and induces M2 polarization  in macrophages leads to a marked defect in M2 polarization in response to IL-4, although LPS stimulation induced inflammatory responses in an mTOR-dependent manner , PPAR-γ, and transcription factor found in inflammatory zone 1 (FIZZ1), which is specific of murine M2 macrophages, are necessary for collagen synthesis, further supporting the role of these cells in tissue remodeling  and IL-33. ILC2s are a regulatory subtype of ILCs. These cells were divided into three distinct populations, ILCs 1, 2, and 3 –127. Thepression . Howeverpression . ILCs arpression .Interleukin-33 is constitutively present in humans and mice, mainly in specialized populations of epithelial and endothelial cells , 131. ItInterleukin-33, as well as ILC2s, has been in the spotlight due to their putative contributions in the improvement of obesity-induced insulin resistance. Upon binding to its receptor, IL-33 induces the production of large amounts of anti-inflammatory cytokines by AT ILC2s and also the polarization of ATMs toward an M2 phenotype . This reIn this context, Brestoff et al. demonstrated that IL-33 plays an important role in the maintenance of ILC2s in AT, promoting energy expenditure, and reducing adiposity in mice . This deTaken together, these studies demonstrate the importance of alternatively activated macrophages to maintain the tissue homeostasis, especially in AT. Moreover, the discovery of new alternative pathways for the polarization of ATMs toward an M2 phenotype is necessary to better understand the mechanisms by which insulin sensitivity in obesity.In addition to cytokines, the availability of substrates in tissues orchestrates macrophage function. Cellular metabolism is not static but is rather a dynamic process that allows cells to adapt to the microenvironment . The typM1 macrophages preferentially metabolize glucose as an energy substrate . During In addition, the expression of glucose transporter-1 (GLUT-1) drives the pro-inflammatory phenotype of M1 macrophages, increases glucose uptake, and, subsequently, augments glucose metabolism .2) tension and inflammatory responses increase TLR4 expression in macrophages (One important molecule regulating glycolysis and macrophage activation is hypoxia inducible factor-1α (Hif-1α). Hif-1α induces a pro-inflammatory phenotype in macrophages  via TLR4rophages . Moreoverophages –149. Becrophages .Activation of macrophages with LPS also results in increased levels of succinate and malate . SuccinaNonetheless, the microenvironment rich in LPS and IFN-γ also enhances M1 macrophage polarization and glycolysis activation independently of Hif-1α. This occurs upon 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFK2) induction .In contrast to M1 glycolytic metabolism, M2 macrophages utilize oxidative metabolism . The indBesides CARKL, the coactivator protein PPAR-γ-coactivator-1β (PGC1-β) induces mitochondrial respiration as well as mitochondrial biogenesis. This is a key player in the metabolic switch of macrophages from M1 to M2 phenotype , 144. BlArg1) to amplify the effector phenotype of M2 macrophages (collagen synthesis)  , 156. Otnthesis) , indicatArg1, which is expressed by M2 macrophages (Hypoxia inducible factor-2α has been shown to regulate the transcription of rophages . Howeverrophages , 157. ThIt is still unclear how M2 macrophages metabolism is regulated during obesity and the role of M2 macrophage metabolism for the development of insulin resistance. Nevertheless, in lean state, they have an oxidative metabolism, which may shift to glycolytic metabolism, during obesity, due to a pro-inflammatory environment and further studies are needed to better understand their role in obesity.Macrophages are central mediators of obesity-induced AT inflammation and insulin resistance. They also are key cells for maintenance of AT homeostasis. Recently, several reports described the importance of these cells as regulators of insulin sensitivity, which involves the activation of innate immune receptors, transcription factors, and intracellular metabolism to support the either pro- or anti-inflammatory AT phenotype. Thus, macrophages have a dual role, changing their status to support immune responses, obesity development, and related diseases.AC and CN have contributed equally in the writing of this manuscript. NC and PM-V contributed in the writing and corrections of this manuscript. AC and CN have contributed equally to this review.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Worldwide, chronic infection with HBV and HDV occurs in more than 10 million people with higher rates of developing cirrhosis, hepatic decompensation and hepatocellular carcinoma. Peginterferon (pegIFN) for 48 weeks is one of the current therapies, with poor response rates; prolonging therapy beyond 12 months might be of benefit for the patients.In order to assess the response to a prolonged pegIFN therapy, we present 2 patients with chronic B/D coinfection, treated with pegIFN alpha 2a, 180 mcg weekly for 96 weeks and monitored, at least, every three months with a complete medical evaluation.Patient 1: Female age 30, diagnosed at age 19 with chronic B/D coinfection; treated the first year with oral azathioprine for 12 weeks, then one week every three months; the second year she started interferon alpha 2a for 48 weeks, achieving biochemical and virologic response. At age 23, she became pregnant, gave birth to a healthy baby girl with no viral infection; after pregnancy, she had an ALT 10 times the upper normal limit, HBV-DNA 1,000,000 IU/mL. For two years, she received support therapy: ursodeoxycholic acid, vitamins, silymarin until we decided to start peginterferon alpha 2a 180 mcg weekly for 48 weeks. Well tolerated, we obtained undetectable HBsAg, HBV-DNA and HDV-RNA in the serum and normal ALT. For a sustained response, we decided to continue this treatment for another 48 weeks. Until present she maintained undetectable HBsAg, HBV-DNA and HDV-RNA; anti HBs < 2.00 (limits 2-10 IU/L), anti-HBe: 0.012 COI (reactive).Patient 2: Male, 40 years old, diagnosed at age 36 with chronic B/D coinfection; we started standard peginterferon alpha 2a therapy for 48 weeks; with a major side-effect: Hashimoto thyroiditis in month 3, successfully resolved. We continued the treatment, then – also based on a second-opinion from a French hospital – we indicated another 48 weeks of treatment that resulted in sustained biochemical and virologic response with viral eradication, until present.A long-term peginterferon therapy could be a chance for cure in patients coinfected with HBV/HDV, with permanent and rigorous monitoring and medical evaluation. This approach needs further studies, for the benefit of patients, considering the higher rates of mortality among them.


