In contrast, there was no significant difference in the recovery of the G-MDSC subtype (Lin-HLA-DR-CD33+) following motolimod treatment. Additionally, treatment with either the TLR7 agonist imiquimod, or TLR9 agonist CpG ODN2006, did not result in a loss of the M-MDSC population. The decrease in M-MDSC population was found to be the result of these cells undergoing apoptosis following motolimod treatment. M-MDSCs were also found to increase FAS expression in response to TLR8 activation by motolimod. Studies to determine if motolimod can decrease the immunosuppressive and/or tumor-promoting activities of MDSC sub-populations are ongoing. Our finding, that TLR8-activated m-MDSCs undergo apoptosis, suggests the potential for using motolimod to modulate MDSC numbers and function in cancer patients and enable a more effective, immune response to tumors. This innovative approach for modulating m-MDSC populations holds great promise for augmenting anti-tumor directed immune responses.There is increasing evidence that myeloid-derived suppressor cells (MDSCs) contribute to the progression of cancer by inhibiting tumor-directed immune responses and producing mediators that promote the growth and survival of tumor cells. While present at low numbers in the peripheral blood of healthy individuals, levels of circulating MDSC increase in cancer patients as tumors produce factors that drive both the expansion and recruitment of these cells. MDSCs are a heterogeneous population and include both granulocytic MDSC (G-MDSC) and monocytic MDSC (M-MDSC) sub-populations, each of which can inhibit T cell function through arginase activity and the production of nitric oxide. Novel approaches to decrease MDSC expansion or inhibit their immunosuppressive functions hold great promise for augmenting tumor directed immune responses. In this study, we examined the effect of motolimod (formerly VTX-2337), a selective TLR8 agonist, on MDSC function. Initially TLR8 mRNA levels were assessed in MDSC isolated from the peripheral blood of healthy donors. TLR8 mRNA levels found in the M-MDSC subpopulation were comparable to those found in monocytes and myeloid dendritic cells (mDC), which express high levels of TLR8 protein and activate in response to motolimod. To further assess the M-MDSC response to TLR8 activation, PBMCs from healthy donors were treated with medium alone, low dose (167 nM), or high dose (500 nM) motolimod. Treatment with motolimod resulted in a significant, concentration–dependent loss of the M-MDSC (HLA-DR


BRCAness’, may lend to therapeutic exploitation in breast cancer. To this end, development and clinical evaluation of platforms to identify signatures of BRCAness are of immense interest. In this issue of Breast Cancer Research, Vollebergh and colleagues report that a BRCA-like array comparative genomic hybridization (aCGH) genomic instability signature is associated with benefit from high-dose cyclophosphamide-thiotepa-carboplatin chemotherapy. We discuss the strengths and weaknesses of this study and consider the clinical significance and applicability of this aCGH BRCAness signature in the context of other existing homologous recombination deficiency detection platforms.Inherited and acquired defects in homologous recombination, a phenotype termed ‘ Breast Cancer Research, Vollebergh and colleagues [BRCA1- and BRCA2-mutant breast cancers. The BRCA1 and BRCA2 genes are required for DNA double-strand break (DSB) and interstrand crosslink repair by homologous recombination (HR) [BRCA1 and BRCA2 mutation carriers are particularly responsive to agents that lead to DSBs  polymerase (PARP) inhibitors) [BRCA1 and BRCA2 account for only 5 % to 10 % of all breast cancers, these and other genes which operate in the HR pathway may be altered by mutation, rearrangement, DNA methylation, or attenuated mRNA expression, resulting in impairment of HR in a significant proportion of patients with breast cancer. Inactivation of HR by these mechanisms in BRCA-wild-type cancers (a phenotype termed ‘BRCAness’) may induce a similar hypersensitivity to DNA-damaging agents which can be therapeutically exploited. To this end, development and clinical evaluation of platforms to identify signatures of BRCAness have recently been a subject of intense investigation, especially in triple-negative breast cancer (TNBC), a subtype thought to be enriched for BRCAness [In this issue of lleagues  report oion (HR) . Pre-cliibitors) -6. AlthoBRCAness -9.90C) with four cycles of FE90C followed by high-dose cyclophosphamide-thiotepa-carboplatin (HD-CTC) with autologous stem cell support in patients with at least four positive axillary lymph nodes. The BRCA1- and BRCA2-like aCGH profiles were based on previous work by this group and were generated on 249 HER-2-negative formalin-fixed paraffin-embedded (FFPE) tumor samples [BRCA-likecgh profile was detected in 32 %  of the study cohort and predicted benefit from HD-CTC in both TNBC and hormone receptor-positive breast cancer. HD-CTC was associated with improved overall survival compared with FE90C in patients with BRCA-likecgh tumors but not in patients with non-BRCA-likecgh tumors. As expected, a significant proportion (76 %) of TNBCs harbored the BRCA-likecgh profile. Interestingly, 28 % of hormone receptor-positive tumors also demonstrated the BRCA-likecgh profile. This aCGH platform is attractive as it allows detection of genomic instability resulting from HR deficiency without requiring identification of the precise molecular etiology. If validated in other cohorts, this assay could potentially identify patients destined to benefit from intensified DNA-damaging therapy.Vollebergh and colleagues retrospectively assessed a subset of HER-2-negative tumor specimens from a randomized clinical trial that compared five cycles of 5-fluorouracil-epirubicin-cyclophosphamide , neither the experimental (HD-CTC) nor the control (taxane-devoid chemotherapy) regimens are considered standard for node-positive breast cancer in this era. Evaluation of this assay in cohorts treated with contemporary neoadjuvant chemotherapy regimens is encouraged.Despite the future potential of this aCGH platform as a predictive marker, the present study has important limitations. First, the HD-CTC regimen included both carboplatin and intensified alkylating chemotherapy (cyclophosphamide and thiotepa) and therefore it remains unclear whether the benefit observed in the BRCAness has not reached routine clinical application, but several functional assays besides the aCGH platform described by Vollebergh and colleagues have emerged. The homologous recombination deficiency (HRD) assay developed by Myriad Genetics, Inc.  evaluates tumor genome loss of heterozygosity, telomeric allelic imbalance (TAI), and large-scale state transitions, which are all indirect measures of tumor genomic instability. High HRD scores are highly correlated with defects in BRCA1/2 and are associated with sensitivity to neoadjuvant platinum-based chemotherapy in TNBC [BRCAness may exhibit characteristic gene expression patterns. Mulligan and colleagues [BRCA1/2 and Fanconi anemia mutations. The DDRDS is enriched for immune response-related genes and predicted favorable response to FE90C chemotherapy in both TNBC and hormone receptor-positive breast cancer. The aCGH, HRD, TAI, and DDRDS platforms are all compatible with FFPE tissues, making them suitable for prospective studies. Furthermore, all of these HR deficiency platforms suggest that the BRCAness phenotype exists in at least half of patients with TNBC and one quarter of patients with hormone receptor-positive breast cancer, implying future predictive applicability to a large fraction of patients with breast cancer.Currently, a standard platform for detecting  in TNBC ,11. Anot in TNBC . In addilleagues  recentlyThere is now renewed interest in the efficacy of platinum agents for the treatment of TNBC as recent randomized trials have demonstrated improvement in pathological complete response rates with the addition of neoadjuvant carboplatin to anthracycline/taxane-based chemotherapy -14. HR dEmploying functional measures of HR pathway deficiency rather than relying on documented changes in specific genes should capture more patients who might benefit from DNA-damaging therapies, and Vollebergh and colleagues should be congratulated on developing one such assay. However, before HR deficiency assays can be used for medical utility, the recognized steps for incorporation of predictive biomarkers into clinical care  have to be fulfilled . If apprBRCAness as a guide to personalize treatment with DNA-damaging agents in patients with breast cancer.In conclusion, considerable strides have been made in analytical development and early clinical validation of functional HR deficiency assays and these efforts bring us one step closer to the eventual utilization of


Drosophila olfactory learning, distinct subsets of dopaminergic neurons appear to assign either positive or negative value to odor representations in mushroom body neurons [Dopaminergic neurons provide value signals in mammals and insects . During  neurons . However neurons . Moreove •Flies trained with unpalatable sugar learn the sweet-, nutrient, and bad-taste qualities•Distinct dopamine neurons reinforce the positive and negative memories in parallel•Early conditioned aversion switches to longer-lasting nutrient-memory-guided attraction•Flies remember individual qualities of a complex food source Das et al. show that flies trained with unpalatable sugar learn both the nutritional and bad-taste qualities. Opposing memories are reinforced in parallel by distinct dopaminergic neurons. Conditioned behavior is initially aversive but soon switches to approach led by the longer-lasting nutrient-reinforced appetitive memory. DEET has been reported to drive aversive behavior in flies through olfactory  and gustFlies trained with only the sugar carrier showed a significant appetitive memory A. In conGr66a-GAL4-directed expression of cell-death genes [dTrpA1  and shaken to create a suspension and spread onto filter paper. Dried papers were rolled into training tubes. Training paradigms are indicated in the figure legends.A performance index (PI) was calculated as the number of flies approaching (appetitive) or avoiding (aversive) the conditioned odor minus the number going the other direction, divided by the total number of flies in the experiment. A single PI is the average of two experiments in which identical genotype flies are trained with the reciprocal reinforced/non-reinforced odor combination. Odors were 3-octanol and 4-methylcyclohexanol.Adult, food-deprived UAS-GCaMP3; THGAL4 flies were waxed to a custom-built imaging chamber, and the head capsule was removed under ice-cold sugar-free saline. Two-photon imaging was performed with a custom-made imaging setup as described .Statistical analyses were performed with PRISM (GraphPad Software) and are specifically referenced in the figure legends.


