Genetics of food allergy.
Food allergy, a growing clinical and public health problem in the United States and worldwide, is likely determined by multiple environmental and genetic factors. The purpose of this review is to summarize recent advances in food allergy genetic research. There is compelling evidence that genetic factors may play a role in food allergy. However, the specific genetic loci that may modulate individual risk of food allergy remain to be identified. To date, only a limited number of candidate gene association studies of food allergy have been reported. Polymorphism(s) in nine genes have been associated with the incidence of food allergy or food allergy severity in at least one study. But most of these findings remain to be replicated in independent populations. In contrast, there are considerable advances in genetics of other allergic diseases such as asthma and atopic dermatitis. Although asthma and atopic dermatitis often coexist with food allergy, the relevance of their candidate genes to food allergy remains to be evaluated. Genetics in food allergy is a promising research area but is still in its infancy. More studies are needed to dissect susceptible genes of food allergy. A genome-wide association approach may serve as a powerful tool to identify novel genes related to food allergy. Furthermore, the role of gene-environment interaction, gene-gene interaction, and epigenetics in food allergy remains largely unexplored. Given the complex nature of food allergy, future studies need to integrate environment, genomics, and epigenomics in order to better understand the multifaceted etiology and biological mechanisms of food allergy.


Genonets server-a web server for the construction, analysis and visualization of genotype networks.
A genotype network is a graph in which vertices represent genotypes that have the same phenotype. Edges connect vertices if their corresponding genotypes differ in a single small mutation. Genotype networks are used to study the organization of genotype spaces. They have shed light on the relationship between robustness and evolvability in biological systems as different as RNA macromolecules and transcriptional regulatory circuits. Despite the importance of genotype networks, no tool exists for their automatic construction, analysis and visualization. Here we fill this gap by presenting the Genonets Server, a tool that provides the following features: (i) the construction of genotype networks for categorical and univariate phenotypes from DNA, RNA, amino acid or binary sequences; (ii) analyses of genotype network topology and how it relates to robustness and evolvability, as well as analyses of genotype network topography and how it relates to the navigability of a genotype network via mutation and natural selection; (iii) multiple interactive visualizations that facilitate exploratory research and education. The Genonets Server is freely available at http://ieu-genonets.uzh.ch.


Astrocytes prevent abnormal neuronal development in the fragile x mouse.
Astrocytes are now distinguished as major regulators of neuronal growth and synaptic development. Recently, they have been identified as key players in the progression of a number of developmental disorders; however, in fragile X syndrome (FXS), the role of astrocytes is not known. Using a coculture design, we found that hippocampal neurons exhibited abnormal dendritic morphology and a decreased number of presynaptic and postsynaptic protein aggregates when they were grown on astrocytes from a fragile X mouse. Moreover, we found that normal astrocytes could prevent the development of abnormal dendrite morphology and preclude the reduction of presynaptic and postsynaptic protein clusters in neurons from a fragile X mouse. These experiments are the first to establish a role for astrocytes in the altered neurobiology of FXS. Our results support the notion that astrocytes contribute to abnormal dendrite morphology and the dysregulated synapse development in FXS.


Female fertility preservation: past, present and future.
Anti-cancer therapy, particularly chemotherapy, damages ovarian follicles and promotes ovarian failure. The only pharmacological means for protecting the ovaries from chemotherapy-induced injury is gonadotrophin-releasing hormone agonist, but its efficiency remains controversial; ovarian transposition is used to shield the ovary from radiation when indicated. Until the late 1990s, the only option for fertility preservation and restoration in women with cancer was embryo cryopreservation. The development of other assisted reproductive technologies such as mature oocyte cryopreservation and in vitro maturation of oocytes has contributed to fertility preservation. Treatment regimens to obtain mature oocytes/embryos have been modified to overcome various limitations of conventional ovarian stimulation protocols. In the last decades, several centres have begun cryopreserving ovarian samples containing primordial follicles from young patients before anti-cancer therapy. The first live birth following implantation of cryopreserved-thawed ovarian tissue was reported in 2004; since then, the number has risen to more than 130. Nowadays, ovarian tissue cryopreservation can be combined with in vitro maturation and vitrification of oocytes. The use of cryopreserved oocytes eliminates the risk posed by ovarian implantation of reseeding the cancer. Novel methods for enhancing follicular survival after implantation are presently being studied. In addition, researchers are currently investigating agents for ovarian protection. It is expected that the risk of reimplantation of malignant cells with ovarian grafts will be overcome with the putative development of an artificial ovary and an efficient follicle class- and species-dependent in vitro system for culturing primordial follicles.


The axon terminal of goldfish retinal horizontal cells: a low membrane conductance measured in solitary preparations and its implication to the signal conduction from the soma.
1. Mechanical dissociation of the enzyme-treated goldfish retina yielded somata and axon terminals of horizontal cells. The membrane properties of these solitary axon terminals were investigated using the whole-cell patch-clamp technique. 2. Axon terminals had a large input resistance, comparable to the seal resistance (approximately 30 G omega). Most axon terminals (greater than 80%) showed a nearly linear current-voltage relation between -60 and +10 mV, where the slope conductance was as small as 5 muS/cm2. Some axon terminals showed a shallow negative slope conductance in the same potential range. 3. The membrane current consisted of two components: transient and sustained. The transient component was carried by sodium ions, and the sustained component was a mixture of calcium and potassium currents. The sodium current (INa) was activated by depolarization of beyond -45 mV and was maximal (approximately 60 pA) at -10 mV. It was blocked by 5 microM tetrodotoxin and disappeared in Na+-free medium. The maximum amplitude of INa was less than 10% of INa of the soma. 4. A small calcium current (less than 6 pA) was isolated in a small proportion of cells, with an amplitude approximately 5% of the calcium current evoked in the soma under the identical recording conditions. 5. A small amount of potassium current through the anomalous rectifier was induced in the axon terminal when the membrane potential was below -60 mV. Its conductance was 15-20 muS/cm2, only 1/20 of the estimate in the soma. Other types of potassium currents were not detected. 6. It is concluded that the soma and the axon terminal have a similar set of membrane currents, but the specific membrane conductance of the axon terminal is extremely low. The signal conductivity from soma to axon terminal was assessed using a passive cable model together with numerical values obtained from the present experiments. Although the membrane conductance of the connecting axon was not measurable directly, the calculation strongly suggests that low conductance of the axon terminal membrane minimizes the leakage of signals arriving electrotonically through the thin connecting axon, even if the membrane conductance of the axon was overestimated as being identical to the soma membrane. 7. These results can explain why light-evoked responses recorded from the axon terminal are similar in amplitude as well as in waveform to those recorded from the soma, despite the lack of direct inputs from photoreceptors.


Meta-analysis of the risk of gastric stump cancer: detection of high risk patient subsets for stomach cancer after remote partial gastrectomy for benign conditions.
Controversy about gastric cancer risk after partial gastrectomy exists, especially in the United States. Therefore, we performed a meta-analysis to determine overall relative risk and weighted mean relative risk for subsets of postgastrectomy patients, define possible high risk patients suitable for surveillance, and assess for publication bias which would overestimate risk. If 2 studies were excluded because of heterogeneity, overall relative risk (RR) for gastric stump cancer in 22 studies analyzed was 1.66 [95% confidence limits (CL), 1.54-1.79]. With these 2 studies included, the RR summarized with a random effects model to account for study heterogeneity was 1.46 (95% CL, 1.18-1.82). No obvious evidence of publication bias was detected. Patients 15 years or more postoperative had a weighted mean RR of 1.48 (95% CL, 1.31-1.67) and patients 5-14 years postoperative had a RR of 0.91 (95% CL, 0.71-1.17) (P = 0.026). Patients operated upon for gastric ulcer had a weighted mean RR of 2.12 (95% CL, 1.73-2.59) and patients with duodenal ulcers had a RR of 0.84 (95% CL, 0.66-1.05) (P = 0.001). The weighted mean RR for females was 1.79 (95% CL, 1.39-2.29) and for males 1.43 (95% CL, 1.27-1.62) (P = 0.074). For Billroth II gastrectomy the weighted mean RR was 1.60 (95% CL, 1.15-2.18) and for Billroth I gastrectomy 1.20 (95% CL, 1.01-1.42) (P = 0.220). Although differences in risk between subsets of postagastrectomy patients seen to exist, recommendations concerning endoscopic surveillance await further studies of cost-benefit analysis.


Out of the mouths of babes: vocal production in infant siblings of children with ASD.
Younger siblings of children with autism spectrum disorders (ASD) are at higher risk for acquiring these disorders than the general population. Language development is usually delayed in children with ASD. The present study examines the development of pre-speech vocal behavior in infants at risk for ASD due to the presence of an older sibling with the disorder. Infants at high risk (HR) for ASD and those at low risk, without a diagnosed sibling (LR), were seen at 6, 9, and 12 months as part of a larger prospective study of risk for ASD in infant siblings. Standard clinical assessments were administered, and vocalization samples were collected during play with mother and a standard set of toys. Infant vocal behavior was recorded and analyzed for consonant inventory, presence of canonical syllables, and of non-speech vocalizations, in a cross-sectional design. Children were seen again at 24 months for provisional diagnosis. Differences were seen between risk groups for certain vocal behaviors. Differences in vocal production in the first year of life were associated with outcomes in terms of autistic symptomotology in the second year. Early vocal behavior is a sensitive indicator of heightened risk for autistic symptoms in infants with a family history of ASD.