Rhodococcus opacus strain R7 (CIP107348) degrades several mono- and polycyclic aromatic hydrocarbons. Here, we present the high-quality draft genome sequence of strain R7, consisting of 10,118,052 bp, with a G+C content of 67.0%, 9,602 protein-coding genes, and 62 RNAs genes. Rhodococcus demonstrate a remarkable ability to degrade a wide range of natural organic and xenobiotic compounds (Rhodococcus opacus R7 (CIP107348) was isolated from a polycyclic aromatic hydrocarbon-contaminated site in Italy because of its ability to grow on naphthalene and o-xylene as the only carbon and energy source . The total numbers of sequence reads were 312,384 from one shotgun library and 380,920 from one paired-end library. All the reads were assembled using Newbler 2.6 into 223 contigs, with an N50 length of 184,729 bp and an average genome coverage of 17×. Based on paired-end directional information, the contigs were further ordered into 6 scaffolds, giving a total genome size of 10.1 Mb with a G+C content of 66.0%.The genome sequencing of Rhodococcus plasmids.The annotation was performed by using the RAST (Rapid Annotation using Subsystem Technology) server . Five scRhodococcus jostii RHA1 (score 501), Rhodococcus opacus B4 (score 409) are the closest neighbors of strain R7. Subsystem (457) categories representing the metabolisms of carbohydrates, amino acids and cofactors, vitamins, prosthetic groups, or pigments are the most abundant, and they account for 1,236, 995, or 665 proteins, respectively. A total of 745 ORFs are involved in metabolism of fatty acids, lipids, and isoprenoids, while 267 ORFs participate in metabolism of aromatic compounds. A total of 129 oxygenases/hydroxylases among the 134 annotated are predicted to initiate oxidation of organic compounds with industrial and environmental relevance, such as linear alkanes, cyclic ketones, aromatic compounds , aminopolycarboxilic acids, nitroalkanes, and phenylalkanoic acids. Forty-five ORFs encode cytochrome P450 monooxygenases that catalyze regio- and stereospecific oxidation of a vast number of substrates , 62 RNAs genes, 9 rRNAs and 53 tRNAs, were predicted. RAST annotation indicates that strains CP008947, CP008948, CP008949, CP008950, CP008951, and CP008952. The versions described in this paper are the first versions.This whole-genome shotgun project has been deposited at DDBJ/EMBL/GenBank under the accession numbers


Apoptosis is the process of programmed cell death (PCD) that occurs in multicellular organisms. This process of normal cell death is required to maintain the balance of homeostasis. In addition, some diseases, such as obesity, cancer, and neurodegenerative diseases, can be cured through apoptosis, which produces few side effects. An effective comprehension of the mechanisms underlying apoptosis will be helpful to prevent and treat some diseases. The identification of genes related to apoptosis is essential to uncover its underlying mechanisms. In this study, a computational method was proposed to identify novel candidate genes related to apoptosis. First, protein-protein interaction information was used to construct a weighted graph. Second, a shortest path algorithm was applied to the graph to search for new candidate genes. Finally, the obtained genes were filtered by a permutation test. As a result, 26 genes were obtained, and we discuss their likelihood of being novel apoptosis-related genes by collecting evidence from published literature. TNFR-1, Fas/CD95, and the TRAIL receptors DR-4 and DR-5 [Apoptosis, an efficient cell death program, plays an important role in maintaining strictly regulated organismal homeostasis and involves the interaction of multiple factors. Since the mid-nineteenth century, cell death has been widely studied, and researchers have learned that all physiological processes of multicellular organisms involve cell death, particularly during embryogenesis and metamorphosis , 2. The and DR-5 . The firand DR-5 . A caspaOrganisms regulate their development and maintain through sophisticated interplay between cells. During development, organisms produce excess cells that finally go through PCD and contribute to the formation of organic structures . In inteApoptosis contributes to maintaining the balance of homeostasis by normal cell death . Necrosihttp://www.genome.jp/dbget-bin/www_bget?hsa04210. The names of these genes are available in Supplementary Material I available online at http://dx.doi.org/10.1155/2015/715639.Previously known apoptosis-related genes were obtained from KEGG , a databhttp://www.string-db.org/) database [To identify the novel candidate genes related to apoptosis, we used protein-protein interaction information that was retrieved from the STRING . Additionally, a permutation test excluded false discoveries that had high betweenness and little relationship with apoptosis by calculating permutation FDR for each of 114 candidate genes and setting 0.05 as the threshold. We finally obtained 26 genes, which are listed in We finally obtained 26 significant candidate genes for apoptosis. The following paragraphs gave detailed discussion on the relationships between these genes and apoptosis.TRAF6  . Understanding the TNFR2-mediated apoptotic and JNK signaling pathways may offer novel therapeutic targets to treat vascular diseases in EC [ TRAF6 plays an important role in the signaling mediated by the TNF receptor superfamily. Therefore, we hypothesize that these two genes may have synergistic effects on apoptosis regulation, which requires further validation.es in EC . This obIQGAP1  and NFAT2 short isoform (CA-NFAT2/A) mutants localize in the nucleus, bind DNA with high affinity, and activate endogenouset genes , 42. Rem AKAP5 is directly related to apoptosis. We hypothesize that AKAP5 may be involved in cell apoptosis by forming complexes with protein kinases, phosphatases, or scaffold proteins. The assembly and localization of signaling complexes are coordinated by scaffold, anchoring, and adaptor proteins to provide efficiency and specificity in signal transduction [ AKAP5 is also linked to apoptosis. In addition, AKAP5 can interact with ADCY8 to regulate Ca2+-dependent c-AMP synthesis in pancreatic and neuronal systems [2+, resulting in the expression of genes such as CALM and CAMK that cause cell proliferation and apoptosis. PKA also inhibits phosphorylation of BAD and suppresses apoptosis. Further experimental verification is necessary to test these hypotheses.Betweenness of this gene was 195 and its permutation FDR was <0.002 refer to , row 7. sduction . It ther systems . In the DIABLO  and xTAK1 mRNAs injected into early embryos can result in cell death [β1 (TGF-β1), TAK1 activates JNK1 and p38 as an upstream MAP3 kinase. The XIAP/TAK1-mediated activation of JNK1 depends on TAB1, and the XIAP/TAK1-mediated activation of JNK1 is involved in protection against apoptosis [κB activation [ TAB1 plays a vital role in regulating apoptosis and survival, consistent with our expectation.The betweenness of this gene was 120 and its permutation FDR was <0.002 refer to , row 9. and JNKs , which cll death . Many inpoptosis . The propoptosis . In TRAItivation . Taken t CAPNS1 may indirectly be involved in the regulation of apoptosis. In addition, some recent studies have demonstrated that CAPNS1 participates in signaling pathways as a partner. In the Ras signaling pathway, CAPNS1 binds the RasGAP-SH3 domain in K-Ras (V12) oncogenic cells, modulating migration and cell survival, and the interaction between CAPNS1 and PP2A-Akt affects FoxO3A-dependent cell death [ CAPNS1 mutation. The muscular biopsy specimens of LGMD2A patients show that lack of calpain 3 causes IκBα accumulation and prevents NF-κB nuclear translocation, ultimately resulting in apoptosis. Moreover, deficiency in CAPN3 (betweenness: 86, permutation FDR: <0.002; refer to  CAPNS1 and CAPN3 interact or coordinately regulate apoptosis still must be studied.CAPNS1 (betweenness: 86, permutation