In a clinical study on ischemic heart disease, reverse redistribution of 99mTc-MIBI was evident after direct percutaneous transluminal coronary angioplasty. The presence of increased washout of 99mTc-MIBI was associated with the infarct-related artery and preserved left ventricular function. In non-ischemic cardiomyopathy, an increased washout rate of 99mTc-MIBI, which correlated inversely with left ventricular ejection fraction, was observed in patients with congestive heart failure. Increased 99mTc-MIBI washout was also observed in mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes (MELAS) and in doxorubicin-induced cardiomyopathy. Noninvasive assessment of cardiac mitochondrial function could be greatly beneficial in monitoring possible cardiotoxic drug use and in the evaluation of cardiac damage in clinical medicine.Mitochondria play an important role in energy production for the cell. The proper function of a myocardial cell largely depends on the functional capacity of the mitochondria. Therefore it is necessary to establish a novel and reliable method for a non-invasive assessment of mitochondrial function and metabolism in humans. Although originally designed for evaluating myocardial perfusion,  Mitochondria play an important role in energy metabolism and are integrally involved in embryonic development, cell signaling activities, cell-cycle control and cell death -2. Hence99mTc-MIBI) is a class of alkylisonitrile technetium compounds designed for noninvasive myocardial perfusion imaging. 99mTc-MIBI concentrates in mitochondria according to membrane potential  technetium-99m , a mitochondrial uncoupler, rapidly depletes cellular content of 99mTc-MIBI in the presence of tetraphenylborate   . Severalhy (DCM) . ElectromTc-MIBI . Therefo99mTc-MIBI single photon emission computed tomography (SPECT) in acute myocardial infarction patients with successful percutaneous transluminal coronary angioplasty. Reverse redistribution of 99mTc-MIBI was evident after direct percutaneous transluminal coronary angioplasty  average count ratio, and washout rate was calculated , a tracer to detect cardiac sympathetic nerve function, was associated with higher washout rate of 99mTc-MIBI in patients with heart failure (99mTc-MIBI to provide complementary information on severity of patients with heart failure (Both dilated and hypertrophic cardiomyopathies are often accompanied by changes in oxidative phosphorylation or respiratory enzyme activities in cardiac tissues . In non-stigated . A 740-Mlculated . Planar  results . An incr failure . Matsuo entricle . The was failure . MIBG sc failure .Quantitative gated single-photon emission computed tomography (QGS) can provide accurate and reproducible values for ejection fraction, regional wall motion, wall thickening and dyssynchrony -20. Alth99mTc-MIBI washout, which correlated inversely with left ventricular ejection fraction, was observed, whereas increased 123I-beta-methyl iodophenyl-pentadecanoic acid (123I-BMIPP) uptake was observed in the region of decreased 99mTc-MIBI uptake.Mitochondrial disorders are a heterogeneous group of diseases resulting from abnormalities in mitochondrial deoxyribonucleic acid and function. Cardiac involvement manifesting as hypertrophic  or dilated cardiomyopathy is frequently observed in mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes (MELAS) . For ins99mTc-MIBI washout rates were higher after the chemotherapy compared to before the chemotherapy in a malignant bone tumor patient with doxorubicin-induced cardiomyopathy  is one of the original anthracyclines isolated in the early 1960s from the pigment-producing bacterium Streptomyoces percetius, together with daunorubicin. However, doxorubicin has been known to exert many biochemical effects on myocytes, including damage to mitochondria, alteration of nucleic acids and protein synthesis, and lipid peroxidation after free radical generation. Its cardiotoxicity is a dose-dependent process resulting in myocyte damage that may cause congestive heart failure. Monitoring of patients and diagnostic procedures are important because its cardiotoxicity can be lethal. We reported that the myopathy . Quantitmyopathy . 99mTc-M without . Furtherpatients . Not onl99mTc-MIBI wash out rate needs to be established by further investigations.There may be several factors that affect the values of washout, including age, sex and sympathetic nerve function. The cut-off value between normal and abnormal mitochondrial function is not yet determined. Quantification of the 99mTc-MIBI outlined in this review suggest utility in assessment of cardiac mitochondrial function. Further studies are needed to fully establish nuclear imaging techniques as a noninvasive assessment of mitochondrial function.Noninvasive assessment of cardiac mitochondrial function could be of great benefit in monitoring effective drug use and in the evaluation of cardiac damage in clinical medicine. Although there are a limited number of clinical studies, important features of


Excessively activated microglia produce several pro-inflammatory enzymes and pro-inflammatory cytokines, leading to damage to surrounding neurons and eventually inducing neurodegeneration. Therefore, the inhibition of microglial overactivation may be a potential therapeutic strategy to prevent the further progression of PD. β-Hydroxybutyric acid (BHBA) has been shown to suppress lipopolysaccharide (LPS)-induced inflammation in BV-2 cells and to protect dopaminergic neurons in previous studies, but the underlying mechanisms remain unclear. Thus, in this study, we further investigated this mechanism in LPS-induced in vitro experiments, primary mesencephalic neuron-glia cultures were pretreated with BHBA and stimulated with LPS. [3H]dopamine (DA) uptake, tyrosine hydroxylase-immunoreactive (TH-ir) neurons and morphological analysis were evaluated and analyzed in primary mesencephalic neuron-glia cultures. In vivo, microglial activation and the injury of dopaminergic neurons were induced by LPS intranigral injection, and the effects of BHBA treatment on microglial activation and the survival ratio and function of dopaminergic neurons were investigated. Four our in vitro mechanistic experiment, primary microglial cells were pretreated with BHBA and stimulated with LPS; the cells were then assessed for the responses of pro-inflammatory enzymes and pro-inflammatory cytokines, and the NF-κB signaling pathway was evaluated and analyzed.For the 3H]DA uptake and loss of TH-ir neurons in the primary mesencephalic neuron/glia mixed culture. BHBA treatment significantly improved the motor dysfunction of the PD model rats induced by intranigral injection of LPS, and this beneficial effect of BHBA was attributed to the inhibition of microglial overactivation and the protection of dopaminergic neurons in the substantia nigra (SN). Our in vitro mechanistic study revealed that the inhibitory effect of BHBA on microglia was mediated by G-protein-coupled receptor 109A (GPR109A) and involved the NF-κB signaling pathway, causing the inhibition of pro-inflammatory enzyme (iNOS and COX-2) and pro-inflammatory cytokine  production.We found that BHBA concentration-dependently attenuated the LPS-induced decrease in [In conclusion, the present study supports the effectiveness of BHBA in protecting dopaminergic neurons against inflammatory challenge. Parkinson’s disease (PD) is the second most prevalent neurodegenerative disorder, affecting millions of people worldwide . A majorAccumulating evidence suggests that neuroinflammation plays an important role in the progression of PD ,10. Postin vitro and in vivo models of inflammation-mediated dopaminergic neurodegeneration are powerful tools in mechanistic studies and the identification of potential therapeutic agents.In mesencephalic neuron-glia cultures, the stimulation of microglia with inflammagen lipopolysaccharide (LPS) induces the production of factors, including TNF-α, IL-1β, IL-6, iNOS, and COX-2 ,17. Stud+) dopamine (DA) in Krebs-Ringer buffer . After washing three times with ice-cold Krebs-Ringer buffer, the cells were lysed in 1 N NaOH. A liquid scintillation counter  was used for measuring radioactivity. Nonspecific DA uptake observed in the presence of mazindol (10 μM) was subtracted.Cultures were incubated for 20 min at 37°C with 1 μM DA uptake. Morphologically, the remaining tyrosine hydroxylase-immunoreactive (TH-ir) neurons in the LPS-treated cultures had significantly fewer dendrites, and shorter or evenly truncated axons  compared with the vehicle-treated control cultures  treatment reduced the uptake capacity by approximately 70% of the vehicle control (P <0.01), and this LPS-induced reduction in [3H]DA was abated by pretreatment with BHBA in a concentration-dependent manner , suggesting that BHBA is devoid of obvious toxicity  before treatment with 10 ng/ml LPS. Seven days later, the degeneration of dopaminergic neurons was assessed by TH immunostaining and DA uptake and loss of TH-ir neurons in a primary mesencephalic neuron/glia mixed culture. In the current in vivo study, we investigated the motor dysfunction of these PD model rats using a rotational behavior assay. Because LPS was injected on one side of the SN, apomorphine-induced rotation to the lesioned side was used to evaluate the degree of damage to the dopaminergic system. Apomorphine-induced rotation significantly increased in the LPS-induced PD model rats, and BHBA showed therapeutic effects on this behavioral dysfunction. Further experiments demonstrated that BHBA inhibited LPS-induced microglial overactivation, pro-inflammatory factor release and dopaminergic neuronal damage. These data suggest that BHBA plays a neuroprotective role through inhibiting microglial overactivation.BHBA is an important intermediate of amino and fatty acid catabolism that has been reported to be effective in the treatment of a variety of inflammatory and autoimmune diseases, such as colonic inflammation and experimental allergic encephalomyelitis (EAE) ,46. A prtoxicity . In vivooduction . Soyeon 6/2 mice . To elucIn vitro experiments have demonstrated that BHBA inhibits pro-inflammatory cytokine production, LDL uptake, and chemotaxis in macrophages via activating GPR109A [In vivo experiments have shown that GPR109A mediates the therapeutic effects of DMF in EAE [GPR109A (PUMA-G in mice and HM74A in humans) is a seven-transmembrane G-protein-coupled receptor of the Gi family that is expressed mainly in white adipocytes and immune cells, such as monocytes and neutrophils . BHBA ha GPR109A . Moreove GPR109A . In vivoF in EAE . In thisWe further assessed the mechanism underlying the anti-inflammatory effect of BHBA in primary rat microglial cells and found that it significantly inhibited LPS-induced proinflammatory mediator production. The knockdown of GPR109A with siRNA resulted in the loss of this anti-inflammatory effect in primary rat microglial cells. Because NF-κB is clearly one of the most important regulators of pro-inflammatory gene expression , we examin vitro and in vivo. Several lines of evidence presented in this study demonstrate that BHBA provides potent neuroprotection to dopaminergic neurons against LPS-induced neurotoxicity through the regulation of GPR109A-mediated signaling pathways. Thus, GPR109A-mediated signaling pathways may represent potential targets for therapeutic intervention to prevent or slow the progression of PD.In conclusion, this study demonstrates that BHBA treatment improves LPS-induced behavioral dysfunction and protects dopaminergic neurons through inhibiting microglia-mediated neuroinflammation both