Recombinant adeno-associated virus type 2-mediated gene delivery into the Rpe65-/- knockout mouse eye results in limited rescue.
Leber's congenital amaurosis (LCA) is a severe form of retinal dystrophy. Mutations in the RPE65 gene, which is abundantly expressed in retinal pigment epithelial (RPE) cells, account for approximately 10-15% of LCA cases. In this study we used the high turnover, and rapid breeding and maturation time of the Rpe65-/- knockout mice to assess the efficacy of using rAAV-mediated gene therapy to replace the disrupted RPE65 gene. The potential for rAAV-mediated gene treatment of LCA was then analyzed by determining the pattern of RPE65 expression, the physiological and histological effects that it produced, and any improvement in visual function. rAAV.RPE65 was injected into the subretinal space of Rpe65-/- knockout mice and control mice. Histological and immunohistological analyses were performed to evaluate any rescue of photoreceptors and to determine longevity and pattern of transgene expression. Electron microscopy was used to examine ultrastructural changes, and electroretinography was used to measure changes in visual function following rAAV.RPE65 injection. rAAV-mediated RPE65 expression was detected for up to 18 months post injection. The delivery of rAAV.RPE65 to Rpe65-/- mouse retinas resulted in a transient improvement in the maximum b-wave amplitude under both scotopic and photopic conditions (76% and 59% increase above uninjected controls, respectively) but no changes were observed in a-wave amplitude. However, this increase in b-wave amplitude was not accompanied by any slow down in photoreceptor degeneration or apoptotic cell death. Delivery of rAAV.RPE65 also resulted in a decrease in retinyl ester lipid droplets and an increase in short wavelength cone opsin-positive cells, suggesting that the recovery of RPE65 expression has long-term benefits for retinal health. This work demonstrated the potential benefits of using the Rpe65-/- mice to study the effects and mechanism of rAAV.RPE65-mediated gene delivery into the retina. Although the functional recovery in this model was not as robust as in the dog model, these experiments provided important clues about the long-term physiological benefits of restoration of RPE65 expression in the retina.


Neoadjuvant Chemotherapy in High-Risk Soft Tissue Sarcomas: Final Results of a Randomized Trial From Italian (ISG), Spanish (GEIS), French (FSG), and Polish (PSG) Sarcoma Groups.
To determine whether the administration of histology-tailored neoadjuvant chemotherapy (HT) was superior to the administration of standard anthracycline plus ifosfamide neoadjuvant chemotherapy (A+I) in high-risk soft tissue sarcoma (STS) of an extremity or the trunk wall. This was a randomized, open-label, phase III trial. Patients had localized high-risk STS (grade 3; size, ≥ 5 cm) of an extremity or trunk wall, belonging to one of the following five histologic subtypes: high-grade myxoid liposarcoma (HG-MLPS); leiomyosarcoma (LMS), synovial sarcoma (SS), malignant peripheral nerve sheath tumor (MPNST), and undifferentiated pleomorphic sarcoma (UPS). Patients were randomly assigned in a 1:1 ratio to receive three cycles of A+I or HT. The HT regimens were as follows: trabectedin in HG-MLPS; gemcitabine plus dacarbazine in LMS; high-dose prolonged-infusion ifosfamide in SS; etoposide plus ifosfamide in MPNST; and gemcitabine plus docetaxel in UPS. Primary and secondary end points were disease-free survival (DFS) and overall survival (OS), estimated using the Kaplan-Meier method and compared using Cox models adjusted for treatment and stratification factors. The study is registered at ClinicalTrials.gov (identifier NCT01710176). Between May 2011 and May 2016, 287 patients (UPS: n = 97 [33.8%]; HG-MLPS: n = 65 [22.6%]; SS: n = 70 [24.4%]; MPNST: n = 27 [9.4%]; and LMS: n = 28 [9.8%]) were randomly assigned to either A+I or HT. At the final analysis, with a median follow-up of 52 months, the projected DFS and OS probabilities were 0.55 and 0.47 (log-rank P = .323) and 0.76 and 0.66 (log-rank P = .018) at 60 months in the A+I arm and HT arm, respectively. No treatment-related deaths were observed. In a population of patients with localized high-risk STS, HT was not associated with a better DFS or OS, suggesting that A+I should remain the regimen to choose whenever neoadjuvant chemotherapy is used in patients with high-risk STS.


Measuring relative lysosomal volume for monitoring lysosomal storage diseases.
Biomarkers are important tools in medicine, which can be used for monitoring disease progression and response to therapy. One of the main problems in rare lysosomal storage diseases is that there are over 70 different diseases, all with different biochemical storage profiles. Developing biochemical biomarkers therefore requires an individual assay per disease/subgroup of diseases. An alternative approach is to develop an assay that is independent of the specific macromolecules stored. This chapter discusses an assay that may serve as a universal biomarker for these diseases and measures the expansion of the late endosomal/lysosomal system. We have developed an assay that takes advantage of a commercially available late endosomal/lysosomal probe, LysoTracker, which becomes trapped in the acidic compartment of cells and emits a fluorescent signal that can be detected using flow cytometry. In this chapter, we detail the methodology behind this assay and discuss the factors that need to be considered when establishing this assay in clinical and research settings.


Fish oil supplementation: evidence for health benefits.
Many health claims for fish oil (which contains omega-3 fatty acids) in conditions from Alzheimer disease to Zellweger syndrome are based on indirect evidence. But the evidence is direct for a benefit in coronary heart disease prevention, and the American Heart Association recently issued guidelines for the intake of omega-3 oils. This article answers a series of questions that health care professionals often ask regarding fish oil, such as what are proper dosages, and are there risks of ingesting pollutants by eating more fish or using supplements?


Three-dimensional reconstruction of thick filaments from Limulus and scorpion muscle.
We have produced three dimensional reconstructions, at a nominal resolution of 5 nm, of thick filaments from scorpion and Limulus skeletal muscle, both of which have a right-handed four-stranded helical arrangement of projecting subunits. In both reconstructions there was a distinct division of density within projecting subunits consistent with the presence of two myosin heads. Individual myosin heads appeared to be curved, with approximate dimensions of 16 X 5 X 5 nm and seemed more massive at one end. Our reconstructions were consistent with the two heads in a projecting subunit being arranged either antiparallel or parallel to each other and directed away from the bare zone. Although we cannot exclude the second of these interpretations, we favor the first as being more consistent with both filament models and also because it would enable easy phosphorylation of light chains. The antiparallel interpretation requires that the two heads within a subunit derive from different myosin molecules. In either interpretation, the two heads have different orientations relative to the thick filament shaft.


Peritoneal and subperitoneal stroma may facilitate regional spread of ovarian cancer.
Epithelial ovarian cancer (EOC) is characterized by early peritoneal involvement ultimately contributing to morbidity and mortality. To study the role of the peritoneum in fostering tumor invasion, we analyzed differences between the transcriptional repertoires of peritoneal tissue lacking detectable cancer in patients with EOC versus benign gynecologic disease. Specimens were collected at laparotomy from patients with benign disease (b) or malignant (m) ovarian pathology and comprised primary ovarian tumors, paired bilateral specimens from adjacent peritoneum and attached stroma (PE), subjacent stroma (ST), peritoneal washes, ascites, and peripheral blood mononuclear cells. Specimens were immediately frozen. RNA was amplified by in vitro transcription and cohybridized with reference RNA to a custom-made 17.5k cDNA microarray. Principal component analysis and unsupervised clustering did not segregate specimens from patients with benign or malignant pathology. Class comparison identified differences between benign and malignant PE and ST specimens deemed significant by permutation test (P = 0.027 and 0.012, respectively). A two-tailed Student's t test identified 402 (bPE versus mPE) and 663 (mST versus bST) genes differentially expressed at a significance level of P2 < or = 0.005 when all available paired samples from each patient were analyzed. The same comparison using one sample per patient reduced the pool of differentially expressed genes but retained permutation test significance for bST versus mST (P = 0.031) and borderline significance for bPE versus mPE (P = 0.056) differences. The presence of EOC may foster peritoneal implantation and growth of cancer cells by inducing factors that may represent molecular targets for disease control.


Regulation of intracellular calcium levels by polyunsaturated fatty acids, arachidonic acid and docosahexaenoic acid, in astrocytes: possible involvement of phospholipase A2.
Pathological conditions in the brain, such as ischemia, trauma and seizure are accompanied by increased levels of free n-6 and n-3 polyunsaturated fatty acids (PUFA), mainly arachidonic acid (AA, 20:4n-6) and docosahexaenoic acid (DHA, 22:6n-3). A neuroprotective role has been suggested for PUFA. For investigation of the potential molecular mechanisms involved in neuroprotection by PUFA, we studied the regulation of the concentration of intracellular Ca2+ ([Ca2+]i) in rat brain astrocytes. We evaluated the presence of extracellular PUFA and the release of intracellular PUFA. Interestingly, only the constitutive brain PUFA AA and DHA, but not eicosapentaenoic acid (EPA) had prominent effects on intracellular Ca2+. AA and DHA suppressed [Ca2+]i oscillation, inhibited store-operated Ca2+ entry, and reduced the amplitudes of Ca2+ responses evoked by agonists of G protein-coupled receptors. Moreover, prolonged exposure of astrocytes to AA and DHA brought the cells to a new steady state of a moderately elevated [Ca2+]i level, where the cells became virtually insensitive to external stimuli. This new steady state can be considered as a mechanism of self-protection. It isolates disturbed parts of the brain, because AA and DHA reduce pathological overstimulation in the tissue surrounding the damaged area. In inflammation-related events, frequently AA and DHA exhibit opposite effects. However, in astrocytes AA and DHA exerted comparable effects on [Ca2+]i. Extracellularly added AA and DHA, but not EPA, were also able to induce the release of [3H]AA from prelabeled astrocytes. Therefore, we also suggest the involvement of phospholipase A2 activation and lysophospholipid generation in the regulation of intracellular Ca2+ in astrocytes.


Unexpected σ bond rupture during the reaction of N-methyl-1,2,4-triazoline-3,5-dione with acenaphthylene and indene.
The reaction of N-methyl-1,2,4-triazoline-3,5-dione (MeTAD) with acenaphthylene and indene leads not only to the formation of the expected [2 + 2] diazetidine cycloadducts but also to unexpected 2:1 adducts of MeTAD with substrate. The structures of the products derived from acenaphthylene were confirmed by X-ray crystallography. A similar distribution of products was afforded from indene. The 2:1 adducts appear to derive from a diradical intermediate, the radical centers of which are strongly stabilized by the bridging urazoyl ring and benzylic delocalization. The triplet states of these diradical intermediates may be trapped via exposure to molecular oxygen to afford oxygen-containing adducts. Computational studies at the (U)B3LYP/6-31G* level provide additional support for the conclusions of our experimental work.