FDR: <0.002; refer to ll death . In addi ADCY8 may serve as a bridge between extracellular stimuli and apoptosis.The betweenness of this gene was 167 and its permutation FDR was 0.002 refer to , row 12. NPRL3 in apoptosis in humans requires further research.The betweenness of this gene was 83 and its permutation FDR was 0.008 refer to , row 13. SMAD family. SMAD proteins can mediate multiple signaling pathways through their roles as signal transducers and transcriptional modulators and negatively regulate BMP and TGF-beta/activin-signaling. Members of the TGF-beta family regulate multiple cellular processes, including cell proliferation, differentiation, organization, migration, and death. In addition, SMAD6 and SMAD2 predict overall survival in oral squamous cell carcinoma patients. However, the role of aberrant TGF-beta signaling is not clear [ SMAD6 upregulates the plasminogen activator inhibitor-1 and phosphorylates SMAD2/3, finally activating TGF-beta signaling. Furthermore, because of the SMAD6 knockdown, the JNK pathway is also activated and the phosphorylation of Rb-1 is reduced, causing G0-G1 cell apoptosis and arrest [β, Bcl-xl and Bcl-2 were downregulated, whereas Bax was upregulated. Furthermore, the mRNAs of SMAD6 and SMAD7 displayed significant upregulation [β pathway, can be induced by TGF-β1; however, this process can be negatively regulated through the SMAD6 but not SMAD7. K63-linked poly-ubiquitination of TRAF6 can be abolished through the TGF-β1-induced SMAD6 in primary hepatocytes and AML-12 mouse liver cells. In addition, in cell culture or animal models, phosphorylation of TAK1 and p38 MAPK/JNK is maintained and apoptosis increased after knockdown of SMAD6 or A29, suggesting an important role of the SMAD6-A20 axis in negative regulation of the TGF-β1-TRAF6-TAK1-p38 MAPK/JNK pathway [β receptor/SMAD pathway in HepG2 cells. In this process, SMAD6 and SMAD7 expression levels both decreased [The betweenness of this gene was 25 and its permutation FDR was 0.014 refer to , row 14.ot clear . In addid arrest . Accordigulation . These r pathway . Recent ecreased . Taken t IRS-1 affects the PI3-kinase/Akt survival pathway, which in turn results in resistance to the antiapoptotic effects of insulin. In addition, the Arg972 polymorphism also impairs human β-cell survival [The betweenness of this gene was 238 and its permutation FDR was 0.016 refer to , row 15.survival . A reporsurvival , finally PKD2 (betweenness: 3, permutation FDR: 0.016; refer to  PKD2 is also responsible for transporting calcium signaling in renal epithelial cells. Calcium concentration changes can induce a series of cell biology processes to occur, such as activation of the MAPK signaling pathway [ PKD2 may regulate calcium cations through opening and closing calcium channels, thereby triggering physiological and pathological change and finally deciding cell fate.The protein encoded by pathway , eventua CSF2 (betweenness: 65, permutation FDR: 0.02; refer to  CSF2 is associated with apoptosis. CSF2 can block apoptosis in bovine embryos through interaction with genes controlling apoptosis [ GM-CSF promotes macrophage apoptosis and plaque necrosis through IL-23 signaling [The protein encoded bypoptosis . In addiignaling .BBC3 (betweenness: 8, permutation FDR: 0.02; refer to  BCL-2 family. These family members are also in the BH3-only proapoptotic subclass. This protein induces mitochondrial outer membrane permeabilization and apoptosis through cooperating with direct activator proteins. As mentioned above, DIABLO is released from mitochondria, which can be increased through mitochondrial outer membrane permeabilization enhancement. In addition, BBC3 was identified 12 years ago, mediates p53-dependent and p53-independent apoptosis, and is also involved in the intrinsic apoptosis pathway [ pathway . In the  pathway . NOTCH1 encodes a member of the NOTCH family. NOTCH signaling participates in maintaining the balance of cell proliferation, differentiation, and apoptosis; therefore, disorders in NOTCH signaling may induce tumorigenesis. The active form of NOTCH1, NOTCH1-ICN, is involved in many cell processes, such as T/B cell development and apoptosis, progression, and deterioration of various cancers. NOTCH1 induces resistance to glucocorticoid-induced apoptosis in developing thymocytes through downregulation of SRG3 expression [The betweenness of this gene was 249 and its permutation FDR was 0.022 refer to , row 19.pression . In addipression . Apoptospression . Moreovepression , 81. PTPN1 (betweenness: 93, permutation FDR: 0.024; refer to  PTP1B. PTP1B deficiency is also involved in protecting against Fas-induced hepatic failure [The protein encoded by failure . p53 in vitro. The apoptosis of primary cell lines is induced by overexpression of c-Maf via a p53-dependent mechanism [The betweenness of this gene was 54 and its permutation FDR was 0.026 refer to , row 21.echanism . In addiechanism . Taken t NFKB2 encodes a subunit of the transcription factor complex NF-κB. NF-κB significantly functions in regulating the immune response; however, in almost the same manner, it also induces proliferation, inflammation, and regulation of apoptosis [κB activity results in mitochondrial apoptosis after infecting cells with pathological prion proteins. In addition, NFKB2, as a subunit of NF-κB, is involved in the MAPK signaling pathway, which finally regulates proliferation, inflammation, and antiapoptosis. Nevertheless, these hypotheses require further validation.The betweenness of this gene was 8 and its permutation FDR was 0.034 refer to , row 22.poptosis . For exa CALM1 and apoptosis; however, calcium concentration changes can induce a series of cell biological processes to happen. Because calcium activates the MAPK and JNK signaling pathways, we presume that calcium activates CALM1, which in turn activates downstream gene expression, potentially including apoptosis-related genes, such as BCL-2, finally regulating cell survival and apoptosis. In addition, in the STRING analysis, CALM1 has interactions with IQGAP1 and ADCY8. As mentioned above, IQGAP1 and ADCY8 have been well characterized to participate in apoptosis. In summary, these arguments all support our results.CALM1 (betweenness: 244, permutation FDR: 0.038; refer to  CAMK2B is a member of the Ca2+/calmodulin-dependent protein kinase subfamily. CAMK2B is involved in several pathways, such as the melanogenesis pathway and the neurotrophin signaling pathway. Because CAMK2B is downstream of the gene CALM1, CALM1 phosphorylates and thereby activates CAMK2B. In addition, CAMK2B is also involved in the Wnt/Ca2+ pathway, which dephosphorylates the NFAT transcription factor family, which in turn induces the expression of genes such as CD40L, CTLA-4, and FasL, finally participating in the process of cell fate decision. Recently, research has demonstrated that CAMK2B protects neurons from homocysteine-induced apoptosis with the involvement of the HIF-1α signal pathway [The betweenness of this gene was 179 and its permutation FDR was 0.04 refer to , row 24. pathway . Cbl-b by shRNA resulted in strongly activating ERK, JNK, and p38 MAPK [CBLB (betweenness: 5, permutation FDR: 0.042; refer to p38 MAPK  and upreThe betweenness of this gene was 211 and its permutation FDR was 0.044 refer to , row 26.This contribution attempted to provide a better comprehension of apoptosis by identifying novel apoptosis-related genes. An existing computational method was applied with a weighted graph, constructed by protein-protein interaction information, to search for possible genes related to apoptosis. The analyses of the obtained genes further suggest that they are related to apoptosis.The Supplementary Material contains two files. In detail, the Supplementary Material I lists 86 human genes that are related to apoptosis; the Supplementary Material II lists 114 candidate genes discovered by our method and their betweenness and permutation FDRs.