In contrast to donor T cells, natural killer (NK) cells are known to mediate anti-cancer effects without the risk of inducing graft-versus-host disease (GvHD). In order to improve cytotoxicity against resistant cancer cells, auspicious efforts have been made with chimeric antigen receptor (CAR) expressing T- and NK cells. These CAR-modified cells express antigen receptors against tumor-associated surface antigens, thus redirecting the effector cells and enhancing tumor-specific immunosurveillance. However, many cancer antigens are also expressed on healthy tissues, potentially leading to off tumor/on target toxicity by CAR-engineered cells. In order to control such potentially severe side effects, the insertion of suicide genes into CAR-modified effectors can provide a means for efficient depletion of these cells. While CAR-expressing T cells have entered successfully clinical trials, experience with CAR-engineered NK cells is mainly restricted to pre-clinical investigations and predominantly to NK cell lines. In this review we summarize the data on CAR expressing NK cells focusing on the possible advantage using these short-lived effector cells and discuss the necessity of suicide switches. Furthermore, we address the compliance of such modified NK cells with regulatory requirements as a new field in cellular immunotherapy. NK cells are of great clinical interest because they contribute to the graft-versus-leukemia/tumor (GvL/GvT) effect but are not responsible for the GvHD. However, the strong cytotoxicity of NK cells can be hampered by various tumor immune escape mechanism  or organ transplantation. Beside hematopoietic stem cells, dendritic cells, mesenchymal stromal cells, unselected T lymphocytes, and antigen-specific or regulatory T cells, alloreactive NK cells are currently getting into the focus of interest as suitable and powerful effector cells for cellular therapy of cancer . NK cellechanism . In ordeExtensive pre-clinical studies over the last decades have led to successful clinical phase I/II studies with CAR expressing T cells in hematological malignancies, including lymphoma, chronic lymphocytic leukemia (CLL), and acute lymphoblastic leukemia ALL; . Especiaex vivo expansion and NK differentiation of UCB-derived CD34+ cells has been successfully translated to the clinics (+ hematopoietic precursors to treat acute myeloid leukemia (AML) in elderly patients . In the last decade effective methods for clinical grade purification and expansion of donor NK cells from PB and PBSC have been established successfully in order to obtain large numbers of NK cells , unstimulated leukapheresis products (PBSC), umbilical cord blood (UCB), bone marrow (BM), human embryonic stem cells (hESCs) or induced pluripotent stem cells iPCSs; . While t clinics . An ongoNK cells . In thisNK cells  or donorNK cells . Dependi studies .Next to primary human NK cells, cell lines can also be useful for allogeneic NK cell therapy. Several human NK cell lines have been established, i.e., NK-92, HANK-1, KHYG-1, NK-YS, NKG, YT, YTS, NKL reviewed in Ex vivo expanded primary human NK cells produce a different spectrum of cytokines compared to T cells including γ-Interferon, IL-3 and the granulocyte macrophage colony stimulating factor . GM-CSF; . CAR-modSeveral of the pre-clinical studies with CAR-expressing NK cells focus on anti-CD19 and anti-CD20 CARs targeting B cell malignancies . Other pImportantly, also non-cancer targets can be eliminated by redirected NK cells. For example, an anti-EBNA3C CAR was used for the NK cell-mediated destruction of EBV positive T cells  and an aStable gene transfer is required to enable sustained CAR expression in expanding and persisting effector cells . CAR endRetro- or lentivirus-based vectors have mostly been used for CAR-engineered NK cells with various transduction efficacies. In this respect, numerous clinical trials using LTR -driven gamma-retroviral vectors for transduction of T cells have failed to show severe adverse events related to insertional mutagenesis reviewed in  underlinDifferent safety concerns are associated with CAR-engineered effector cells, which include induction of (acute) GvHD as well as on-target/off-tumor effects, tumor lysis syndrome and cytotoxicity to normal tissues due to limited selectivity of the chosen target antigen . To dateSuicide gene therapy was first introduced in the context of human SCT performed in the 1990s using the herpes simplex thymidine kinase (HSV-TK)/Ganciclovir (GCV) suicide system . AlthougdimKIR+CD62L+ cells have a lifespan up to months. Worth mentioning would also the finding, that IL15/4-1BBL activated NK cells could contribute to an acute GvHD caused by inflammation or other factors up-regulating activating ligand expression on non-malignant tissues . In this study, the donor-derived NK cells are expanded by co-culture with the irradiated K562 cell line expressing membrane bound IL-15 and 41BB ligand (K562-mb15-41BBL). Another study for pediatric and adult patients with refractory ALL is the recently approved study at the National University Hospital in Singapore . In this study, haploidentical NK cells will be activated by incubation with IL-2 and transduced with the same construct as employed in the St. Jude trial . A numbeAccording to their biological and pharmacological complexity CAR/suicide gene expressing NK cells will be classified as advanced therapy medicinal products (ATMP) in Europe and regulated either centralized or under the hospital exemption by the member states . Although the primary therapeutic effect remains NK cell-specific the significant alteration of targeting by introduction of CAR will result in a gene therapy medicinal product (GTMP), whereas lifespan control by engineered suicidality only would result in a somatic cell therapy medicinal product (CTMP) .Figure 1) will have to focus on the transduction efficacy, as well as on the safety and efficacy of the CAR and/ or the suicide constructs introduced. Therefore quality aspects related to CTMP and GTMP as defined in guidelines  will apply to the identity, potency, and activity including the conditional suicidality, purity, and safety of vectors and genetically modified product. The establishment of correspondingly adequate in-process and quality controls as well as of process target values and product specifications will have to take into account the variability of the primary effector cell as the starting material (For successful clinical translation of gene modified NK cells, the preclinical and clinical development (material .dimKIR+NKG2A-CD62L-NK cells, suicide genes might not be necessary, but this could be different for more immature CD56dimKIR+CD62L+ NK cells derived from cord blood or iPS cells. While theoretical risk-benefit considerations, only argue for the incorporation of suicide switches in the context of CAR-modified and/ or more immature NK cells, careful preclinical investigations are needed to provide a final answer in the future.To date suicide gene-modified T cells and CAR expressing T cells have successfully entered clinical trials. For future clinical applications, a combination of both modifications might lead to an improved safety strategy. In contrast to gene-modified T cells, experiments with modified primary human NK cells are mainly restricted to the pre-clinical setting with promising results. Importantly, the first two clinical studies using CAR-expressing NK cells have started very recently. Due to the limited lifespan of mature CD56The 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.


Mast cell infiltrates are associated with tumors, though their role in the tumor microenvironment remains unclear. Mast cells express high levels of KIT throughout differentiation and as mature cells. Mast cells in tumors have been shown to release proinflammatory cytokines and promote angiogenesis, increasing tumor growth and metastasis. KIT signaling and mast cells also appear to modulate myleloid-derived suppressor cell (MDSC) development, recruitment and activity in tumors. Furthermore, treatment of tumor-bearing mice or cancer patients with small molecule KIT inhibitors or an anti-mouse KIT antibody decreased tumoral MDSCs and other immunosuppressive cells including regulatory T cells. KTN0158 is a humanized anti-KIT IgG1 monoclonal antibody that specifically binds KIT and is being developed as a potential therapy for cancer and mast cell-related diseases such as neurofibromatosis type 1 (NF1). It binds canine, feline, non-human primate and human KIT with high affinity, but does not bind mouse or rat KIT. KTN0158 inhibits KIT signaling and function in vitro and in vivo and has shown antitumor activity in dogs with mast cell tumors expressing either wild-type or mutant KIT. Immune checkpoint inhibitors targeting the CTLA-4 and PD-1 pathways have demonstrated single agent efficacy in cancer patients, and the combinations have shown superior efficacy in some preclinical and clinical settings. To test the effects of KIT inhibition on immune tolerance in syngeneic mouse tumor models, the anti-mouse KIT monoclonal antibody ACK2 was used as a surrogate for KTN0158 and tested alone and in combination with CTLA-4 and PD-1 inhibitors. In the Colon26 model, single agent treatment with the anti-KIT antibody had little antitumor activity. However, the combination of anti-KIT and anti-CTLA-4 had substantial antitumor effects, comparable to those observed for anti-CTLA-4 plus anti-PD1. These antitumor effects were dose-dependent. Pharmacodynamic assessments will also be presented. Collectively, these data suggest that the combination of immunotherapies targeting KIT on innate immune cells and checkpoint pathway inhibitors may have clinical benefit.


The current special issue is the 2014 issue which includes 7 interesting papers.As one of the modalities of traditional oriental medicine, acupuncture has been widely used to treat many disorders and diseases including chronic pain, stroke, and insomnia as well as depression, while its mechanisms remain unclear. Enhanced production of reactive oxygen species causes oxidative stress leading to damage in lipids, proteins, and nucleic acids. Recent experimental studies have demonstrated that acupuncture could attenuate oxidative stress, which seems possible to explore the physiological antioxidative mechanism of acupuncture in various diseases.This special issue contains 7 papers, of which 5 articles study the antioxidative mechanism of acupuncture in some diseases by animal models. These studies suggested that acupuncture may result from antioxidation, anti-inflammation, and antiapoptosis effects in several kinds of diseases. Among these, one study is related to the effect of laser acupuncture on memory impairment, oxidative stress status, and the functions of both cholinergic and dopaminergic systems in hippocampus of animal model of Parkinson's disease. One study explores whether electroacupuncture reduces myocardial ischemia-reperfusion (I/R) injury and inflammatory responses through inhibiting early growth response (Egr)-1 expression via the extracellular signal-regulated protein kinase-1 and kinase-2 (ERK1/2) pathway in a mouse model of myocardial ischemia reperfusion. Besides, one study compares the effects of antioxidant interventions on the electrical potential difference between acupoints along the stomach meridian on human. This paper suggests a possible underlying mechanism of acupuncture involving superoxide removal. One study focuses on the emerging links between acupuncture and redox modulation in vascular dementia, Alzheimer's vascular dementia, Parkinson's disease and hypertension, which represents an important step forward in the research of acupuncture antioxidative effect.We are excited to explore the studies on the specific oxidative stress biomarkers and redox signaling cascades using oxidative stress-related assessments techniques should be particularly useful in generating new hypotheses to enhance our understanding of the mechanism of antioxidative effects of acupuncture.Cun-Zhi LiuCun-Zhi LiuShu-Feng ZhouShu-Feng ZhouSergio-Botelho GuimarãesSergio-Botelho GuimarãesWilliam Chi-shing ChoWilliam Chi-shing ChoGuang-Xia ShiGuang-Xia Shi


Over-expression of endosialin/CD248 (herein referred to as CD248) has been associated with increased tumor microvasculature in various tissue origins which makes it an attractive anti-angiogenic target. In an effort to target CD248, we have generated a human CD248 knock-in mouse line and MORAb-004, the humanized version of the mouse anti-human CD248 antibody Fb5. Here, we report that MORAb-004 treatment significantly impacted syngeneic tumor growth and tumor metastasis in the human CD248 knock-in mice. In comparison with untreated tumors, MORAb-004 treated tumors displayed overall shortened and distorted blood vessels. Immunofluorescent staining of tumor sections revealed drastically more small and dysfunctional vessels in the treated tumors. The CD248 levels on cell surfaces of neovasculature pericytes were significantly reduced due to its internalization. This reduction of CD248 was also accompanied by reduced α-SMA expression, depolarization of pericytes and endothelium, and ultimately dysfunctional microvessels. These results suggest that MORAb-004 reduced CD248 on pericytes, impaired tumor microvasculature maturation and ultimately suppressed tumor development.