S-nitrosothiols signal hypoxia-mimetic vascular pathology.
NO transfer reactions between protein and peptide cysteines have been proposed to represent regulated signaling processes. We used the pharmaceutical antioxidant N-acetylcysteine (NAC) as a bait reactant to measure NO transfer reactions in blood and to study the vascular effects of these reactions in vivo. NAC was converted to S-nitroso-N-acetylcysteine (SNOAC), decreasing erythrocytic S-nitrosothiol content, both during whole-blood deoxygenation ex vivo and during a 3-week protocol in which mice received high-dose NAC in vivo. Strikingly, the NAC-treated mice developed pulmonary arterial hypertension (PAH) that mimicked the effects of chronic hypoxia. Moreover, systemic SNOAC administration recapitulated effects of both NAC and hypoxia. eNOS-deficient mice were protected from the effects of NAC but not SNOAC, suggesting that conversion of NAC to SNOAC was necessary for the development of PAH. These data reveal an unanticipated adverse effect of chronic NAC administration and introduce a new animal model of PAH. Moreover, evidence that conversion of NAC to SNOAC during blood deoxygenation is necessary for the development of PAH in this model challenges conventional views of oxygen sensing and of NO signaling.


'Double or quits': perceptions and management of organ transplantation by adults with cystic fibrosis.
Medical sociologists have often considered lay perceptions of the risks of medical interventions, yet in many empirical studies respondents are people who are not likely to be exposed to a particular intervention. Furthermore, it has been well documented that risk perceptions may change over time and with diminishing health state. This paper explores perceptions and management of the risks of organ transplantation amongst adults with cystic fibrosis (CF), the most common autosomal recessive genetic disease in the UK. Although the focus of medical research is now on providing gene replacement therapy to this group, transplantation is currently the last treatment that an adult with CF can be offered when all other treatment has failed to maintain their health. Thirty-one respondents with varying degrees of health state from a specialist CF centre were interviewed as part of a larger study concerning perceptions of health and risks of treatment. Interviews were audiotaped, transcribed and analysed using ATLAS-ti. During analysis respondents' transcripts were divided into two groups: firstly those who did not anticipate needing a transplant in the near future (if at all) and secondly those who were currently being considered for transplantation, on the transplant list, or who had already received donor organs. The paper focuses on themes arising from interview transcripts and finds that although the focus of risk differs between the two groups, the influence of luck is perceived as strong for both groups and emotion work features heavily in those undergoing the transplant process. Contrary to previous research, fears of inheriting donor characteristics are not found amongst adults with CF, but rather body components are commodified when talking of both giving and receiving organs.


A new iridescent tarantula of the genus Thrigmopoeus Pocock, 1899 from Western Ghats, India.
A distinctive new species of ground burrowing tarantula from Western Ghats endemic genus Thrigmopoeus is described from Kerala State, India. Thrigmopoeus psychedelicus sp. nov. differs from putative species of the genus in the adults being black overall with a metallic blue lustre on the carapace and abdomen. Females of Thrigmopoeus psychedelicus sp. nov. exhibit polychromatism. Juveniles and sub-adults are paler with vibrant maroon colouration on its abdomen whereas adult females are much darker and lack vibrant colouration as sub-adults.


Simultaneous Electrochemical Evaluation of Ascorbic Acid, Epinephrine and Uric Acid at Disposable Pencil Graphite Electrode: Highly Sensitive Determination in Pharmaceuticals and Biological Liquids by Differential Pulse Voltammetry.
As is known, AA, EP and UA can also coexist in biological fluids. Therefore, the determination of the levels of these compounds in biological fluids is extremely important both for the diagnosis and treatment of the related diseases. In the presence of many interfering substances in biological fluids such as blood and urine samples, it is very important that these compounds can be selectively analyzed. All electrochemical experiments were performed using an Autolab PGSTAT 128N potentiostat. Before beginning the electrochemical measurements, the PGE was activated. The electrochemical pretreatment of PG was exercised by anodically +1.40 V for 60 s. Then, measurements were performed with CV (-0.4 V to 1.2 V) and DPV (-0.2 V to 0.7 V) for single and simultaneous voltammetric behaviour of AA, EP, and UA in the electrochemical method. The anodic peak potentials of AA and UA were observed at about +0.32 V and +0.62 V, respectively. On the other hand, for EP, while anodic peak potential was observed at about +0.53 V, in the reverse scan, cathodic peak potentials were observed at about +0.41 V and +0.007 V. The reduction peak observed at +0.3 V with the oxidation peak observed at +0.53 V are the reversible peaks. In the method developed for the electrochemical simultaneous determination of AA, EP and UA using PGE with DPV technique in BR buffer solution (pH 4.0), the anodic peak potentials are sufficiently separated from each other. A voltammetric method was developed for the simultaneous determination of AA, EP and UA with PGE for the first time. Here, the most important thing is that the simultaneous determination of AA, EP and UA was successfully achieved with that targeted voltammetric method which was sensitive, low-cost, practical and well-repeated; and that these were proven to be selectively applicable in pharmaceutical products and biological liquids.


Progress in the preclinical discovery and clinical development of class I and dual class I/IV phosphoinositide 3-kinase (PI3K) inhibitors.
The phosphoinositide 3-kinases (PI3Ks) constitute an important family of lipid kinase enzymes that control a range of cellular processes through their regulation of a network of signal transduction pathways, and have emerged as important therapeutic targets in the context of cancer, inflammation and cardiovascular diseases. Since the mid-late 1990s, considerable progress has been made in the discovery and development of small molecule ATP-competitive PI3K inhibitors, a number of which have entered early phase human trials over recent years from which key clinical results are now being disclosed. This review summarizes progress made to date, primarily on the discovery and characterization of class I and dual class I/IV subtype inhibitors, together with advances that have been made in translational and clinical research, notably in cancer.


Creation of a palliative care advanced practice registered nurse peer-review process.
The mark of a true profession is the ability to self-regulate. As such, advanced practice registered nurses (APRNs) are challenged by their professional organisations to participate in self-evaluation and peer review. Peer review is a method for evaluating the care provided by the APRN to both ensure quality nursing care and promote professional growth. Despite guidelines to participate in a formal peer-review process, there is little information within the nursing profession on how to accomplish peer review. A comprehensive literature review failed to provide a framework for peer review that is practice focused, fosters a learning environment and encompasses the thought process and clinical decision-making of the palliative care advanced practice nurse. A group of palliative care APRNs set out to create a process that encompassed the breadth of clinical decision-making in palliative care advanced nursing practice. Using the eight domains of palliative care, a narrative case review structure was created. The resulting process both assisted the APRNs in professional growth and provided timely feedback for the annual performance evaluation.


Mortality attributable to hot and cold ambient temperatures in India: a nationally representative case-crossover study.
Most of the epidemiological studies that have examined the detrimental effects of ambient hot and cold temperatures on human health have been conducted in high-income countries. In India, the limited evidence on temperature and health risks has focused mostly on the effects of heat waves and has mostly been from small scale studies. Here, we quantify heat and cold effects on mortality in India using a nationally representative study of the causes of death and daily temperature data for 2001-2013. We applied distributed-lag nonlinear models with case-crossover models to assess the effects of heat and cold on all medical causes of death for all ages from birth (n = 411,613) as well as on stroke (n = 19,753), ischaemic heart disease (IHD) (n = 40,003), and respiratory diseases (n = 23,595) among adults aged 30-69. We calculated the attributable risk fractions by mortality cause for extremely cold (0.4 to 13.8°C), moderately cold (13.8°C to cause-specific minimum mortality temperatures), moderately hot (cause-specific minimum mortality temperatures to 34.2°C), and extremely hot temperatures (34.2 to 39.7°C). We further calculated the temperature-attributable deaths using the United Nations' death estimates for India in 2015. Mortality from all medical causes, stroke, and respiratory diseases showed excess risks at moderately cold temperature and hot temperature. For all examined causes, moderately cold temperature was estimated to have higher attributable risks (6.3% [95% empirical confidence interval (eCI) 1.1 to 11.1] for all medical deaths, 27.2% [11.4 to 40.2] for stroke, 9.7% [3.7 to 15.3] for IHD, and 6.5% [3.5 to 9.2] for respiratory diseases) than extremely cold, moderately hot, and extremely hot temperatures. In 2015, 197,000 (121,000 to 259,000) deaths from stroke, IHD, and respiratory diseases at ages 30-69 years were attributable to moderately cold temperature, which was 12- and 42-fold higher than totals from extremely cold and extremely hot temperature, respectively. The main limitation of this study was the coarse spatial resolution of the temperature data, which may mask microclimate effects. Public health interventions to mitigate temperature effects need to focus not only on extremely hot temperatures but also moderately cold temperatures. Future absolute totals of temperature-related deaths are likely to depend on the large absolute numbers of people exposed to both extremely hot and moderately cold temperatures. Similar large-scale and nationally representative studies are required in other low- and middle-income countries to better understand the impact of future temperature changes on cause-specific mortality.


AggreCount: an unbiased image analysis tool for identifying and quantifying cellular aggregates in a spatially defined manner.
Protein quality control is maintained by a number of integrated cellular pathways that monitor the folding and functionality of the cellular proteome. Defects in these pathways lead to the accumulation of misfolded or faulty proteins that may become insoluble and aggregate over time. Protein aggregates significantly contribute to the development of a number of human diseases such as amyotrophic lateral sclerosis, Huntington's disease, and Alzheimer's disease. In vitro, imaging-based, cellular studies have defined key biomolecular components that recognize and clear aggregates; however, no unifying method is available to quantify cellular aggregates, limiting our ability to reproducibly and accurately quantify these structures. Here we describe an ImageJ macro called AggreCount to identify and measure protein aggregates in cells. AggreCount is designed to be intuitive, easy to use, and customizable for different types of aggregates observed in cells. Minimal experience in coding is required to utilize the script. Based on a user-defined image, AggreCount will report a number of metrics: (i) total number of cellular aggregates, (ii) percentage of cells with aggregates, (iii) aggregates per cell, (iv) area of aggregates, and (v) localization of aggregates (cytosol, perinuclear, or nuclear). A data table of aggregate information on a per cell basis, as well as a summary table, is provided for further data analysis. We demonstrate the versatility of AggreCount by analyzing a number of different cellular aggregates including aggresomes, stress granules, and inclusion bodies caused by huntingtin polyglutamine expansion.