Determining the driving factors and molecular flow-through that define the switch from favorable to aggressive high-risk disease is critical to the betterment of neuroblastoma cure.In this study, we examined the cytogenetic and tumorigenic physiognomies of distinct population of metastatic site- derived aggressive cells (MSDACs) from high-risk tumors, and showed the influence of acquired genetic rearrangements on poor patient outcomes.LOC100288142, CD1C, CFHR3, FOXP2, MDFIC, RALYL, CSMD3, SAMD12-AS1, and MAL2, and a loss in ADAM5, LOC400927, APOBEC3B, RPL3, MGAT3, SLC25A17, EP300, L3MBTL2, SERHL, POLDIP3, A4GALT, and TTLL1. QPCR analysis and immunoblotting showed a definite association between DNA-copy number changes and matching transcriptional/translational expression in clones of MSDACs. Further, MSDACs exert a stem-like phenotype. Under serum-free conditions, MSDACs demonstrated profound tumorosphere formation ex vivo. Moreover, MSDACs exhibited high tumorigenic capacity in vivo and prompted aggressive metastatic disease. Tissue microarray analysis coupled with automated IHC revealed significant association of RALYL to the tumor grade in a cohort of 25 neuroblastoma patients. Clinical outcome association analysis showed a strong correlation between the expression of CFHR3, CSMD3, MDFIC, FOXP2, RALYL, POLDIP3, SLC25A17, SERHL, MGAT3, TTLL1, or LOC400927 and overall and relapse-free survival in patients with neuroblastoma.Karyotyping in SH-SY5Y and MSDACs revealed trisomy of 1q, with additional non-random chromosomal rearrangements on 1q32, 8p23, 9q34, 15q24, 22q13 (additions), and 7q32 (deletion). Array CGH analysis of individual clones of MSDACs revealed genetic alterations in chromosomes 1, 7, 8, and 22, corresponding to a gain in the copy numbers of Together, these data highlight the ongoing acquired genetic rearrangements in undifferentiated tumor-forming neural crest cells, and suggest that these alterations could switch favorable neuroblastoma to high-risk aggressive disease, promoting poor clinical outcomes.The online version of this article (doi:10.1186/s12885-015-1463-y) contains supplementary material, which is available to authorized users. Neuroblastoma (NB) is the most common cancer of infancy . It origMYCN proto-oncogene amplification [NB is characterized by non-random chromosomal abnormalities with diagnostic and prognostic significance, including large-scale chromosomal imbalances –14. Tradfication , 21. MYCfication . Recent fication . The frefication , 25. IdeThis study used spontaneous and reproduced mouse models of aggressive human NB to document acquired genetic alterations in the NB cells, and further identified the gene manipulations orchestrated as a cause effect. We established clones of distinct populations with aggressive physiognomy (MSDACs), using tumors derived from multiple metastatic sites of various animals. These clones were examined for genetic rearrangements, cancer stem cell (CSC) status, and ability to prompt aggressive disease with systemic metastasis. Clinical outcome association studies in cohorts of neuroblastoma patients showed a strong association of these acquired genetic rearrangements with poor overall and relapse-free survival. For the first time, this study demonstrated the ongoing acquisition of genetic rearrangements and the subsequent switch from favorable NB to high-risk disease, identifying an association between genetic rearrangement, the switch to high-risk disease, and poor clinical outcomes.2 humidified incubator.The SH-SY5Y human neuroblastoma cell line was obtained from ATCC  and was cultured and maintained as described earlier . For pashttp://r2.amc.nl) to demonstrate the significance of altered genes in high-risk disease and their relevance to clinical outcomes. Neuroblastoma xenograft and/or aggressive metastatic disease development was performed as described earlier [2 asphyxiation. The tumors from metastatic sites and non-metastatic xenografts were harvested and prepared as single-cell suspensions as described earlier [All animal experiments conformed to American Physiological Society standards for animal care and were carried out in accordance with guidelines laid down by the National Research Council. All protocols were approved by the University of Oklahoma Health Sciences Center - Institutional Animal Care and Use Committee. However, Human data used were obtained from public database . Cells were maintained at 37 °C, 5 % CO2 for 72 h. We assessed formation of well-organized tumorospheres using phase contrast light microscopy. In parallel, we examined 1000 cells plated in 96-well culture plates with high-content real-time fluorescent time-lapsed video imaging as described earlier [We plated a total of 10 earlier .All cell preparations for cytogenetic analysis, karyotyping, and array CGH were performed in the Cytogenetic Molecular division of the University of Oklahoma Health Sciences Center Clinical Genetics Core. We harvested parental SH-SY5Y cells and MSDACs according to our laboratory standard protocols. Chromosomes were treated and stained by trypsin-Giemsa banding (GTG-banding). A total of 50 cells were analyzed and karyotyped (in a blinded fashion) from each clone of the cell line. For array CGH, genomic DNA was extracted from the parental SH-SY5Y, and aggressive MSDACs by the phenol-chloroform method with slight modifications, as described previously . A totalADAM5, A4GALT, APOBEC3B, CD1C, EP300, FOXP2, SLC25A17, L3MBTL2, MAL2, NBPF20, POLDIP3, RALYL, and SERHL (corresponding genes for the observed copy number variation) in SH-SY5Y and in clones of MSDACs grown ex vivo as described earlier [-actin as a positive control. A negative control without template RNA was also included. Each experiment was carried out in triplicate. The ΔΔCt values were calculated by normalizing the gene expression levels to β-actin. The relative expression level was expressed as a fold change over parental SH-SY5Y cells. Group-wise comparisons were performed with two-way ANOVA with Tukey’s post-hoc correction .We used real-time QPCR to analyze the transcriptional alterations of  earlier , 30. We Total protein extraction and immunoblotting were performed as described in our earlier studies , 24. ForGrade 1 or well-differentiated - Cells appear normal and are not growing rapidly; (ii) Grade 2 or moderately-differentiated - Cells appear slightly different than normal; (iii) Grade 3 or poorly differentiated - Cells appear abnormal and tend to grow and spread more aggressively. H & E stained TMA was reviewed for pathology. For this study, TMA-IHC staining was performed with Rabbit polyclonal RALYL (Abgent). The slides were micro-digitally scanned using an Aperio Scanscope  slide scanner and analyzed using integrated Spectrum software. RALYL nuclear positivity for the cores was then correlated with tumor grades. Group-wise comparisons were performed with GraphPad Prism.All tissue microarrays (TMA) IHC staining were performed in the Stephenson Cancer Center Cancer Tissue Pathology Core. To better characterize the correlation between acquired alterations of RALYL to the neuroblastoma progression in clinical subjects, we used a commercially available human neuroblastoma tissue array . The 5 μm thick human TMA is equipped with duplicate 1.5 mm cores of neuroblastoma tissues from various sites including the retroperitoneum, mediastinum, abdominal and pelvic cavities, and the adrenal glands of 25 patients. Further, the TMA is armed with clinical variables including sex, age, site/organ, diagnosis, and tumor grading. Pathology diagnosis classification includes: (i) http://r2.amc.nl) created by Dr. Jan Koster at the Academic Medical Center (AMC), Amsterdam, to examine the association of the observed genetic alterations with overall and relapse-free survival. This web-based application correlates a select gene expression profile with clinical outcomes for samples from various cohorts of patients.We used Ingenuity Pathway Analysis software  to examine the intermolecular interactions and the role that the genes with altered copy numbers and expression played in cancer progression. In this way, we were able to characterize the genetic rearrangements observed to accompany the functional biological response, defined here as tumor progression. We also used the R2: microarray analysis and visualization platform . This process is presumed to simulate the events of tissue regeneration and maintenance from cells that survive suspension conditions. In this process, an initial phase of symmetric expansion of the seeding stem cells precedes a phase of asymmetric division, which gives rise to the differentiated progeny that comprise the sphere bulk. Real-time high-content observation of MSDACs under controlled conditions showed an aggressive aggregation and tumorosphere formation within 18 h  xenografts as reported earlier [To better characterize the established high-risk aggressive disease model and to underscore the enrichment of select clones from the parental line or ongoing acquisition of genetic rearrangements, MSDAC clones were discretely characterized by karyotyping, whole genome array CGH analysis, and tumorosphere-forming capacity. MSDACs are relatively small and spherical with thin neurites. More importantly, every investigated clone of MSDACs exhibited intrinsic CSC characteristics per the ability to readily grow ospheres 8. This p earlier . The mic+, and compared these with the parental cells. All karyotyping was performed in double blinded fashion. We investigated at least 20 cells per clone. SH-SY5Y cells exhibited the 47,XX, add(1)(q32), +del(7)?