High-density lipoprotein (HDL) is a lipoprotein which has anti-atherogenic property by reversing cholesterol transport from the peripheral tissues to liver. Low HDL-cholesterol (HDL-C) as well as high low-density lipoprotein-cholesterol (LDL-C) is associated with the development of coronary heart diseases (CHD). Various epidemiological studies have suggested that the development of CHD increase in individuals with less than 40 mg/dL of HDL-C. In spite of accumulation of evidences suggesting a significant association between low HDL-C and CHD, effects of dietary factors on HDL metabolism remained largely unknown. We reviewed published articles about effects of dietary fat intake on HDL metabolism. The substitution of fatty acids (FA) for carbohydrates is beneficially associated with HDL metabolism. Monounsaturated FA intake may not affect HDL-C. Trans-FA is significantly associated with reduction of HDL-C, and is also adversely related with total cholesterol/HDL-C. Fish oils consumption, especially docosahexaenoic acid consumption, may be favorably associated with HDL metabolism. Although plant sterols and stanols may not affect HDL-C, policosanol intake is associated with a clinically significant decrease in the LDL/HDL ratio. Atherogenic dyslipidemia is characterized as elevated serum levels of triglyceride (TG) and low-density lipoprotein-cholesterol (LDL-C), and low serum levels of high-density lipoprotein-cholesterol (HDL-C). Since HDL is an anti-atherogenic lipoprotein which plays a role in reversing cholesterol transport from the peripheral tissues to the liver, low HDL-C level induces the development of coronary heart diseases (CHD) , 2, and A significant influence of low HDL-C on CHD and CVD has been widely known; however, roles of dietary factors for HDL metabolism remained largely unknown. We reviewed published articles about effects of dietary factors, especially dietary fat intake on HDL metabolism. We regarded systematic reviews and meta-analyses as important articles in this review.Mensink et al calculated the effect of changes in carbohydrate and FA intake on serum lipids, reviewing 27 controlled trials published between 1970 and 1991  4-8]. T. T4-8]. Salas-Salvado et al reviewed the clinical trials on humans that evaluate how mixtures of conjugated linoleic acid (CLA) isomers administered as supplements or CLA-enriched products can affect plasma lipids, and they found that some of studies have observed that various CLA isomers decreases HDL-C . WendlanSchwingshackl et al analyzed long-term, randomized controlled trials (RCTs) to investigate the effects of MUFA on cardiovascular risk factors  9-11]. . 9-11]. Recently, TFA has been reported to adversely affect cardiovascular health, and Mozaffarian et al reviewed the evidence for effects of TFA consumption on CHD risks. They found that reduction of HDL-C by TFA consumption was observed in both controlled trials and observational studies . These eFish oils have been widely reported to be a useful supplement to reduce serum TG levels in individuals with hyperlipidemia; however, effects of fish oils on serum HDL-C remained obscure. Lewis et al reviewed all RCTs from 1994 to 2003 which addressed the efficacy of long-chain omega-3 FA for dyslipidemia, and they found that 10 studies reported long-chain omega-3 FA to be effective in the treatment of hypertriglyceridemia  12-16]..12-16]. Plant sterols and stanols have a similar chemical structure and function to cholesterol. Therefore, plant sterols and stanols ingestion are sometimes suggested to beneficially modify serum lipids. Plant sterols have a higher degree of absorption as compared with plant stanols. Talati et al performed a meta-analysis of RCTs to compare the effect of plant sterols with plant stanols on serum lipid levels in healthy individuals and also patients with hypercholesterolemia  17-20]..17-20]. The substitution of FA for carbohydrates is beneficially associated with HDL metabolism. MUFA intake may not affect HDL-C. TFA is significantly associated with reduction of HDL-C, and is also adversely related with TC/HDL-C, coronary risks. Fish oils consumption, especially DHA consumption, may be favorably associated with HDL metabolism. Although plant sterols and stanols may not affect HDL-C, policosanol intake is associated with a clinically significant decrease in the LDL/HDL ratio.