A CT-based Radiomics Signature Is Associated with Response to Immune Checkpoint Inhibitors in Advanced Solid Tumors.
Background Reliable predictive imaging markers of response to immune checkpoint inhibitors are needed. Purpose To develop and validate a pretreatment CT-based radiomics signature to predict response to immune checkpoint inhibitors in advanced solid tumors. Materials and Methods In this retrospective study, a radiomics signature was developed in patients with advanced solid tumors (including breast, cervix, gastrointestinal) treated with anti-programmed cell death-1 or programmed cell death ligand-1 monotherapy from August 2012 to May 2018 (cohort 1). This was tested in patients with bladder and lung cancer (cohorts 2 and 3). Radiomics variables were extracted from all metastases delineated at pretreatment CT and selected by using an elastic-net model. A regression model combined radiomics and clinical variables with response as the end point. Biologic validation of the radiomics score with RNA profiling of cytotoxic cells (cohort 4) was assessed with Mann-Whitney analysis. Results The radiomics signature was developed in 85 patients (cohort 1: mean age, 58 years ± 13 [standard deviation]; 43 men) and tested on 46 patients (cohort 2: mean age, 70 years ± 12; 37 men) and 47 patients (cohort 3: mean age, 64 years ± 11; 40 men). Biologic validation was performed in a further cohort of 20 patients (cohort 4: mean age, 60 years ± 13; 14 men). The radiomics signature was associated with clinical response to immune checkpoint inhibitors (area under the curve [AUC], 0.70; 95% CI: 0.64, 0.77; P < .001). In cohorts 2 and 3, the AUC was 0.67 (95% CI: 0.58, 0.76) and 0.67 (95% CI: 0.56, 0.77; P < .001), respectively. A radiomics-clinical signature (including baseline albumin level and lymphocyte count) improved on radiomics-only performance (AUC, 0.74 [95% CI: 0.63, 0.84; P < .001]; Akaike information criterion, 107.00 and 109.90, respectively). Conclusion A pretreatment CT-based radiomics signature is associated with response to immune checkpoint inhibitors, likely reflecting the tumor immunophenotype. © RSNA, 2021 Online supplemental material is available for this article. See also the editorial by Summers in this issue.


A novel combination: ranpirnase and rosiglitazone induce a synergistic apoptotic effect by down-regulating Fra-1 and Survivin in cancer cells.
Accumulating evidence supports the idea that two known phosphatidylinositol 3'-kinase (PI3K) downstream proteins, Fra-1 and Survivin, are potential targets for cancer therapy. Increased expression of Fra-1, a Fos family member of the transcription factor activator protein-1, has been implicated in both the maintenance and the progression of the transformed state of several cancer cells. In addition, high Survivin expression in tumors correlates with more aggressive behavior, lower response to chemotherapeutic drugs, and shortened survival time. Previously, we reported that, in malignant mesothelioma cells with increased PI3K activity, small-molecule inhibitors of the PI3K/AKT pathway acted cooperatively with the amphibian RNase chemotherapeutic drug ranpirnase to inhibit cell growth. Because the thiazolidinedione antidiabetic drug rosiglitazone targets the PI3K/AKT pathway, we investigated the effect of the combination of these two drugs in cell survival in several cancer cell lines. We show here that the combination of ranpirnase and rosiglitazone synergistically decreases cell viability and increases cell apoptosis in several cancer cell lines. Cell killing is associated with decreased Fra-1 and Survivin expression and knockdown of Fra-1 increases cell killing by ranpirnase in a dose-dependent manner but not by rosiglitazone. The drug combination does not have a synergistic effect on killing in Fra-1 knockdown cells, showing that Fra-1 modulation accounts in part for the synergism. The novel drug combination of ranpirnase and rosiglitazone is a promising combination to treat cancers with increased PI3K-dependent Fra-1 expression or Survivin.


Regional brain metabolism in schizophrenia: The influence of antipsychotics.
Schizophrenia has been associated with a plethora of metabolic changes in the brain that vary with duration and type of psychoses. Additionally, it has been observed that antipsychotics can further alter cerebral glucose metabolism. These changes resulting from antipsychotics have been postulated to be reflective of the duration and mechanism of action of the medication. We aimed to examine the influence of antipsychotics on brain metabolism in individuals with schizophrenia in a naturalistic setting. A cross-sectional study was carried out by the psychiatry department of a tertiary care hospital in collaboration with the Radiation Medicine Centre. Eighteen male patients with schizophrenia in different phases of treatment underwent an 18F-deoxyglucose positron emission tomography scan in a resting state 12 hours after the last dose of antipsychotic. The types and duration of treatment were then compared with the regional glucose uptake in 14 predetermined regions of interest. The relative Uptake Values were further compared using SPSS 11.0. An immediate increase followed by a decrease in cortical uptake was noted while the basal ganglia uptake remained high, albeit with a decreasing trend. Typical antipsychotics were associated with lower frontal cortical and higher basal ganglia and cerebellar uptake as compared to atypical antipsychotics. The differential influence of the type and duration of antipsychotic on glucose uptake suggests a possible trend towards long-term side effects with typical medications that were not noted on clinical examination. This however needs to be confirmed with larger, controlled studies.


Phase I and immunomodulatory study of a muramyl peptide, muramyl tripeptide phosphatidylethanolamine.
Muramyl tripeptide phosphatidylethanolamine (MTP-PE; CGP 19835A from Ciba Geigy) is a synthetic muramyl tripeptide structurally related to bacterial cell wall constituents. MTP-PE activates monocytes in vitro to a tumoricidal state and has in vivo antitumor effects in animal models. We studied the toxicity and immunomodulatory effects of once weekly i.v. administration of liposomal-encapsulated MTP-PE for 8 weeks in 27 patients with advanced malignancies. Doses ranged from 0.1 to 2.7 mg/m2. No major tumor responses were seen; 11 patients had stable disease after 8 weeks of therapy and 3 continued on maintenance therapy because of minor tumor regressions and/or clinical improvement. MTP-PE at these doses was well tolerated. Shaking chills and fevers were the most common toxicities and occurred at all dose levels. There was no treatment-induced loss of performance status. Immunomodulatory studies revealed evidence of a biological effect on monocytes. C-reactive protein levels rose in the majority of patients with end-of-treatment values 2 to 10 times higher than baseline. Serum neopterin levels were consistently increased 24 h after MTP-PE administration and significant decreases in expression of two different types of Fc receptors on peripheral blood monocytes were noted 6 h after treatment. Although no major tumor responses were seen in this group of patients with advanced malignancies, MTP-PE was well tolerated and exerted biological effects on monocytes. Serum neopterin levels may be a useful marker for the biological effects of MTP-PE.


Office laboratory procedures, office economics, patient and parent education, and urinary tract infection.
This section updates the reader on four important areas of office practice: office laboratory procedures, office economics, patient and parent education, and urinary tract infections. Dr. Michael Aldous reviews the recent literature about office laboratory procedures, including the continued impact of the Clinical Laboratory Improvement Ammendments, what is new in the diagnosis of streptococcal pharyngitis, urinalysis improvements, the diagnosis of anemia, and which patients should undergo cholesterol screening. Dr. Rickey Williams discusses the literature on office economics, including new technology for billing and charting, whether pediatricians should bill for telephone calls, and the latest information on health care policy and the changes offices are facing with the growing managed care market. Dr. Burris Duncan reviews patient and parent education, including new apporaches to infant colic, sleep positioning for the prevention of sudden infant death, the need for the hepatitis B vaccine (which has been slowly implemented), and finally ways that pediatricians can help with parenting. Dr. John Ey discusses the recent literature on urinary tract infections in children, including better ways of making the diagnosis, whether there are any new treatment approaches for urinary tract infections, useful investigational studies for evaluating the urinary system, and how best to follow up children with infected urinary tracts. We hope that this review will help the practicing pediatrician to better care for patients and provide each of you with a greater satisfaction in delivering health care in an office setting.


Knowledge of hepatocellular carcinoma screening guidelines and clinical practices among gastroenterologists.
Screening of high-risk patients for hepatocellular carcinoma (HCC) may result in early diagnosis and improved outcomes. Our aim was to assess gastroenterologists' knowledge of HCC management guidelines established by the American Association for the Study of Liver Diseases (AASLD) and usual clinical practice. We surveyed gastroenterologists attending two gastroenterology board review courses regarding their knowledge of HCC screening guidelines and usual practice of screening for HCC. Practices were compared and adherence to the 2005 published HCC guidelines was assessed. The median age of gastroenterology attending physicians (n = 160) was 41 years, and 75% were men with a median of 11.5 years of practice. A total of 79% of respondents correctly identified the high-risk patients who qualify for HCC screening. Most gastroenterologists correctly identified the screening methods (88.5%) and screening interval (98%). Among those who knew guideline recommendations (i.e., correct identification and certainty of guideline recommendations), 100% reported that they followed the guideline recommendation in their own practices. Regarding the management of abnormal test, 31% of gastroenterologists did not identify that referral for liver transplantation is the recommended management strategy for small HCC in a Child B patient with cirrhosis. The number of years in clinical practice (p = 0.30) and involvement in a malpractice suit (p = 0.34) did not affect the practice patterns. Most gastroenterologists correctly identified the common high-risk scenarios, methods, and interval of HCC screening as recommended by AASLD. Gastroenterologists who knew the HCC guidelines applied them in their own practice. However, approximately one-quarter do not know the appropriate management of a positive result, thereby likely hampering the overall effectiveness of screening.


The comparative release of FGF1 by hypoxia and temperature stress.
The signal peptide-less FGF gene family prototype, FGF1 is released in response to temperature stress in vitro as a latent reducing agent-sensitive homodimer non-covalently complexed with the extravesicular p40 domain of p65 synaptotagmin (Syt)1. Because FGF1 is well recognized as an angiogenesis factor in vivo and angiogenesis is known to be induced by hypoxia, we examined the release of FGF1 and p40 Syt1 under conditions of hypoxia and temperature stress using a chemostatic microcarrier cell culture system. We report that like the pathway used by FGF1 and p40 Syt1 release under temperature stress, hypoxia also induces the release of FGF1 and p40 Syt1 with similar kinetic and pharmacologic properties including the requirement for functional cysteine residues. Lastly, FGF1 and p40 Syt1 release in response to hypoxia and temperature stress is sensitive to lipoxygenase and cyclooxygenase inhibitors suggesting that arachidonic acid metabolism may play an important role in the mechanism of FGF1 release in vitro.