(q32), add(8)(p23), add(9)(q34), add(15)(q24), add(22)(q13) [22Cancer cells are typically characterized by intricate karyotypes, including both structural and numerical changes. To determine and illustrate that the aggressive tumors developing in multiple metastatic sites were derived from the parental human SH-SY5Y cells, we karyotyped MSDACs, with and without characterized CD133CD1C, CFHR3, FOXP2, MDFIC, ADAM5, RALYL, CSMD3, SAMD12-AS1, MAL2, OR52N5, LOC400927, APOBEC3B, RPL3, MGAT3, SLC25A17, EP300, L3MBTL2, SERHL, POLDIP3, A4GALT, and TTLL1 genes.  roles in cancer progression related bio-functions including Cancer Cell Morphology, Progression of tumor, Cell Cycle-replicative senescence, Cellular Assembly DNA Replication, Cell Cycle arrest, Cell Death and Survival, Cellular Function and Maintenance, Post-Translational Modification, Cell-To-Cell Signaling, Cellular Assembly/Organization, Cellular Growth and Proliferation, Cellular Movement, Cellular Response to Therapeutics etc.,  that correlates a select gene expression profile with clinical outcomes for samples from multiple cohorts of patients with neuroblastoma. Kaplan-Meier plots showed a significant association between increased expression of CFHR3, MDFIC, CSMD3, FOXP2, or RALYL (genes with gains in coding regions) and poor OS in patients with neuroblastoma  demonstrated a definite association with their loss and poor OS  and relapse-free survival in patients with neuroblastoma. We utilized a web-based microarray analysis and visualization platform  that were common across the highly malignant MSDACs. Over the last two decades, identification of numerous oncogenes and tumor suppressors has aided the study of genetic alterations in cancer cells and helped us understand tumor progression and metastasis , 35. Outex vivo. Further, the results provide evidence of aggressive tumor-forming potential with multiple metastases of MSDACs in vivo. These outcomes illustrate the clonal enrichment of a select genetically modified, highly malignant sub-population disseminating to distant sites and promoting aggressive disease. To understand the acquired genetic rearrangements in these cells, array CGH coupled with QPCR and immunoblotting are ideal tools. Since cancer stem cells play an instrumental role in cancer relapse and tumor progression [Cytogenetic analysis now extends beyond the simple description of the chromosomal status of a genome, and allows the study of in-depth essential biological questions, including the nature of inherited syndromes, genomic changes involved in tumorigenesis, and three-dimensional organization of the human genome . The resgression , 41, exhCFHR3, MDFIC, CSMD3, FOXP2, or RALYL and loss in SLC25A17, POLDIP3, SERHL, LOC400927, MGAT3, or TTLL1 drive poor patient outcomes.Substantiating our findings of acquired genetic rearrangements in the development of aggressive disease, both G-banding and array CGH results confirmed the parental SH-SY5Y cell derivation. Human SH-SY5Y cells are a unique neuroblastoma line composed of N-type and S-type cells . Since tTo better understand how cancer cells acquire aggressive metastatic potentials, we must clarify the causative genetic alterations unique to cancer cells with metastatic ability , 38. ResCD1C, CFHR3, FOXP2, MDFIC, RALYL, CSMD3, SAMD12-AS1, MAL2, and OR52N5, and CNV loss in coding regions and associated regulation of ADAM5, LOC400927, APOBEC3B, RPL3, MGAT3, SLC25A17, EP300, L3MBTL2, SERHL, POLDIP3, A4GALT, and TTLL1. Highly malignant MSDACs that were derived from metastatic sites exhibited CSC status and exerted robust tumorosphere formations ex vivo. These MSDACs also initiated and reproduced high-risk aggressive metastatic disease in vivo. Clinical outcome association analysis recognized and identified a strong association with the gain in CFHR3, FOXP2, MDFIC, RALYL, or CSMD3 and loss in SLC25A17, SERHL, POLDIP3 LOC400927, MGAT3, or TTLL1 and poor overall and relapse-free survival. This study described the novel genetic alterations that accumulate during neuroblastoma progression, and defined the role of acquired genetic rearrangements in the clinical outcomes of children with high-risk aggressive metastatic neuroblastoma.In conclusion, the results of this study show an accumulation of genetic rearrangements in neuroblastoma cells that drive high-risk aggressive metastatic disease. Specifically, there is CNV gain in the coding regions and conforming expressions of


Coleoptera of New Brunswick have generated interest among entomologists for over a century. The first records of Coleoptera from New Brunswick were the adventive Carabusgranulatus Linnaeus and Carabusnemoralis Müller collected by W.H. Harrington in Saint John during 1891 : William McIntosh, Phillip R. McIntosh, A. Gordon Leavitt, and George Morrisey, mostly between 1898 and 1909  holdings in 2010, many apparently were either sent to other people or were lost to insect pests (Quediusmesomelinus (Marsham) (Staphylinidae) (Attagenusunicolorjaponicas Reitter (Dermestidae) (Ernobiusmollis (Linnaeus) (Ptiliidae) (Brachyperazoilus (Scopoli) (Curculionidae) (The ing 1891 . The firand 1909 . Most ofand 1909 . By 1914leoptera . Howeverct pests . Among tlinidae) , Attagenestidae) , Ernobiuiliidae) , Brachypionidae) , and othColeoptera were collected from the 1920s through 1940s by R.P. Gorham of the Dominion Entomological Laboratories. Another significant source of Coleoptera specimens was the Forest Insect and Disease Survey (FIDS) of the Canadian Forest Service (CFS) in Fredericton. This survey began in 1936 and was discontinued in 1996. Most of these specimens are in the collection at the Atlantic Forestry Centre (AFC) in Fredericton and the Canadian National Collection of Insects, Arachnids and Nematodes (CNC) in Ottawa. During the summer months of 1977 and 1978, scientists from the Biosystematics Research Center  collected Coleoptera during a survey of the invertebrate fauna at Kouchibouguac National Park. A report of this major undertaking summarized the most significant findings in the families Carabidae, Dytiscidae, Staphylinidae, Silphidae, Scarabaeidae, Lampyridae, Coccinellidae, and Chrysomelidae  at the Jacquet River Gorge Protected Natural Area (PNA), the Caledonia Gorge PNA, and the Grand Lake Lowlands PNA CFS - AFC) that used pitfall traps to investigate the effects of silvicultural practices on diversity and abundance of ground beetles in red spruce stands, 58 new provincial records and seven species of Staphylinidae new to science were discovered  in the following publications, adding 259 new records for New Brunswick, including eight new Canadian records, and one new North American record [number of new provincial records (NPR) or new Canadian records (NCR) in brackets]: Gyrinidae [4 NPR] . In 2003, Anne-Sophie Bertrand, then a graduate student at the Université de Moncton, collected runswick . Gaétan ands PNA . A signirunswick . In anotscovered . Many Cohis area . Majka e [4 NPR] , Carabid [6 NPR] , Halipli [6 NPR] , Histeri [2 NPR] , Ptiliidg 1 NCR] , Leiodid [8 NPR] , Lucanid [3 NPR] , Eucinet [1 NPR] , Clambid [4 NPR] , Eucnemi [1 NPR] , Throsciroscidae , Elateri[13 NPR] , Derodon [3 NPR] , Trogoss [3 NPR] , Clerida [3 NPR] , Melyrid [3 NPR] , Sphindi [2 NPR] , Erotyli [2 NPR] , Monotom [1 NPR] , Cryptopg 1 NCR] , Silvani [2 NPR] , Phalacr [2 NPR] , Kateret[28 NPR] , Cerylonylonidae , Endomyc [2 NPR] , 2009, Cnellidae , Corylop [2 NPR] , Latridi record] , Mycetop [3 NPR] , Tetrato [3 NPR] , Mordell [1 NPR] , Zopheripheridae , Tenebri[13 