Genetic profiling represents the future of neuro-oncology but suffers from inadequate biopsies in heterogeneous tumors like Glioblastoma (GBM). Contrast-enhanced MRI (CE-MRI) targets enhancing core (ENH) but yields adequate tumor in only ~60% of cases. Further, CE-MRI poorly localizes infiltrative tumor within surrounding non-enhancing parenchyma, or brain-around-tumor (BAT), despite the importance of characterizing this tumor segment, which universally recurs. In this study, we use multiple texture analysis and machine learning (ML) algorithms to analyze multi-parametric MRI, and produce new images indicating tumor-rich targets in GBM.We recruited primary GBM patients undergoing image-guided biopsies and acquired pre-operative MRI: CE-MRI, Dynamic-Susceptibility-weighted-Contrast-enhanced-MRI, and Diffusion Tensor Imaging. Following image coregistration and region of interest placement at biopsy locations, we compared MRI metrics and regional texture with histologic diagnoses of high- vs low-tumor content (≥80% vs <80% tumor nuclei) for corresponding samples. In a training set, we used three texture analysis algorithms and three ML methods to identify MRI-texture features that optimized model accuracy to distinguish tumor content. We confirmed model accuracy in a separate validation set.We collected 82 biopsies from 18 GBMs throughout ENH and BAT. The MRI-based model achieved 85% cross-validated accuracy to diagnose high- vs low-tumor in the training set . The model achieved 81.8% accuracy in the validation set .Multi-parametric MRI and texture analysis can help characterize and visualize GBM’s spatial histologic heterogeneity to identify regional tumor-rich biopsy targets. Glioblastoma (GBM) represents one of the most genetically heterogeneous, resistant, and lethal of all human cancers ,2. WhileYet, securing tumor-rich biospecimens for genomic profiling remains a significant challenge. In their initial report, TCGA found that only 35% of submitted biopsy samples contained adequate tumoral content and/or genetic material . This loAdvanced MRI offers an adjunct to conventional imaging and should help delineate tumor-rich biopsy targets. Image-based features such as tumor cell density on diffusion-weighted imaging (DWI), white matter infiltration on diffusion tensor imaging (DTI), and microvessel morphology on perfusion MRI (pMRI) reflect key biophysical characteristics associated with tumor pathogenesis ,7–10. AdAlthough prior studies suggest benefits from combining imaging techniques, there is currently no system that integrates the diverse image-based phenotypes and rich textural data of multi-parametric MRI to quantify tumor content within biopsy targets. To address this need, we assess the feasibility of using machine learning (ML) to integrate an array of image-based texture features from pre-operative MRI to predict tumor-rich biopsies from both enhancing core and non-enhancing BAT in a cohort of GBM patients. Our overarching goal is to develop non-invasive correlates of histology that can facilitate image-guided biopsy and genomic profiling within the framework of individualized oncology.The unique contributions of this study are: 1) a new method to produce images of histologic tumor content within an individual GBM; 2) a new protocol that uses multiple machine learning and texture algorithms to analyze co-registered multi-parametric MRI; 3) model construction and cross-validation in a training cohort of patients, followed by validation in a de-novo group of patients; and, 4) a model with high accuracy predicting histologic tumor content in the non-enhancing, and traditionally problematic, brain-around-tumor (BAT) zone.We recruited patients with clinically suspected GBM undergoing preoperative stereotactic MRI for surgical resection. We confirmed the absence of previous treatment (including steroid administration) and excluded subjects with an estimated glomerular filtration rate <60 mg/min/1.72 m2. We obtained approval from the institutional review boards at Barrow Neurological Institute (BNI) and Mayo Clinic in Arizona (MCA) and obtained written and informed consent from each subject prior to enrollment.Our group used pre-operative conventional MRI, including T1-Weighted contrast-enhanced (T1+C) and T2-Weighted sequences (T2W), to guide stereotactic biopsies as previously described . In shorTissue specimens (target volume of 125mg) were flash frozen in liquid nitrogen within 1–2 min from collection in the operating suite and stored in -80°C freezer until subsequent processing. Tissue was retrieved from the freezer and embedded frozen in optimal cutting temperature (OCT) compound. Tissue was cut at 4 um sections in a -20 degree C cryostat (Microm-HM-550) utilizing microtome blade. Tissue sections were stained with hematoxylin and eosin (H&E) for neuropathology review. H&E slides were reviewed blinded to diagnosis by a neuropathologist and assessed for tumor content. Taking into account all visible cells , the percent tumor nuclei were estimated. Based on the TCGA criteria, we used the threshold of at least 80% tumor nuclei content to define tumor-rich  biopsy samples . The original DTI image DICOM files were converted to a FSL recognized NIfTI file format, using MRIConvert  of 0.1 mmol/kg was administered to minimize T1W leakage errors. After PLD, we employed Gradient-echo (GE) EPI  for 3 minutes. At 45 sec after the start of the DSC sequence, we administered another 0.05 mmol/kg i.v. bolus Gd-DTPA ,17. The www.itk.org) and IB Suite (Imaging Biometrics) as previously described [For image coregistration, we employed tools from ITK (escribed . BecauseA board-certified neuroradiologist (LH) placed square (8x8 voxel) regions of interest (ROIs) at the locations corresponding to each biopsy site. For all analysis methods, we extracted ROIs from original images and analyzed each independently. Prior to texture analysis, we acquired first order statistics from raw image signals: average and standard deviation (SD) of gray-level intensities. Next, intensity values within each extracted ROI were mapped onto the range 0–255. This step helped standardize intensities between ROIs and reduced effects of intensity non-uniformity on features extracted during subsequent texture analysis. In this study, we incorporate 3 separate but complementary texture algorithms with multi-parametric MRI to characterize GBM’s regional variability in histologic tumor content –14. Mult0, 450, 900, and 1350) to produce a set of 13 rotationally invariant features: entropy (matrix randomness), energy/angular second moment , homogeneity (uniformity of co-occurrence matrix), dissimilarity (measurement of how different each matrix element is), and correlation (measurement of gray-tone linear dependencies) [GLCM provides detailed gray scale data by describing the angular relationships and distances between neighboring image voxels with similar gray scale intensities . Commonldencies) . In totaLBP provides highly discriminatory rotational and illumination invariant structural information by labeling each image voxel (in binary fashion) as higher or lower intensity compared with neighboring voxels . Highly DOST directly measures local spatial frequencies, which have shown specific applicability to neurologic diseases such as glioma ,14. The With a total of 256 image-based features from each ROI and corresponding biopsy sample, the dimensionality of the image-based features far exceeded the sample size, increasing the risk of overfitting artifacts and creating challenges for classification. To address this, we used Principal Component Analysis (PCA) which identifies the linear combination of features, called Principal Components (PCs), to reduce the dimensionality of the imaging data –21. HereS1 Appendix). Thus, we henceforth report the results from the DLDA analysis.We used classification algorithms with sequential forward feature selection to identify the subset of image-based PCs (determined from PCA above) with the greatest combined discrimination for biopsy tumor content (high- vs low-tumor). To represent the spectrum of classification methodologies, we separately applied 3 distinct but commonly described classification algorithms: Diagonal Linear Discriminate Analysis (DLDA), Diagonal Quadratic Discriminate Analysis (DQDA) and Support Vector Machines (SVM) –24. As tTable 1. The vast majority of biopsies (90%) were separated by >1cm. Overall, 59.2% of tissue samples from ENH demonstrated high tumor content, similar to the frequency of adequate tumor samples reported by the TCGA (60.2%). When samples originated from BAT, a much lower percentage (21.2%) of samples demonstrated high tumor content.In total, we collected 82 biopsy samples from 18 GBM patients. We describe the distribution of training  and validation  biopsy samples throughout enhancing core (ENH) and BAT in Table 2lists selected features and the cross-validation (CV) accuracy for DLDA, DQDA and SVM. Specifically, DLDA with sequential forward feature selection, identified 3 MRI-based texture features that optimized the classification model to distinguish high- vs. low-tumor biopsy samples (85% accuracy). The three features are: raw measure of rCBV, GLCM feature from EPI+C and LBP feature from T1+C. In general, rCBV provides non-invasive measures of microvascular volume, which link closely to histologic tumor content, malignant potential, and prognosis [rognosis ,16,17. Trognosis . Finallyrognosis .Table 1for both enhancing core (ENH) and BAT. In the validation set, model classification achieved an overall accuracy (81.8%) similar to that in the training dataset. Importantly, the model achieved a high degree of accuracy in non-enhancing BAT , which represents a problematic area for conventional CE-MRI-guided biopsies [Fig 1. Ten percent of training set biopsies (n = 6) were located within 5-10mm of adjacent biopsies. To test the potential confounding effects on accuracy, we performed a subanalysis that excluded these samples, as shown in S2 Appendix. With 54 total samples, the model achieved an accuracy of 85.2%, nearly identical to the main analysis, suggesting that the effects are negligible.After completing the MRI feature selection on the training dataset (n = 60), we applied the optimized model  to classify high- vs. low-tumor content biopsies in a separate, retrospectively collected validation set . The CV accuracy of the training set and the accuracy of applying the training model on the separate validation set are listed in biopsies ,5,7,25. Genetic profiling represents the future of neuro-oncology but hinges on the recovery of adequate biopsy material from representative tumor segments ,6. WhilePrevious glioma studies have used advanced MRI-guided stereotactic biopsies to validate the correlations with spatially matched regional histology. Barajas et al. compared DWI and pMRI measurements with tumor cell density, among other histologic indices , in ENH and BAT biopsies . CompareBased on the correlations in our training set, we have used machine learning (ML) to identify three MRI-based features that optimize classification accuracy for high- vs. low-tumor content . Mean rCFig 2. Also, the NPV  of the ML model would suggest a high degree of confidence in excluding tumor-rich populations in tumoral subregions, thereby improving surgical safety by preventing unnecessary biopsies. Besides biopsy guidance, the ML-based MRI texture model also presents applicability for improving the extent of maximal surgical resection, which correlates strongly with patient survival [Overall model accuracy  suggests that the ML-based MRI texture model can guide surgical biopsies to improve recovery of tumor-rich biospecimens compared to current CE-MRI methods. Particularly encouraging is the high model accuracy for non-enhancing BAT . Even in the subgroup analysis, which excludes 6 samples within 5–10 mm of each other, the ML-based model achieves high accuracy in BAT (90.5%). Based on current limitations of CE-MRI and low prevalence of tumor-rich foci in BAT , neurosurgeons would need approximately 5–7 biopsy attempts to recover one adequate tumor-rich sample (≥80% tumor nuclei) . The ML-survival . CurrentFig 2).A number of studies have shown the utility of intraoperative fluorescence microscopy with 5-aminolevulinic acid (5-ALA) to facilitate gross total resection (GTR) of enhancing core (ENH), which currently represents the primary target of surgical biopsy and resection ,28. The In this study, we define high tumor content (≥80% tumor) based on published TCGA criteria for sample adequacy . This thWe used Principal Component Analysis (PCA) to help interpret the high dimensional data that results from combining multiple texture algorithms and complementary MRI contrasts. Specifically, PCA identifies linear combinations of features, called Principal Components (PCs) that account for most variability across the original features –21. ThisS2 Appendix) excluded biopsies located within 5-10mm of adjacent biopsies and results remained consistent suggesting a negligible impact of this limitation.We recognize potential limitations. First, while this study demonstrates proof of concept that multi-parametric MRI and texture analysis can facilitate tumor localization, we recognize the need to validate these findings in a larger patient population. Second, we classify the adequacy of tumor content based on well-established guidelines, which ensure the quality of DNA isolation and the integrity of genetic analysis. Nonetheless, the requirements for tumor content may change as genetic sequencing technology improves, and future studies may prove necessary to evaluate the accuracy of MRI texture based on other histologic criteria and/or tumor thresholds. Third, image distortions and brain shift following craniotomy could also lead to misregistration errors. To compensate, neurosurgeons used small craniotomy sizes to minimize brain shift and also visually validated stereotactic image location with intracranial neuroanatomic landmarks to help correct for random brain shifts. Rigid-body coregistration of stereotactic and DSC-MR imaging also helped reduce possible geometric distortions ,16,17. OS1 AppendixShown are classification accuracies and selected MRI features based on DLDA, DQDA, and SVM classification methods. Incremental accuracy gains are listed for each MRI feature and for each classification method.(DOCX)Click here for additional data file.S2 AppendixAfter excluding 6 biopsy samples (within 5–10 mm of each other) from the training dataset, the table shows the distribution of biopsy samples by tumor content (high- vs. low-) for enhancing core (ENH) and non-enhancing BAT in both training (n = 54) and validation (n = 22) datasets. Test accuracies  for the optimized model (using the 3 MRI-based features in (DOCX)Click here for additional data file.


Single-shot (SSH) pulse sequences in CMR are beneficial for rapid image acquisition that is robust to motion, especially in arrhythmic patients or poor breath-holders. However, this fast scanning technique trades scan time for a lower signal-to-noise ratio compared to conventional multi-shot acquisitions. Here we propose a motion-compensated denoising technique that improves the image quality from multiple free-breathing single-shot acquisitions.Acquisition - In 7 patients with suspected cardiovascular disease, 4 repetitions of SSH Late Gadolinium Enhancement (LGE) were acquired in free-breathing 7-10 minutes after Gd injection [Reconstruction - The proposed method consists of 3 steps: 1) SENSE reconstruction of the undersampled data, 2) extraction of the motion that occurred between the repetitions with a non-rigid registration, and 3) motion compensated joint denoising of the whole dataset with Beltrami regularization, which offers an ideal compromise between feature preservation and staircasing reduction [Example reconstructions results are shown in Fig We introduced a new joint denoising and motion correction algorithm designed for fast, robust cardiac imaging. By iteratively incorporating the estimated motion into the reconstruction process, we increase the robustness of the model and exhibit good quality imaging. A limitation to the method is that potential through-plane motion cannot be corrected, although it remains negligible compared to the slice thickness. Ultimately, this method could be beneficial to both cardiac exam workflow and patient comfort since it allows robust image quality without the need for breath-holds.This work was supported by the European Commission, through Grant 605162.


Activator protein-1 (AP1) is a transcription factor that consists of the Jun and Fos family proteins. It regulates gene expression in response to a variety of stimuli and controls cellular processes including proliferation, transformation, inflammation, and innate immune responses. AP1 binds specifically to 12-O-tetradecanoylphorbol-13-acetate (TPA) responsive element 5′-TGAG/CTCA-3′ (AP1 site). It has been found constitutively active in breast, ovarian, cervical, and lung cancers. Numerous studies have shown that inhibition of AP1 could be a promising strategy for cancer therapeutic applications. The present in silico study provides insights into the inhibition of Jun-Fos-DNA complex formation by curcumin derivatives. These derivatives interact with the amino acid residues like Arg155 and Arg158 which play a key role in binding of Jun-Fos complex to DNA (AP1 site). Ala151, Ala275, Leu283, and Ile286 were the residues present at binding site which could contribute to hydrophobic contacts with inhibitor molecules. Curcumin sulphate was predicted to be the most potent inhibitor amongst all the natural curcumin derivatives docked. C-fos and c-jun genes are autoregulated; the transcription of c-jun is stimulated by its own product, and in contrast c-fos is negatively autoregulated isoxazol-6-yl)methoxy)phenyl)propanoic acid] bound to Jun-Fos complex with Δectively . The binectively . When doectively . When doectively .We observed that curcumin derivatives form polar contacts preferentially with residues like Arg155, Arg158, Lys276, Arg279, Lys280, and Lys282 when docked to DBR of Jun-Fos complex amongst which Arg155 and Arg158 are the key residues by which Jun-Fos complex binds to DNA. The results suggested that interaction of curcumin derivatives with residues like Arg155 and Arg158 could be the possible mechanism by which curcumin derivatives inhibit Jun-Fos-DNA complex formation. Ala151, Ala275, Leu283, and Ile286 were the hydrophobic residues present at binding site contributing to hydrophobic contacts with inhibitor molecules. The present molecular docking study provides insights into the inhibition of Jun-Fos-DNA complex formation by curcumin derivatives. The involvement of residues like Arg155, Arg158, Lys276, Lys280, and Lys282 seems to play a key role in binding of curcumin derivatives to Jun-Fos complex through polar contacts which prevents its binding to DNA (AP1 site). Ala151, Ala275, Leu283 and Ile286 were the important hydrophobic residues present at binding site. Most of the curcumin derivatives were predicted to be more potent than inhibitors like resveratrol and dihydroguaiaretic acid. Curcumin sulphate was predicted to be the most potent inhibitor amongst all the natural curcumin derivatives docked.