Segregating variation in the polycomb group gene cramped alters the effect of temperature on multiple traits.
The phenotype produced by a given genotype can be strongly modulated by environmental conditions. Therefore, natural populations continuously adapt to environment heterogeneity to maintain optimal phenotypes. It generates a high genetic variation in environment-sensitive gene networks, which is thought to facilitate evolution. Here we analyze the chromatin regulator crm, identified as a candidate for adaptation of Drosophila melanogaster to northern latitudes. We show that crm contributes to environmental canalization. In particular, crm modulates the effect of temperature on a genomic region encoding Hedgehog and Wingless signaling effectors. crm affects this region through both constitutive heterochromatin and Polycomb silencing. Furthermore, we show that crm European and African natural variants shift the reaction norms of plastic traits. Interestingly, traits modulated by crm natural variants can differ markedly between Drosophila species, suggesting that temperature adaptation facilitates their evolution.


Vibrational Modes Promoting Exciton Relaxation in the B850 Band of LH2.
Exciton relaxation dynamics in multichromophore systems are often modeled using Redfield theory, where bath fluctuations mediate the relaxation among the exciton eigenstates. Identifying the vibrational or phonon modes that are implicated in exciton relaxation allows more detailed understanding of exciton dynamics. Here we focus on a well-studied light-harvesting II complex (LH2) isolated from the photosynthetic purple bacterium Rhodoblastus acidophilus strain 10050. Using two synchronized mode-locked lasers, we carried out a polarization-dependent two-dimensional electronic spectroscopy (2DES) study of an ultrafast exciton relaxation in the B850 band of LH2. 2DES data with different polarization configurations enable us to investigate the exciton relaxation between the k = ±1 exciton states. Then, we identify vibrational modes coupled to the exciton relaxation by analyzing the coherent wavepackets in the 2DES signals. Focusing on the coherent vibrational wavepackets, the data suggest that certain symmetry-breaking modes of monomeric units play a key role in exciton relaxation.


Williams syndrome: recent advances in our understanding of cognitive, social and psychological functioning.
Since the last review of Williams syndrome in Current Opinion (2001) there have been many advances in knowledge about the cognitive, social and psychological impairments that characterize the disorder. The present review focuses on current research in these areas. Williams syndrome is associated with a wide range of cognitive, linguistic, social and other difficulties. When young, these deficits may appear relatively mild - for example, many children are highly sociable and talkative - but with age the impact of these difficulties becomes more evident. Thus, inappropriate social behaviours can significantly increase the risk of social exclusion and vulnerability to abuse. Their superficially good speech can lead to educational and other services failing to understand the true extent of impairments or the need for specialist support. Mental health problems, especially related to anxiety, often become an increasing challenge from adolescence onwards. The core difficulties associated with Williams syndrome have a cascading effect on many areas of development over time. However, specialist provision is rare and intervention trials are almost nonexistent. Longitudinal research is needed to identify factors associated with cognitive, social and emotional problems and to develop more effective ways of minimizing and treating difficulties.


Artificial light at night decreases metamorphic duration and juvenile growth in a widespread amphibian.
Artificial light at night (ALAN) affects over 20% of the earth's surface and is estimated to increase 6% per year. Most studies of ALAN have focused on a single mechanism or life stage. We tested for indirect and direct ALAN effects that occurred by altering American toads' (Anaxyrus americanus) ecological interactions or by altering toad development and growth, respectively. We conducted an experiment over two life stages using outdoor mesocosms and indoor terraria. In the first phase, the presence of ALAN reduced metamorphic duration and periphyton biomass. The effects of ALAN appeared to be mediated through direct effects on toad development, and we found no evidence for indirect effects of ALAN acting through altered ecological interactions or colonization. In the second phase, post-metamorphic toad growth was reduced by 15% in the ALAN treatment. Juvenile-stage ALAN also affected toad activity: in natural light, toads retreated into leaf litter at night whereas ALAN toads did not change behaviour. Carry-over effects of ALAN were also present; juvenile toads that had been exposed to larval ALAN exhibited marginally increased activity. In this time frame and system, our experiments suggested ALAN's effects act primarily through direct effects, rather than indirect effects, and can persist across life stages.


Prevention of vertebral fractures by strontium ranelate in postmenopausal women with osteoporosis.
The antifracture efficacy of strontium ranelate, a compound with a novel mechanism of action on bone, has been assessed in two large, randomized, controlled trials conducted in postmenopausal women. Strontium ranelate was given at a daily dose of 2 g, and all women received calcium and vitamin D supplements. In women with established osteoporosis there was a 41% reduction in vertebral fractures over 3 years' treatment [relative risk (RR) 0.59; 95% confidence interval (CI) 0.48-0.73; P<0.001]; significant reductions were also seen after only 1 year of treatment. The beneficial effect was also seen for clinical vertebral fractures: over 3 years there was a significant reduction in new clinical vertebral fractures (RR 0.62; 95% CI 0.47-0.83; P<0.001); this reduction was also observed during the first year of treatment (RR 0.48; 95% CI 0.29-0.80; P=0.003). Over the 3-year treatment period significantly fewer patients had height loss and fewer patients reported new or worsening back pain in the treated group than in the control group. These results demonstrate that strontium ranelate is a new therapeutic option in the prevention of osteoporotic vertebral fractures in postmenopausal women.


The evolution of index signals to avoid the cost of dishonesty.
Animals often convey useful information, despite a conflict of interest between the signaller and receiver. There are two major explanations for such 'honest' signalling, particularly when the size or intensity of signals reliably indicates the underlying quality of the signaller. Costly signalling theory (including the handicap principle) predicts that dishonest signals are too costly to fake, whereas the index hypothesis predicts that dishonest signals cannot be faked. Recent evidence of a highly conserved causal link between individual quality and signal growth appears to bolster the index hypothesis. However, it is not clear that this also diminishes costly signalling theory, as is often suggested. Here, by incorporating a mechanism of signal growth into costly signalling theory, we show that index signals can actually be favoured owing to the cost of dishonesty. We conclude that costly signalling theory provides the ultimate, adaptive rationale for honest signalling, whereas the index hypothesis describes one proximate (and potentially very general) mechanism for achieving honesty.


Rab35 GTPase: A Central Regulator of Phosphoinositides and F-actin in Endocytic Recycling and Beyond.
Rab35 is one of the first discovered members of the large Rab GTPase family, yet it received little attention for 10 years being considered merely as a Rab1-like GTPase. In 2006, Rab35 was recognized as a unique Rab GTPase localized both at the plasma membrane and on endosomes, playing essential roles in endocytic recycling and cytokinesis. Since then, Rab35 has become one of the most studied Rabs involved in a growing number of cellular functions, including endosomal trafficking, exosome release, phagocytosis, cell migration, immunological synapse formation and neurite outgrowth. Recently, Rab35 has been acknowledged as an oncogenic GTPase with activating mutations being found in cancer patients. In this review, we provide a comprehensive summary of known Rab35-dependent cellular functions and detail the few Rab35 effectors characterized so far. We also review how the Rab35 GTP/GDP cycle is regulated, and emphasize a newly discovered mechanism that controls its tight activation on newborn endosomes. We propose that the involvement of Rab35 in such diverse and apparently unrelated cellular functions can be explained by the central role of this GTPase in regulating phosphoinositides and F-actin, both on endosomes and at the plasma membrane.


The Health and Retirement Study Harmonized Cognitive Assessment Protocol Project: Study Design and Methods.
The Harmonized Cognitive Assessment Protocol (HCAP) Project is a substudy within the Health and Retirement Study (HRS), an ongoing nationally representative panel study of about 20,000 adults aged 51 or older in the United States. The HCAP is part of an international research collaboration funded by the National Institute on Aging to better measure and identify cognitive impairment and dementia in representative population-based samples of older adults, in the context of ongoing longitudinal studies of aging in high-, middle-, and low-income countries around the world. The HCAP cognitive test battery was designed to measure a range of key cognitive domains affected by cognitive aging (including attention, memory, executive function, language, and visuospatial function) and to allow harmonization and comparisons to other studies in the United States and around the world. The HCAP included a pair of in-person interviews, one with the target HRS respondent (a randomly selected HRS sample member, aged 65+) that lasted approximately 1 h and one with an informant nominated by the respondent that lasted approximately 20 min. The final HRS HCAP sample included 3,496 study subjects, representing a 79% response rate among those invited to participate. Linking detailed HCAP cognitive assessments to the wealth of available longitudinal HRS data on cognition, health, biomarkers, genetics, health care utilization, informal care, and economic resources and behavior will provide unique and expanded opportunities to study cognitive impairment and dementia in a nationally representative US population-based sample. The fielding of similar HCAP projects in multiple countries around the world will provide additional opportunities to study international differences in the prevalence, incidence, and outcomes of dementia globally with comparable data. Like all HRS data, HCAP data are publicly available at no cost to researchers.


A soluble, single-chain Kd molecule produced by yeast selects a peptide repertoire indistinguishable from that of cell-surface-associated Kd.
Peptide binding to a soluble, single-chain Kd protein produced by the yeast strain Kluyveromyces lactis, and to Kd molecules on Kd-expressing cells (P815) was studied using radiolabeled Kd-restricted peptides. The stability of the peptide-Kd complexes formed was monitored in the absence and presence of unlabeled competitor peptides. Radioiodination of the Tyr anchor residue in position 2 of the peptide interferes with binding. A Kd-biased peptide library and a modified antigenic peptide in which a second Tyr was added in positions 6 and 8, respectively, were therefore used to assay binding. Recombinant and cell-associated Kd molecules are very similar in the following respects: the ease with which the proteins can be loaded with labeled peptide; the spectrum of peptides selected from a peptide library; the stability of the labeled peptide-Kd complex formed; and the ability to partially dissociate the class I-peptide complex with exogenous, unlabeled peptides. These results imply that measurements of peptide binding to soluble Kd molecules are a reliable indicator of the peptide-binding properties of Kd proteins on living cells. The large quantities of soluble recombinant Kd protein currently available represent an invaluable tool not only for dissecting the molecular mechanisms of antigen presentation but also for vaccinations and the design of T cell-specific toxins.