NPR] , Stenotr [1 NPR] , Oedemer [2 NPR] , Boridae [1 NPR] , Anthici [3 NPR] , AderidaAderidae , Scraptiaptiidae , Ischali [1 NPR] , Ceramby [2 NPR] , Chrysom [2 NPR] , Nemonycg 2 NCR] , 2008c.Carabidae, but later broadening his efforts to the Dytiscidae and other families in the early 2000s. Sampling, using a variety of methods such as sifting litter, hand sampling, sweeping, and light trapping, was done throughout the province, but was concentrated in the Fredericton and Charters Settlement area in York Co. and the Grand Lake area in Queens Co. Between 2006 and 2008, Webster in partnership with Stephen Clayden, NBM Curator of Botany, examined the beetle and lichen communities of old-growth New Brunswick cedar stands. This work revealed numerous species among both groups that were new to the region or that were new to science and led directly to the protection of several sites under the provincial Protected Natural Areas Act  to develop improved methods for survey and detection of exotic and potentially invasive bark and wood-boring beetles , many Coleoptera specimens were collected in Lindgren funnel traps. Vincent Webster, Chantelle Alderson, Colin MacKay, Marie-Andrée Giguère, Cory Hughes, Michelle Roy, and others collected and processed many of those samples. Experiments were conducted between 2009 and 2015 at sites throughout the province in most forest types, often in old or old-growth stands in Protected Natural Areas.Reginald Webster began intensively sampling beetles in New Brunswick in the early 1990s, initially focusing on the reas Act . WebsterGyrinidae [2 NPR]  to the provincial list. Dwayne Sabine reported the rare Cicindelamarginipennis Dejean for the first time from Canada from New Brunswick  in the following publications, adding 448 new records for New Brunswick, including nine new Canadian records:  [2 NPR] , Carabid[54 NPR]  Dytiscidg 1 NCR] ; Histeri[18 NPR] ; Geotrup[12 NPR] ; Eucinetg 1 NCR] ; Buprest [9 NPR] ; Dryopid [1 NPR] ; Eucnemi [9 NPR] ; Elateri[22 NPR] ; Lycidae [8 NPR] ; Dermest [5 NPR] ; Trogoss [2 NPR] ; Silvani [3 NPR] ; Sphindi [6 NPR] ; Kateret [8 NPR] ; Mycetop[10 NPR] ; Mordell [1 NPR] ; Tenebri [2 NPR] ; Stenotr [3 NPR] ; Cerambyg 4 NCR] ; Megalop[28 NPR] ; Anthribg 1 NCR] . In thesrunswick  and BousStaphylinidae of New Brunswick received relatively little attention prior to the publication of the first edition of the “Checklist of the beetles of Canada and Alaska”. Only 166 species of Staphylinidae, including 19 species in the subfamily Aleocharinae, were recorded from the province by Aleocharinae alone, Reginald Webster and coauthors Jan Klimaszewski, Christopher Majka, and others published a series of papers between 2001 and 2012 that included 183 new records from New Brunswick. Among these were 29 new Canadian records and 15 species new to science (NS), described, in many cases, from material from New Brunswick. These generic treatments are as follows: Placusa [1 NPR] Tinotus (1 NCR) (Silusa [2 NPR including 1 NCR)](Leptusa [1 NS] (Oxypoda [4 NPR] (Athetaacadiensis Klimaszewski and Majka [1 NS] (Amarochara  ] ] Schistoglossa  , 2 NS]  , Silusa g 1 NCR)], Leptusaa [1 NS] , Oxypoda [4 NPR] , Athetaa [1 NS] , Amaroch), 1 NS] , CaloderR, 2 NS] , DiglottR, 2 NS] , Gyropha [2 NPR] , Alisali), 2 NS] , Aleocha [1 NPR] , AleochaR, 4 NS] , AleochaAmerica] , Aleocha[12 NPR] , Aleochag 6 NCR] , 2012c.Staphylinidae, Chris Majka contributed 19 new records for New Brunswick in the following papers: introduced species [10 NPR]  (NCR) (Quediusspelaeus Horn [NPR] (Philonthushepaticus Erichson (NPR)  (NCR) , Quediusrn [NPR] , Philonton (NPR) . Webster [3 NPR] , Pselaphg 2 NCR] , Tachypog 1 NCR] , Scaphid [6 NPR] , Oxypori [5 NPR] , Paederi[17 NPR] , and Sta[44 NPR] .CNC, the number of species reported from New Brunswick nearly doubled from the 1,365 species reported in the first Checklist of the beetles of Canada and Alaska by Coleoptera fauna of New Brunswick.As a result of the above publications and additional data from material in the Aleocharinae in the genera Atheta, Clusiota, Dinaraea, Gnathusa, Mniusa, Ocyusa, and Mocyta to the faunal list of New Brunswick, based on new species descriptions and new records. Euaesthetus  added nine species to the provincial list, including three that were new to science, based in part on material collected in New Brunswick. Ochthephilus (Oxytelinae), described Ochthephilusashei Makanczy, based in part on a specimen from NB, and reported Ochthephilusforticornis (Hochhuth) and Ochthephilusplanus (LeConte) from the province, both of which were new provincial records. Paratenetus of North America, described Paratenetusexutus Bousquet and Bouchard (Tenebrionidae) from New Brunswick and included many localities from the province. Carabusauratusauratus Linnaeus (Carabidae) was newly recorded for Canada from New Brunswick by Buprestisconsularis Gory (Buprestidae) was added by Dryocoeteskrivolutzkajae Mandelshtam (Curculionidae) was reported for the first time for North America by Cryptophagussaginatus Sturm and Cryptophagussubfumatus Kraatz (Cryptophagidae) in a review of the adventive Cucujoidea of Canada. Most recently, Cerambycidae.Since the publication of In this special issue, 303 species and one new subspecies are newly recorded for New Brunswick. Among the new records are 32 species new to science, four new North American records, 21 new Canadian records, 270 new provincial records, and 45 adventive species. Three species were removed from the provincial list and one species was re-instated that was erroneously not included for New Brunswick by Coleoptera occurring in New Brunswick. This work would not have been possible to complete without the enthusiasm, determination, and professionalism of a small number of dedicated individuals who are acknowledged in the papers in this special issue. We hope that this special issue will generate a positive response and further interest in the Coleoptera fauna of New Brunswick and Canada, as many new discoveries await.It is important to remind ourselves that the understanding of biological diversity is not possible without taxonomic research, which is thought by many to be the foundation of biological science. Data on the mega-diversity of life and knowledge on species identity and distribution require discovery, description, cataloguing, and organization in order to be made accessible to a wide audience. This information constitutes a baseline of biological knowledge that is critical to support other branches of science. The present work provides these baseline data for the


Acute lymphoblastic leukemia (ALL) is a disseminated malignancy of B- or T-lymphoblasts which imposes a rapid and accurate diagnostic process to support an optimal risk-oriented therapy and thus increase the curability rate. The need for a precise diagnostic algorithm is underlined by the awareness that both ALL therapy and related success rates may vary greatly between ALL subsets, from standard chemotherapy in patients with standard-risk ALL, to allotransplantation (SCT) and targeted therapy in high-risk patients and cases expressing suitable biological targets, respectively. This review summarizes how best to identify ALL and the most relevant ALL subsets. Current standards for acute lymphoblastic leukemia (ALL) diagnosis integrate the study of cell morphology, immunophenotype and genetics/cytogenetics as detailed in the 2008 WHO classification of lymphoid neoplasms.A morphological bone marrow assessment represents the first step in the diagnostic pathway, for the primary diagnosis of ALL and for the differentiation from acute myeloid leukemia (AML),Table 1.7From the morphological point of view, there are no reproducible criteria to distinguish between B- and T-lineage ALL. It can also be difficult to distinguish B-lineage lymphoblasts from normal B-lineage lymphoid precursors, known as hematogones, which are observed in the peripheral blood in various conditions, including primary myelofibrosis and in children in the phase of recovery following chemotherapy(The bone marrow morphology of ALL is however quite variable as previously indicated in the FAB classification –2. Rare  and otheImmunophenotyping by means of multi-channel flow cytometry (MFC) has become the standard procedure for ALL diagnosis and subclassification, and was also developed as useful tool for the detection and monitoring of minimal residual disease . The consensus by European Group for the Immunological characterization of leukaemias (EGIL) is that a threshold of 20% should be used to define a positive reaction of blast