We can predict how an object would look like if we were to see it from different viewpoints. The brain network governing mental rotation (MR) has been studied using a variety of stimuli and tasks instructions. By using activation likelihood estimation (ALE) meta-analysis we tested whether different MR networks can be modulated by the type of stimulus (body vs. non-body parts) or by the type of tasks instructions (motor imagery-based vs. non-motor imagery-based MR instructions). Testing for the bodily and non-bodily stimulus axis revealed a bilateral sensorimotor activation for bodily-related as compared to non-bodily-related stimuli and a posterior right lateralized activation for non-bodily-related as compared to bodily-related stimuli. A top-down modulation of the network was exerted by the MR tasks instructions with a bilateral  network for motor imagery- vs. non-motor imagery-based MR instructions and the latter activating a preferentially posterior right occipito-temporal-parietal network. The present quantitative meta-analysis summarizes and amends previous descriptions of the brain network related to MR and shows how it is modulated by top-down and bottom-up experimental factors. Imagining scenes, sounds and actions, in the absence of appropriate stimuli for the relevant perception, takes place through mental imagery  or the viewer's position . We considered also the reference frame effect, where it was possible, i.e., as indicated by the authors' instructions. For instance, all the studies in which a mental change of the whole body position in space (a self-rotation) is required  meta-analyses . Based on previous neuropsychological studies , the Web of Knowledge database (www.webofknowledge.com), and the Sleuth on-line database (http://brainmap.org), for functional neuroimaging experiments that investigated MR processes. Moreover, the literature cited in the selected papers and reviews was also searched for additional neuroimaging studies on MR. The included studies were PET or fMRI experiments carried out on healthy subjects. Studies involving pharmacological trials or clinical populations were excluded. They were analyzed by means of a random-effects analysis. Analyses based on regions of interest (ROIs) of functional localizers were excluded. All single-cases studies were excluded, too. In addition, only studies which reported the coordinates in a standard reference space  were considered. Differences in coordinate spaces  were accounted for by transforming coordinates reported in Talairach space into MNI coordinates using a linear transformation , pathological subjects were included (N = 39), pharmacological treatment was performed (N = 10), ROI analyses were carried out (N = 17), children (under 18) were included (N = 9), single cases were included (N = 5) and other  through verbal instructions).Accordingly we included studies involving bodily-related stimuli  and non-bodily related stimuli , motor imagery based MR  and visuo-spatial imagery based MR . Studies included for each of these categories are indicated in Table The reported coordinates for functional activation in each category were analyzed for topographic convergence using the ALE method.p < 0.05 (cluster level corrected for multiple comparisons) , the reported studies were grouped as i) bodily-  and non-bodily  related stimuli. In addition, we included an analysis comparing MR of hands and MR of body stimuli. In Table In a third analysis, we explored how the MR network is influenced by egocentric/motor-imagery based strategies and allocentric/visual-imagery based strategies. In addition, we included an analysis by distinguishing the strategy from the reference frame variable. We thus compared motor-imagery based MR, see in Table Lastly, we included an additional analysis comparing MR of single stimulus or pairs of stimuli  inferior and superior parietal lobule bilaterally; (ii) left precentral gyrus; (iii) inferior frontal gyrus bilaterally; (iii) middle frontal gyrus bilaterally; (iv) SMA; (v) left insula; (vi) inferior and middle occipital gyrus bilaterally, and (vii) cerebellum bilaterally  cerebellum bilaterally and left calcarine cortex, (ii) left inferior parietal lobe, right angular gyrus, left superior parietal lobe and right postcentral gyrus Areas 2, 3b, and 4p), (iii) right insula, left superior frontal gyrus and middle cingulate cortex. The reverse contrast (non-bodily related > bodily-related stimuli) included exclusively right-lateralized activations in the: (i) middle occipital gyrus, (ii) cuneus, and (iii) superior parietal cortex  cerebellum, middle and inferior occipital and calcarine gyrus, (ii) superior parietal lobule, postcentral gyrus Area 2) bilaterally, left postcentral gyrus (Area 1), left inferior parietal lobe and right supramarginal gyrus, (iii) left precentral gyrus and inferior frontal gyrus (pars opercularis) bilaterally, left superior frontal gyrus, right middle frontal gyrus and posterior frontal gyrus medially, in addition to the right insula (see Table  bilateraMR of non-bodily related stimuli included activations in the: (i) middle occipital gyrus bilaterally, left inferior occipital gyrus, left cerebellum and left inferior temporal gyrus, (ii) right superior and inferior parietal lobe, and inferior frontal gyrus (pars opercularis and triangularis) bilaterally, in insula and the middle frontal gyrus in addition to the right precentral gyrus and the posterior frontal gyrus medially  right postcentral gyrus , (ii) left postcentral gyrus  extending to the inferior parietal lobe, (iii) left superior parietal lobe and iv) the right angular gyrus. The reverse contrast  included activations in the: (i) left middle occipital gyrus, left inferior occipital gyrus, left inferior temporal gyrus, and right superior occipital gyrus, (ii) right precuneus, (iii) left superior parietal lobe, and (iv) right posterior medial frontal gyrus and right superior frontal gyrus  cerebellum bilaterally, (ii) right inferior occipital gyrus and middle occipital gyrus bilaterally, (iii) superior parietal lobe and postcentral gyrus Area 2) bilaterally, (iv) left precentral gyrus, (v) inferior frontal gyrus (pars opercularis) bilaterally, (vi) left superior frontal gyrus and middle frontal gyrus bilaterally, (vii) posterior medial frontal gyrus, and (viii) right insula  cerebellum bilaterally, (ii) middle occipital gyrus bilaterally, right inferior occipital gyrus and left inferior temporal gyrus, (iii) superior and inferior parietal lobe, (iv) left inferior parietal lobe, (v) right postcentral gyrus (Area 2), (vi) precentral gyrus bilaterally, (vii) superior frontal gyrus bilaterally, (viii) posterior medial frontal gyrus, (ix) right inferior frontal gyrus (pars opercularis), and (x) left insula  included activations in the: (i) left inferior and in the middle frontal gyrus, (ii) left superior parietal lobule, and (iii) middle occipital gyrus  and left cerebellum. MR of single stimulus (vs. pair of stimuli) included activations in the: (i) right inferior frontal gyrus and in the middle frontal gyrus , (ii) right and left inferior, in the right superior parietal lobule, and precuneus bilaterally, (iii) left paracentral lobule , in addition to the left superior frontal gyrus and middle cingulate cortex. These findings are consistent with the TMS and neuropsychological literature on MR of hand shapes which seem to be related to the parietal lobe  included exclusively right-lateralized activations in the middle occipital gyrus, cuneus and superior parietal cortex. The literature on MR has shown that the key areas supporting MR of 3D cubes seem to be the superior parietal lobule (BA 7), together with the inferior frontal gyrus (BA 44/45)  included bilateral activations involving the posterior occipital-temporal-parietal cortex. In addition, we performed an analysis by distinguishing the strategy from the reference frame variable. Motor-imagery based MR (vs. egocentric MR) activated the left superior parietal lobule, the right postcentral gyrus  and the precentral gyrus/middle and superior frontal gyrus bilaterally, and the left inferior occipital gyrus. Egocentric MR (vs. motor-imagery based MR) activated the left cuneus, the left middle temporal gyrus, the left lingual gyrus and calcarine sulcus, and the right erebellum. In a PET study  modulates the MR processing. When directly comparing the strategy account to the reference frame variable we found that motor-imagery based MR (vs. egocentric MR) activated the left superior parietal lobule, the right postcentral gyrus  and the precentral gyrus/middle and superior frontal gyrus bilaterally, and the left inferior occipital gyrus, whereas the egocentric MR (vs. motor-imagery based MR) activated the left cuneus, the left middle temporal gyrus, the left lingual gyrus and calcarine sulcus, and the right cerebellum. These results indicate that these two mechanisms are different. Motor strategies engage motor behavior covertly whereas the egocentric based MR involves a  judgment from another point of view .The presentation of single stimulus vs. pairs of stimuli also influenced the MR network, however given the strength of the modulation exerted by the type of stimulus and the type of strategy we believe that it would be a very limited account considering the number of stimuli shown alone.bilateral sensorimotor activation for bodily-related (vs. non-bodily-related stimuli) and a posterior right lateralized activation for non-bodily-related (vs. bodily-related stimuli) is consistent, as far as lateralization effects is concerned, with the result related to the type of strategy reporting a bilateral  network for motor imagery- vs. non-motor imagery-based MR and the latter activating a preferentially posterior right occipito-temporal-parietal network.Lastly, the result related to the type of stimuli reporting a physically rotated by the subject. This operation can be triggered implicitly by hands, as confirmed by our meta-analysis. Indeed, in our meta-analysis M1 was found to be activated in the MR of bodily vs. non-bodily stimuli contrast (cluster 2) and it was found to be sensitive to the cognitive strategy used since it was again found in the motor-imagery/egocentric based strategy vs. visuo-spatial imagery/allocentric based strategy (cluster 1). We suggest that the type of stimulus  may implicitly trigger one strategy or the other , and that the left M1 supports the former. Results of the present meta-analysis are also consistent with neuropsychological data showing that MR can be impaired in patients with a tumor affecting the hand area, providing that they imagined the rotation as a consequence of their own hand action. By contrast, lesions in the left M1 sparing the hand area did not lead to an MR deficit  were:Can the bottom up modulation shed light on MR-related hemispheric lateralization issue? Our results showed that bodily- (vs. non-bodily) stimuli activate a bilateral sensorimotor network, whereas non-bodily (vs. bodily)-related stimuli) included exclusively right-lateralized activations.Can the top down modulation shed light on the debate about whether the sensorimotor cortex is involved in MR? Our results showed that when we directly compared the motor to the visual strategy, we found bilateral activations in the sensorimotor areas.May the M1 activation depend upon the type of stimuli and strategy used? Our results showed that M1 was activated for motor  strategy and bodily- (vs. non-bodily) stimuli.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.