The control of Hyalomma ticks, vectors of the Crimean-Congo hemorrhagic fever virus: Where are we now and where are we going?
At a time of major global, societal, and environmental changes, the shifting distribution of pathogen vectors represents a real danger in certain regions of the world as generating opportunities for emergency. For example, the recent arrival of the Hyalomma marginatum ticks in southern France and the concurrent appearance of cases of Crimean-Congo hemorrhagic fever (CCHF)-a disease vectored by this tick species-in neighboring Spain raises many concerns about the associated risks for the European continent. This context has created an urgent need for effective methods for control, surveillance, and risk assessment for ticks and tick-borne diseases with a particular concern regarding Hyalomma sp. Here, we then review the current body of knowledge on different methods of tick control-including chemical, biological, genetical, immunological, and ecological methods-and the latest developments in the field, with a focus on those that have been tested against ticks from the genus Hyalomma. In the absence of a fully and unique efficient approach, we demonstrated that integrated pest management combining several approaches adapted to the local context and species is currently the best strategy for tick control together with a rational use of acaricide. Continued efforts are needed to develop and implement new and innovative methods of tick control.


(Electro)Sensing of Phenicol Antibiotics-A Review.
The presence of residues from frequent antibiotic use in animal feed can cause serious health risks by contaminating products meant for human consumption such as meat and milk. The present paper gives an overview of the electrochemical methods developed for the detection of phenicol antibiotic residues (chloramphenicol, thiamphenicol, and florfenicol) in different kinds of foodstuffs. Electrochemical sensors based on different biomolecules and nanomaterials are described. The detection limit of various developed methods with their advantages and disadvantages will be highlighted.


Temporal gene regulation during HIV-1 infection of human CD4+ T cells.
CD4(+) T-cell depletion is a characteristic of human immunodeficiency virus type 1 (HIV-1) infection. In this study, modulation of mRNA expression of 6800 genes was monitored simultaneously at eight time points in a CD4(+) T-cell line (CEM-GFP) during HIV infection. The responses to infection included: (1) >30% decrease at 72 h after infection in overall host-cell production of monitored mRNA synthesis, with the replacement of host-cell mRNA by viral mRNA, (2) suppression of the expression of selected mitochondrial and DNA repair gene transcripts, (3) increased expression of the proapoptotic gene and its gene p53-induced product Bax, and (4) activation of caspases 2, 3, and 9. The intense HIV-1 transcription resulted in the repression of much cellular RNA expression and was associated with the induction of apoptosis of infected cells but not bystander cells. This choreographed host gene response indicated that the subversion of the cell transcriptional machinery for the purpose of HIV-1 replication is akin to genotoxic stress and represents a major factor leading to HIV-induced apoptosis.


Lipidomic Analysis Reveals Serum Alteration of Plasmalogens in Patients Infected With ZIKA Virus.
Zika virus (ZIKV) is an arthropod-borne virus (arbovirus) in the Flavivirus genus of the Flaviviridae family. Since the large outbreaks in French Polynesia in 2013-2014 and in Brazil in 2015, ZIKV has been considered a new public health threat. Similar to other related flavivirus, ZIKV is associated with mild and self-limiting symptoms such as rash, pruritus, prostration, headache, arthralgia, myalgia, conjunctivitis, lower back pain and, when present, a short-term low grade fever. In addition, ZIKV has been implicated in neurological complications such as neonatal microcephaly and Guillain-Barré syndrome in adults. Herein, serum lipidomic analysis was used to identify possible alterations in lipid metabolism triggered by ZIKV infection. Patients who presented virus-like symptoms such as fever, arthralgia, headache, exanthema, myalgia and pruritus were selected as the control group. Our study reveals increased levels of several phosphatidylethanolamine (PE) lipid species in the serum of ZIKV patients, the majority of them plasmenyl-phosphatidylethanolamine (pPE) (or plasmalogens) linked to polyunsaturated fatty acids. Constituting up to 20% of total phospholipids in humans, plasmalogens linked to polyunsaturated fatty acids are particularly enriched in neural membranes of the brain. The biosynthesis of plasmalogens requires functional peroxisomes, which are important sites for viral replication, including ZIKV. Thus, increased levels of plasmalogens in serum of ZIKV infected subjects suggest a link between ZIKV life cycle and peroxisomes. Our data provide important insights into specific host cellular lipids that are likely associated with ZIKV replication and may serve as platform for antiviral strategy against ZIKV.


Distinguishing Fast and Slow Processes in Accuracy - Response Time Data.
We investigate the relation between speed and accuracy within problem solving in its simplest non-trivial form. We consider tests with only two items and code the item responses in two binary variables: one indicating the response accuracy, and one indicating the response speed. Despite being a very basic setup, it enables us to study item pairs stemming from a broad range of domains such as basic arithmetic, first language learning, intelligence-related problems, and chess, with large numbers of observations for every pair of problems under consideration. We carry out a survey over a large number of such item pairs and compare three types of psychometric accuracy-response time models present in the literature: two 'one-process' models, the first of which models accuracy and response time as conditionally independent and the second of which models accuracy and response time as conditionally dependent, and a 'two-process' model which models accuracy contingent on response time. We find that the data clearly violates the restrictions imposed by both one-process models and requires additional complexity which is parsimoniously provided by the two-process model. We supplement our survey with an analysis of the erroneous responses for an example item pair and demonstrate that there are very significant differences between the types of errors in fast and slow responses.


The usefulness of preoperative biopsy in unilateral nasal masses.
Unilateral nasal masses are considered suspicious for proliferative diseases. Several tools are routinely used to investigate unilateral lesions such as imaging and nasal biopsy. This study investigated the usefulness of nasal biopsy in predicting the actual nature of unilateral lesions. Preoperative nasal biopsy pathological results were compared with the final pathology obtained during an operation. Forty-six patients with unilateral nasal masses were included in the study group. In 40 patients the final pathology was similar to the preoperative nasal biopsy. In three patients the biopsy specimen was a benign polyp and the final pathology was of an inverted papilloma in two patients and hemangiopericytoma in one patient. In two patients the biopsy specimen was suspicious for an inverted papilloma and the final pathology was a benign polyp. In one patient the biopsy specimen was chordoma and the final pathology was osteosarcoma. The total agreement was 86.9%. The kappa value was 81.2%. Preoperative nasal biopsy is important and useful in evaluating unilateral nasal masses.


Generalizing the dynamic field theory of spatial cognition across real and developmental time scales.
Within cognitive neuroscience, computational models are designed to provide insights into the organization of behavior while adhering to neural principles. These models should provide sufficient specificity to generate novel predictions while maintaining the generality needed to capture behavior across tasks and/or time scales. This paper presents one such model, the dynamic field theory (DFT) of spatial cognition, showing new simulations that provide a demonstration proof that the theory generalizes across developmental changes in performance in four tasks-the Piagetian A-not-B task, a sandbox version of the A-not-B task, a canonical spatial recall task, and a position discrimination task. Model simulations demonstrate that the DFT can accomplish both specificity-generating novel, testable predictions-and generality-spanning multiple tasks across development with a relatively simple developmental hypothesis. Critically, the DFT achieves generality across tasks and time scales with no modification to its basic structure and with a strong commitment to neural principles. The only change necessary to capture development in the model was an increase in the precision of the tuning of receptive fields as well as an increase in the precision of local excitatory interactions among neurons in the model. These small quantitative changes were sufficient to move the model through a set of quantitative and qualitative behavioral changes that span the age range from 8 months to 6 years and into adulthood. We conclude by considering how the DFT is positioned in the literature, the challenges on the horizon for our framework, and how a dynamic field approach can yield new insights into development from a computational cognitive neuroscience perspective.


Can the Positive and Negative Syndrome scale (PANSS) differentiate treatment-resistant from non-treatment-resistant schizophrenia? A factor analytic investigation based on data from the Pattern cohort study.
Treatment-Resistant Schizophrenia (TRS) and Non-Treatment-Resistant Schizophrenia (NTRS) may represent different subtypes of schizophrenia. However, few studies have investigated their PANSS symptom dimensions by Exploratory (EFA) or Confirmatory (CFA). Data from the present study are derived from 1429 patients of the Pattern study. TRS was defined by the use of clozapine in the previous year whereas NTRS by the use of non-clozapine antipsychotics ("by proxy"). Factors were chosen based on the Kaiser criterion and considered valid when loadings were greater than or equal to 0.5. The fit to the data was evaluated by CFA in comparison with well-established PANSS models, using fit indexes. The EFA yielded similar five-factor model in both groups: Negative, Positive, Anxiety/Depression, Cognitive and Excited. CFA showed a satisfactory, but not perfect, fit to the data, as compared with the previous PANSS factor analytic models. Despite the limitations regarding the 'by proxy' definition of TRS, the results of the present study show that there are no differences in the factorial structure of PANSS in patients with TRS and NTRS.


Sleep and circadian rhythm disruption alters the lung transcriptome to predispose to viral infection.
Sleep and circadian rhythm disruption (SCRD), as encountered during shift work, increases the risk of respiratory viral infection including SARS-CoV-2. However, the mechanism(s) underpinning higher rates of respiratory viral infection following SCRD remain poorly characterized. To address this, we investigated the effects of acute sleep deprivation on the mouse lung transcriptome. Here we show that sleep deprivation profoundly alters the transcriptional landscape of the lung, causing the suppression of both innate and adaptive immune systems, disrupting the circadian clock, and activating genes implicated in SARS-CoV-2 replication, thereby generating a lung environment that could promote viral infection and associated disease pathogenesis. Our study provides a mechanistic explanation of how SCRD increases the risk of respiratory viral infections including SARS-CoV-2 and highlights possible therapeutic avenues for the prevention and treatment of respiratory viral infection.