cells to a given monoclonal antibody, except for MPO, CD3, CD79a and TdT, which are considered positive at the 10% level of expression.In B-lineage ALL the most important markers for diagnosis, differential diagnosis and subclassification are CD19, CD20, CD22, CD24, and CD79a. The earliest B-lineage markers are CD19, CD22 (membrane and cytoplasm) and CD79a.Among other B-cell markers, B-I and B-II ALL are often CD24 positive and 4G7 (pro- and pre-B surrogate light chain specific MoAb) positive;17T-cell ALL constitutes approximately 25% of all adult cases of ALL. T-cell markers are CD1a, CD2, CD3 (membrane and cytoplasm), CD4, CD5, CD7 and CD8. CD2, CD5 and CD7 antigens are markers of the most immature T-cell cells, but none of them is absolutely lineage-specific, so that the unequivocal diagnosis of T-ALL rests on the demonstration of surface/cytoplasmic CD3. In T-ALL the expression of CD10 is quite common (25%) and not specific; CD34 and myeloid antigens CD13 and/or CD33 can be expressed too. Recognized T-ALL subsets are the following: pro-T EGIL T-I , pre-T EGIL T-II , cortical T EGIL T-III  and mature-T EGIL T-IV . Finally, a novel subgroup that was recently characterized is represented by the so called ETP-ALL (Early-T Precursor), which shows characteristic immunophenotypic features, namely lack of CD1a and CD8 expression, weak CD5 expression, and expression of at least one myeloid and/or stem cell marker.18With currently refined diagnostic techniques the occurrence of acute leukemia of ambiguous cell lineage, i.e. mixed phenotype acute leukemia (MPAL) is relatively rare (<4%).CD56, a marker of natural killer (NK) cell differentiation, defines a rare subgroup of about 3% of adult ALL cases which often display other early T-cell antigens, CD7 CD2 CD5, and sometimes cCD3.With few exceptions, ALL is readily identified by morphological marrow assessment and MFC evaluation, with no need for additional tests, since genetics/cytogenetics and genomics are available at a later stage and cannot be employed for purely diagnostic purposes, even if they add very useful clinical-prognostic information. Differentiation between ALL and AML is initially obtained by excluding reactivity to SBB or MPO stains in ALL cells (<3% positive). On cytochemical evaluation, some rare ALL cases are SBB positive but MPO and chloroacetate esterase are negative. True ALL cases that are immunoreactive to MPO or express detectable levels of MPO mRNA have been described. This can occur in Ph+ ALL and occasionally in T-lineage ALL.25Cytogenetics represents an important step in ALL classification. Conventional karyotyping can be helpful in the identification of recurrent translocations, as well as gain and loss of gross chromosomal material; however, the major limitation of this technique is that in some cases leukemic cells fail to enter metaphase. However, fluorescence in situ hybridization (FISH) can enable the detection and direct visualization of virtually all investigated chromosomal abnormalities in ALL, with a sensitivity of around 99%. Finally, array-comparative genomic hybridization  and single nucleotide polymorphisms (SNP) arrays can permit the identification of cryptic and/or submicroscopic changes in the genome. Karyotypic changes found in ALL include both numerical and structural alterations which have profound prognostic significance.Among the good prognosis aberrations, it is worth mentioning del(12p) or t(12p)/t in B-lineage ALL, and t in T-ALL. These abnormalities are relatively rare in adults compared with childhood ALL.Aberrations associated with an intermediate-risk comprise the normal diploid subset plus cases with hyperdiploidy and several other recurrent or random chromosomal abnormalities.CRLF2.38Other aberrations, i.e. those with isolated trisomy 21, trisomy 8, and perhaps del(6q) and t/E2A-PBX1 may constitute an intermediate-high risk group; recent evidence suggests that the dismal outcome previously reported for the t/E2A-PBX1 is overcome by current therapeutic approaches.th decade of life.MLL1-rearranged ALL, for which outcome is very poor unless allogeneic transplantation is adopted.46Finally, patients with tor BCR-ABL1 rearrangements or a positive FISH test (Ph+ ALL), t or MLL rearrangements at 11q23, monosomy 7, hypodiploidy/low hypodiploidy (and the strictly related near triploid group) fall into the poor-risk cytogenetic category, with an overall disease-free survival (DFS) rate of about 25%, or 10% in the case of Ph+ ALL prior to the introduction of tyrosine kinase inhibitors (TKI).(MYC gene), such as t (90% of cases), t(10% of cases), and t (rarely observed), are virtually present in 100% of cases of mature B-ALL with L3/Burkitt morphology and clonal surface immunoglobulins. Typical cytogenetic aberrations are also found in T-lineage ALL.49Some other karyotypes are unique to specific ALL syndromes. Translocations involving chromosome 8 The integration of results of several techniques, i.e. gene expression profiling (GEP), SNP array analysis, and currently next-generation sequencing (NGS), have permitted a better definition of the molecular scenario of ALL and the identification of a constellation of novel mutations; as for the latter, however, caution must be shown, since while the biological role has been elucidated for some, while further investigation is required for others. These findings are detailed below , 4.IKZF1, encoding for the transcription factor Ikaros, is frequently disrupted in BCR/ABL+ ALL (80% of cases). IKZF1 deletions, that can be different in size, are predictors of poor outcome in Ph+ ALL,55CRLF2 (Δ–CRLF2), found exclusively in 5–10% B-ALL cases without known molecular rearrangementsCRLF2 and the Ig heavy chain locus (IGH@-CRLF2) or an interstitial PAR1 deletion that juxtaposes intron 1 of P2RY8 to the coding region of CRLF2 itself. More rarely, CRLF2 mutations can be detected. Δ-CRLF2 can be detected together with IKZF1 deletion in Ph-negative ALL patients and with JAK mutations (JAK1 or JAK2) or IL7R mutations; furthermore, they are identified in roughly 50% of children with Down syndrome;55Deregulated overexpression of IKZF1 deletions and CRLF2 rearrangements and adismal outcome. NGS has revealed the presence of mutations and/or rearrangements activating tyrosine kinases, i.e IGH-CRLF2, NUP214-ABL1 rearrangements, in-frame fusions of EBF1-PDGFRB, BCR-JAK2 or STRN3-JAK2 and cryptic IGH-EPOR rearrangements.65By the integration of genome -wide technologies, the “BCR/ABL-like” subgroup has been suggested/identified in both the adultNRAS, KRAS, FLT3 and NF1) can be detected in up to 70% of near haploid cases, whereas low hypodiploid cases are characterized by lesions involving members of the Ikaros family, particularly IKZF2, and by TP53 disruptions, that can be identified in 91.2% of these cases. In adult ALL, these cases are characterized by nonrandom chromosomal losses and the CDKN2A/B locus deletion as sole recurrent abnormality; as already reported in children, these cases frequently harbor TP53 mutations.67Hypodiploid ALL, regarded as a poor prognosis group, has been extensively evaluated in pediatric ALL:TP53 disruption has been also recently evaluated in childhood and adult ALL. In childrenTP53 mutations are identified at diagnosis in 8.2% of cases (11.1% T-ALL and 6.4% BALL), and are preferentially identified in cases without molecular aberrations, where they are detected in 14% of cases, and are associated with refractoriness to chemotherapy.CREBBP and its paralogue, EP300 (p300),NT5C2 mutations, which confer increased enzymatic activity on the NT5C2 protein, which normally dephosphorylates nucleoside analogs, such as mercaptopurine, used in consolidation and maintenance therapy, have been described. Other lesions identified by NGS in B-lineage ALL, are represented by mutations in PICALM-MLLT10, NUP214-ABL1 fusion formed on episomes, EML-ABL1, SET-NUP214 fusion and MLL gene rearrangements with numerous different partners. The prognostic significance of these lesions is uncertain.In T-ALL, well-recognized aberrations include the T-cell receptor (TCR) gene rearrangements, chromosomal deletions, and focal gene deletions .75–83 MoCEBPA, CEBPB, CEBPD), and a set of micro-RNAs . NGS has highlighted the presence of mutations usually found in acute myeloid leukemia ,ETV6 gene. Finally, these cases rarely harbor NOTCH1 mutations.Furthermore, the ETP subgroup and/or myeloid-like subgroup emerged as a grey zone between AML and TALL by applying genome-wide technologies.NOTCH1, FBW7, BCL11B, JAK1, PTPN2, IL7R and PHF6, beyond those identified