Large controlled clinical trials have demonstrated reductions with statin therapy in cardiovascular events in patients presenting with acute coronary syndromes and stable coronary heart disease and individuals at high risk of a cardiovascular event. In trials of acute coronary syndromes and stable coronary heart disease, high-intensity statin therapy is more effective in the prevention of recurrent cardiovascular events than low-intensity statin therapy. Thus, evidence-based guidelines recommend in-hospital initiation of high-intensity statin therapy for all acute coronary syndrome patients. Clinical trials report high adherence to and low discontinuation of high-intensity statin therapy; however, in clinical practice, high-intensity statins are prescribed to far fewer patients, who often discontinue their statin after the first refill. A coordinated effort among the patient, provider, pharmacist, health system, and insurer is necessary to improve utilization and persistence of prescribed medications. The major cause for statin discontinuations reported by patients is perceived adverse events. Evaluation of potential adverse events requires validated tools to distinguish between statin-associated adverse events versus non-specific complaints. Treatment options for statin-intolerant patients include the use of a different statin, often at a lower dose or frequency. In order to lower LDL cholesterol, lower doses of statins may be combined with ezetimibe or bile acid sequestrants. Newer treatment options for patients with statin-associated muscle symptoms may include proprotein convertase subtilisin kexin 9 (PCSK9) inhibitors. In a meta-analysis, high-intensity statin therapy was more effective than control or low-moderate intensity for reducing the risk of vascular death  and all-cause mortality 2. Thus, the American College of Cardiology/American Heart Association (ACC/AHA) guidelines for the treatment of ACS and secondary prevention of CHD recommend the initiation of high-intensity statin therapy in patients with clinical atherosclerotic CVD (ASCVD) regardless of baseline low-density lipoprotein (LDL) cholesterol levels2. In contrast, European Society of Cardiology/European Atherosclerosis Society (ESC/EAS) guidelines recommend reductions in LDL cholesterol to less than 1.8 mmol/L or by 50% or more3. The National Lipid Association (NLA) and International Atherosclerosis Society adopted a similar LDL cholesterol-centric perspective and recommend LDL cholesterol levels less than 1.8 mmol/L regardless of the statin dosage needed to achieve this target5. From the perspective of randomized clinical trials, the mandate for empiric high-intensity statins is particularly relevant for ACS patients in whom initiation of this therapy is recommended before hospital discharge6.Among patients hospitalized for an acute coronary syndrome (ACS) and stable coronary heart disease (CHD), randomized controlled trials have demonstrated that high-dose/high-intensity atorvastatin therapy is more effective than placebo, low-intensity therapy with pravastatin, moderate-intensity therapy with simvastatin, or low-dose atorvastatin therapy in the reduction of recurrent cardiovascular disease (CVD) events7. However, few prior studies have reported the percentage of patients who filled prescriptions for high-intensity statins following CHD events. In ACS registries conducted from 2003 to 2008, utilization of statins ranged from 80 to 91%, while only 23–38% were prescribed high-intensity statins. In a real-world analysis of Medicare beneficiaries, which included hospitalizations for CHD between 2007 and 2011, only 27% with insurance coverage for medications were prescribed high-intensity statins after hospitalization for a coronary event8. The principal factor associated with being discharged and remaining on high-intensity statin therapy for 365 days was prior use of a high-intensity statin. These data suggest that clinicians focus on the LDL cholesterol rather than the clinical trial evidence supporting high-intensity statins that encompasses other atherothrombotic properties9. Utilization of high-intensity statins in Medicare beneficiaries diminished progressively during the ensuing year, such that an additional 24% of participants reduced their statin dosage or discontinued high-intensity statins8. Among patients hospitalized for a non-cardiovascular illness, who then have an in-hospital myocardial infarction, the use of high-intensity statins is lower10. At the time of this survey, simvastatin 80 mg was the only generic statin. Due to the safety concerns with this dosage of simvastatin, particularly in the elderly and in those patients taking multiple medications, simvastatin 40 mg daily was the more commonly prescribed dosage.Data from ACS registries suggest that over 80% of patients are prescribed statins following a myocardial infarction (MI) or coronary revascularization6; however, the clinical consequences of non-adherence to high-intensity statin therapy have been less well studied. In preliminary data from a 5% sample Medicare population, poor adherence and discontinuation of high-intensity statin therapy after the first prescription fill was accompanied by higher rates of hospitalization for cardiovascular and non-cardiovascular causes and more deaths during the ensuing 5 years11.Utilization and persistence of high-intensity statins represents an important performance measure12. Thus, discontinuation of statins and other evidence-based secondary preventive therapies has implications for the patient’s future health as well as economic costs to the patient, their family, and society.Since hospitalizations for recurrent cardiovascular events increase more rapidly in patients hospitalized for a MI than age- and sex-matched controls hospitalized for other causes, short-term and long-term secondary preventive measures are crucial to minimize the risk of recurrent events13. However, clinical trials select individuals with lower risk for muscle events based on age, prior musculoskeletal complaints, renal function, and concomitant non-drug and drug therapies that interact with drug elimination pathways. Discontinuation of statins is more common among patients with side effects, which were reported by 60% of former users and 25% of current users14. Since statin intolerance is often symptom based, it is important to develop and administer validated measures of statin intolerance. The NLA proposed a clinical tool for the assessment of SAMS15 that was based on a retrospective analysis of the STOMP trial, which investigated the effects of statin medication on muscle performance16. In order to improve the accuracy of diagnosis of an adverse event, this index incorporates the fundamental process of de-challenge and re-challenge with either the same statin at a lower dosage or an alternate statin that has different drug elimination pathways that may be genetically based.Several reasons contribute to statin down-titration or discontinuation. In clinical trials, statin-associated adverse events (statin-associated muscle symptoms [SAMS]) are no different between participants assigned to statins or placebo19. The mean LDL cholesterol reduction was 53–56% with evolocumab compared with 15–18% with ezetimibe18. Alirocumab reduced LDL cholesterol by 45% compared to 15% with ezetimibe19. Bempedoic acid (ETC-1002) inhibits ATP citrate lyase (ACL), a key enzyme that supplies substrate for cholesterol and fatty acid synthesis in the liver. Since bempedoic acid inhibits an enzyme earlier in the cholesterol synthetic pathway than statins, it is possible that adverse muscle symptoms caused by HMG-CoA reductase inhibition would also occur with drugs that inhibit metabolites such as mevalonate even earlier in the synthetic pathway. This agent was evaluated in a subgroup of 56 SAMS participants (37 ETC-1002 group and 19 placebo group) enrolled in a larger randomized, placebo-controlled trial; the mean difference in LDL cholesterol was 28.7% versus placebo20. Adverse muscle complaints were similar in the placebo and ETC-1002 treatment groups.Non-statin LDL cholesterol-lowering therapies have been evaluated in patients who report SAMS. These studies have randomized individuals who experienced SAMS with two statins that included one agent at the lowest approved dosages. Second-line LDL cholesterol-lowering agents often used in patients who experience SAMS include ezetimibe, bile acid sequestrants, and niacin. However, these agents have modest LDL cholesterol-lowering efficacy. In trials with fully human monoclonal antibodies to proprotein convertase subtilisin kexin 9 (PCSK9) inhibitors , SAMS were reported in fewer individuals and no more often than when treated with ezetimibe19. In the second phase, continuing participants were randomized to double-blind treatment (2:2:1) with alirocumab, ezetimibe 10 mg daily, or atorvastatin 20 mg daily. On re-exposure to statin therapy, nearly 50% tolerated atorvastatin. SAMS were 39% less frequent with alirocumab versus atorvastatin. The GAUSS III trial design incorporated the fundamental concept for evaluation of adverse drug reactions through a randomized challenge and de-challenge treatment with placebo and atorvastatin 20 mg daily21. Thus, this trial design may serve as a state-of-the-art model for future such trials in SAMS patients. GAUSS III enrolled 511 patients with uncontrolled LDL cholesterol and history of intolerance to two or more statins22. These trials have not incorporated a placebo-controlled challenge and re-challenge phase with statin therapy to ensure appropriate identification of SAMS individuals. The ODYSSEY ALTERNATIVE included a single-blind placebo run-in and excluded participants who reported SAMS with placebo23. The first large cardiovascular outcome trial with a PCSK9 inhibitor is expected to announce results in the second half of 201624. These trials have enrolled patients with cardiovascular disease who have elevated LDL cholesterol levels on moderate- to high-intensity statins and therefore do not address clinical outcomes in patients treated with PCSK9 monotherapy.Currently, the phase III clinical outcome trials with PCSK9 inhibitors have not been reportedNon-adherence to evidence-based secondary preventive LDL cholesterol-lowering statin therapies increases the risk for recurrent cardiovascular and non-cardiovascular events, and all-cause mortality. Coordinated efforts to improve adherence involve patient factors, provider behavior, and health system factors. Many patients who discontinue their medications perceive that non-specific complaints are drug related and then decide to terminate treatment on their own initiative without the input of healthcare professionals. Thus, it is important to engage in a dialogue concerning major adverse reactions with any medication and evaluate reported side effects with an objective clinical tool, such as the statin muscle index. Non-statin approaches are second-line therapies to LDL cholesterol, but they are ineffective in either lowering LDL cholesterol levels by more than 50% or achieving LDL cholesterol levels that meet established targets proposed by several consensus documents. Anti-PCSK9 antibodies are more effective LDL cholesterol-lowering agents than ezetimibe in multiple short-term studies. The efficacy, safety, tolerability, and long-term persistence of PCSK9 inhibitors await completion of large ongoing clinical trials. These trials have enrolled patients with cardiovascular disease, including ACS, who have LDL cholesterol levels ≥70 mg/dL on maximally tolerated moderate- to high-intensity statins. These trials did not specifically include patients with SAMS. Future trials with PCSK9 inhibitor monotherapy will be required to address the unmet need in high-risk patients who refuse or cannot tolerate statin treatment.