Characterization of protein transacetylase from human placenta as a signaling molecule calreticulin using polyphenolic peracetates as the acetyl group donors.
We have earlier shown that a unique membrane-bound enzyme mediates the transfer of acetyl group(s) from polyphenolic peracetates (PA) to functional proteins, which was termed acetoxy drug: protein transacetylase (TAase) because it acted upon several classes of PA. Here, we report the purification of TAase from human placental microsomes to homogeneity with molecular mass of 60 kDa, exhibiting varying degrees of specificity to several classes of PA confirming the structure-activity relationship for the microsome-bound TAase. The TAase catalyzed protein acetylation by a model acetoxy drug, 7,8-diacetoxy-4-methyl coumarin (DAMC) was established by the demonstration of immunoreactivity of the acetylated target protein with anti-acetyl lysine antibody. TAase activity was severely inhibited in calcium-aggregated microsomes as well as when Ca2+ was added to purified TAase, suggesting that TAase could be a calcium binding protein. Furthermore, the N-terminal sequence analysis of purified TAase (EPAVYFKEQFLD) using Swiss Prot Database perfectly matched with calreticulin (CRT), a major microsomal calcium binding protein of the endoplasmic reticulum (ER). The identity of TAase with CRT was substantiated by the observation that the purified TAase avidly reacted with commercially available antibody raised against the C-terminus of human CRT (13 residues peptide, DEEDATGQAKDEL). Purified TAase also showed Ca2+ binding and acted as a substrate for phosphorylation catalyzed by protein kinase C (PKC), which are hallmark characteristics of CRT. Further, purified placental CRT as well as the commercially procured pure CRT yielded significant TAase catalytic activity and were also found effective in mediating the acetylation of the target protein NADPH cytochrome P-450 reductase by DAMC as detected by Western blot using anti-acetyl lysine antibody. These observations for the first time convincingly attribute the transacetylase function to CRT. Hence, this transacetylase function of CRT is designated calreticulin transacetylase (CRTAase). We envisage that CRTAase plays an important role in protein modification by way of acetylation independent of Acetyl CoA.


In vivo 13C NMR assessment of brain glycogen concentration and turnover in the awake rat.
Brain glycogen metabolism was recently observed in vivo and found to be very slow in the lightly alpha-chloralose anesthetized rat [J. Neurochem. 73 (1999) 1300]. Based on that slow turnover, the total glycogen content in the awake rat brain and its turnover time were assessed after administering 13C-labeled glucose for 48 h. Label incorporation into glycogen, glucose, amino acid, and N-acetyl-aspartate (NAA) resonances was observed. The amount of 13C label incorporated into glycogen was variable and did not correlate with that in glutamate (r=-0.1, P>0.86). However, the amount of 13C label incorporated into glycogen was very similar to that in NAA (r=0.93), implying similar turnover times between brain glycogen and NAA (approximately 10 h). Absolute quantification of the total concentration of brain glycogen in the awake, normoglycemic rat yielded 3.3+/-0.8 micromol/g (n=6, mean+/-S.D.).


Transcranial Brain Stimulation Techniques For Major Depression: Should We Extend TMS Lessons to tDCS?
Transcranial magnetic stimulation (TMS) and transcranial direct current stimulation (tDCS) are non-invasive brain stimulation techniques that, by means of magnetic fields and low intensity electrical current, respectively, aim to interefere with and modulate cortical excitability, at the level of dorsolateral prefrontal cortex, in patients with major depression and poor response to standard antidepressants. While the clinical efficacy of TMS in major depression has been extensively investigated over the last 10 years, tDCS has attracted research interest only in the last years, with fewer randomized clinical trials (RCTs) in the field. Nevertheless, in spite of the different rationale and mechanism of action of the two techniques, tDCS recent acquisitions, in relation to the treatment of major depression, seem to parallel those previously obtained with TMS, in terms of treatment duration to achieve optimal benefit and patient's history of drug-resistance. After briefly introducing the two techniques, the article examines possible common pathways of clinical use for TMS and tDCS, emerging from recent RCTs and likely orienting future investigation with non invasive brain stimulation for the treatment of major depression.


Viscoelastic properties of isomeric alkylglucoside surfactants studied by surface light scattering.
Surface light scattering (SLS) by capillary waves was used to investigate the adsorption behavior of non-ionic sugar surfactants at the air/liquid interface. SLS by the subphase (water) followed predictions from hydrodynamic theory. The viscoelastic properties (surface elasticity and surface viscosity) of monolayers formed by octyl beta-glucoside, octyl alpha-glucoside, and 2-ethylhexyl alpha-glucoside surfactants were quantified at submicellar concentrations. It is further concluded that a diffusional relaxation model describes the observed trends in high-frequency, nonintrusive laser light scattering experiments. The interfacial diffusion coefficients that resulted from fitting this diffusional relaxation model to surface elasticity values obtained with SLS reflect the molecular dynamics of the subphase near the interface. However, differences from the theoretical predictions indicate the existence of effects not accounted for such as thermal convection, molecular rearrangements, and other relaxation mechanisms within the monolayer. Our results demonstrate important differences in molecular packing at the air-water interface for the studied isomeric surfactants.


Neighbourhood exposure to point-of-sale price promotions for cigarettes is associated with financial stress among smokers: results from a population-based study.
To examine the association between neighbourhood exposure to point-of-sale (POS) cigarette price promotions and financial stress among smokers in a Midwestern metropolitan area in the USA. Survey data from 888 smokers provided information on sociodemographic and smoking related variables. Financial stress was measured with the question: 'In the last six months, because of lack of money, was there a time when you were unable to buy food or pay any important bills on time, such as electricity, telephone, credit card, rent or your mortgage? (Yes/No).' Using audit data from 504 tobacco retailers, we estimated a score of POS price promotions for each respondent by summing the different types of promotion in each store in their neighbourhood, as defined by a 1-km roadway buffer. Adjusted results provided strong support for an association between higher scores of neighbourhood POS cigarette price promotions and a higher probability of financial stress (p=0.007). Exposure to POS cigarette price promotions is associated with financial stress. This finding, coupled with previous reports that smokers with financial stress are less likely to attempt to quit or succeed in quitting smoking, suggests that POS cigarette price promotions may act as an impediment to smoking cessation.


The Association between Noncommercial Partnerships and Risk of HIV among Female Sex Workers: Evidences from a Cross-Sectional Behavioral and Biological Survey in Southern India.
This paper examines the association between female sex workers' (FSWs) noncommercial partnerships with risk of HIV in Andhra Pradesh, India. Data were drawn from a cross-sectional behavioral and biological survey conducted in 2009 among 3225 FSWs from Andhra Pradesh. Participants were asked about their sexual partnerships, condom use, and vulnerability factors and tested for HIV and sexually transmitted infections. The key independent variables considered were presence of a noncommercial sexual partner (no, yes) and the nature of such partnerships (regular, nonregular). FSWs who reported husband as noncommercial partner were considered to have a regular partner, while the rest were defined as having nonregular partners. Adjusted odds ratios (OR) and 95% confidence intervals (CI) were estimated to measure the associations between variables of interest. Almost three-fourths (74.8%) of FSWs reported having noncommercial partners (regular: 55.6%; nonregular: 19.3%). FSWs in nonregular partnerships were more likely to be HIV positive (13.1% versus 10.9%, adjusted OR: 1.4, 95% CI: 1.1-1.8), have syphilis (10.3% versus 4.2%, adjusted OR: 2.3, 95% CI: 1.6-3.3), use condoms inconsistently with occasional clients (21.0% versus 16.5%, adjusted OR: 1.5, 95% CI: 1.2-1.9), and report forced sex (25.1% versus 14.1%, adjusted OR: 1.9, 95% CI: 1.5-2.4) as compared to those in regular partnerships. HIV prevention programs need to emphasize safe sex behaviors, particularly among FSWs who have nonregular partners.


Selecting strong Brønsted acid zeolites through screening from a database of hypothetical frameworks.
The database of prospective zeolites () has been screened in search of feasible zeolites with the condition of having at least one strong Brønsted site. Several criteria of zeolite feasibility have been tested using energetic and structural concepts, allowing a fast elimination of unsuitable candidates. Based on improved definitions to count and enumerate rings in zeolites, Brønsted acidity has been assessed in a fast albeit inaccurate way, by calculating a structural descriptor related to ammonia desorption energy. In each zeolite, the value of this descriptor was calculated for all the possible centres where a Brønsted acid site can be located. Ranking each zeolite through the value of the strongest candidate acid site allowed obtaining a selection of potentially strong acid zeolites. With further selection criteria, a final short list of 12 structures was obtained, where accurate calculations using periodic DFT indicate that 6 of them must contain a Brønsted site of very strong acidity.


Delayed intracranial hemorrhage following cerebral infarction in sickle cell anemia.
Clinical and necropsy findings in 11 patients with sickle cell anemia (SS) indicate that intracranial hemorrhage (IH) is a delayed sequela of the same vasculopathy that causes cerebral infarction during childhood. Evidence of prior cerebral infarction during childhood included hemiparesis, seizures, an episode of coma, or mental retardation. Computerized tomography (CT) scans showed cerebral infarcts with lucent areas and dilated ventricles or cerebral atrophy. CT or magnetic resonance imaging (MRI) scans after the intracranial hemorrhage demonstrated intraventricular or intracerebral hemorrhages. Angiography or autopsy in seven patients showed widespread vascular occlusion and narrowing of arterial vessels. Moyamoya with internal carotid artery occlusion was identified in two cases. At the time of the IH, three patients were being treated with prophylactic transfusion regimens. We hypothesize that the central nervous system vasculopathy progresses over time and that arterial narrowing in both large and small vessels secondary to endothelial hyperplasia is followed by neovascularization and hemorrhage. Recognition of this pattern of delayed intracranial hemorrhage following cerebral infarction should encourage more intensive evaluation aimed at developing rational interventional therapy prior to a terminal intracranial hemorrhage.


Effects of melatonin and Pycnogenol on small artery structure and function in spontaneously hypertensive rats.
It was suggested that oxidative stress has a key role in the development of endothelial dysfunction, as well as microvascular structural alterations. Therefore, we have investigated 2 substances with antioxidant properties: melatonin and Pycnogenol. We treated 7 spontaneously hypertensive rats (SHRs) with melatonin and 7 with Pycnogenol for 6 weeks. We compared results obtained with those observed in 7 SHRs and 7 Wistar-Kyoto normotensive control rats kept untreated. Mesenteric small resistance arteries were dissected and mounted on a wire myograph, and a concentration-response curve to acetylcholine was performed. Aortic contents of metalloproteinase 2, Bax, inducible NO synthase, and cyclooxygenase 2 were evaluated, together with the aortic content of total collagen and collagen subtypes and apoptosis rate. A small reduction in systolic blood pressure was observed. A significant improvement in mesenteric small resistance artery structure and endothelial function was observed in rats treated with Pycnogenol and melatonin. Total aortic collagen content was significantly greater in untreated SHRs compared with Wistar-Kyoto control rats, whereas a full normalization was observed in treated rats. Apoptosis rate was increased in the aortas of untreated SHRs compared with Wistar-Kyoto control rats; an even more pronounced increase was observed in treated rats. Bax and metalloproteinase 2 expressions changed accordingly. Cyclooxygenase 2 and inducible NO synthase were more expressed in the aortas of untreated SHRs compared with Wistar-Kyoto control rats; this pattern was normalized by both treatments. In conclusion, our data suggest that treatment with Pycnogenol and melatonin may protect the vasculature, partly independent of blood pressure reduction, probably through their antioxidant effects.