in ETPs; some of them have recognized prognostic significance, whereas for others further studies are required. In fact, NOTCH1 and/or FBW7 mutations, which occur in more than 60% and about 20% of cases, respectively, are usually associated with a favorable outcome. In the light of this, a prognostic model has been recently proposed, defining as low-risk patients those who harbor NOTCH1 and FBW7 mutations, and as high risk those without these mutations or with lesions involving RAS/PTEN.JAK1 mutations, which increase JAK activity and alter proliferation and survival have been associated with chemotherapy refractoriness and should be considered as poor prognostic markers.94A large set of mutations  has beenBCL11B lesions, which can induce a developmental arrest and aberrant self-renewal activity;PTPN2- a negative regulator of tyrosine kinases-, mutations, often detected in TLX1 overexpressing cases, T-ALL, NUP214-ABL+ patients and JAK1 mutated cases;IL7Ralpha, that lead to constitutive JAK1 and JAK3 activation and enhancement of cell cycle progression;PHF6 mutations; PTPRC, encoding the protein tyrosine phosphatase CD45, usually detected in combination with activating mutations of IL7R, JAK1 or LCK, and associated with downregulation of CD45 expression;CNOT3, presumed to be a tumor suppressor; 7) mutations of RPL5 and RPL10, which impair ribosomal activity.NT5C2 mutations. 105Finally, another group of mutations/lesions is possibly involved in leukemogenesis, but their prognostic impact is either unknown or absent. They include: 1) Due to the reviewed evidence and the complexity of all the issues at play, it is recommended that adult patients with ALL should be treated within prospective clinical trials, which is the best way to ensure both diagnostic accuracy and therapeutic efficacy. In the context of a modern risk- and subset-oriented therapy, the early diagnostic work-up is of the utmost importance and therefore needs to be carried out by well trained and highly experienced personnel . As a fi


Transcript 1 encoded 362-amino acids (aa) wild type pGPR120-WT, which shared 88% homology with human short form GPR120. Transcript 1 was the mainly expressed transcript of pGPR120. It was expressed predominantly in ileum, jejunum, duodenum, spleen, and adipose. Transcript 3 (coding 320-aa isoform) was detected in spleen, while the transcript 2 (coding 310-aa isoform) was only slightly expressed in spleen. A selective agonist for human GPR120 (TUG-891) and PUFAs activated SRE-luc and NFAT-luc reporter in HEK293T cells transfected with construct for pGPR120-WT but not pGPR120-V2. However, 320-aa isoform was not a dominant negative isoform. The extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation levels in cells transfected with construct for pGPR120-WT were well activated by PUFAs, especially n-3 PUFA. These results showed that although pGPR120 had 3 transcripts, transcript 1 which encoded pGPR120-WT was the mainly expressed transcript. TUG-891 and PUFAs, especially n-3 PUFA, well activated pGPR120-WT. The current study contributed to dissecting the molecular regulation mechanisms of n-3 PUFA in pigs.The polyunsaturated fatty acids (PUFAs) receptor GPR120 exerts a significant impact on systemic nutrient homeostasis in human and rodents. However, the porcine GPR120 (pGPR120) has not been well characterized. In the current study, we found that  Free fatty acids (FFAs) are well known in serving as important nutrients to provide energy for the body and play important roles in various physiological regulation as signaling molecules –3. RecenIn human, mouse, and rat, GPR120 is expressed abundantly in intestinal tract where GPR120 mediates FFA-induced the secretion of the gut hormones (glucagon-like peptide-1 and cholecystokinin), thereby regulating appetite , 11. Addαq/11 is well known as a coupled G protein which can improve the cytoplasmic calcium released from endoplasmic reticulum and result in activation of pathways dependent on the calcium [αq/11 signaling in various cell types [2+]i and ERK1/2 activation are used as indexes to reflect the function of GPR120 i and ERK1/2 signaling. As expected, the specific agonist for human and mouse GPR120, TUG-891, can activate pGPR120-WT. However, the pGPR120-V2 almost showed negligible or no response to the small molecular agonist. It has been showed that, upon the stimulation of another human agonist GW9508, the hGPR120L also showed the poor response to the treatment while the hGPR120S can be well activated by this agonist [μM).The distinction between the same GPCRs from various species might cause the pharmacological variation . Therefo agonist . As show agonist . Compare agonist . This diAlternative splicing acts as a main mechanism for regulating the expression of GPCR genes and allowing for the protein synthesis of truncated isoforms . The truAs previously mentioned, in human and mouse, activation of GPR120 under the stimulus of the PUFAs could stimulate the ERK1/2 phosphorylation signal transduction cascade , 10, 17.Additionally, pGPR120-WT was also highly expressed in the adipose tissue and mature adipocyte, but not in the preadipocyte. This result matched well with the murine phenotype . Knockdo pGPR120 has 3 spliced variants, transcript 1 which encoded pGPR120-WT was the mainly expressed transcript of pGPR120. In addition, pGPR120 was also the only functional isoform, which was activated by synthetic agonist (TUG-891) and PUFAs, especially n-3 PUFA. Our study contributed to understanding the molecular regulation mechanisms of PUFA in pigs.In the current study, we cloned and characterized pGPR120 for the first time. We showed that pGPR120 was expressed predominantly in ileum, jejunum, duodenum, spleen, and adipose in pigs. Although


Staphylococcus aureus (MRSA) clinical isolates, hospital-associated perirectal isolate 32S (ST 239) from a colitis tracheostomy patient and community-associated MRSA isolate 42S (ST 772) from a hepatic-splenomegaly patient in Rawalpindi, Pakistan.Here, we report the draft genome sequences of two methicillin-resistant  Staphylococcus aureus is one of the leading human pathogens that causes a broad variety of infections, ranging from minor skin infections to life-threatening conditions, such as endocarditis, pneumonia, toxic shock syndrome, and bloodstream infections (–S. aureus (MRSA) isolates will enhance the understanding of their evolution and aid in rapid diagnosis, detection, and development of better therapeutic intervention strategies. The isolates described here were highly resistant to oxacillin  and ciprofloxacin , and also exhibited resistance to ampicillin, tetracycline, penicillin, erythromycin, gentamicin, and kanamycin. Sequence analysis of the quinolone resistance determinant region of the target gene gyrA indicated a double point mutation (S84L and G106D) in 32S and a single point mutation in 42S (S84L). The grlA gene exhibited E84G mutation in 32S and S80Y in 42S. RT-qPCR analysis of the efflux pump genes, norA, norB, norC, and mdeA, indicated an increased efflux pump expression in these isolates , adhesin genes , and virulence genes . Multilocus sequence typing (MLST) as described in Enright et al. (spa gene with the online spa-typing program (http://spatyper.fortinbras.us) revealed the spa type t030 for 32S and t5414 for 42S.Both of the isolates were positive for staphylococcal cassette chromosome type SCCt et al.  indicateS. aureus were extracted using a Master Pure Gram-positive DNA purification kit (Epicentre Biotechnologies). The library was prepared using a TruSeq DNA library preparation kit (Illumina) and the TruSeq paired-end cluster kit was used for the DNA preparation and cluster generation. The Illumina HiSeq 2500 system was used for sequencing. CLC Genomics Workbench 6.5.1 (Qiagen) was used for the de novo assembly of high-quality 100 bp paired-end reads and further genome annotation was performed using an annotation method GeneMarkS+ in the NCBI Prokaryotic Genome Annotation Pipeline  with 3,082 genes, 2,912 CDs, 110 pseudo genes, 37 frameshifted genes, 3 rRNAs , and 56 tRNAs. The draft genome sequence of S. aureus 42S has 32.7% G+C content, is 2,766,469 bp in length which is distributed in 86 contigs  with 2,816 genes, 35 frame-shifted genes, 63 pseudo genes, 2,692 CDs, 6 rRNAs , and 55 tRNAs.The draft genome sequence of JTJX00000000 and JTFJ00000000 for 32S and 42S, respectively. The versions described in this paper are versions JTJX02000000 and JTFJ01000000.This whole-genome shotgun project was deposited at DDBJ/EMBL/GenBank under the accession numbers