Fusarium solani species complex (FSSC) is a clade of environmentally ubiquitous fungi that includes plant, animal, and insect associates. Here, we report the draft genome sequence of the undescribed species FSSC 6 (isolate MYA-4552), housed in the gut of the wood-boring cerambycid beetle Anoplophora glabripennis.The  Fusarium solani species complex (FSSC) is a ubiquitous and functionally diverse group of fungi consisting of plant pathogens . Following extraction, genomic DNA (gDNA) concentration and integrity were assessed via agarose gel electrophoresis and using a NanoDrop ND-8000 UV-Vis spectrophotometer (Thermo Fisher Scientific). Three 454 FLX+ pyrosequencing libraries  were prepared using the GS FLX Titanium sequencing kit XLR70, according to the manufacturer’s specifications. The three 454 FLX+ runs provided a total of 7.138 Gb of single-end sequencing data. Low-quality base calls and 454-specific sequencing adapters were removed using Trimmomatic . Sequencdenovo , and Veldenovo  and the  species ) genome Bowtie 2  to both  genomes , 17, the genomes  was usedLWBZ00000000. The version described in this paper is the first version of the assembly.The whole-genome shotgun sequencing project for the FSSC 6 isolate MYA-4552 has been deposited at DDBJ/EMBL/GenBank under the accession no.


Background. HIV-infected adolescents may be at higher risk for high-grade cervical lesions than HIV-uninfected adolescents. The purpose of this study was to compare the prevalence of high-risk HPV (HR-HPV) infections and Pap smear abnormalities between these two groups. Methods.In this cross-sectional study, we compared the HPV DNA and Pap smear results between 35 HIV-infected and 50 HIV-uninfected adolescents in order to determine the prevalence of HR-HPV genotypes and cervical cytological abnormalities. Comparisons were made using Pearson χ2 and independent-samples t-tests analyses, and associations between demographic and behavioral characteristics and HPV infections were examined. Results. HIV-infected participants were more likely to be infected with any HPV  and with at least one HR-HPV , and to have multiple concurrent HPV infections . HPV 16 and 18 were relatively underrepresented among HR-HPV infections. Abnormal Pap test results were more common among HIV-infected participants . A history of smoking was associated with HR-HPV infection. Conclusions. HIV-infected adolescents have an increased risk of infection with HR-HPV and of Pap test abnormalities. The majority of HR-HPV infections among our participants would not be prevented by the currently available vaccinations against HPV. Human papillomaviruses (HPV) cause cervical cancer . The gloWomen who are infected with HIV are at increased risk of dying from cervical cancer . They arScreening guidelines also differ between adults and adolescents. Due to the high rate of spontaneous clearance of low-grade cervical lesions and the risks associated with over-treatment among adolescents, it is recommended that screening does not begin until the age of 21 , 12. HIVWe compared the HPV DNA and Pap smear results between 50 HIV-uninfected and 35 HIV-infected adolescent females from Masiphumelele, South Africa, in order to determine the prevalence of HR-HPV, multiple concurrent HPV infections, and cervical cytological abnormalities. We examined associations between these results and potential risk factors including sexual activity, contraception use, and smoking.M = 19.06 years; S.D. = 1.48; IQR = 18.00–20.00). While definitions of the age range that constitutes adolescence vary, we defined it in accordance with the American Academy of Pediatrics that categorizes late adolescence to extend to the age of 21 [Between October 2012 and February 2014 we conducted a cross-sectional prevalence study in which cervical specimens for Pap smear and self-collected vaginal swabs for HPV DNA testing were collected from 50 HIV-uninfected and 35 HIV-infected sexually active South African adolescent females aged 17–21 (ge of 21 . ExclusiAll participants signed informed consent (aged 18 and older) or signed adolescent assent documents (aged 17) to accompany parental consent forms in order to participate in the study. This study was approved by the Research Subjects Review Board at the University of Rochester and the Human Research Ethics Committee at the University of Cape Town.For self-sampling, patients were instructed to insert a Dacron swab high into the vagina and twirl it for 10 seconds. Self-sampling was conducted in private. Samples were stored in Digene transport medium and DNA was extracted using the MagNA Pure Compact Nucleic Acid Isolation Kit (Roche Diagnostics). HPV genotyping was conducted using Roche's Linear Array HPV Test. This kit detects 37 HPV genotypes  includin Trichomonas) were referred for appropriate treatment. Most recent CD4 test results were obtained from the local government clinic where HIV-infected participants receive care.Pap smear results were reported in accordance with the Bethesda system . All posχ2 and independent-samples t-tests analyses, and associations between demographic/behavioral characteristics and HPV infections were examined using Spearman correlations. All analyses were performed using IBM SPSS 22.0.HIV-infected and HIV-uninfected youth were compared using Pearson We found a large burden of HPV infection in our study cohort. Overall 64.7% of participants had at least one HPV infection and 38.8% had at least one HR-HPV infection. Multiple concurrent HPV infections were common with 41.2% of all participants testing positive for two or more HPV genotypes (63.6% of HPV-infected participants were infected with greater than one genotype). Multiple concurrent HR-HPV infections were found in 24.7% of all participants and 63.6% of HR-HPV-positive participants were infected with two or more HR-HPV genotypes. Eleven out of the 85 participants (12.9%) had five or more concurrent HPV infections detected. Likewise, 24.2% of HR-HPV positive participants had three or more concurrent HR-HPV infections.Non-vaccine HR-HPV genotypes (those other than 16 and 18) were notably common among our cohort. Among HR-HPV positive participants, only 12.1% were infected with exclusively vaccine genotypes, while 51.5% were infected with exclusively non-vaccine genotypes (36.4% were infected with a combination of vaccine and non-vaccine genotypes).P = 0.049), among all participants with one or more HR-HPV infections, HIV-infected individuals were more likely to be infected with a non-vaccine HR-HPV genotype (P = 0.006). Numerous significant differences in HPV infection were identified between the HIV-infected and HIV-uninfected groups. HIV-infected participants were more likely to be infected with any HPV, to be infected with at least one HR-HPV, and to have multiple concurrent HPV infections. Although HPV 16 was more prevalent in the HIV-infected group .Overall, 18.8% of our cohort had abnormal Pap tests. Pap smear abnormalities were found to be more common among HIV-infected participants than their HIV-uninfected counterparts. No study participants were found to have high-grade squamous intraepithelial lesions (HSIL). There were two HIV-infected youths observed with atypical squamous cells—cannot rule out HSIL (ASC-H) and six with low-grade squamous intraepithelial lesions (LSIL) compared to zero and two, respectively, among HIV-uninfected youth. 3 . The average CD4 for those on ARVs was 301 and for those not on ARVs was 557 (P = 0.012). We did not identify any significant relationship between ARVs and HPV infection. Likewise, CD4 count was not found to be significantly associated with HR-HPV infection or abnormal Pap test.Among the HIV-infected participants, 22.9% were receiving antiretroviral medications (ARVs) at the time of testing. The average CD4 count among all HIV-infected participants was 490/mmP = 0.049). A history of smoking was positively associated with HR-HPV infection (P = 0.024). Methods of contraception used, number of lifetime sexual partners, and number of sexual partners reported in the six months prior to enrollment were not significantly associated with HPV infection.Among our bivariate analyses, we found the older members of our cohort to have a greater number of multiple concurrent HR-HPV infections (P = 0.001). Marais et al. have reported similar differences in HR-HPV prevalence between HIV-infected and HIV-uninfected South African women from an older cohort (mean age: 35) [Our results show an increased risk of infection with HR-HPV and associated Pap smear abnormalities among HIV-infected adolescents compared to their HIV-uninfected counterparts. This is consistent with previous data from South Africa. A recent report comparing the distribution of HPV genotypes between HIV-infected and HIV-uninfected South African women aged 17–65 years found differences in HR-HPV prevalence across all age groups including 17–19-year-olds who were found to have an HR-HPV prevalence of 75% and 60%, respectively [age: 35) . The lonAlthough identifying the HR-HPV genotypes associated with cases of invasive cervical cancer provides the most critical information regarding the proportion of cases preventable by vaccination, the high relative prevalence of non-vaccine HR-HPV compared to the vaccine genotypes in our cohort does demonstrate that the currently available vaccines against HPV would not prevent the majority of HR-HPV infections among our study population. Still, without longitudinal data demonstrating incident and/or persistent cervical disease associated with non-vaccine HR-HPV infection, the significance of this observation is uncertain.HIV-infected women in sub-Saharan Africa are among the groups at highest risk of developing cervical cancer. Although adolescents are generally at low risk for cervical cancer , those wAll cervical cancer screening strategies endeavor to identify cervical dysplasia prior to it reaching an invasive stage while also protecting women from the risks of overtesting and overtreatment. Tremendous efforts have been made to identify cost-effective approaches to the follow-up of negative Pap tests and low-grade Pap smear abnormalities, resulting in recommendations that utilize HPV DNA testing to extend intervals between Pap tests , 12. TheIn this study, self-collected vaginal swabs were used to provide samples for HPV DNA testing. Agreement between the results from clinician-collected and self-collected specimens has been reported to be high in several studies , even amOur study setting has very high density of HIV and HPV infection, providing a unique opportunity to investigate cervical cancer risk among HIV-infected adolescents. Still, our study has several limitations. Although numerous significant data were found, our sample size is relatively small and may be insufficient to identify real differences between study groups. Our current sample size limits the types of multivariate analyses that could be performed to accompany the descriptive statistics and bivariate correlations presented. Subsequent research should seek to examine differences across HIV status while accounting for demographic and behavioral variables. Only 22.9% of our HIV-infected participants were on ARVs and our cohort had relatively high CD4 counts so we are unable to correlate our findings with levels of immunosuppression. Previous research has shown that more advanced HIV disease is strongly associated with increased cumulative HPV prevalence, more advanced cervical dysplasia, and an increased rate of progression of cervical disease , 28–30. Our results underscore the overall high-risk for HPV-related cervical disease in our study population and the significant potential difference in risk for cervical cancer between HIV-infected and HIV-uninfected young women in South Africa. The majority of HR-HPV infections among our study participants would not be prevented by the currently available vaccinations against HPV although the impact of these infections on incident advanced cervical disease is not clear. An optimal, evidence-based approach to cervical cancer screening for HIV-infected adolescents may include the use of HPV testing which has the potential to allow for an increase in the interval between Pap tests. This would have particular benefits in low-resource settings such as Masiphumelele. Longitudinal studies with this and similar cohorts are required to quantify the difference in risk for high-grade cervical lesions between HIV-infected and HIV-uninfected adolescents and to determine if HPV testing can be effectively integrated into a cervical cancer-screening program for adolescents with HIV.