Socioeconomic development, health interventions and mortality decline in Costa Rica.
Costa Rica, whose life expectancy was 74 years by 1985, has reached a health level comparable to a developed country. The health achievements of this country are product of political and socioeconomic circumstances as well as of right public health policies. Until about 1970 the features of Costa Rica mortality, although somewhat better than the Latin American average, evolved in a similar way to the rest of the region. In particular, the decades of 1940s and 1950s saw dramatic improvements in life expectancy, thanks mainly to the import of low-cost, high-effectiveness health technologies. In the 1970s, however, Costa Rica departed from a regional pattern of stagnation and managed to close the gap with developed countries in terms of mortality levels. A dramatic decline in the infant mortality rate from 60 to 19 per 1,000 took place in this decade. The main determinants of this breakthrough were health interventions, notably a primary health care program, even though favorable socioeconomic conditions and a reduced fertility also played a role. Ecological data and other evidence suggest that up to three fourths of the mortality decline was accounted for contemporary improvements in public health services, with about 40 percent attributable to primary health care interventions. Furthermore, by targeting interventions on the less privileged population, these interventions had the merit of reducing geographic and socioeconomic differentials in child mortality.


Atrial arrhythmias and pacing after orthotopic heart transplantation: bicaval versus standard atrial anastomosis.
Right and left atrial configuration is more normal when the donor left atrium is anastomosed to a recipient left atrial cuff with direct anastomoses of the donor and recipient vena cavas on the right side. The right atrium and sinus node may be less disturbed by the technique of bicaval anastomosis than by the standard procedure. To compare the incidence of atrial arrhythmias and pacing after bicaval and standard anastomoses. 75 patients had heart transplants between January 1991 and December 1993. The notes were reviewed. Nine patients who died within the first 30 days were excluded from further analysis (seven patients with standard anastomoses, one with bicaval anastomosis, and one with a hybrid technique). 66 patients survived for more than 30 days. Thirty five patients had standard anastomoses and 31 bicaval anastomoses. Atrial tachyarrhythmias (atrial fibrillation, atrial flutter, atrial tachycardia, or supraventricular tachycardia) occurred on four days in three patients in the bicaval group compared with 27 days in 13 patients in the standard group (P = 0.009). The relative risk of atrial tachyarrhythmias with standard anastomosis was 5.52 (P = 0.015) compared with that of bicaval anastomosis. Atrial tachyarrhythmias requiring treatment occurred less often in the bicaval group (four episodes in three patients in the bicaval group and eight episodes in four patients in the standard group), and fewer patients with a bicaval anastomosis required temporary pacing (pacing on 20 days in 10 patients in the bicaval group, but pacing on 49 days in 16 patients in the standard group) and late permanent pacing (no patients in the bicaval group and three patients in the standard group), although these differences were not statistically significant. Patients in the bicaval group were discharged from hospital sooner than those in the standard group (mean 24.1 v 29.1 days, P = 0.024). The technique of bicaval anastomosis, in addition to theoretical advantages from maintaining a more normal atrial configuration, has a lower incidence of postoperative atrial tachyarrhythmias, may reduce the need for pacing, and allows earlier discharge from hospital.


Selection bias: maintaining less-differentiated T cells for adoptive immunotherapy.
The clinical application of T cell immunotherapy depends on ex vivo modification and expansion of T cells for adoptive transfer. In preclinical models, the use of a purified, naive T cell subset enhances persistence and antitumor immunity; however, the majority of clinical studies rely on modification of mixed populations of T cells that contain only a small subset of highly functional T cells with less-differentiated phenotype. In this month's issue of the JCI, Klebanoff and colleagues uncover a Fas-mediated interaction between naive T cells and antigen-experienced T cells that drives differentiation and impairs adoptive immunotherapy. Further, they show that blockade of Fas signaling enhances antitumor immunity and increases survival in a mouse model of melanoma. Their work supports a growing body of evidence that the use of naive T cells enhances the efficacy of adoptive T cell therapy and suggests a new therapeutic strategy for preserving less-differentiated T cell populations.


Human Dendritic Cells: Their Heterogeneity and Clinical Application Potential in Cancer Immunotherapy.
Dendritic cells (DC) are professional antigen presenting cells, uniquely able to induce naïve T cell activation and effector differentiation. They are, likewise, involved in the induction and maintenance of immune tolerance in homeostatic conditions. Their phenotypic and functional heterogeneity points to their great plasticity and ability to modulate, according to their microenvironment, the acquired immune response and, at the same time, makes their precise classification complex and frequently subject to reviews and improvement. This review will present general aspects of the DC physiology and classification and will address their potential and actual uses in the management of human disease, more specifically cancer, as therapeutic and monitoring tools. New combination treatments with the participation of DC will be also discussed.


Lipoxygenation of arachidonic acid by differentiated and undifferentiated human promyelocytic HL-60 cells.
The human promyelocytic leukemia cell line HL-60 can be induced to differentiate into mature granulocytes by exposure to dimethyl sulfoxide (DMSO), as well as into cells of monocyte-macrophage lineage by exposure to 12-0-tetra-decanoylphorbol-13-acetate (TPA). Incubation of the promyelocytic leukemic HL-60 cells and the two chemically induced differentiated cells with carbon 14-labeled arachidonic acid overnight resulted in the incorporation of label mainly into cellular phospholipids. Stimulation of the labeled cells with Ca2+ ionophore (A23187) resulted in the generation of lipoxygenase products in all the cells. Notably, the DMSO-induced differentiated granulocytic cells biosynthesized predominantly the peptidoeicosatetraenoic acids LTC4 and LTD4 compared with the dihydroxyeicosatetraenoic acid LTB4. On the other hand, the TPA-induced differentiated monocytic-macrophage cells biosynthesized predominantly LTB4 compared with LTC4 and LTD4. The undifferentiated promyelocytic cells lacked the capacity to biosynthesize LTB4, as evidenced in the chemically induced differentiated cells. The monohydroxyeicosatetraenoic acid (HETE) 5-HETE, a metabolite of the 5-lipoxygenase pathway, was a minor component in these cells, and 12-HETE and 15-HETE were barely detectable. These results suggest that the two models of DMSO-induced and TPA-induced differentiated cells may be useful systems for further investigations of arachidonic acid metabolites in granulocytic and monocytic-macrophage functions.


Targeted induction of a silent fungal gene cluster encoding the bacteria-specific germination inhibitor fumigermin.
Microorganisms produce numerous secondary metabolites (SMs) with various biological activities. Many of their encoding gene clusters are silent under standard laboratory conditions because for their activation they need the ecological context, such as the presence of other microorganisms. The true ecological function of most SMs remains obscure, but understanding of both the activation of silent gene clusters and the ecological function of the produced compounds is of importance to reveal functional interactions in microbiomes. Here, we report the identification of an as-yet uncharacterized silent gene cluster of the fungus Aspergillus fumigatus, which is activated by the bacterium Streptomyces rapamycinicus during the bacterial-fungal interaction. The resulting natural product is the novel fungal metabolite fumigermin, the biosynthesis of which requires the polyketide synthase FgnA. Fumigermin inhibits germination of spores of the inducing S. rapamycinicus, and thus helps the fungus to defend resources in the shared habitat against a bacterial competitor.


Task and spatial frequency modulations of object processing: an EEG study.
Visual object processing may follow a coarse-to-fine sequence imposed by fast processing of low spatial frequencies (LSF) and slow processing of high spatial frequencies (HSF). Objects can be categorized at varying levels of specificity: the superordinate (e.g. animal), the basic (e.g. dog), or the subordinate (e.g. Border Collie). We tested whether superordinate and more specific categorization depend on different spatial frequency ranges, and whether any such dependencies might be revealed by or influence signals recorded using EEG. We used event-related potentials (ERPs) and time-frequency (TF) analysis to examine the time course of object processing while participants performed either a grammatical gender-classification task (which generally forces basic-level categorization) or a living/non-living judgement (superordinate categorization) on everyday, real-life objects. Objects were filtered to contain only HSF or LSF. We found a greater positivity and greater negativity for HSF than for LSF pictures in the P1 and N1 respectively, but no effects of task on either component. A later, fronto-central negativity (N350) was more negative in the gender-classification task than the superordinate categorization task, which may indicate that this component relates to semantic or syntactic processing. We found no significant effects of task or spatial frequency on evoked or total gamma band responses. Our results demonstrate early differences in processing of HSF and LSF content that were not modulated by categorization task, with later responses reflecting such higher-level cognitive factors.


The Saccharomyces cerevisiae calponin/transgelin homolog Scp1 functions with fimbrin to regulate stability and organization of the actin cytoskeleton.
Calponins and transgelins are members of a conserved family of actin-associated proteins widely expressed from yeast to humans. Although a role for calponin in muscle cells has been described, the biochemical activities and in vivo functions of nonmuscle calponins and transgelins are largely unknown. Herein, we have used genetic and biochemical analyses to characterize the budding yeast member of this family, Scp1, which most closely resembles transgelin and contains one calponin homology (CH) domain. We show that Scp1 is a novel component of yeast cortical actin patches and shares in vivo functions and biochemical activities with Sac6/fimbrin, the one other actin patch component that contains CH domains. Purified Scp1 binds directly to filamentous actin, cross-links actin filaments, and stabilizes filaments against disassembly. Sequences in Scp1 sufficient for actin binding and cross-linking reside in its carboxy terminus, outside the CH domain. Overexpression of SCP1 suppresses sac6Delta defects, and deletion of SCP1 enhances sac6Delta defects. Together, these data show that Scp1 and Sac6/fimbrin cooperate to stabilize and organize the yeast actin cytoskeleton.